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The Medicare Health Outcomes Survey (HOS) is the first Medicare managed care outcomes measure. CMS, in collaboration with NCQA, launched the Medicare HOS in the 1998 Health Plan Employer Data and Information Set (HEDIS?). The measure includes the most recent advances in summarizi...
Beyond ubiquitination: proteolytic and nonproteolytic roles of HOS1.
Jung, Jae-Hoon; Lee, Hyo-Jun; Park, Mi-Jeong; Park, Chung-Mo
The E3 ubiquitin ligase HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENES 1 (HOS1) functions as a cold signaling attenuator by degrading the INDUCER OF CBF EXPRESSION 1 transcription factor, which is a key regulator of the cold-induced transcriptome and freezing tolerance in plants. Recent studies demonstrate that HOS1 also plays nonproteolytic roles in gene expression regulation. HOS1 acts as a chromatin remodeling factor that modulates FLOWERING LOCUS C chromatin in cold regulation of flowering time. It associates with the nuclear pore complex to facilitate nucleocytoplasmic mRNA export to maintain circadian periodicity over a range of light and temperature conditions. In this review, we summarize recent advances in molecular mechanisms underlying HOS1 function during plant development in response to fluctuating environmental conditions. PMID:24768209
Alternative splicing and expression analysis of High expression of osmotically responsive genes1 (HOS1) in Arabidopsis.
Lee, Jeong Hwan; Kim, Soo Hyun; Kim, Jae Joon; Ahn, Ji Hoon
High expression of osmotically responsive genes1 (HOS1), a key regulator of low temperature response and flowering time, encodes an E3 ubiquitin ligase in Arabidopsis. Here, we report characterization of a newly identified splice variant (HOS1-L) of HOS1. Comparative analyses revealed that HOS1-L has a longer 5' nucleotide sequence than that of the previously identified HOS1 (HOS1-S) and that its protein sequence was more conserved than that of HOS1-S in plants. HOS1-L transcripts were spatio-temporally more abundant than those of HOS1-S. The recovery rate of HOS1-S expression was faster than that of HOS1-L after cold treatment. Diurnal oscillation patterns of HOS1-L revealed that HOS1-L expression was affected by photoperiod. An in vitro pull-down assay revealed that the HOS1-L protein interacted with the ICE1 protein. HOS1-L overexpression caused delayed flowering in wild-type plants. Collectively, these results suggest regulation of HOS1 expression at the post-transcriptional level. PMID:23010172
Functional characterization of Candida albicans Hos2 histone deacetylase
Karthikeyan, G; Paul-Satyaseela, Maneesh; Dhatchana Moorthy, Nachiappan; Gopalaswamy, Radha; Narayanan, Shridhar
Candida albicans is a mucosal commensal organism capable of causing superficial (oral and vaginal thrush) infections in immune normal hosts, but is a major pathogen causing systemic and mucosal infections in immunocompromised individuals. Azoles have been very effective anti-fungal agents and the mainstay in treating opportunistic mold and yeast infections. Azole resistant strains have emerged compromising the utility of this class of drugs. It has been shown that azole resistance can be reversed by the co-administration of a histone deacetylase (HDAC) inhibitor, suggesting that resistance is mediated by epigenetic mechanisms possibly involving Hos2, a fungal deacetylase. We report here the cloning and functional characterization of HOS2 (High Osmolarity Sensitive) , a gene coding for fungal histone deacetylase from C. albicans. Inhibition studies showed that Hos2 is susceptible to pan inhibitors such as trichostatin A (TSA) and suberoylanilide hydroxamic acid (SAHA), but is not inhibited by class I inhibitors such as MS-275. This in vitro enzymatic assay, which is amenable to high throughput could be used for screening potent fungal Hos2 inhibitors that could be a potential anti-fungal adjuvant. Purified Hos2 protein consistently deacetylated tubulins, rather than histones from TSA-treated cells. Hos2 has been reported to be a putative NAD+ dependent histone deacetylase, a feature of sirtuins. We assayed for sirtuin activation with resveratrol and purified Hos2 protein and did not find any sirtuin activity. PMID:25110576
Objective Pathological Voice Quality Assessment Based on HOS Features
NASA Astrophysics Data System (ADS)
Lee, Ji-Yeoun; Jeong, Sangbae; Choi, Hong-Shik; Hahn, Minsoo
This work proposes new features to improve the pathological voice quality classification performance. They are the means, the variances, and the perturbations of the higher-order statistics (HOS) such as the skewness and the kurtosis. The HOS-based features show meaningful differences among normal, grade 1, grade 2, and grade 3 voices classified in the GRBAS scale. The jitter, the shimmer, the harmonic-to-noise ratio (HNR), and the variance of the short-time energy are utilized as the conventional features. The performances are measured by the classification and regression tree (CART) method. Specifically, the CART-based method by utilizing both the conventional features and the HOS-based ones shows its effectiveness in the pathological voice quality measurement, with the classification accuracy of 87.8%.
Genetic heterogeneity in the cysA-fus region of the Bacillus subtilis chromosome: identification of the hos gene.
Matsuzaki, S; Kobayashi, Y
We identified a new gene, hos, which exerts different sporulation phenotypes in Bacillus subtilis strains with different genetic backgrounds. The hos+ gene showed normal sporulation in the genetic background of JH642 but showed temperature-sensitive sporulation in that of the Tano-oka W. The hos gene was mapped between cysA and rpoB. PMID:3922952
HOS1-mediated activation of FLC via chromatin remodeling under cold stress.
Jung, Jae-Hoon; Park, Chung-Mo
The Arabidopsis E3 ubiquitin ligase HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE 1 (HOS1) has been shown to act as a negative regulator of cold responses by degrading the INDUCER OF CBF EXPRESSION 1 (ICE1) transcription factor through the ubiquitin/proteasome pathway. Notably, loss-of-function hos1 mutants exhibit early flowering, and the transcript level of the floral repressor FLOWERING LOCUS C (FLC) is downregulated in the mutants. However, it is largely unknown how HOS1 regulates FLC transcription. We found that HOS1 activates FLC transcription by inhibiting the activity of histone deacetylase 6 (HDA6) under cold stress. Cold temperatures induce the binding of HOS1 to FLC chromatin in an FVE-dependent manner. Cold-activated HOS1 promotes the dissociation of HDA6 from FLC chromatin, and the cold effects disappear in both hos1 and fve mutants. It is therefore clear that HOS1 regulates FLC transcription via chromatin remodeling, providing new insights into the signaling crosstalks between cold response and flowering time control. PMID:24390058
HOS1, a genetic locus involved in cold-responsive gene expression in arabidopsis.
Ishitani, M; Xiong, L; Lee, H; Stevenson, B; Zhu, J K
Low-temperature stress induces the expression of a variety of genes in plants. However, the signal transduction pathway(s) that activates gene expression under cold stress is poorly understood. Mutants defective in cold signaling should facilitate molecular analysis of plant responses to low temperature and eventually lead to the identification and cloning of a cold stress receptor(s) and intracellular signaling components. In this study, we characterize a plant mutant affected in its response to low temperatures. The Arabidopsis hos1-1 mutation identified by luciferase imaging causes superinduction of cold-responsive genes, such as RD29A, COR47, COR15A, KIN1, and ADH. Although these genes are also induced by abscisic acid, high salt, or polyethylene glycol in addition to cold, the hos1-1 mutation only enhances their expression under cold stress. Genetic analysis revealed that hos1-1 is a single recessive mutation in a nuclear gene. Our studies using the firefly luciferase reporter gene under the control of the cold-responsive RD29A promoter have indicated that cold-responsive genes can be induced by temperatures as high as 19 degrees C in hos1-1 plants. In contrast, wild-type plants do not express the luciferase reporter at 10 degrees C or higher. Compared with the wild type, hos1-1 plants are l ess cold hardy. Nonetheless, after 2 days of cold acclimation, hos1-1 plants acquired the same degree of freezing tolerance as did the wild type. The hos1-1 plants flowered earlier than did the wild-type plants and appeared constitutively vernalized. Taken together, our findings show that the HOS1 locus is an important negative regulator of cold signal transduction in plant cells and that it plays critical roles in controlling gene expression under cold stress, freezing tolerance, and flowering time. PMID:9668134
HOS1 Regulates Argonaute1 by Promoting the Transcription of the MicroRNA Gene MIR168b in Arabidopsis
Wang, Bangshing; Duan, Cheng-Guo; Wang, Xingang; Hou, Yueh-Ju; Yan, Jun; Gao, Caiqiu; Kim, Jin-Hong; Zhang, Huiming; Zhu, Jian-Kang
Summary Proper accumulation and function of miRNAs is essential for plant growth and development. While core components of the miRNA biogenesis pathway and miRNA-induced silencing complex have been well characterized, cellular regulators of miRNAs remain to be fully explored. Here we report that HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENES1 (HOS1) is a regulator of an important miRNA, mi168a/b, that targets the ARGONAUTE 1 (AGO1) gene in Arabidopsis. HOS1 functions to regulate plant cold stress responses as an ubiquitin E3 ligase, associate with the nuclear pores to regulate mRNA export and circadian clock and flowering time by binding to chromatin of the flowering regulator FLC. We isolated a loss-of-function Arabidopsis mutant of HOS1 in a genetic screen for enhancers of sic-1, which is defective in miRNA biogenesis. Like other hos1 mutant alleles, the new hos1-7 mutant allele flowered early and was smaller in stature than the wild type. Dysfunction in HOS1 reduced the abundance of miR168a/b but not of other miRNAs. In hos1 mutants, pri-MIR168b and pre-MIR168b levels were decreased, and RNA polymerase II occupancy was reduced at the promoter of MIR168b but not MIR168a. Chromatin immunoprecipitation assays revealed HOS1 protein is enriched at the chromatin of the MIR168b promoter. The reduced miR168a/b level in hos1 mutants results in an increase in the mRNA and protein levels of its target gene, AGO1. Our results reveal that HOS1 regulates miR168a/b and AGO1 levels in Arabidopsis by maintaining proper transcription of MIR168b. PMID:25619693
HOS-ocean: Open-source solver for nonlinear waves in open ocean based on High-Order Spectral method
NASA Astrophysics Data System (ADS)
Ducrozet, Guillaume; Bonnefoy, Félicien; Le Touzé, David; Ferrant, Pierre
HOS-ocean is an efficient High-Order Spectral code developed to solve the deterministic propagation of nonlinear wavefields in open ocean. HOS-ocean is released as open-source, developed and distributed under the terms of GNU General Public License (GPLv3). Along with the source code, a documentation under wiki format is available which makes easy the compilation and execution of the source files. The code has been shown to be accurate and efficient.
HOS cell adhesion on Ti6Al4V surfaces texturized by laser engraving
NASA Astrophysics Data System (ADS)
Sandoval Amador, A.; Carreño Garcia, H.; Escobar Rivero, P.; Peña Ballesteros, D. Y.; Estupiñán Duran, H. A.
The cell adhesion of the implant is determinate by the chemical composition, topography, wettability, surface energy and biocompatibility of the biomaterial. In this work the interaction between human osteosarcoma HOS cells and textured Ti6Al4V surfaces were evaluated. Ti6Al4V surfaces were textured using a CO2 laser in order to obtain circular spots on the surfaces. Test surfaces were uncoated (C1) used as a control surface, and surfaces with points obtained by laser engraving, with 1mm spacing (C2) and 0.5mm (C3). The HOS cells were cultured in RPMI-1640 medium with 10% fetal bovine serum and 1% antibiotics. No cells toxicity after one month incubation time occurred. The increased cell adhesion and cell spreading was observed after 1, 3 and 5 days without significant differences between the sample surfaces (C2 and C3) and control (uncoated) at the end of the experiment.
A theoretical investigation of HSO/HOS and their positive ions
Plummer, P.L.M. Department of Physics, University of Missouri-Columbia, Columbia, MO )
The formation and eventual fate of sulfur-containing aerosols play a central role in global pollution. An understanding of the oxidation paths for sulfur species and of the formation and stability of radical and ionic intermediates is required for optimum control of acid deposition. To gain insight into these processes {ital ab} {ital initio} calculations were performed for ground and first excited electronic states of the isomers HSO/HOS and for the ground and first two excited states for their positive ions, HSO{sup +}/HOS{sup +}. A variety of basis sets were used for calculations at the self-consistent field (SCF) level. (Full details available in supplementary material for this article.) These calculations included full optimization of the geometry and examination of the potential surface for transition states. Post Hartree--Fock calculations were performed and included Moller--Plesset calculations through fourth order with quadruple excitations as well as configuration interaction including all double excitations. The effect of geometry optimization at the MP4 level and the effect of freezing the core orbitals in the calculations were examined. Five transition states for the isomerization of HSO to HOS were located for the ground and excited electronic states of the neutral and ion systems. For all transition states in both systems, the geometry resembles an isosceles triangle. The barriers to rearrangement are quite high, ranging from {similar to}20 to {similar to}60 kcal/mol depending on the state. Two of the barriers were approximately symmetric while three were asymmetric with a lower barrier for HSO to HOS isomerization.
Anticancer Activity Studies of Ruthenium(II) Complex Toward Human Osteosarcoma HOS Cells.
Zhu, Jian-Wei; Liu, Si-Hong; Zhang, Gui-Qiang; Xu, Hui-Hua; Wang, Yu-Xuan; Wu, Yong; Liu, Ya-Min; Wang, Yan; Liang, Jun-Bo; Guo, Qi-Feng
A new Ru(II) complex [Ru(dmp)2(NMIP)](ClO4)2 (dmp = 2,9-dimethyl-1,10-phenanthroline, NMIP = 2'-(2″-nitro-3″,4″-methylenedioxyphenyl)imidazo[4',5'-f][1,10]-phenanthroline) was synthesized and characterized by elemental analysis, ESI-MS and (1)H NMR. The cytotoxic activity of the complex against MG-63, U2OS, HOS, and MC3T3-e1 cell lines was investigated by MTT method. The complex shows moderate cytotoxicity toward HOS (IC50 = 35.6 ± 2.6 µM) and MC3T3-e1 (IC50 = 41.6 ± 2.8 µM) cell lines. The morphological studies show that the complex can induce apoptosis in HOS cells and cause an increase of reactive oxygen species levels and a decrease in the mitochondrial membrane potential. The cell cycle distribution demonstrates that the complex inhibits the cell growth at S phase. Additionally, the antitumor activity in vivo reveals that the complex can induce a decrease in tumor weight. PMID:27007877
The Hos2 Histone Deacetylase Controls Ustilago maydis Virulence through Direct Regulation of Mating-Type Genes
Elías-Villalobos, Alberto; Fernández-Álvarez, Alfonso; Moreno-Sánchez, Ismael; Helmlinger, Dominique; Ibeas, José I.
Morphological changes are critical for host colonisation in plant pathogenic fungi. These changes occur at specific stages of their pathogenic cycle in response to environmental signals and are mediated by transcription factors, which act as master regulators. Histone deacetylases (HDACs) play crucial roles in regulating gene expression, for example by locally modulating the accessibility of chromatin to transcriptional regulators. It has been reported that HDACs play important roles in the virulence of plant fungi. However, the specific environment-sensing pathways that control fungal virulence via HDACs remain poorly characterised. Here we address this question using the maize pathogen Ustilago maydis. We find that the HDAC Hos2 is required for the dimorphic switch and pathogenic development in U. maydis. The deletion of hos2 abolishes the cAMP-dependent expression of mating type genes. Moreover, ChIP experiments detect Hos2 binding to the gene bodies of mating-type genes, which increases in proportion to their expression level following cAMP addition. These observations suggest that Hos2 acts as a downstream component of the cAMP-PKA pathway to control the expression of mating-type genes. Interestingly, we found that Clr3, another HDAC present in U. maydis, also contributes to the cAMP-dependent regulation of mating-type gene expression, demonstrating that Hos2 is not the only HDAC involved in this control system. Overall, our results provide new insights into the role of HDACs in fungal phytopathogenesis. PMID:26317403
Higher Order Suppressor (HOS) for the PolLux Microspectroscope Beamline at the Swiss Light Source SLS
Frommherz, U.; Stefani, R.; Ellenberger, U.; Raabe, J.; Watts, B.
The mechanical design and performance of a device to suppress higher orders of a spherical grating monochromator at a constant deviation angle is described. The higher order suppressor (HOS) is used for a scanning transmission x-ray microspectroscope beamline (PolLux) at a bending magnet of the Swiss Light Source (SLS). The instruments allow microspectroscopy in polymer science, of biological samples in the water window as well as the study of magnetic materials with circular or linear polarized light in a photon energy range of 200 eV to 1400 eV. The HOS uses three mirrors acting as a low pass filter for soft x-rays to improve the absorption spectroscopy of carbon, oxygen and nitrogen 1s core levels. The successful installation and operation of the HOS located after the monochromator is reported. First results obtained using samples from materials research and environmental sciences exemplify the improved spectroscopy capabilities of the instrument.
Higher Order Suppressor (HOS) for the PolLux Microspectroscope Beamline at the Swiss Light Source SLS
NASA Astrophysics Data System (ADS)
Frommherz, U.; Raabe, J.; Watts, B.; Stefani, R.; Ellenberger, U.
The mechanical design and performance of a device to suppress higher orders of a spherical grating monochromator at a constant deviation angle is described. The higher order suppressor (HOS) is used for a scanning transmission x-ray microspectroscope beamline (PolLux) at a bending magnet of the Swiss Light Source (SLS). The instruments allow microspectroscopy in polymer science, of biological samples in the water window as well as the study of magnetic materials with circular or linear polarized light in a photon energy range of 200 eV to 1400 eV. The HOS uses three mirrors acting as a low pass filter for soft x-rays to improve the absorption spectroscopy of carbon, oxygen and nitrogen 1s core levels. The successful installation and operation of the HOS located after the monochromator is reported. First results obtained using samples from materials research and environmental sciences exemplify the improved spectroscopy capabilities of the instrument.
The negative regulator of plant cold responses, HOS1, is a RING E3 ligase that mediates the ubiquitination and degradation of ICE1.
Dong, Chun-Hai; Agarwal, Manu; Zhang, Yiyue; Xie, Qi; Zhu, Jian-Kang
Plant responses to cold stress are mediated by a transcriptional cascade, in which the transcription factor ICE1 and possibly related proteins activate the expression of C-repeat (CRT)-binding factors (CBFs), leading to the transcription of downstream effector genes. The variant RING finger protein high expression of osmotically responsive gene (HOS)1 was identified genetically as a negative regulator of cold responses. We present evidence here that HOS1 is an E3 ligase required for the ubiquitination of ICE1. HOS1 physically interacts with ICE1 and mediates the ubiquitination of ICE1 both in vitro and in vivo. We found that cold induces the degradation of ICE1 in plants, and this degradation requires HOS1. Consistent with enhanced cold-responsive gene expression in loss-of-function hos1 mutant plants, overexpression of HOS1 represses the expression of CBFs and their downstream genes and confers increased sensitivity to freezing stress. Our results indicate that cold stress responses in Arabidopsis are attenuated by a ubiquitination/proteasome pathway in which HOS1 mediates the degradation of the ICE1 protein. PMID:16702557
HosA, a MarR Family Transcriptional Regulator, Represses Nonoxidative Hydroxyarylic Acid Decarboxylase Operon and Is Modulated by 4-Hydroxybenzoic Acid.
Roy, Ajit; Ranjan, Akash
Members of the Multiple antibiotic resistance Regulator (MarR) family of DNA binding proteins regulate transcription of a wide array of genes required for virulence and pathogenicity of bacteria. The present study reports the molecular characterization of HosA (Homologue of SlyA), a MarR protein, with respect to its target gene, DNA recognition motif, and nature of its ligand. Through a comparative genomics approach, we demonstrate that hosA is in synteny with nonoxidative hydroxyarylic acid decarboxylase (HAD) operon and is present exclusively within the mutS-rpoS polymorphic region in nine different genera of Enterobacteriaceae family. Using molecular biology and biochemical approach, we demonstrate that HosA binds to a palindromic sequence downstream to the transcription start site of divergently transcribed nonoxidative HAD operon and represses its expression. Furthermore, in silico analysis showed that the recognition motif for HosA is highly conserved in the upstream region of divergently transcribed operon in different genera of Enterobacteriaceae family. A systematic chemical search for the physiological ligand revealed that 4-hydroxybenzoic acid (4-HBA) interacts with HosA and derepresses HosA mediated repression of the nonoxidative HAD operon. Based on our study, we propose a model for molecular mechanism underlying the regulation of nonoxidative HAD operon by HosA in Enterobacteriaceae family. PMID:26818787
Butyl benzyl phthalate suppresses the ATP-induced cell proliferation in human osteosarcoma HOS cells
Liu, P.-S.; Chen, C.-Y.
Butyl benzyl phthalate (BBP), an endocrine disruptor present in the environment, exerts its genomic effects via intracellular steroid receptors and elicits non-genomic effects by interfering with membrane ion-channel receptors. We previously found that BBP blocks the calcium signaling coupled with P2X receptors in PC12 cells (Liu and Chen, 2006). Osteoblast P2X receptors were recently reported to play a role in cell proliferation and bone remodeling. In this present study, the effects of BBP on ATP-induced responses were investigated in human osteosarcoma HOS cells. These receptors mRNA had been detected, named P2X4, P2X7, P2Y2, P2Y4, P2Y5, P2Y9, and P2Y11, in human osteosarcoma HOS cells by RT-PCR. The enhancement of cell proliferation and the decrease of cytoviability had both been shown to be coupled to stimulation via different concentrations of ATP. BBP suppressed the ATP-induced calcium influx (mainly coupled with P2X) and cell proliferation but not the ATP-induced intracellular calcium release (mainly coupled with P2Y) and cytotoxicity in human osteosarcoma HOS cells. Suramin, a common P2 receptor's antagonist, blocked the ATP-induced calcium signaling, cell proliferation, and cytotoxicity. We suggest that P2X is mainly responsible for cell proliferation, and P2Y might be partially responsible for the observed cytotoxicity. BBP suppressed the calcium signaling coupled with P2X, suppressing cell proliferation. Since the importance of P2X receptors during bone metastasis has recently become apparent, the possible toxic risk of environmental BBP during bone remodeling is a public problem of concern.
77 FR 40142 - Applications for Exemption: Commercial Driver's License (CDL) and Hours-of-Service (HOS) of Drivers
Federal Register 2010, 2011, 2012, 2013, 2014
... motor carriers on behalf of their commercial motor vehicle (CMV) drivers. The Agency reviewed each... maintain a record of duty status (RODS) on board the CMV they are operating (49 CFR 395.8). However, operators of ``utility service vehicles'' (USVs) are exempt from all the HOS rules , including the...
Red Light-Mediated Degradation of CONSTANS by the E3 Ubiquitin Ligase HOS1 Regulates Photoperiodic Flowering in Arabidopsis
Lazaro, Ana; Mouriz, Alfonso; Piñeiro, Manuel; Jarillo, José A.
The regulation of CONSTANS (CO) gene expression is crucial to accurately measure changes in daylength, which influences flowering time in Arabidopsis thaliana. CO expression is under both transcriptional and posttranslational control mechanisms. We previously showed that the E3 ubiquitin ligase HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENES1 (HOS1) physically interacts with CO in Arabidopsis. This interaction is required to precisely modulate the timing of CO accumulation and, consequently, to maintain low levels of FLOWERING LOCUS T expression during the first part of the day. The data presented here demonstrate that HOS1 is involved in the red light-mediated degradation of CO that takes place in the early stages of the daylight period. Our results show that phytochrome B (phyB) is able to regulate flowering time, acting in the phloem companion cells, as previously described for CO and HOS1. Moreover, we reveal that phyB physically interacts with HOS1 and CO, indicating that the three proteins may be present in a complex in planta that is required to coordinate a correct photoperiodic response in Arabidopsis. PMID:26373454
Expression, Functional Characterization and X-ray Analysis of HosA, A Member of MarR Family of Transcription Regulator from Uropathogenic Escherichia coli.
Roy, Ajit; Reddi, Ravikumar; Sawhney, Bhavik; Ghosh, Debasish Kumar; Addlagatta, Anthony; Ranjan, Akash
Regulators belonging to multiple antibiotic resistance regulator (MarR) family are widespread in prokaryotes and are involved in regulation of genes that are responsible for virulence and pathogenicity in most of the clinically important pathogens. Here we report the transcriptional, biophysical, and X-ray analyses of homologue of SlyA (HosA), a member of MarR family that is predominantly present in the pathogenic strains of Enterobacteriaceae family. The initiation of hosA transcription was observed to occur at two independent start sites and subsequent binding study has revealed that the purified HosA interacts with its upstream region suggesting a probable autoregulation. The secondary structure analysis through circular dichroism spectroscopy demonstrated that HosA is predominantly composed of the alpha helix with higher thermal stability. To further understand the three-dimensional structure, HosA was crystallized and the crystals were diffracted to maximum of 2.9 Ǻ on exposure to X-rays. Analysis of the X-ray crystallographic data suggested a primitive space group (P 6 ? 2 2), with unit cell parameters a = b = 64.19 Å and c = 244.25 Å. The solvent content and Matthews coefficient were 41 % and 2.11 Å(3) Da(-1), respectively, which indicated the existence of two molecules of HosA in the asymmetric unit of crystal. PMID:27325406
HOS3, an ELO-Like Gene, Inhibits Effects of ABA and Implicates a S-1-P/Ceramide Control System for Abiotic Stress Responses in Arabidopsis thaliana
Quist, Tanya M.; Sokolchik, Irina; Shi, Huazhong; Joly, Robert J.; Bressan, Ray A.; Maggio, Albino; Narsimhan, Meena; Li, Xia
A hyper-osmotically sensitive mutant of Arabidopsis thaliana, designated hos3-1 (high expression of osmotically responsive genes), was identified based on its hyper-luminescence of RD29A:LUC promoter fusion plants upon treatment with NaCl and ABA. These responses implicate the disrupted gene as a direct or indirect negative regulator of the RD29A stress-responsive pathway. By sequencing the flanking regions of the T-DNA borders, it was determined that the disrupted gene is at locus At4g36830, annotated as encoding a putative protein with high homology to CIG30 (ELO2/FEN1). CIG30 has been implicated in synthesis of very long chain fatty acids (VLCFA), which are essential precursors for sphingolipids and ceramides. Altered stress responses characteristic of ABA-hypersensitivity, including reduced root growth inhibition and reduced germination with ABA treatment and reduced water loss from leaves, were exhibited by allelic hos3-1 and hos3-2 mutants. The hos3-2 mutant is partially suppressed in its transcript abundance and is inherited as a recessive trait. Further, the HOS3 ORF under the control of the 35SCaMV promoter restored wild-type NaCl- and ABA-root growth sensitivity as well as RD29A:LUC luminescence in mutant plants. We also show here that the HOS3 wild-type gene functionally complements the sensitivity of elo2 and elo3 yeast mutants to monensin. Furthermore, both hos3-1 and hos3-2 alleles shared increased sensitivity to the herbicide Metolachlor, which inhibits acyl chain elongation in synthesis of VLCFA, and HOS3 functionally complemented both elo2 and elo3 and restored levels of VLCFA. Together, these data establish that HOS3 inhibits ABA-mediated stress responses and implicate the VLCFA pathway and products as control points for several aspects of abiotic stress signaling and responses. The results also provide support for a role of ceramide in the control of stomatal behavior. PMID:19529829
Development and Validation of Written Exam Items for the Agricultural Hazardous Occupations Orders (AgHOs) Certification Training Program.
Mann, A J; Field, W E; Tormoehlen, R; French, B F
Research was conducted to develop and validate a pool of exam items that can be used to test the readiness of youth, ages 14-15 years, to be certified under the current federally mandated Agricultural Hazardous Occupations Orders (AgHOs). The AgHOs require training prior to employment in agricultural workplaces that the Secretary of Labor has determined are especially hazardous for youth within the prescribed age range. Under the current provisions of the AgHOs certification process, non-exempt youth seeking employment in agriculture are required to pass a written exam concentrating on safe work practices as partial satisfaction to receive certification of eligibility for employment to perform certain tasks. However, the regulations provide little guidance concerning the format of the exam, subject matter to be covered, degree of difficulty, or minimum passing score. As part of the USDA-sponsored Hazardous Occupations Safety Training in Agriculture (HOSTA) initiative, efforts have been made to develop consistent and evidence-based testing methods for disseminating the test protocols to instructors. The goal was to expand, enhance, and maintain the reliability of the exam item pool for the AgHOs certification process. Item development was based on the HOSTA-supported Gearing Up for Safety: Production Agriculture Safety Training for Youth curriculum. To ensure adequate item availability, the current item pool was expanded to include a minimum of two test items for each of the 157 cognitive-based core competencies developed as part of the Gearing Up curriculum design process. Administering 70-item exams that were generated from the item pool to 568 youth, ages 13-18 years, provided evidence of item validity. The result was a pool of 367 validated exam items. PMID:27373061
Rice root curling, a response to mechanosensing, is modulated by the rice E3-ubiquitin ligase HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE1 (OsHOS1).
Lourenço, T F; Serra, T S; Cordeiro, A M; Swanson, S J; Gilroy, S; Saibo, N J M; Oliveira, M M
Plant development depends on the perception of external cues, such as light, gravity, touch, wind or nutrients, among others. Nevertheless, little is known regarding signal transduction pathways integrating these stimuli. Recently, we have reported the involvement of a rice E3-ubiquitin ligase (OsHOS1, HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE1), previously associated with abiotic stress response, in root responses to mechanical stimuli. We showed that OsHOS1 is involved in the regulation of root curling after mechanosensing and that RNAi::OsHOS1 plants failed to exhibit the root curling phenotype observed in WT. Interestingly, the straight root phenotype of these transgenics correlated with the up-regulation of rice ROOT MEANDER CURLING (OsRMC, a negative regulator of rice root curling) and was reverted by the exogenous application of jasmonic acid. Altogether, our results highlight the role of the proteasome modulating plant responses to mechanical stimuli and suggest that OsHOS1 is a hub integrating environmental and hormonal signaling into plant growth and development. PMID:27467198
Computer-aided diagnosis of Myocardial Infarction using ultrasound images with DWT, GLCM and HOS methods: A comparative study.
Vidya, K Sudarshan; Ng, E Y K; Acharya, U Rajendra; Chou, Siaw Meng; Tan, Ru San; Ghista, Dhanjoo N
Myocardial Infarction (MI) or acute MI (AMI) is one of the leading causes of death worldwide. Precise and timely identification of MI and extent of muscle damage helps in early treatment and reduction in the time taken for further tests. MI diagnosis using 2D echocardiography is prone to inter-/intra-observer variability in the assessment. Therefore, a computerised scheme based on image processing and artificial intelligent techniques can reduce the workload of clinicians and improve the diagnosis accuracy. A Computer-Aided Diagnosis (CAD) of infarcted and normal ultrasound images will be useful for clinicians. In this study, the performance of CAD approach using Discrete Wavelet Transform (DWT), second order statistics calculated from Gray-Level Co-Occurrence Matrix (GLCM) and Higher-Order Spectra (HOS) texture descriptors are compared. The proposed system is validated using 400 MI and 400 normal ultrasound images, obtained from 80 patients with MI and 80 normal subjects. The extracted features are ranked based on t-value and fed to the Support Vector Machine (SVM) classifier to obtain the best performance using minimum number of features. The features extracted from DWT coefficients obtained an accuracy of 99.5%, sensitivity of 99.75% and specificity of 99.25%; GLCM have achieved an accuracy of 85.75%, sensitivity of 90.25% and specificity of 81.25%; and HOS obtained an accuracy of 93.0%, sensitivity of 94.75% and specificity of 91.25%. Among the three techniques presented DWT yielded the highest classification accuracy. Thus, the proposed CAD approach may be used as a complementary tool to assist cardiologists in making a more accurate diagnosis for the presence of MI. PMID:25912990
Recycling of Volatiles in Subduction Zones: Evidence from H-O-S Isotopic Signatures of Indonesian Arc Basalts
NASA Astrophysics Data System (ADS)
de Hoog, J. C.; Taylor, B. E.; van Bergen, M. J.
Subaerial lavas from eight volcanoes, situated in different sectors of the Indonesian arc system, have been analyzed for hydrogen, oxygen and sulfur isotopic compositions. The lavas are mostly basaltic and cover a range from low-K tholeiitic to high-K calc-alkaline affinities. The following whole-rock compositions were observed: δ D = -109 to -57‰ V-SMOW (avg = -89‰ ), δ 18O = +5.3 to +6.7‰ V-SMOW (avg = +5.9‰ ), δ 34S = +2.0 to +7.8‰ V-CDT (avg = +4.7‰ ). All samples are strongly degassed, as average water contents are ~0.15 wt.% and average sulfur concentrations are ~14 ppm. Because degassing-induced shifts in sulfur isotopes are limited at magmatic temperatures and oxidation states typical of basalts, the data suggest that, in all cases, the magma source in the sub-arc mantle is significantly enriched in 34S compared to MORB-source mantle. Hydrogen isotopes fractionate strongly, and our data are most consistent with multi-stage degassing of magmatic water. Based on primary H-O-S isotopic signatures inferred for the Indonesian arc basalts and on assumed compositions of the pre-subduction mantle, we performed mass-balance calculations to estimate volatile concentrations in the slab component. The results indicate that addition of about 1-1.5 wt.% aqueous fluid containing 1.5% sulfur will produce the H-O-S isotopic compositions of primary basaltic melts of the volcanoes studied. Alternatively, the slab-derived material may consist of a mix of melts and fluids, in which melts can be more voluminous, but fluids dominate the budgets of sulfur, hydrogen, and other hydrophile elements. Irrespective of the preferred model, S and H2O contents must have increased considerably in the sub-arc mantle (to ~350 ppm and ~1.3 wt.%, respectively). The S value is consistent with estimates based on primary sulfur contents in arc basalts ( ~2000 ppm), but the H2O concentration is higher than the mantle can accommodate, as it will induce melting. We argue, however, that
Computer-aided diagnosis of psoriasis skin images with HOS, texture and color features: A first comparative study of its kind.
Shrivastava, Vimal K; Londhe, Narendra D; Sonawane, Rajendra S; Suri, Jasjit S
Psoriasis is an autoimmune skin disease with red and scaly plaques on skin and affecting about 125 million people worldwide. Currently, dermatologist use visual and haptic methods for diagnosis the disease severity. This does not help them in stratification and risk assessment of the lesion stage and grade. Further, current methods add complexity during monitoring and follow-up phase. The current diagnostic tools lead to subjectivity in decision making and are unreliable and laborious. This paper presents a first comparative performance study of its kind using principal component analysis (PCA) based CADx system for psoriasis risk stratification and image classification utilizing: (i) 11 higher order spectra (HOS) features, (ii) 60 texture features, and (iii) 86 color feature sets and their seven combinations. Aggregate 540 image samples (270 healthy and 270 diseased) from 30 psoriasis patients of Indian ethnic origin are used in our database. Machine learning using PCA is used for dominant feature selection which is then fed to support vector machine classifier (SVM) to obtain optimized performance. Three different protocols are implemented using three kinds of feature sets. Reliability index of the CADx is computed. Among all feature combinations, the CADx system shows optimal performance of 100% accuracy, 100% sensitivity and specificity, when all three sets of feature are combined. Further, our experimental result with increasing data size shows that all feature combinations yield high reliability index throughout the PCA-cutoffs except color feature set and combination of color and texture feature sets. HOS features are powerful in psoriasis disease classification and stratification. Even though, independently, all three set of features HOS, texture, and color perform competitively, but when combined, the machine learning system performs the best. The system is fully automated, reliable and accurate. PMID:26830378
Overexpression of an Orchid (Dendrobium nobile) SOC1/TM3-Like Ortholog, DnAGL19, in Arabidopsis Regulates HOS1-FT Expression.
Liu, Xiao-Ru; Pan, Ting; Liang, Wei-Qi; Gao, Lan; Wang, Xiao-Jing; Li, Hong-Qing; Liang, Shan
Flowering in the appropriate season is critical for successful reproduction in angiosperms. The orchid species, Dendrobium nobile, requires vernalization to achieve flowering in the spring, but the underlying regulatory network has not been identified to date. The MADS-box transcription factor DnAGL19 was previously identified in a study of low-temperature treated D. nobile buds and was suggested to regulate vernalization-induced flowering. In this study, phylogenetic analysis of DnAGL9 and the MADS-box containing proteins showed that DnAGL19 is phylogenetically closely related to the SOC1-like protein from orchid Dendrobium Chao Parya Smile, DOSOC1. The orchid clade closed to but is not included into the SOC1-1/TM3 clades associated with either eudicots or monocots, suggesting that DnAGL19 is an SOC1-1/TM3-like ortholog. DnAGL19 was found to be highly expressed in pseudobulbs, leaves, roots, and axillary buds but rarely in flowers, and to be substantially upregulated in axillary buds by prolonged low-temperature treatments. Overexpression of DnAGL19 in Arabidopsis thaliana resulted in a small but significantly reduced time to bolting, suggesting that flowering time was slightly accelerated under normal growth conditions. Consistent with this, the A. thaliana APETELA1 (AP1) gene was expressed at an earlier stage in transgenic lines than in wild type plants, while the FLOWERING LOCUS T (FT) gene was suppressed, suggesting that altered regulations on these transcription factors caused the weak promotion of flowering. HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE 1 (HOS1) was slightly activated under the same conditions, suggesting that the HOS1-FT module may be involved in the DnAGL19-related network. Under vernalization conditions, FT expression was significantly upregulated, whereas HOS1 expression in the transgenic A. thaliana has a level similar to that in wild type. Taken together, these results suggest that DnAGL19 controls the action of the HOS1-FT module
Overexpression of an Orchid (Dendrobium nobile) SOC1/TM3-Like Ortholog, DnAGL19, in Arabidopsis Regulates HOS1-FT Expression
Liu, Xiao-Ru; Pan, Ting; Liang, Wei-Qi; Gao, Lan; Wang, Xiao-Jing; Li, Hong-Qing; Liang, Shan
Flowering in the appropriate season is critical for successful reproduction in angiosperms. The orchid species, Dendrobium nobile, requires vernalization to achieve flowering in the spring, but the underlying regulatory network has not been identified to date. The MADS-box transcription factor DnAGL19 was previously identified in a study of low-temperature treated D. nobile buds and was suggested to regulate vernalization-induced flowering. In this study, phylogenetic analysis of DnAGL9 and the MADS-box containing proteins showed that DnAGL19 is phylogenetically closely related to the SOC1-like protein from orchid Dendrobium Chao Parya Smile, DOSOC1. The orchid clade closed to but is not included into the SOC1-1/TM3 clades associated with either eudicots or monocots, suggesting that DnAGL19 is an SOC1-1/TM3-like ortholog. DnAGL19 was found to be highly expressed in pseudobulbs, leaves, roots, and axillary buds but rarely in flowers, and to be substantially upregulated in axillary buds by prolonged low-temperature treatments. Overexpression of DnAGL19 in Arabidopsis thaliana resulted in a small but significantly reduced time to bolting, suggesting that flowering time was slightly accelerated under normal growth conditions. Consistent with this, the A. thaliana APETELA1 (AP1) gene was expressed at an earlier stage in transgenic lines than in wild type plants, while the FLOWERING LOCUS T (FT) gene was suppressed, suggesting that altered regulations on these transcription factors caused the weak promotion of flowering. HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE 1 (HOS1) was slightly activated under the same conditions, suggesting that the HOS1-FT module may be involved in the DnAGL19-related network. Under vernalization conditions, FT expression was significantly upregulated, whereas HOS1 expression in the transgenic A. thaliana has a level similar to that in wild type. Taken together, these results suggest that DnAGL19 controls the action of the HOS1-FT module
CO Substitution in HOs3(CO)10(l-SC6H4Me-4) by the Diphosphine 4,5-Bis(diphenylphosphino)-4-cyclopentadiene-1,3-dione (bpcd): Structural and DFT Evaluation of the Isomeric Clusters HOs3(CO)8(bpcd)(mu-SC6H4Me-4)
Yang, Li; Nesterov, Vladimir; Wang, Xiaoping; Richmond, Michael G.
The reaction of the cluster HOs{sub 3}(CO){sub 10}({mu}-SC{sub 6}H{sub 4}Me-4) (1) with the diphosphine 4,5-bis(diphenylphosphino)-4-cyclopentadiene-1,3-dione (bpcd) has been investigated. 1 reacts with bpcd at room temperature in the presence of Me{sub 3}NO to give the isomeric clusters 1,2-HOs{sub 3}(CO)8(bpcd)({mu}-SC{sub 6}H{sub 4}Me-4) (2a) and 1,1-HOs{sub 3}(CO)8(bpcd)({mu}-SC{sub 6}H{sub 4}Me-4) (2b). Clusters 2a and 2b have been isolated, and the molecular structure of each compound has been established by X-ray crystallography. The X-ray structure of 2a confirms the coordination of one of the non-hydride-bridged Os-Os vectors by the bpcd ligand, while the structure of 2b exhibits a chelating bpcd ligand that is bound to one of the osmium centers ligated by the thiolate and hydrido ligands. 2a and 2b are stable in refluxing toluene and show no evidence for bridge-to-chelate isomerization of the ancillary bpcd ligand. DFT calculations on 2a and 2b indicate that the former cluster is the thermodynamically more stable isomer. Near-UV irradiation of 2b leads to CO loss and ortho metalation of the thiolate moiety, yielding the dihydride cluster H{sub 2}Os{sub 3}(CO)7(bpcd)({mu},{sigma}-SC{sub 6}H{sub 3}Me-4) (3). The conversion of 2b to 3 and free CO is computed to be endothermic by 14.1 kcal/mol and the reaction is driven by the entropic release of CO. The photochemically promoted ortho-metalation reaction is isomer dependent since cluster 2a is inert under identical conditions.
The Set3/Hos2 Histone Deacetylase Complex Attenuates cAMP/PKA Signaling to Regulate Morphogenesis and Virulence of Candida albicans
Hnisz, Denes; Majer, Olivia; Frohner, Ingrid E.; Komnenovic, Vukoslav; Kuchler, Karl
Candida albicans, like other pleiomorphic fungal pathogens, is able to undergo a reversible transition between single yeast-like cells and multicellular filaments. This morphogenetic process has long been considered as a key fungal virulence factor. Here, we identify the evolutionarily conserved Set3/Hos2 histone deacetylase complex (Set3C) as a crucial repressor of the yeast-to-filament transition. Cells lacking core components of the Set3C are able to maintain all developmental phases, but are hypersusceptible to filamentation-inducing signals, because of a hyperactive cAMP/Protein Kinase A signaling pathway. Strikingly, Set3C-mediated control of filamentation is required for virulence in vivo, since set3Δ/Δ cells display strongly attenuated virulence in a mouse model of systemic infection. Importantly, the inhibition of histone deacetylase activity by trichostatin A exclusively phenocopies the absence of a functional Set3C, but not of any other histone deacetylase gene. Hence, our work supports a paradigm for manipulating morphogenesis in C. albicans through alternative antifungal therapeutic strategies. PMID:20485517
TNF-alpha-dependent activation of NF-kappa B in human osteoblastic HOS-TE85 cells is repressed in vector-averaged gravity using clinostat rotation.
Kobayashi, K; Kambe, F; Kurokouchi, K; Sakai, T; Ishiguro, N; Iwata, H; Koga, K; Gruener, R; Seo, H
Effects of vector-averaged gravity on tumor necrosis factor (TNF)-alpha-dependent activation of nuclear factor kappa B (NF-kappa B) in human osteoblastic HOS-TE85 cells were investigated by culturing the cells using clinostat rotation (clinorotation). Cell cultures were rotated for 72 h at 40 rpm in a clinostat. At the end of clinorotation, the cells were treated with TNF-alpha for 30 min under stationary conditions. Electrophoretic mobility shift assays revealed that TNF-alpha-dependent activation of NF-kappa B was markedly reduced in the clinorotated cells when compared with the cells in control stationary cultures or after horizontal rotation (motional controls). The NF-kappa B-dependent transactivation was also impaired in the clinorotated cells, as evidenced by a transient transfection assay with a reporter plasmid containing multimerized NF-kappa B sites. Consistent with these findings, the TNF-alpha-dependent induction of endogenous NF-kappa B-responsive genes p105, I kappa B-alpha, and IL-8, was significantly attenuate in clinorotated cells. These results demonstrate that vector-averaged gravity inhibits the responsiveness of osteoblasts to TNF-alpha by repressing NF-kappa B activation. PMID:11112449
Isomerization of the diphosphine ligand 3,4-bis(diphenylphosphino)-5-methoxy-2(5H)-furanone (bmf) at a triosmium cluster and P C bond cleavage in the unsaturated cluster 1,1-Os3(CO)9(bmf): Synthesis and X-ray diffraction structures of the isomeric Os3(CO)10(bmf) clusters and HOs3(CO)8( -C6H4)[ -PhPCC(Ph2P)CH(OMe)OC(O)
Kandala, Srikanth; Yang, Li; Campana, Charles F.; Nesterov, Vladimir; Richmond, Michael G.
The labile cluster 1,2-Os3(CO)10(MeCN)2 (1) reacts with the chiral diphosphine ligand 3,4-bis(diphenylphosphino)-5-methoxy-2(5H)-furanone (bmf) to furnish 1,2-Os3(CO)10(bmf) (2a) in high yield. Heating cluster 2a over the temperature range 358 383 K under CO leads to isomerization of the bmf ligand and formation of the diphosphine-chelated cluster 1,1-Os3(CO)10(bmf) (2b) and an equilibrium mixture consisting of 2a and 2b in a 15:85 ratio. Extended thermolysis of an equilibrium mixture of Os3(CO)10(bmf) is accompanied by CO loss and ortho-metalation of an aryl ring to afford an inseparable mixture of three diastereomeric hydride clusters HOs3(CO)9(C29H23O3P2) (3a c). Thermolysis of HOs3(CO)9(C29H23O3P2) (3a c) in refluxing toluene leads to P C bond cleavage and formation of the benzyne-substituted clusters HOs3(CO)8( -C6H4)( -C23H19O3P2) (4a,b) as a 4:1 mixture of diastereomers. The unequivocal identity of the major benzyne-substituted cluster has been determined by X-ray diffraction analysis, where the activation of one of the phenyl groups situated to the furanone carbonyl group in the bmf ligand has been established. The isomerization and activation of the bmf ligand are contrasted with other Os3(CO)10(diphosphine) derivatives prepared by our groups.
New osmium cluster compounds containing the heterocyclic ligand 2,3-bis-(diphenylphosphino)quinoxaline (dppq): Ligand isomerization and crystal structures of dppq, the isomeric clusters Os3(CO)10(dppq), and HOs3(CO)9[ -2,3-PhP(h1-C6H4)(Ph2P)quinoxaline
Hunt, Sean W; Yang, Li; Wang, Xiaoping; Richmond, Michael G.
Treatment of the labile cluster 1,2-Os{sub 3}(CO){sub 10}(MeCN){sub 2} (1) with the diphosphine ligand 2,3-bis(diphenylphosphino)quinoxaline (dppq) at room temperature affords 1,2-Os{sub 3}(CO){sub 10}(dppq) (2b) as the kinetic product of ligand substitution in 84% yield. 2b isomerizes to the thermodynamically more stable dppq-chelated cluster 1,1-Os{sub 3}(CO){sub 10}(dppq) (2c) as the sole observable product under CO at temperatures below 358 K. The kinetics for the conversion of 2b {yields} 2c have been investigated by NMR spectroscopy in CDCl{sub 3} over the temperature range 323-353 K, and the reaction was found to exhibit a rate law that is first order in 2b. The calculated activation parameters [{Delta}H{sup {ne}} = 25.4(4) kcal/mol; {Delta}S{sup {ne}} = -3(1) eu] support an intramolecular isomerization scenario, one that involves the migration of phosphine and CO groups about the cluster polyhedron. The disposition of the dppq ligand in the isomeric Os{sub 3}(CO){sub 10}(dppq) clusters has been established by X-ray crystallography and {sup 31}P NMR spectroscopy. Photolysis of 2c at 366 nm leads to CO loss and ortho metalation of one of the aryl groups on the Ph{sub 2}P moiety to furnish the hydride cluster HOs{sub 3}(CO){sub 9}[{mu}-PhP({eta}{sup 1}-C{sub 6}H{sub 4})(Ph{sub 2}P)quinoxaline] (3). The isomerization behavior exhibited by 2b follows that of related diphosphine-substituted Os{sub 3} clusters prepared by us.
Materials Data on HOs3C10ClO10 (SG:2) by Materials Project
Kristin Persson
Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations
HOS network-based classification of power quality events via regression algorithms
NASA Astrophysics Data System (ADS)
Palomares Salas, José Carlos; González de la Rosa, Juan José; Sierra Fernández, José María; Pérez, Agustín Agüera
This work compares seven regression algorithms implemented in artificial neural networks (ANNs) supported by 14 power-quality features, which are based in higher-order statistics. Combining time and frequency domain estimators to deal with non-stationary measurement sequences, the final goal of the system is the implementation in the future smart grid to guarantee compatibility between all equipment connected. The principal results are based in spectral kurtosis measurements, which easily adapt to the impulsive nature of the power quality events. These results verify that the proposed technique is capable of offering interesting results for power quality (PQ) disturbance classification. The best results are obtained using radial basis networks, generalized regression, and multilayer perceptron, mainly due to the non-linear nature of data.
Atheromatic™: symptomatic vs. asymptomatic classification of carotid ultrasound plaque using a combination of HOS, DWT & texture.
Acharya, U Rajendra; Faust, Oliver; Sree, S Vinitha; Alvin, Ang Peng Chuan; Krishnamurthi, Ganapathy; Seabra, José C R; Sanches, João; Suri, Jasjit S
Quantitative characterization of carotid atherosclerosis and classification into either symptomatic or asymptomatic is crucial in terms of diagnosis and treatment planning for a range of cardiovascular diseases. This paper presents a computer-aided diagnosis (CAD) system (Atheromatic™, patented technology from Biomedical Technologies, Inc., CA, USA) which analyzes ultrasound images and classifies them into symptomatic and asymptomatic. The classification result is based on a combination of discrete wavelet transform, higher order spectra and textural features. In this study, we compare support vector machine (SVM) classifiers with different kernels. The classifier with a radial basis function (RBF) kernel achieved an accuracy of 91.7% as well as a sensitivity of 97%, and specificity of 80%. Encouraged by this result, we feel that these features can be used to identify the plaque tissue type. Therefore, we propose an integrated index, a unique number called symptomatic asymptomatic carotid index (SACI) to discriminate symptomatic and asymptomatic carotid ultrasound images. We hope this SACI can be used as an adjunct tool by the vascular surgeons for daily screening. PMID:22255336
76 FR 37876 - Hours of Service (HOS) of Drivers; Renewal of American Pyrotechnics Association (APA) Exemption...
Federal Register 2010, 2011, 2012, 2013, 2014
... published in the Federal Register on January 17, 2008 (73 FR 3316) or you may visit http://edocket.access..., FMCSA granted a limited exemption to 14 new APA-member motor carriers (74 FR 29266, June 19, 2009) and renewed 61exemptions of APA-member motor carriers (74 FR 29264, June 19, 2009) for their...
76 FR 30232 - Hours of Service (HOS) of Drivers; Application of American Pyrotechnics Association (APA) for...
Federal Register 2010, 2011, 2012, 2013, 2014
... review DOT's complete Privacy Act Statement in the Federal Register published on January, 17, 2008 (73 FR... FR 29264, June 19, 2009). The renewal would cover 54 of the 61 member motor carriers listed in the... 14 new APA- member motor carriers (74 FR 29266, June 10, 2009) and renewed 61 exemptions of...
Materials Data on HoS (SG:225) by Materials Project
Kristin Persson
Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations
76 FR 37880 - Hours of Service (HOS) of Drivers; Granting of Exemption; American Pyrotechnics Association (APA)
Federal Register 2010, 2011, 2012, 2013, 2014
.... In 2009, FMCSA granted the same limited exemption to 14 new APA-member motor carriers (74 FR 29266, June 19, 2009) and renewed 61exemptions of APA-member motor carriers (74 FR 29264, June 19, 2009) for..., FMCSA published a notice in the Federal Register (76 FR 30232) announcing APA's application...
Materials Data on HoS2 (SG:227) by Materials Project
Kristin Persson
Computed materials data using density functional theory calculations. These calculations determine the electronic structure of bulk materials by solving approximations to the Schrodinger equation. For more information, see https://materialsproject.org/docs/calculations
77 FR 38378 - Hours of Service (HOS) of Drivers; Revision of Exemption; American Pyrotechnics Association (APA)
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... carriers and added 9 new APA-member carriers (76 FR 37876 and 76 FR 37880, June 28, 2011) for their... published on June 28, 2011 (76 FR 37876-37882). Issued on: June 21, 2012. Larry W. Minor, Associate... Mandeville, LA 70471........ 548303 38. Rainbow Fireworks, Inc 76 Plum Ave Inman, KS 67546 1139643 39....
76 FR 16852 - Hours of Service (HOS) of Drivers; Assessing the Safety Impact of the Exemption From the 14-Hour...
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78 FR 26690 - Hours of Service (HOS) of Drivers; Application for Renewal and Expansion of American Pyrotechnics...
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Higher order software - A methodology for defining software
NASA Technical Reports Server (NTRS)
Hamilton, M.; Zeldin, S.
Higher order software (HOS) is concerned only with computable functions and relationships. The HOS methodology can be used for the definition of software for multiprogrammed, multiprocessor, or multicomputer systems. A description of HOS methodology is presented, giving attention to questions of formulation, interface correctness, specification language principles, and HOS analyzers. Aspects of system design are considered, and details of software management are discussed. Attention is given to modularity as defined by HOS, frozen module management, the assembly control supervisor, and aspects of reliability and efficiency.
The Cold Signaling Attenuator HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE1 Activates FLOWERING LOCUS C Transcription via Chromatin Remodeling under Short-Term Cold Stress in Arabidopsis[C][W
Jung, Jae-Hoon; Park, Ju-Hyung; Lee, Sangmin; To, Taiko Kim; Kim, Jong-Myong; Seki, Motoaki; Park, Chung-Mo
Exposure to short-term cold stress delays flowering by activating the floral repressor FLOWERING LOCUS C (FLC) in Arabidopsis thaliana. The cold signaling attenuator HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE1 (HOS1) negatively regulates cold responses. Notably, HOS1-deficient mutants exhibit early flowering, and FLC expression is suppressed in the mutants. However, it remains unknown how HOS1 regulates FLC expression. Here, we show that HOS1 induces FLC expression by antagonizing the actions of FVE and its interacting partner histone deacetylase 6 (HDA6) under short-term cold stress. HOS1 binds to FLC chromatin in an FVE-dependent manner, and FVE is essential for the HOS1-mediated activation of FLC transcription. HOS1 also interacts with HDA6 and inhibits the binding of HDA6 to FLC chromatin. Intermittent cold treatments induce FLC expression by activating HOS1, which attenuates the activity of HDA6 in silencing FLC chromatin, and the effects of intermittent cold are diminished in hos1 and fve mutants. These observations indicate that HOS1 acts as a chromatin remodeling factor for FLC regulation under short-term cold stress. PMID:24220632
High order software - A methodology for defining software
NASA Technical Reports Server (NTRS)
Hamilton, M.; Zeldin, S.
Higher order software (HOS) is a formal methodology for reliable systems specification and development. HOS is concerned only with computable functions and their relationships for any given system. Questions of methodology are considered, taking into account aspects of formulation meta-language principles, and HOS analyzers. Details of system design are discussed, giving attention to aspects of immediate self-control and indirect self-control. A description is given of the approaches used for software management.
Visual performance modeling in the human operator simulator
NASA Technical Reports Server (NTRS)
Strieb, M. I.
A brief description of the history of the development of the human operator simulator (HOS) model is presented. Features of the HOS micromodels that impact on the obtainment of visual performance data are discussed along with preliminary details on a HOS pilot model designed to predict the results of visual performance workload data obtained through oculometer studies on pilots in real and simulated approaches and landings.
HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENES1 Is Required for Circadian Periodicity through the Promotion of Nucleo-Cytoplasmic mRNA Export in Arabidopsis[W][OPEN
MacGregor, Dana R.; Gould, Peter; Foreman, Julia; Griffiths, Jayne; Bird, Susannah; Page, Rhiannon; Stewart, Kelly; Steel, Gavin; Young, Jack; Paszkiewicz, Konrad; Millar, Andrew J.; Halliday, Karen J.; Hall, Anthony J.; Penfield, Steven
Cold acclimation has been shown to be attenuated by the degradation of the INDUCER OF CBF EXPRESSION1 protein by the E3 ubiquitin ligase HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENES1 (HOS1). However, recent work has suggested that HOS1 may have a wider range of roles in plants than previously appreciated. Here, we show that hos1 mutants are affected in circadian clock function, exhibiting a long-period phenotype in a wide range of temperature and light environments. We demonstrate that hos1 mutants accumulate polyadenylated mRNA in the nucleus and that the circadian defect in hos1 is shared by multiple mutants with aberrant mRNA export, but not in a mutant attenuated in nucleo-cytoplasmic transport of microRNAs. As revealed by RNA sequencing, hos1 exhibits gross changes to the transcriptome with genes in multiple functional categories being affected. In addition, we show that hos1 and other previously described mutants with altered mRNA export affect cold signaling in a similar manner. Our data support a model in which altered mRNA export is important for the manifestation of hos1 circadian clock defects and suggest that HOS1 may indirectly affect cold signaling through disruption of the circadian clock. PMID:24254125
77 FR 47850 - Agency Information Collection Activities: Submission for OMB Review; Comment Request
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... notice published on April 27, 2012, (77 FR 25181). Subsequently, the HOS Questionnaire collection..., Physical Activity in Older Adults, Fall Risk Management, and Osteoporosis Testing in Older Women...
76 FR 81133 - Hours of Service of Drivers
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... notice of proposed rulemaking (NPRM) to revise the HOS rules (75 FR 82170). The Agency sought comment on... period to accept comments on four studies related to the HOS proposal (76 FR 26681). C. Final Rule... history of the earlier provisions (65 FR 25540) and the December 29, 2010, NPRM of this final rule for...
How In-Service Science Teachers Integrate History and Nature of Science in Elementary Science Courses
ERIC Educational Resources Information Center
Hacieminoglu, Esme
The purpose of this study is to investigate how the in-service science teachers' (IST) perceptions and practices about curriculum and integration of the history of science (HOS) and the nature of science (NOS) affect their science courses. For this aim, how ISTs integrated the NOS and HOS in their elementary science courses for understanding…
Using a Professional Development Program for Enhancing Chilean Biology Teachers' Understanding of Nature of Science (NOS) and Their Perceptions about Using History of Science to Teach NOS
ERIC Educational Resources Information Center
Pavez, José M.; Vergara, Claudia A.; Santibañez, David; Cofré, Hernán
A number of authors have recognized the importance of understanding the nature of science (NOS) for scientific literacy. Different instructional strategies such as decontextualized, hands-on inquiry, and history of science (HOS) activities have been proposed for teaching NOS. This article seeks to understand the contribution of HOS in enhancing…
Bilateral Scapulohumeral Ankylosis after Prolonged Mechanical Ventilation.
van Lotten, Manon L; Schreinemakers, J Rieneke; van Noort, Arthur; Rademakers, Maarten V
This case demonstrates a rarely reported bilateral scapulohumeral bony ankylosis. A young woman developed extensive heterotopic ossifications (HOs) in both shoulder joints after being mechanically ventilated for several months at the intensive care unit in a comatose status. She presented with a severe movement restriction of both shoulder joints. Surgical resection of the bony bridges was performed in 2 separate sessions with a significant improvement of shoulder function afterwards. No postoperative complications, pain, or recurrence of HOs were noted at 1-year follow-up. Mechanical ventilation, immobilization, neuromuscular blockage, and prolonged sedation are known risk factors for the development of HOs in the shoulder joints. Relatively early surgical resection of the HOs can be performed safely in contrary to earlier belief. Afterwards, nonsteroidal anti-inflammatory drugs and/or radiation therapy can be possible treatment modalities to prevent recurrence of HOs. PMID:27583120
Bilateral Scapulohumeral Ankylosis after Prolonged Mechanical Ventilation
Schreinemakers, J. Rieneke; van Noort, Arthur; Rademakers, Maarten V.
This case demonstrates a rarely reported bilateral scapulohumeral bony ankylosis. A young woman developed extensive heterotopic ossifications (HOs) in both shoulder joints after being mechanically ventilated for several months at the intensive care unit in a comatose status. She presented with a severe movement restriction of both shoulder joints. Surgical resection of the bony bridges was performed in 2 separate sessions with a significant improvement of shoulder function afterwards. No postoperative complications, pain, or recurrence of HOs were noted at 1-year follow-up. Mechanical ventilation, immobilization, neuromuscular blockage, and prolonged sedation are known risk factors for the development of HOs in the shoulder joints. Relatively early surgical resection of the HOs can be performed safely in contrary to earlier belief. Afterwards, nonsteroidal anti-inflammatory drugs and/or radiation therapy can be possible treatment modalities to prevent recurrence of HOs. PMID:27583120
Heme oxygenases from Arabidopsis thaliana reveal different mechanisms of carbon monoxide binding
NASA Astrophysics Data System (ADS)
Gisk, Björn; Molitor, Bastian; Frankenberg-Dinkel, Nicole; Kötting, Carsten
Heme oxygenases (HO) are widely distributed enzymes involved in the degradation of heme to biliverdin, carbon monoxide and Fe2+. The model plant Arabidopsis thaliana possesses three functional HOs (HY1, HO3 and HO4) which are thus far biochemically indistinguishable. Here, we investigate binding of the reaction product and putative inhibitor CO to these three HOs with various spectroscopic techniques: Nanosecond time-resolved absorption, millisecond time-resolved multi-wavelength absorption and Fourier-transform-infrared difference spectroscopy. Kinetics of CO rebinding were found to differ substantially among the HOs. At low CO concentrations a novel intermediate was identified for HO3 and HO4, substantially slowing down rebinding. All HOs show relatively slow geminate rebinding of CO indicating the existence of an additional transient binding niche for CO. The positions found for the IR absorptions of νCO and νFeC suggest a nonpolar distal binding site for all three HOs. The frequency of the νFeC vibration was calculated by a combination band on which we report here for the first time. Another band in the FTIR difference spectrum could be assigned to a histidine residue, probably the proximal ligand of the heme-iron. The observed different rebinding kinetics among the HOs could indicate adaptation of the HOs to different environments.
Heme oxygenases from Arabidopsis thaliana reveal different mechanisms of carbon monoxide binding.
Gisk, Björn; Molitor, Bastian; Frankenberg-Dinkel, Nicole; Kötting, Carsten
Heme oxygenases (HO) are widely distributed enzymes involved in the degradation of heme to biliverdin, carbon monoxide and Fe(2+). The model plant Arabidopsis thaliana possesses three functional HOs (HY1, HO3 and HO4) which are thus far biochemically indistinguishable. Here, we investigate binding of the reaction product and putative inhibitor CO to these three HOs with various spectroscopic techniques: Nanosecond time-resolved absorption, millisecond time-resolved multi-wavelength absorption and Fourier-transform-infrared difference spectroscopy. Kinetics of CO rebinding were found to differ substantially among the HOs. At low CO concentrations a novel intermediate was identified for HO3 and HO4, substantially slowing down rebinding. All HOs show relatively slow geminate rebinding of CO indicating the existence of an additional transient binding niche for CO. The positions found for the IR absorptions of ν(CO) and ν(FeC) suggest a nonpolar distal binding site for all three HOs. The frequency of the ν(FeC) vibration was calculated by a combination band on which we report here for the first time. Another band in the FTIR difference spectrum could be assigned to a histidine residue, probably the proximal ligand of the heme-iron. The observed different rebinding kinetics among the HOs could indicate adaptation of the HOs to different environments. PMID:22204880
A systematic review of studies performing the hypo-osmotic swelling test to evaluate the quality of canine spermatozoa.
Karger, S; Arlt, S; Haimerl, P; Heuwieser, W
The hypo-osmotic swelling test (HOS test) is a simple and inexpensive test to evaluate the functional integrity of sperm cell membranes. According to the existing literature, its simple applicability has turned it into a valuable additional parameter to standard canine semen analysis. In the recent years, much research has been conducted in this field. The aim of this systematic review was to evaluate the quality of published literature in canine reproduction concerning the HOS test. Using two distinguished databases, 38 articles were detected and analysed subsequently according to various aspects, for example study design, population, semen sampling and implementation concerning the HOS test. Although there are numerous articles available, the diagnostic value of the HOS test remains ambiguous. Until now, neither a recognized test protocol nor reliable reference values have been defined. Most of the trials evaluated show serious methodological flaws and therefore do not permit drawing reliable conclusions. According to our results, approximately half of the studies (n = 20) included a sample size of five or less animals. None of the studies examined the inter- or intraobserver agreement for the HOS test. Further research is warranted including appropriate statistical methods and a sufficient number of animals to establish a standardized test protocol as well as reliable reference values. Most importantly, it is required to clarify a correlation between the HOS test and the fertilizing capacity to determine the diagnostic value of the HOS test. PMID:23931704
Atypical Carpal Tunnel Syndrome in a Holt Oram Patient: A Case Report and Literature Review
Mace, James; Reddy, Srikanth; Mohil, Randeep
We present a case report of a patient diagnosed with Holt-Oram syndrome (HOS) presenting with clinical and electrophysiologically confirmed carpal tunnel syndrome. Pre-operative Magnetic resonance imaging revealed an abnormal course of the median nerve; as such an atypical incision and approach were carried out to decompress the nerve to excellent post operative clinical effect. To our knowledge this is the first description of abnormal nervous course in a patient with HOS leading to peripheral entrapment. A literature surrounding the important aspects of HOS to the orthopaedic surgeon is presented concomitantly. PMID:25621081
Using a Professional Development Program for Enhancing Chilean Biology Teachers' Understanding of Nature of Science (NOS) and Their Perceptions About Using History of Science to Teach NOS
NASA Astrophysics Data System (ADS)
Pavez, José M.; Vergara, Claudia A.; Santibañez, David; Cofré, Hernán
A number of authors have recognized the importance of understanding the nature of science (NOS) for scientific literacy. Different instructional strategies such as decontextualized, hands-on inquiry, and history of science (HOS) activities have been proposed for teaching NOS. This article seeks to understand the contribution of HOS in enhancing biology teachers' understanding of NOS, and their perceptions about using HOS to teach NOS. These teachers ( N = 8), enrolled in a professional development program in Chile are, according to the national curriculum, expected to teach NOS, but have no specific NOS and HOS training. Teachers' views of NOS were assessed using the VNOS-D+ questionnaire at the beginning and at the end of two modules about science instruction and NOS. Both the pre- and the post-test were accompanied by interviews, and in the second session we collected information about teachers' perceptions of which interventions had been more significant in changing their views on NOS. Finally, the teachers also had to prepare a lesson plan for teaching NOS that included HOS. Some of the most important study results were: significant improvements were observed in teachers' understanding of NOS, although they assigned different levels of importance to HOS in these improvements; and although the teachers improved their understanding of NOS, most had difficulties in planning lessons about NOS and articulating historical episodes that incorporated NOS. The relationship between teachers' improved understanding of NOS and their instructional NOS skills is also discussed.
A gene for Holt-Oram syndrome maps to chromosome 12q24.1
Bonnet, D.; Pelet, A.; Sidi, D.
Originally described in 1960, Holt-Oram syndrome (HOS, MIM:142900) is a rare autosomal dominant disorder of unknown origin (1/100,000 live births) characterized by congenital septal heart defects with associated malformations of upper limbs. We have reported on the mapping of a gene causing HOS to the distal long arm of chromosome 12 (12q21-qter) by linkage analysis in 9 multiplex families (Zmax=8.19 at the D12S354 locus). In addition, multipoint linkage analysis provided evidence for mapping of the disease locus to the genetic interval (7cM) defined by loci D12S105 and D12S79. In situ hybridization of YACs containing the flanking loci D12S105 and D12S79 demonstrates that the HOS locus maps to 12q24.1 thus exluding the candidate genes KOX20 and KOX1. We tested three HOS multiplex families with polydactily or without heart defect and showed that they do not map to chromosome 12q (homog-test: {chi}{sup 2}=13.28, p=0.0001). This observation supports the view that genetic heterogeneity holds true for typical HOS only. The mapping of a gene for HOS is, to our knowledge, the first chromosomal localization of a gene responsible for congenital septal defect in human. The characterization of the disease causing gene will hopefully shed light on the molecular mechanisms that govern heart septation and limb development in the early stages of embryogenesis.
Holt-Oram syndrome and diaphragmatic hernia associate with paracentric inversion of chromosome 8
Eswara, M.S.; Batanian, J.R.
Holt-Oram syndrome (HOS) consists of congenital heart disease, usually atrial septal defect, along with thumb anomalies and occasionally more extensive limb defects. Inheritance is autosomal dominant. Previous reports have associated HOS with cytogenetic abnormalities on chromosomes 4, 14 and 20. Recently a linkage study has suggested a HOS locus on chromosome 12. We describe another case of HOS with a de novo cytogenetic abnormality. On prenatal ultrasound, IUGR, oligohydramnios and left diaphragmatic hernia were noted. Following delivery, patient was placed on extra-corporeal membrane oxygenation because of severe lung hypoplasia; diaphragmatic hernia was repaired with mesh graft. He expired on day 17 of life. On exam, he had subtle dysmorphic features with hypotelorism and abnormal folding of the ear lobes. He had bilateral radial aplasia, aplasia of thumbs, index and middle fingers, along with the metacarpals. On autopsy he was found to have atrial septal defect of the ostium secundum type, right side aortic arch with vascular ring formation, bicuspid pulmonic valve and severe lung hypoplasia worse on the left. Cytogenetic analysis on blood and skin showed 48,XX,inv(8)(q24.2q13). Chromosome fragility study was negative. Parental chromosomes were normal. Our observation of inv(8)q with HOS and diaphragmatic hernia may indicate genetic heterogeneity with this condition. Regulation of morphogenesis is likely under the control of a hierarchy of genes; multiple loci for conditions such as HOS would not be surprising.
Antiproliferative effect of Tualang honey on oral squamous cell carcinoma and osteosarcoma cell lines
Background The treatment of oral squamous cell carcinomas (OSCC) and human osteosarcoma (HOS) includes surgery and/or radiotherapy which often lead to reduced quality of life. This study was aimed to study the antiproliferative activity of local honey (Tualang) on OSCC and HOS cell lines. Methods Several concentrations of Tualang honey (1% - 20%) were applied on OSCC and HOS cell lines for 3, 6, 12, 24, 48 and 72 hours. Morphological characteristics were observed under light and fluorescent microscope. Cell viability was assessed using MTT assay and the optical density for absorbance values in each experiment was measured at 570 nm by an ELISA reader. Detection of cellular apoptosis was done using the Annexin V-FITC Apoptosis Detection Kit. Results Morphological appearance showed apoptotic cellular changes like becoming rounded, reduction in cell number, blebbed membrane and apoptotic nuclear changes like nuclear shrinkage, chromatin condensation and fragmented nucleus on OSCC and HOS cell lines. Cell viability assay showed a time and dose-dependent inhibitory effect of honey on both cell lines. The 50% inhibitory concentration (IC50) for OSCC and HOS cell lines was found to be 4% and 3.5% respectively. The maximum inhibition of cell growth of ≥80% was obtained at 15% for both cell lines. Early apoptosis was evident by flow cytometry where percentage of early apoptotic cells increased in dose and time dependent manner. Conclusion Tualang honey showed antiproliferative effect on OSCC and HOS cell lines by inducing early apoptosis. PMID:20840769
Wave-breaking onset in a High-Order Spectral model
NASA Astrophysics Data System (ADS)
Seiffert, Betsy; Ducrozet, Guillaume
We examine the implementation of a wave-breaking onset parameter into two numerical models, HOS-ocean and HOS-NWT, which are computationally efficient, open source codes that solve for highly nonlinear wave fields in the open ocean and a numerical wave tank, respectively, using the High-Order Spectral (HOS) method. Due to the assumptions of solving for potential flow, the HOS solvers assume a single-valued free surface and therefore cannot produce breaking waves. The goal of implementing a wave-breaking mechanism into the HOS models is to approximate the broken free surface as a single value. By doing this, we can increase the application range of the models including calculating more extreme sea states, which is important when predicting dynamics of offshore vessels, predicting loads on marine structures and the general physics of ocean waves, for example. In nature, a steep wave may grow to surpass some limiting threshold leading to a collapse of the water surface as a broken wave. Energy from the wave is transferred through the generation of currents and turbulence, and changes occur in the spectral distribution of energy before and after a wave breaks. To implement a wave-breaking mechanism into the HOS models, first a wave-breaking onset parameter needs to be identified, and second, a strategy for dissipating and distributing the energy after the wave has broken needs to be determined. To calculate wave breaking onset in the HOS solvers, a dynamic wave-breaking criteria is proposed, following the work of Barthelemy, et al. (submitted). The criteria assumes that if the ratio of local energy flux velocity to the local crest velocity surpasses a limiting threshold, the wave will break. Calculation of the local crest velocity is non-trivial, and therefore partial Hilbert transforms are used. When calculating this wave-breaking parameter in the HOS model, it successfully distinguishes between an "unbroken" wave and a "broken" wave, evidenced by the manifestation of
A conserved histone deacetylase with a role in the regulation of cytokinesis in Schizosaccharomyces pombe
Background In Schizosaccharomyces pombe the SET domain protein, Set3p - together with its interacting partners, Snt1p, and Hif2p - form a complex that aids in preventing cell division failure upon mild cytokinetic stress. Intriguingly, the human orthologs of these proteins (MLL5, NCOR2, and TBL1X) are also important for the faithful completion of cytokinesis in tissue culture cells. Since MLL5, NCOR2, and TBL1X form a complex with the histone deacetylase, HDAC3, we sought to determine if an orthologous counterpart played a regulatory role in fission yeast cytokinesis. Results In this report we identify the hos2 gene as the fission yeast HDAC3 ortholog. We show that Hos2p physically interacts with Set3p, Snt1p, and Hif2p, and that hos2∆ mutants are indeed compromised in their ability to reliably complete cell division in the presence of mild cytokinetic stresses. Furthermore, we demonstrate that over-expression of hos2 causes severe morphological and cytokinetic defects. Lastly, through recombinase mediated cassette exchange, we show that expression of human HDAC3 complements the cytokinetic defects exhibited by hos2∆ cells. Conclusions These data support a model in which Hos2p functions as an essential component of the Set3p-Snt1p-Hif2p complex with respect to the regulation of cytokinesis. The ability of human HDAC3 to complement the cytokinesis defects associated with the deletion of the hos2 gene suggests that further analysis of this system could provide insight into the role of HDAC3 in both the regulation of cell division, as well as other biological processes influenced by HDAC3 deacetylation. PMID:22559741
Psychometric Evaluation of the Lower Extremity Computerized Adaptive Test, the Modified Harris Hip Score, and the Hip Outcome Score
Hung, Man; Hon, Shirley D.; Cheng, Christine; Franklin, Jeremy D.; Aoki, Stephen K.; Anderson, Mike B.; Kapron, Ashley L.; Peters, Christopher L.; Pelt, Christopher E.
Background: The applicability and validity of many patient-reported outcome measures in the high-functioning population are not well understood. Purpose: To compare the psychometric properties of the modified Harris Hip Score (mHHS), the Hip Outcome Score activities of daily living subscale (HOS-ADL) and sports (HOS-sports), and the Lower Extremity Computerized Adaptive Test (LE CAT). The hypotheses was that all instruments would perform well but that the LE CAT would show superiority psychometrically because a combination of CAT and a large item bank allows for a high degree of measurement precision. Study Design: Cohort study (diagnosis); Level of evidence, 2. Methods: Data were collected from 472 advanced-age, active participants from the Huntsman World Senior Games in 2012. Validity evidences were examined through item fit, dimensionality, monotonicity, local independence, differential item functioning, person raw score to measure correlation, and instrument coverage (ie, ceiling and floor effects), and reliability evidences were examined through Cronbach alpha and person separation index. Results: All instruments demonstrated good item fit, unidimensionality, monotonicity, local independence, and person raw score to measure correlations. The HOS-ADL had high ceiling effects of 36.02%, and the mHHS had ceiling effects of 27.54%. The LE CAT had ceiling effects of 8.47%, and the HOS-sports had no ceiling effects. None of the instruments had any floor effects. The mHHS had a very low Cronbach alpha of 0.41 and an extremely low person separation index of 0.08. Reliabilities for the LE CAT were excellent and for the HOS-ADL and HOS-sports were good. Conclusion: The LE CAT showed better psychometric properties overall than the HOS-ADL, HOS-sports, and mHHS for the senior population. The mHHS demonstrated pronounced ceiling effects and poor reliabilities that should be of concern. The high ceiling effects for the HOS-ADL were also of concern. The LE CAT was superior
MicroRNA 19a replacement partially rescues fin and cardiac defects in zebrafish model of Holt Oram syndrome.
Chiavacci, Elena; D'Aurizio, Romina; Guzzolino, Elena; Russo, Francesco; Baumgart, Mario; Groth, Marco; Mariani, Laura; D'Onofrio, Mara; Arisi, Ivan; Pellegrini, Marco; Cellerino, Alessandro; Cremisi, Federico; Pitto, Letizia
Holt-Oram Syndrome (HOS) is an autosomal dominant heart-hand syndrome caused by mutations in the TBX5 gene, a transcription factor capable of regulating hundreds of cardiac-specific genes through complex transcriptional networks. Here we show that, in zebrafish, modulation of a single miRNA is sufficient to rescue the morphogenetic defects generated by HOS. The analysis of miRNA-seq profiling revealed a decreased expression of miR-19a in Tbx5-depleted zebrafish embryos compared to the wild type. We revealed that the transcription of the miR-17-92 cluster, which harbors miR-19a, is induced by Tbx5 and that a defined dosage of miR-19a is essential for the correct development of the heart. Importantly, we highlighted that miR-19a replacement is able to rescue cardiac and pectoral fin defects and to increase the viability of HOS zebrafish embryos. We further observed that miR-19a replacement shifts the global gene expression profile of HOS-like zebrafish embryos towards the wild type condition, confirming the ability of miR-19a to rescue the Tbx5 phenotype. In conclusion our data demonstrate the importance of Tbx5/miR-19a regulatory circuit in heart development and provide a proof of principle that morphogenetic defects associated with HOS can be rescued by transient miRNA modulation. PMID:26657204
A comparative prospective study using matched samples to determine the influence of subnormal hypo-osmotic test scores of spermatozoa on subsequent fertilization and pregnancy rates following in-vitro fertilization.
Check, J H; Stumpo, L; Lurie, D; Benfer, K; Callan, C
The achievement of pregnancies in vivo is rare in couples where the male partner has defective sperm membranes as shown by hypo-osmotic swelling (HOS) test scores of < 50%. However, there have been mixed reports on the value of the HOS test in predicting outcome following invitro fertilization; some studies suggest reduced fertilization rates and others find little, if any, predictability of decreased fertilization. The assumption has been made that fertilization rates are proportional to pregnancy rates; however, this may not necessarily be true since defective spermatozoa could lead to a less viable pre-embryo and therefore a decreased viable pregnancy rate. We performed a comparative prospective study using matched controls to evaluate fertilization rates and to determine subsequent pregnancy rates. The mean HOS scores were 70.0 and 36.7% respectively, with mean motile sperm concentrations of 35.7 and 34.0 x 10(6)/ml in 27 matched pairs. There was no difference in the mean number of oocytes retrieved, fertilization rates or number of embryos transferred between the two groups by HOS score. The clinical and viable pregnancy rates and implantation rates were 25.9, 18.5 and 9.9% for normal versus 3.7, 3.7 and 1.1% for subnormal groups. These data suggest that low HOS scores may be associated with the formation of defective embryos, leading to low pregnancy rates but normal fertilization rates. PMID:7657765
Investigating the mechanical properties of zona pellucida of whole human oocytes by atomic force spectroscopy.
Andolfi, Laura; Masiero, Elena; Giolo, Elena; Martinelli, Monica; Luppi, Stefania; Dal Zilio, Simone; Delfino, Ines; Bortul, Roberta; Zweyer, Marina; Ricci, Giuseppe; Lazzarino, Marco
The role of mechanics in numerous biological processes is nowadays recognized, while in others, such as the fertilization process, it is still neglected. In the case of oocytes the description of their mechanical properties could improve the comprehension of the oocyte-spermatozoon interaction and be helpful for application in in vitro fertilization (IVF) clinics. Herein the mechanical properties of whole human oocytes (HOs) immediately after retrieval are investigated by indentation measurements with atomic force spectroscopy under physiological conditions. Measurements are performed on immature (metaphase I - MI) and mature (metaphase II - MII) HOs. According to their morphological characteristics MII-HOs are classified as "suitable" and "rejected"; these latter would be usually rejected for intracytoplasmic sperm injection (ICSI). For all maturation stages we observe that the elastic response of the zona pellucida (ZP) outer layer was different and distinguishable from the rest of the ZP-HO. The elasticity of this ZP outer layer varies with maturation and quality: stiffness decreases from immature MI to good quality MII, up to poor-quality rejected MII. An indirect analysis with IVF outcome indicates that the ZP outer layer of analysed HOs donated by women who achieved pregnancy is stiffer than that of HOs from women with negative outcome. Our findings suggest that mechanical properties can represent important oocyte quality indicators that may be exploited for the design of innovative ICSI dedicated cell sorters. PMID:27476747
Isolation and Characterization of High-Osmolarity-Sensitive Mutants of Fission Yeast
Aiba, Hirofumi; Kawaura, Ryosuke; Yamamoto, Eiji; Yamada, Hisami; Takegawa, Kaoru; Mizuno, Takeshi
For the fission yeast Schizosaccharomyces pombe, adaptation to high-osmolarity medium is mediated by a mitogen-activated protein (MAP) kinase cascade, involving the Wis1 MAP kinase kinase and the Sty1 MAP kinase. The MAP kinase pathway transduces an osmotic signal and accordingly regulates the expression of the downstream target gene (gpd1+) that encodes NADH-dependent glycerol-3-phosphate dehydrogenase, in order to adaptively accumulate glycerol inside the cells as an osmoprotectant. We previously characterized a set of high-osmolarity-sensitive S. pombe mutants, including wis1, sty1, and gpd1. In this study, we attempted to further isolate novel osmolarity-sensitive mutants. For some of the mutants isolated, profiles of glycerol production in response to the osmolarity of the growth medium were indistinguishable from that of the wild-type cells, suggesting that they are novel types. They were classified into three distinct types genetically and, thus, were designated hos1, hos2, and hos3 (high osmolarity sensitive) mutants. One of them, the hos1 mutant, was characterized in detail. The hos1 mutant was demonstrated to have a mutational lesion in the known ryh1+ gene, which encodes a small GTP-binding protein. Disruption of the ryh1+ gene results not only in osmosensitivity but also in temperature sensitivity for growth. It was also found that the Δryh1 mutant is severely sterile. These results are discussed with special reference to the osmoadaptation of S. pombe. PMID:9748434
MicroRNA 19a replacement partially rescues fin and cardiac defects in zebrafish model of Holt Oram syndrome
Chiavacci, Elena; D'Aurizio, Romina; Guzzolino, Elena; Russo, Francesco; Baumgart, Mario; Groth, Marco; Mariani, Laura; D'Onofrio, Mara; Arisi, Ivan; Pellegrini, Marco; Cellerino, Alessandro; Cremisi, Federico; Pitto, Letizia
Holt-Oram Syndrome (HOS) is an autosomal dominant heart-hand syndrome caused by mutations in the TBX5 gene, a transcription factor capable of regulating hundreds of cardiac-specific genes through complex transcriptional networks. Here we show that, in zebrafish, modulation of a single miRNA is sufficient to rescue the morphogenetic defects generated by HOS. The analysis of miRNA-seq profiling revealed a decreased expression of miR-19a in Tbx5-depleted zebrafish embryos compared to the wild type. We revealed that the transcription of the miR-17-92 cluster, which harbors miR-19a, is induced by Tbx5 and that a defined dosage of miR-19a is essential for the correct development of the heart. Importantly, we highlighted that miR-19a replacement is able to rescue cardiac and pectoral fin defects and to increase the viability of HOS zebrafish embryos. We further observed that miR-19a replacement shifts the global gene expression profile of HOS-like zebrafish embryos towards the wild type condition, confirming the ability of miR-19a to rescue the Tbx5 phenotype. In conclusion our data demonstrate the importance of Tbx5/miR-19a regulatory circuit in heart development and provide a proof of principle that morphogenetic defects associated with HOS can be rescued by transient miRNA modulation. PMID:26657204
Numerical Simulation of a Seaway with Breaking
NASA Astrophysics Data System (ADS)
Dommermuth, Douglas; O'Shea, Thomas; Brucker, Kyle; Wyatt, Donald
The focus of this presentation is to describe the recent efforts to simulate a fully non-linear seaway with breaking by using a high-order spectral (HOS) solution of the free-surface boundary value problem to drive a three-dimensional Volume of Fluid (VOF) solution. Historically, the two main types of simulations to simulate free-surface flows are the boundary integral equations method (BIEM) and high-order spectral (HOS) methods. BIEM calculations fail at the point at which the surface impacts upon itself, if not sooner, and HOS methods can only simulate a single valued free-surface. Both also employ a single-phase approximation in which the effects of the air on the water are neglected. Due to these limitations they are unable to simulate breaking waves and air entrainment. The Volume of Fluid (VOF) method on the other hand is suitable for modeling breaking waves and air entrainment. However it is computationally intractable to generate a realistic non-linear sea-state. Here, we use the HOS solution to quickly drive, or nudge, the VOF solution into a non-linear state. The computational strategies, mathematical formulation, and numerical implementation will be discussed. The results of the VOF simulation of a seaway with breaking will also be presented, and compared to the single phase, single valued HOS results.
Bedomning Av Kommunikativ Formaga Hos Personer Med Utvecklingsstorning. Teknik, Kommunikation, Handikapp Forskningsrapport nr 12 (Assessment of the Communicative Ability in Persons with Mental Retardation. Technology, Communication, Disability Research Report No. 12). TeleCommunity, RACE 2033 (Research in Advanced Communications Technologies in Europe).
ERIC Educational Resources Information Center
Brodin, Jane; Thurfjell, Fredrik
This report was developed as part of a Swedish study to examine whether videotelephones can be used to develop the communicative ability of individuals with mental retardation and whether access to telecommunications increased the possibilities of social integration. Communication was viewed as sharing a feeling, action, or experience with another…
Recent Applications of Higher-Order Spectral Analysis to Nonlinear Aeroelastic Phenomena
NASA Technical Reports Server (NTRS)
Silva, Walter A.; Hajj, Muhammad R.; Dunn, Shane; Strganac, Thomas W.; Powers, Edward J.; Stearman, Ronald
Recent applications of higher-order spectral (HOS) methods to nonlinear aeroelastic phenomena are presented. Applications include the analysis of data from a simulated nonlinear pitch and plunge apparatus and from F-18 flight flutter tests. A MATLAB model of the Texas A&MUniversity s Nonlinear Aeroelastic Testbed Apparatus (NATA) is used to generate aeroelastic transients at various conditions including limit cycle oscillations (LCO). The Gaussian or non-Gaussian nature of the transients is investigated, related to HOS methods, and used to identify levels of increasing nonlinear aeroelastic response. Royal Australian Air Force (RAAF) F/A-18 flight flutter test data is presented and analyzed. The data includes high-quality measurements of forced responses and LCO phenomena. Standard power spectral density (PSD) techniques and HOS methods are applied to the data and presented. The goal of this research is to develop methods that can identify the onset of nonlinear aeroelastic phenomena, such as LCO, during flutter testing.
Users guide: The LaRC human-operator-simulator-based pilot model
NASA Technical Reports Server (NTRS)
Bogart, E. H.; Waller, M. C.
A Human Operator Simulator (HOS) based pilot model has been developed for use at NASA LaRC for analysis of flight management problems. The model is currently configured to simulate piloted flight of an advanced transport airplane. The generic HOS operator and machine model was originally developed under U.S. Navy sponsorship by Analytics, Inc. and through a contract with LaRC was configured to represent a pilot flying a transport airplane. A version of the HOS program runs in batch mode on LaRC's (60-bit-word) central computer system. This document provides a guide for using the program and describes in some detail the assortment of files used during its operation.
The hypo-osmotic swelling test for evaluation of sperm membrane integrity.
Ramu, Sivakumar; Jeyendran, Rajasingam S
A functional membrane is requisite for the fertilizing ability of spermatozoa, as it plays an integral role in sperm capacitation, acrosome reaction, and binding of the spermatozoon to the egg surface. The hypo-osmotic swelling (HOS) test evaluates the functional integrity of the sperm's plasma membrane and also serves as a useful indicator of fertility potential of sperm. The HOS test predicts membrane integrity by determining the ability of the sperm membrane to maintain equilibrium between the sperm cell and its environment. Influx of the fluid due to hypo-osmotic stress causes the sperm tail to coil and balloon or "swell." A higher percentage of swollen sperm indicates the presence of sperm having a functional and intact plasma membrane. Here, we present the detailed protocol for performing the HOS test and explain the results for interpretation. PMID:22992900
Technical decision making with higher order structure data: higher order structure characterization during protein therapeutic candidate screening.
Jiang, Yijia; Li, Cynthia; Li, Jenny; Gabrielson, John P; Wen, Jie
Protein therapeutics differ considerably from small molecule drugs because of the presence of higher order structure (HOS), post-translational modifications, inherent molecular heterogeneity, and unique stability profiles. At early stages of development, multiple molecular candidates are often produced for the same biological target. In order to select the most promising molecule for further development, studies are carried out to compare and rank order the candidates in terms of their manufacturability, purity, and stability profiles. This note reports a case study on the use of selected HOS characterization methods for candidate selection and the role of HOS data in identifying potential challenges that may be avoided by selecting the optimal molecular entity for continued development. PMID:25716705
Psychometric Evaluation of the SF-36® Health Survey in Medicare Managed Care
Gandek, Barbara; Sinclair, Samuel J.; Kosinski, Mark; Ware, John E.
Data quality and scoring assumptions for the SF-36® Health Survey were evaluated among the elderly and disabled, using 1998 Cohort I baseline Medicare HOS data (n=177,714). Missing data rates were low, and scoring assumptions were met. Internal consistency reliability was 0.83 to 0.93 for the eight scales and 0.94 and 0.89, respectively, for the physical (PCS) and mental (MCS) component summary measures. Results declined with increased risk factors (e.g., older age, more chronic conditions), but were well above accepted standards for all subgroups. These findings support using standard algorithms for scoring the SF-36® in the HOS and subgroup analyses of HOS data. PMID:15493441
Psychometric evaluation of the SF-36 health survey in Medicare managed care.
Gandek, Barbara; Sinclair, Samuel J; Kosinski, Mark; Ware, John E
Data quality and scoring assumptions for the SF-36 Health Survey were evaluated among the elderly and disabled, using 1998 Cohort I baseline Medicare HOS data (n=177,714). Missing data rates were low, and scoring assumptions were met. Internal consistency reliability was 0.83 to 0.93 for the eight scales and 0.94 and 0.89, respectively, for the physical (PCS) and mental (MCS) component summary measures. Results declined with increased risk factors (e.g., older age, more chronic conditions), but were well above accepted standards for all subgroups. These findings support using standard algorithms for scoring the SF-36 in the HOS and subgroup analyses of HOS data. PMID:15493441
Validation of the Spanish version of the hip outcome score: a multicenter study
Background The Hip Outcome Score (HOS) is a self-reported questionnaire evaluating the outcomes of treatment interventions for hip pathologies, divided in 19 items of activities of daily life (ADL) and 9 sports' items. The aim of the present study is to translate and validate HOS into Spanish. Methods A prospective and multicenter study with 100 patients undergoing hip arthroscopy was performed between June 2012 and January 2013. Crosscultural adaptation was used to translate HOS into Spanish. Patients completed the questionnaire before and after surgery. Feasibility, reliability, internal consistency, construct validity (correlation with Western Ontario and McMaster Universities Osteoarthritis Index), ceiling and floor effects and sensitivity to change were assessed for the present study. Results Mean age was 45.05 years old. 36 women and 64 men were included. Feasibility: 13% had at least one missing item within the ADL subscale and 17% within the sport subscale. Reliability: the translated version of HOS was highly reproducible with intraclass correlation coefficient of 0.95 for ADL and 0.94 for the sports subscale. Internal consistency was confirmed with Cronbach's alpha >0.90 in both subscales. Construct validity showed statistically significant correlation with WOMAC. Ceiling effect was observed in 6% and 12% for ADL and sports subscale, respectively. Floor effect was found in 3% and 37% ADL and sports subscale, respectively. Large sensitivity to change was shown in both subscales. Conclusion The translated version of HOS into Spanish has shown to be feasible, reliable and sensible to changes for patients undergoing hip arthroscopy. This validated translation of HOS allows for comparisons between studies involving either Spanish- or English-speaking patients. Level of evidence Prognostic study, Level I PMID:24884511
Patient reported outcomes for patients who returned to sport compared with those who did not after hip arthroscopy: minimum 2-year follow-up.
Domb, Benjamin G; Dunne, Kevin F; Martin, Timothy J; Gui, Chengcheng; Finch, Nathan A; Vemula, S Pavan; Redmond, John M
Previous studies assessed elite athletes' return to sport (RTS) after hip arthroscopy, but few investigated a cohort including athletes from all levels of sport. This study compared athletes who returned to sport to those who did not, based on four patient-reported outcome (PRO) scores, including the Hip Outcome Score-Sports Specific Subscale (HOS-SSS). Between September 2008 and April 2012, hip arthroscopies were performed on 157 patients (168 hips) who reported playing a sport preoperatively and indicated their level of sports activity post-operatively. Two-year follow-up was available for 148 (94%) amateur and professional athletes with a total of 158 hips. Of these 60 cases (65 hips) did not return to sports (NRTS) and were in the NRTS group. The remaining 88 cases (93 hips) constituted the RTS group. The modified Harris Hip Score, Non-Arthric Hip Score, Hip Outcome-Activities of Daily Living (HOS-ADL), and HOS-SSS were used to assess outcomes. The HOS-SSS was used to assess specific sport-related movement. Both groups demonstrated significant improvement at 2 years post-operatively in visual analog score and four PRO scores (P < 0.001). There was no significant preoperative differences in HOS-SSS scores between groups; however, the RTS group had significantly higher HOS-SSS scores at 1 and 2 years post-surgery. Post-operatively, the RTS group had significantly better ability to jump, land from a jump, stop quickly and perform cutting/lateral movements (P < 0.05). In summary, patients who indicated RTSs demonstrated significantly higher PRO scores and abilities to perform several sport-related movements, compared with patients who did not. PMID:27583148
Signal transduction and downregulation of C-MET in HGF stimulated low and highly metastatic human osteosarcoma cells
Husmann, Knut; Ducommun, Pascal; Sabile, Adam A.; Pedersen, Else-Marie; Born, Walter; Fuchs, Bruno
The poor outcome of osteosarcoma (OS), particularly in patients with metastatic disease and a five-year survival rate of only 20%, asks for more effective therapeutic strategies targeting malignancy-promoting mechanisms. Dysregulation of C-MET, its ligand hepatocyte growth factor (HGF) and the fusion oncogene product TPR-MET, first identified in human MNNG-HOS OS cells, have been described as cancer-causing factors in human cancers. Here, the expression of these molecules at the mRNA and the protein level and of HGF-stimulated signaling and downregulation of C-MET was compared in the parental low metastatic HOS and MG63 cell lines and the respective highly metastatic MNNG-HOS and 143B and the MG63-M6 and MG63-M8 sublines. Interestingly, expression of TPR-MET was only observed in MNNG-HOS cells. HGF stimulated the phosphorylation of Akt and Erk1/2 in all cell lines investigated, but phospho-Stat3 remained at basal levels. Downregulation of HGF-stimulated Akt and Erk1/2 phosphorylation was much faster in the HGF expressing MG63-M8 cells than in HOS cells. Degradation of HGF-activated C-MET occurred predominantly through the proteasomal and to a lesser extent the lysosomal pathway in the cell lines investigated. Thus, HGF-stimulated Akt and Erk1/2 signaling as well as proteasomal degradation of HGF activated C-MET are potential therapeutic targets in OS. - Highlights: • Expression of TPR-MET was only observed in MNNG-HOS cells. • HGF stimulated the phosphorylation of Akt and Erk1/2 but not of Stat3 in osteosarcoma cell lines. • Degradation of HGF-activated C-MET occurred predominantly through the proteasomal pathway.
Patient reported outcomes for patients who returned to sport compared with those who did not after hip arthroscopy: minimum 2-year follow-up
Domb, Benjamin G.; Dunne, Kevin F.; Martin, Timothy J.; Gui, Chengcheng; Finch, Nathan A.; Vemula, S. Pavan; Redmond, John M.
Previous studies assessed elite athletes' return to sport (RTS) after hip arthroscopy, but few investigated a cohort including athletes from all levels of sport. This study compared athletes who returned to sport to those who did not, based on four patient-reported outcome (PRO) scores, including the Hip Outcome Score—Sports Specific Subscale (HOS-SSS). Between September 2008 and April 2012, hip arthroscopies were performed on 157 patients (168 hips) who reported playing a sport preoperatively and indicated their level of sports activity post-operatively. Two-year follow-up was available for 148 (94%) amateur and professional athletes with a total of 158 hips. Of these 60 cases (65 hips) did not return to sports (NRTS) and were in the NRTS group. The remaining 88 cases (93 hips) constituted the RTS group. The modified Harris Hip Score, Non-Arthric Hip Score, Hip Outcome-Activities of Daily Living (HOS-ADL), and HOS-SSS were used to assess outcomes. The HOS-SSS was used to assess specific sport-related movement. Both groups demonstrated significant improvement at 2 years post-operatively in visual analog score and four PRO scores (P < 0.001). There was no significant preoperative differences in HOS-SSS scores between groups; however, the RTS group had significantly higher HOS-SSS scores at 1 and 2 years post-surgery. Post-operatively, the RTS group had significantly better ability to jump, land from a jump, stop quickly and perform cutting/lateral movements (P < 0.05). In summary, patients who indicated RTSs demonstrated significantly higher PRO scores and abilities to perform several sport-related movements, compared with patients who did not. PMID:27583148
On the measurement of human osteosarcoma cell elastic modulus using shear assay experiments.
Cao, Yifang; Bly, Randy; Moore, Will; Gao, Zhan; Cuitino, Alberto M; Soboyejo, Wole
This paper presents a method for determining the elastic modulus of human osteosarcoma (HOS) cells. The method involves a combination of shear assay experiments and finite element analysis. Following in-situ observations of cell deformation during shear assay experiments, a digital image correlation (DIC) technique was used to determine the local displacement and strain fields. Finite element analysis was then used to determine the Young's moduli of HOS cells. This involved a match of the maximum shear stresses estimated from the experimental shear assay measurements and those calculated from finite element simulations. PMID:17200819
Holt-Oram Syndrome Associated with Aortic Atresia: A Rare Association
Rodagi, Sunil B.; Surana, Snehal S.; Potdar, Vijaykumar R.; Kirdi, Sharanbasav S.
Holt-Oram syndrome (HOS) is a rare autosomal dominant disorder that causes abnormalities of the upper limbs and heart. It is seen in 1:1, 00, 000 live births. It is linked to a single-gene TBX5 "protein-producing" mutation with gene map locus 12q24. Most commonly it is characterized by the cardiac septation defects and pre-axial radial ray abnormalities. We are reporting a case of HOS with aortic atresia which is a rare association. PMID:27293527
Functional health outcomes as a measure of health care quality for Medicare beneficiaries.
Bierman, A S; Lawrence, W F; Haffer, S C; Clancy, C M
OBJECTIVE: the Medicare Health Outcomes Survey (HOS), a new quality measure in the Health Plan Employer Data and Information Set, is designed to assess physical and mental functional health outcomes of Medicare beneficiaries enrolled in Medicare+Choice organizations. We discuss the rationale for the HOS measure together with methodologic challenges in its use and interpretation, using descriptive data from the baseline Medicare HOS to illustrate some of these challenges. DATA SOURCES/STUDY DESIGN: The 1999 Cohort 2 Medicare HOS baseline data were used for a cross-sectional descriptive analysis. A random sample of 1,000 beneficiaries from each health plan with a Medicare+Choice contract was surveyed (N = 156,842; 282 organizations included in these analyses) . PRINCIPAL FINDINGS: The HOS measure is designed to assess a previously unmeasured dimension of quality. Plan-level variation was seen across all baseline measures of sociodemographic characteristics and illness burden. At the individual level socioeconomic position as measured by educational attainment was strongly associated with functional status. The least educated beneficiaries had the highest burden of illness on all measures examined, and there was a consistent and significant gradient in health and functional status across all levels of education. In analyses stratified by race and ethnicity, socioeconomic gradients in f un ct ion persist ed. CONCLUSIONS Despite limitations, by focusing at t en t ion on the need to improve functional health out comes among elderly Medicare beneficiaries enrolled in Medicare+Choice, the HOS can serve as an important new tool to support efforts to improve health care quality. The HOS provides valuable information at the federal, state, and health plan levels that can be used to identify, prioritize, and evaluate quality improvement interventions and monitor progress for the program overall as well as for vulnerable subgroups. To interpret the HOS as a quality measure
Dipsacus asperoides polysaccharide induces apoptosis in osteosarcoma cells by modulating the PI3K/Akt pathway.
Chen, Jun; Yao, Dong; Yuan, Huixin; Zhang, Shaojun; Tian, Jinhong; Guo, Wenjing; Liang, Weizhi; Li, Huiyuan; Zhang, Yong
An alkaline extractable and water-soluble polysaccharide (ADAPW), with an average molecular weight of 16kDa, was purified from the alkaline extraction of the roots of Dipsacus asperoides. Monosaccharide component analysis indicated that ADAPW was composed of glucose, rhamnose, arabinose and mannose in a molar ratio of 8.54:1.83:1.04:0.42. This study aimed to investigate the effect of ADAPW on the viability of human osteosarcoma cell line HOS cells, and explore the possible mechanisms. The results revealed that ADAPW inhibited the proliferation of HOS cells in a dose-dependent manner by inducing apoptosis. Furthermore, treatment with ADAPW caused a loss of mitochondrial membrane potential and accumulation of reactive oxygen species (ROS). In addition, Western blot analysis demonstrated that ADAPW down-regulated the protein expressions of PI3K and phosphorylated Akt (pAkt) in HOS cells. Taken together, induction of apoptosis on HOS cells by ADAPW was mainly associated with ROS production, mitochondrial dysfunction, and inhibition of PI3K/Akt signaling pathway. So this finding suggests that ADAPW may be potentially effective in cancer prevention against human osteosarcoma. PMID:23648042
The pig hos been proposed as a potential animal model for methanol-induced neuro-ocular toxicosis in humans because of its reported low liver tetrahydro folate levels and therefore, slower formate metabolism as compared to humans. o determine the validity of the animal model, min...
The Psychometric Properties of the Agricultural Hazardous Occupations Order Certification Training Program Written Examinations
ERIC Educational Resources Information Center
French, Brian F.; Breidenbach, Daniel H.; Field, William E.; Tormoehlen, Roger
The written certification exam that accompanies the Gearing Up for Safety-Agricultural Production Safety Training for Youth curriculum was designed to partially meet the testing requirements of the Agricultural Hazardous Occupations Order (AgHOs) Certification Training Program. This curriculum and accompanying assessment tools are available for…
75 FR 33664 - Hours of Service of Drivers: RockTenn, Application for Exemption
Federal Register 2010, 2011, 2012, 2013, 2014
... the Federal Register published on April 11, 2000 (65 FR 19476) or you may visit http://DocketInfo.dot... Federal Motor Carrier Safety Administration Hours of Service of Drivers: RockTenn, Application for... RockTenn requesting an exemption from the driver hours-of-service (HOS) provisions in Part 395 of...
78 FR 64265 - Hours of Service of Drivers: U.S. Department of Defense (DOD); Application for Exemption
Federal Register 2010, 2011, 2012, 2013, 2014
...FMCSA announces its decision to grant the U.S. Department of Defense (DOD) Military Surface Deployment and Distribution Command (SDDC) an exemption from the minimum 30-minute rest break provision of the Agency's hours-of-service (HOS) regulations for commercial motor vehicle (CMV) drivers. The exemption enables SDDC's contract motor carriers and their employee-drivers engaged in the......
Bile Acid Responses in Methane and Non-Methane Producers to Standard Breakfast Meals
Technology Transfer Automated Retrieval System (TEKTRAN)
Bile acids and their conjugates are important regulators of glucose homeostasis. Previous research has revealed the ratio of cholic acid to deoxycholic acid to affect insulin resistance in humans. Bile acid de-conjugation and intestinal metabolism depend on gut microbes which may be affected by hos...
Federal Register 2010, 2011, 2012, 2013, 2014
... the FDMS published in the Federal Register on January 17, 2008 (73 FR 3316), or you may visit http... listening sessions concerning the HOS rulemaking (75 FR 285). On January 15, 2010, FMCSA announced its plans to hold a fourth public listening session (75 FR 2467). FMCSA now announces a fifth public...
Federal Register 2010, 2011, 2012, 2013, 2014
... Federal Motor Carrier Safety Administration 49 CFR Part 395 RIN 2126-AB26 Hours of Service AGENCY: Federal...-service (HOS) regulations. Specifically, the Agency wants to know what factors, issues, and data it should... Act Statement for the FDMS published in the Federal Register on January 17, 2008 (73 FR 3316), or...
The role of nitric oxide on the proliferation of a human osteoblast cell line stimulated with hydroxyapatite.
Sosroseno, Wihaskoro; Sugiatno, Erwan; Samsudin, Abdul Rani; Ibrahim, Fikri
The aim of the present study was to test the hypothesis that the proliferation of a human osteoblast cell line (HOS cells) stimulated with hydroxyapatite (HA) may be regulated by nitric oxide (NO). The cells were cultured on the surface of HA. Medium or cells alone were used as controls. L-arginine, D-arginine, 7-NI (an nNOS inhibitor), L-NIL (an iNOS inhibitor), L-NIO (an eNOS inhibitor) or carboxy PTIO, a NO scavenger, was added in the HA-exposed cell cultures. The cells were also precoated with anti-human integrin alphaV antibody. The levels of nitrite were determined spectrophotometrically. Cell proliferation was assessed by colorimetric assay. The results showed increased nitrite production and cell proliferation by HA-stimulated HOS cells up to day 3 of cultures. Anti-integrin alphaV antibody, L-NIO, or carboxy PTIO suppressed, but L-arginine enhanced, nitrite production and cell proliferation of HA-stimulated HOS cells. The results of the present study suggest, therefore, that interaction between HA and HOS cell surface integrin alphaV molecule may activate eNOS to catalyze NO production which, in turn, may regulate the cell proliferation in an autocrine fashion. PMID:18780564
78 FR 32001 - Agency Information Collection Activities; Approval of a New Information Collection Request...
Federal Register 2010, 2011, 2012, 2013, 2014
... electronic onboard recorders (EOBRs) installed in CMVs manufactured on or after June 4, 2012 (75 FR 17208... interstate commerce to use EOBRs to document their drivers' HOS (76 FR 5537, February 1, 2011). Based on... comment on the harassment issue on April 13, 2011 (76 FR 20611). The Agency sought and received...
Genomic and biological characterization of Tomato necrotic streak virus, a novel subgroup 2 ilarvirus infecting tomato in Florida
Technology Transfer Automated Retrieval System (TEKTRAN)
Tomato necrotic streak virus (TomNSV) is a recently described ilarvirus that was detected in tomato in Florida. The full TomNSV genome sequence revealed it to be a novel subgroup 2 ilarvirus with little nucleotide identity to other previously reported tomato-infecting ilarviruses. Experimental hos...
Addressing Nature of Science Core Tenets with the History of Science: An Example with Sickle-Cell Anemia & Malaria
ERIC Educational Resources Information Center
Howe, Erica M.
The history of science (HOS) has proven to be a useful pedagogical tool to help students learn about what has come to be regarded as an agreed upon set of core nature of science (NOS) tenets. The following article illustrates an example of how teachers can instrumentally use the history of research on heterozygote protection in sickle-cell anemia…
The Hematopoietic Organ: A Cornerstone for Wolbachia Propagation Between and Within Hosts
Braquart-Varnier, Christine; Raimond, Maryline; Mappa, Gaëtan; Chevalier, Frédéric D.; Le Clec'h, Winka; Sicard, Mathieu
Wolbachia is an intracellular α-proteobacterium which is transmitted vertically from mother to offspring but also frequently switches horizontally from one host to another. Our hypothesis is based on the role of immune cells and the organs that produce them, the hematopoietic organs (HOs), as primordial niches for the propagation of Wolbachia via hemocytes both (i) within hosts: to initiate and maintain the systemic infection and (ii) between hosts: to promote both vertical and horizontal transmission of Wolbachia. Therefore, we review some fundamental ideas underlying this hypothesis and go further with new empirical data that lead to a first close-up analysis of the potential role of HOs in Wolbachia propagation. The monitoring of the first steps of Wolbachia infection in horizontally infected host organs by transmission electron microscopy and qPCR suggests that (i) HOs are colonized early and extensively as soon as they are in contact with Wolbachia which find in these cells a favorable niche to multiply and (ii) infected HOs which expel hemocytes all lifelong can generate and maintain a systemic infection that could contribute to increase both vertical and horizontal propagation of these symbionts. PMID:26733969
Cytogenetic and Molecular Characterization of Durum Alien Disomic Addition Line with Enhanced Tolerance to Fusarium Head Blight Resistance
Technology Transfer Automated Retrieval System (TEKTRAN)
Fusarium head blight (FHB), or scab, caused by the fungus Fusarium graminearum Schwabe, is a serious disease of durum wheat (Triticum turgidum L., 2n = 4x = 28; AABB genomes). Current durum cultivars have very little or no FHB resistance. A wild relative, diploid wheatgrass Lophopyrum elongatum (Hos...
Epigenetic mechanisms at work: Exploiting nutrient-specific modulation of genetic networks
Technology Transfer Automated Retrieval System (TEKTRAN)
"Epigenetics" was used to describe actions of genes with their environment that bring the phenotype into being". The modern version of epigenetics includes the molecular mechanisms that influence the phenotypic outcome of a gene or genome, in absence of changes to the underlying DNA sequence. A hos...
We have identified several transposable gene-activating elements from Psuedomonas cepacia on the basis of their ability to increase expression of the genes of the broad-hos-range plasmid pGC9l.14. hen introduced into auxotrophic derivatives of P. cepacia 249 (ATCC 17616), this pl...
Hydrologic hydrochemical investigations were conducted to determine the long-term fate of hazardous chemical waste disposed in the Texas Gulf Coast Tertiary formations by deep-well injection. The study focused on the hydrostatic section of the Frio Formation because it is the hos...
76 FR 32388 - Agency Information Collection Activities; Request for Comment; Extension of an Information...
Federal Register 2010, 2011, 2012, 2013, 2014
... Docket Management System published in the Federal Register on January 17, 2008 (73 FR 3316), or you may... duty status in a specified format and verify the accuracy of the HOS of each driver (47 FR 53383). The... public by reducing the number of tired CMV drivers on the highways. Most States receive grants from...
76 FR 50313 - Agency Information Collection Activities; Request for Comment; Extension of an Information...
Federal Register 2010, 2011, 2012, 2013, 2014
... the ICR (76 FR 32388). One comment was received. DATES: Please send your comments by September 12... their duty status in a specified format and verify the accuracy of the HOS of each driver (47 FR 53383... helps FMCSA protect the public by reducing the number of tired CMV drivers on the highways. Most...
78 FR 76392 - Hours of Service of Drivers: Application for Exemption; David Muresan
Federal Register 2010, 2011, 2012, 2013, 2014
... published on January 17, 2008 (73 FR 3316). Public Participation: The Federal eRulemaking Portal is... shifts required for drivers of property-carrying CMVs (68 FR 22456; April 28, 2003). However, drivers of... employ), including ``rest'' and ``sleep.'' Mr. Muresan believes that the Agency's HOS rules do...
77 FR 33331 - Regulatory Guidance on the Applicability of Property-Carrier Hours-of-Service Rules to the...
Federal Register 2010, 2011, 2012, 2013, 2014
...The property-carrier hours-of-service (HOS) rules in 49 CFR 395.3 are applicable to drivers operating commercial motor vehicles designed or used to transport passengers on ``driveaway-towaway'' trips, as defined in 49 CFR 390.5. This notice provides Federal and State enforcement personnel, and the motor carrier industry, with uniform guidance concerning these...
78 FR 76757 - Regulatory Guidance on Hours of Service of Drivers Rest Break Requirement; Drivers Who Become...
Federal Register 2010, 2011, 2012, 2013, 2014
... December 27, 2011 (76 FR 81133), FMCSA published a final rule amending its HOS regulations for drivers of... Service of Drivers Rest Break Requirement; Drivers Who Become Ineligible for the ``Short-Haul'' Exception... why the rest break was not taken within the required 8 hours of the last off-duty break of at least...
Percutaneous Absorption of 2,3,7,8-Tetrachlorodibenzo-P-Dioxin (TCDD) From Soil (Journal Article)
Eight dermal absorption (two in vivo; six in vitro) and one intravenous experiment were conducted using 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) either neat (high dose at ~250 µg/cm2 and low dose at 10 ng/cm2) or sorbed on a low organic (LOS) or high organic (HOS) soil at 1 ppm...
Microarray Comparison of Endophyte-Infected and Endophyte-Free Tall Fescue
Technology Transfer Automated Retrieval System (TEKTRAN)
Many grasses have mutualistic symbioses with fungi of the family Clavicipitaceae: Tall fescue (Festuca arudinacea = Lolium arundinaceum) can harbor the obligate endophyte, Neotyphodium coenophialum that are asexually propagated and transmitted via host seeds. In an effort to begin to dissect the hos...
The fission yeast DASH complex is essential for satisfying the spindle assembly checkpoint induced by defects in the inner-kinetochore proteins.
Kobayashi, Yasuyo; Saitoh, Shigeaki; Ogiyama, Yuki; Soejima, Saeko; Takahashi, Kohta
Spindle assembly checkpoint (SAC) is an evolutionarily conserved surveillance system for chromosome missegregation. We isolated fission yeast Hos2, a component of the Dam1/DASH complex, as a multicopy suppressor of temperature-sensitive (ts) growth of nnf1-495 mutant that exhibits the minichromosome instability (mis) phenotype, producing lethal aneuploids without prominent mitotic delay. It remains elusive why SAC is satisfied in mis mutants despite the occurrence of missegregation. We found that Hos2 binds to the inner-kinetochore regions in both prometaphase and metaphase. Hos2 is essential for kinetochore localization of Dis1, a microtubule (MT) associated Dis1/XMAP215/TOG family protein that is required for proper MT dynamics. Cells lacking DASH exhibit cold-sensitive (cs) growth with the defective in sister-chromatid disjoining (dis) phenotype, which is characterized by hyper-condensed sister-chromatid pairs and elongated spindle MTs. Although DASH-deficient cells are viable at high temperatures, DASH-deletion transforms all the inner-kinetochore mis mutants so far tested into a constitutively active state of SAC, leading to the dis phenotype. We also discovered that Hos2 over-expression commonly suppresses growth retardation in a variety of inner-kinetochore mutants. These genetic interactions highlight the DASH-action(s) in satisfying SAC when aneuploids are formed during mitosis in the inner-kinetochore-defective mis mutants. PMID:17352737
Polydnaviruses: Roles in insect pathology and applications
Technology Transfer Automated Retrieval System (TEKTRAN)
One of the more unusual groups of insect pathogens consists of members of the family Polydnaviridae, DNA insect viruses that live in mutual symbioses with their associated parasitoid wasp (Hymentoptera) carriers until they are injected into specific Lepidopteran hosts. Once inside this secondary hos...
78 FR 48927 - Hours of Service of Drivers: U.S. Department of Defense (DOD); Application for Exemption
Federal Register 2010, 2011, 2012, 2013, 2014
...FMCSA announces that it has received an application from the U.S. Department of Defense (DOD) Military Surface Deployment and Distribution Command (SDDC) for an exemption from the minimum 30-minute rest break provision of the Agency's hours-of-service (HOS) regulations for commercial motor vehicle (CMV) drivers. The exemption would enable SDDC's contract motor carriers and their......
76 FR 8990 - Hours of Service of Drivers; Availability of Supplemental Documents and Corrections to Notice of...
Federal Register 2010, 2011, 2012, 2013, 2014
..., please see the preamble to the December 2010 HOS NPRM (75 FR 82170; December 29, 2010). The docket for... December 29, 2010 (75 FR 82170): ] 1. Page 82177. Correct footnote 17 to read as follows: \\17\\ Balkin, T..., J.A.M. & Wasserman, J.F., ``Improving Heavy-Duty Diesel Truck Ergonomics to Reduce Fatigue...
Unraveling the Slut Narrative: Gender Constraints on Adolescent Girls' Sexual Decision-Making
ERIC Educational Resources Information Center
Summit, Aleza K.; Kalmuss, Debra; DeAtley, Jenifer; Levack, Andrew
Limited research exists on the slut labeling process, a key means of enforcing rules around appropriate female sexuality. This study explores that process through qualitative interviews with 44 adolescent girls in Travis County, Texas. Labeling girls as sluts or hos was pervasive and was based on a number of factors beyond sexual behavior,…
77 FR 7544 - Hours of Service of Drivers: Correction
Federal Register 2010, 2011, 2012, 2013, 2014
.... SUMMARY: FMCSA corrects the hours of service (HOS) final rule published on December 27, 2011 (76 FR 81143... the discussion of the Agency's intentions in the section- by-section analysis on 76 FR 81165 col. 3... December 27, 2011 rule. This notice does not change, in any manner, the regulatory text intended. In FR...
Hyperglycemia is a risk factor for early death and morbidity in extremely low-birth weight infants
Technology Transfer Automated Retrieval System (TEKTRAN)
OBJECTIVES. The objectives of this study were to determine the prevalence of hyperglycemia in extremely low birth-weight infants and to determine whether hyperglycemia increases the risk of early adverse outcomes (death or intraventricular hemorrhage of grade 3 or 4) and/or affects the length of hos...
77 FR 74267 - Agency Information Collection Activities; New Information Collection Request: Driver and Carrier...
Federal Register 2010, 2011, 2012, 2013, 2014
... Docket Management System published in the Federal Register on January 17, 2008 (73 FR 3316), or you may... operating CMVs in interstate commerce to use EOBRs to document their drivers' HOS (76 FR 5537, February 1... requesting public comment on the harassment issue on April 13, 2011 (76 FR 20612). The Agency sought...
Synthesis of Conformationally North-Locked Pyrimidine Nucleosides Built on an Oxa-bicyclo[3.1.0]hexane Scaffold
Ludek, Olaf R.; Marquez, Victor E.
Beginning with a known 3-oxabicyclo[3.1.0]hexane scaffold (I), the relocation of the fused cyclopropane ring bond and the shifting of the oxygen atom to an alternative location engendered a new 2-oxabicyclo[3.1.0]hexane template (II) that mimics more closely the tetrahydrofuran ring of conventional nucleosides. The synthesis of this new class of locked nucleosides involved a novel approach that required the isocyanate II (B = NCO) with a hydroxyl-protected scaffold as a pivotal intermediate that was obtained in eleven steps from a known dihydrofuran precursor. The completion of the nucleobases was successfully achieved by quenching the isocyanate with the lithium salts of the corresponding acrylic amides that led to the uracil and thymidine precursors in a single step. Ring closure of these intermediates led to the target, locked nucleosides. The anti-HIV activity of 29 (uridine analogue), 31 (thymidine analogue), and 34 (cytidine analogue) was explored in human osteosarcoma (HOS) cells or modified HOS cells (HOS-313) expressing the herpes simplex virus 1 thymidine kinase (HSV-1 TK). Only the cytidine analogue showed moderate activity in HOS-313 cells, which means that the compounds are not good substrates for the cellular kinases. PMID:22026578
Bobcat (Lynx rufus) as a new natural intermediate host for Sarcocystis neurona
Technology Transfer Automated Retrieval System (TEKTRAN)
The protozoan Sarcocystis neurona is an important cause of severe clinical disease of horses (called equine protozoal myeloencephalitis, EPM), marine mammals, companion animals, and several species of wildlife animals in the Americas. The Virginia opossum (Didelphis virginiana) is its definitive hos...
Histone deacetylases in fungi: novel members, new facts.
Trojer, Patrick; Brandtner, Eva M; Brosch, Gerald; Loidl, Peter; Galehr, Johannes; Linzmaier, Roland; Haas, Hubertus; Mair, Karin; Tribus, Martin; Graessle, Stefan
Acetylation is the most prominent modification on core histones that strongly affects nuclear processes such as DNA replication, DNA repair and transcription. Enzymes responsible for the dynamic equilibrium of histone acetylation are histone acetyltransferases (HATs) and histone deacetylases (HDACs). In this paper we describe the identification of novel HDACs from the filamentous fungi Aspergillus nidulans and the maize pathogen Cochliobolus carbonum. Two of the enzymes are homologs of Saccharomyces cerevisiae HOS3, an enzyme that has not been identified outside of the established yeast systems until now. One of these homologs, HosB, showed intrinsic HDAC activity and remarkable resistance against HDAC inhibitors like trichostatin A (TSA) when recombinant expressed in an Escherichia coli host system. Phylo genetic analysis revealed that HosB, together with other fungal HOS3 orthologs, is a member of a separate group within the classical HDACs. Immunological investigations with partially purified HDAC activities of Aspergillus showed that all classical enzymes are part of high molecular weight complexes and that a TSA sensitive class 2 HDAC constitutes the major part of total HDAC activity of the fungus. However, further biochemical analysis also revealed an NAD(+)-dependent activity that could be separated from the other activities by different types of chromatography and obviously represents an enzyme of the sirtuin class. PMID:12853613
Technology Transfer Automated Retrieval System (TEKTRAN)
Xylella fastidiosa (Xf) is a xylem-limited bacterial pathogen that causes leaf scorch symptoms in numerous host plant species in urban, agricultural, and natural ecosystems worldwide. The exact mechanism of hydraulic disruption and systemic colonization of xylem by Xf remains elusive across all hos...
76 FR 5537 - Electronic On-Board Recorders and Hours of Service Supporting Documents
Federal Register 2010, 2011, 2012, 2013, 2014
... complete Privacy Act Statement in the Federal Register notice published on April 11, 2000 (65 FR 19476). D... Motor Carrier Safety Regulations--FMCSRs Federal Register--FR Fleet Management System--FMS Global... carriers with significant HOS violations (75 FR 17208). That final rule required a motor carrier that...
"The Use of History of Mechanics in Teaching Motion and Force Units"
ERIC Educational Resources Information Center
Seker, Hayati; Welsh, Laura C.
This paper reports a 4-month study that investigated the effectiveness of curriculum materials incorporating the history of science (HOS) on learning science, understanding the nature of science (NOS), and students' interest in science. With regards to these objectives, three different class contexts were developed with three main types of…
Technical decision-making with higher order structure data: specific binding of a nonionic detergent perturbs higher order structure of a therapeutic monoclonal antibody.
Budyak, Ivan L; Doyle, Brandon L; Weiss, William F
Robust higher order structure (HOS) characterization capability and strategy are critical throughout biopharmaceutical development from initial candidate selection and formulation screening to process optimization and manufacturing. This case study describes the utility of several orthogonal HOS methods as investigational tools during purification process development. An atypically high level of residual detergent in a development drug substance batch of a therapeutic monoclonal antibody triggered a root cause investigation. Several orthogonal biophysical techniques were used to uncover and characterize a specific interaction between the detergent and the antibody. Isothermal titration calorimetry (ITC) was used to quantify the molar ratio and affinity of the binding event, and circular dichroism (CD) spectroscopy and differential scanning calorimetry (DSC) were used to evaluate corresponding impacts on secondary/tertiary structure and thermal stability, respectively. As detergents are used routinely in biopharmaceutical processing, this case study highlights the value and power of HOS data in informing technical investigations and underlines the importance of HOS characterization as a component of overall biopharmaceutical analytical control strategy. PMID:25545760
Proposed performance standards for the Agricultural Hazardous Occupations Order certification training program.
French, B F; Field, W E; Tormoehlen, R
The performance standards that accompany Gearing Up for Safety, the agricultural production safety training curriculum for youth, were designed to meet the certification testing requirements of the Agricultural Hazardous Occupations Order (AgHOs) certification training program. The AgHOs requires that certain youth who are older than 13 and younger than 16 years of age complete a prescribed training program before being eligible for employment on farms to perform certain tasks considered hazardous. These training resources will be available for implementation and can be accessed by contacting the second author. To our knowledge, up until this time, few, if any, standardized testing procedures or instruments have had the necessary psychometric evidence to support use of the resulting scores to assess the readiness of youth to meet the basic certification requirements contained in the AgHOs. This article describes the rationale, methods, and key findings from a standard-setting session held to establish minimum passing scores for a proposed testing process. The test will be used as a component of the Gearing Up for Safety certification process for youth workers seeking employment to perform job functions as specified under the AgHOs. Readers may be able to apply this standardized testing process, or similarly accepted processes, to other or future testing and certification projects involving agricultural safety and health. PMID:17892071
Polarity, selectivity and performance of hydrophilic organic/salt-containing aqueous two-phase system on counter-current chromatography for polar compounds.
Liu, Dan; Hong, Zhilai; Gao, Mingzhe; Wang, Zhixin; Gu, Ming; Zhang, Xiaozhe; Xiao, Hongbin
The essential attributes of a solvent system for separation polar compounds on CCC are polarity, selectively and performance. Here, hydrophilic organic/salt-containing aqueous two-phase system (HO/S TPS) was evaluated as an alternative solvent system for CCC separation of polar compounds. Polarity measurements based on Rohrschneider-Snyder parameter was developed as quantitative assessing the polarity of HO/S TPS and comparing with an organic/aqueous system. All investigated 1-butanol/ethanol/saturated ammonium sulfate solution/water (BEAsWat) and 1-butanol/ethanol/saturated dipotassium hydrogen phosphate solution/water (BEDhpWat) systems with polarity values of organic phase from 4.5 to 6.8, were more polar than chloroform/methanol/water (1/1/1). The considerable water content of BEAsWat and BEDhpWat (0/1/1/1/) was 45.4 and 42.6% (w%) of hydrophilic organic phase, and 66.4 and 51.2% (w%) of salt-containing aqueous phase, respectively, closed to conventional aqueous two-phase system. Therefore, the polarity of HO/S TPS is in the middle of organic/aqueous and aqueous two-phase system. The LogKC values of twenty four polar compounds as model mixture confirmed that the polarities of HO/S TPSs were matched to that of the polar compounds and shown to be a very selective technique capable of separating positional isomers. Moreover, BEAsWat and BEDhpWat systems can be easily retained in CCC column with suitable elution mode. The hydrodynamic behavior reversion of HO/S TPS on hydrodynamic CCC was observed and was tentatively explained based on the density difference. Finally, caffeoylquinic acid isomers and dihydroxybenzoic acid isomers were successfully separated with HO/S TPS on CCC, respectively. Those results demonstrate that HO/S TPS on CCC is a performant and stable way to separate polar compounds from natural products. PMID:27131958
Palmitic acid in the sn-2 position of dietary triacylglycerols does not affect insulin secretion or glucose homeostasis in healthy men and women
Filippou, A; Teng, K-T; Berry, S E; Sanders, T A B
Background/objectives: Dietary triacylglycerols containing palmitic acid in the sn-2 position might impair insulin release and increase plasma glucose. Subjects/Methods: We used a cross-over designed feeding trial in 53 healthy Asian men and women (20–50 years) to test this hypothesis by exchanging 20% energy of palm olein (PO; control) with randomly interesterified PO (IPO) or high oleic acid sunflower oil (HOS). After a 2-week run-in period on PO, participants were fed PO, IPO and HOS for 6 week consecutively in randomly allocated sequences. Fasting (midpoint and endpoint) and postprandial blood at the endpoint following a test meal (3.54 MJ, 14 g protein, 85 g carbohydrate and 50 g fat as PO) were collected for the measurement of C-peptide, insulin, glucose, plasma glucose-dependent insulinotropic polypeptide and glucagon-like peptide-1, lipids and apolipoproteins; pre-specified primary and secondary outcomes were postprandial changes in C-peptide and plasma glucose. Results: Low density lipoprotein cholesterol was 0.3 mmol/l (95% confidence interval (95% CI)) 0.1, 0.5; P<0.001) lower on HOS than on PO or IPO as predicted, indicating good compliance to the dietary intervention. There were no significant differences (P=0.58) between diets among the 10 male and 31 female completers in the incremental area under the curve (0–2 h) for C-peptide in nmol.120 min/l: GM (95% CI) were PO 220 (196, 245), IPO 212 (190, 235) and HOS 224 (204, 244). Plasma glucose was 8% lower at 2 h on IPO vs PO and HOS (both P<0.05). Conclusion: Palmitic acid in the sn-2 position does not adversely impair insulin secretion and glucose homeostasis. PMID:25052227
Anti-inflammatory effects of sucrose-derived oligosaccharides produced by a constitutive mutant L. mesenteroides B-512FMCM dextransucrase in high fat diet-fed mice.
Kang, Min-Gyung; Lee, Hee Jae; Cho, Jae-Young; Kim, Kanghwa; Yang, Soo Jin; Kim, Doman
Oligosaccharide (OS) is used as a sugar replacement as well as an ingredient in functional foods because of its beneficial effects, mainly on reducing calorie content and promoting intestinal health. By contrast, the effects of OS on inflammation are less well investigated. The purpose of this study was to investigate the effects of sucrose-derived OS on glucose control and inflammation in high fat (HF) diet-fed mice. Male C57BL6 mice were randomly assigned to six treatment groups (n = 10-14 mice per group): 1) lean control (CON), 2) HF control, 3) HF-low sucrose (LS, 100 mg/kg/day), 4) HF-high sucrose (HS, 1000 mg/kg/day), 5) HF-low OS (LOS, 100 mg/kg/day), and 6) HF-high OS (HOS, 1000 mg/kg/day). PBS (vehicle), sucrose, and OS were administered by stomach gavage. Body weight, food intake, and markers of liver function (activities of aspartate aminotransferase and alanine aminotransferase) were not affected by the treatments. HOS treatment decreased levels of serum glucose, insulin, and homeostasis model assessment-insulin resistance compared with sucrose treatment. However, serum adiponectin levels of the HOS group were higher than those of the sucrose groups. Serum levels of the pro-inflammatory cytokines interleukin-6 (IL-6) and fetuin-A were lower in the HOS group than in the sucrose groups. Hepatic gene expression levels of pro-inflammatory cytokines and related factors (fetuin-A, NF-κB, TLR4, TNF-alpha, and IL-6) were decreased and the levels of insulin signaling-related molecules (sirtuin 1, insulin receptor, and Akt) were increased in HOS-treated mice as compared with sucrose-treated mice. These results demonstrate that OS treatment is effective in improving glucose control and inflammation in high fat diet-fed mice. PMID:27342664
Molecular cloning and expression analysis of KIN10 and cold-acclimation related genes in wild banana 'Huanxi' (Musa itinerans).
Liu, Weihua; Cheng, Chunzhen; Lai, Gongti; Lin, Yuling; Lai, Zhongxiong
Banana cultivars may experience chilling or freezing injury in some of their cultivated regions, where wild banana can still grow very well. The clarification of the cold-resistant mechanism of wild banana is vital for cold-resistant banana breeding. In this study, the central stress integrator gene KIN10 and some cold-acclimation related genes (HOS1 and ICE1s) from the cold-resistant wild banana 'Huanxi' (Musa itinerans) were cloned and their expression patterns under different temperature treatments were analyzed. Thirteen full-length cDNA transcripts including 6 KIN10s, 1 HOS1 and 6 ICE1s were successfully cloned. Quantitative real-time PCR (qRT-PCR) results showed that all these genes had the highest expression levels at the critical temperature of banana (13 °C). Under chilling temperature (4 °C), the expression level of KIN10 reduced significantly but the expression of HOS1 was still higher than that at the optimal temperature (28 °C, control). Both KIN10 and HOS1 showed the lowest expression levels at 0 °C, the expression level of ICE1, however, was higher than control. As sucrose plays role in plant cold-acclimation and in regulation of KIN10 and HOS1 bioactivities, the sucrose contents of wild banana under different temperatures were detected. Results showed that the sucrose content increased as temperature lowered. Our result suggested that KIN10 may participate in cold stress response via regulating sucrose biosynthesis, which is helpful in regulating cold acclimation pathway in wild banana. PMID:26753116
Anemone altaica Induces Apoptosis in Human Osteosarcoma Cells.
Chang, I-Chang; Chiang, Tsay-I; Lo, Chun; Lai, Yi-Hua; Yue, Chia-Herng; Liu, Jer-Yuh; Hsu, Li-Sung; Lee, Chia-Jen
In the past decade, no significant improvement has been made in chemotherapy for osteosarcoma (OS). To develop improved agents against OS, we screened 70 species of medicinal plants and treated two human OS cell lines with different agent concentrations. We then examined cell viability using the MTT assay. Results showed that a candidate plant, particularly the rhizomes of Anemone altaica Fisch. ex C. A. Mey aqueous extract (AAE), suppressed the viability of HOS and U2OS cells in a concentration-dependent manner. Flow cytometry analysis revealed that AAE significantly increased the amount of cell shrinkage (Sub-G1 fragments) in HOS and U2OS cells. Moreover, AAE increased cytosolic cytochrome c and Bax, but decreased Bcl-2. The amount of cleaved caspase-3 and poly-(ADP-ribose) polymerase-1 (PARP-1) were significantly increased. AAE suppressed the growth of HOS and U2OS through the intrinsic apoptotic pathway. Data suggest that AAE is cytotoxic to HOS and U2OS cells and has no significant influence on human osteoblast hFOB cells. The high mRNA levels of apoptosis-related factors (PPP1R15A, SQSTM1, HSPA1B, and DDIT4) and cellular proliferation markers (SKA2 and BUB1B) were significantly altered by the AAE treatment of HOS and U2OS cells. Results show that the anticancer activity of AAE could up-regulate the expression of a cluster of genes, especially those in the apoptosis-related factor family and caspase family. Thus, AAE has great potential as a useful therapeutic drug for human OS. PMID:26224029
A two-stage approach using Gaussian mixture models and higher-order statistics for a classification of normal and pathological voices
NASA Astrophysics Data System (ADS)
Lee, Ji Yeoun
A two-stage classifier is used to improve the classification performance between normal and pathological voices. A primary classification between normal and pathological voices is achieved by the Gaussian mixture model (GMM) log-likelihood scores. For samples that do not meet the thresholds for normal or disordered voice in the GMM, the final decision is made by a higher-order statistics (HOS)-based parameter. The normalized skewness and kurtosis, and means of the normalized skewness and kurtosis were estimated using a sustained vowel /a/ from 53 normal and 173 pathological voices taken from the Disordered Voice Database. Mel-frequency cepstral coefficients (MFCC)-based GMM, the HOS methods, and a two-stage classifier based on the GMM-HOS were performed for each voice signal. A Mann-Whitney rank sum test was used to detect differences in the means of the HOS-based parameters. A fivefold cross-validation scheme was performed to test the classification method. When 16 Gaussian mixtures were used, the MFCC-based GMM algorithm is performed with 92.0% accuracy. When means of the normalized skewness and kurtosis were used, performances of 82.31 and 83.67% were obtained, respectively. The two-stage classifier with 16 Gaussian mixtures and the mean of the normalized kurtosis classified samples with a 96.96% accuracy were obtained. The proposed two-stage classifier is more accurate than the MFCC-based GMM and HOS methods alone and shows potential for the classification of voices in the clinic.
alpha-Diimine Ligand Coordination and C H Bond Activation in the Reaction of Os3(CO)10(MeCN)2 with 6-R-2,2'-Bipyridine (where R = Et, Ph): X-ray Diffraction Structures of the Ortho-Metalated
Carrano, Carl J.; Wang, Xiaoping; Poola, Bhaskar; Powell, Cynthia B.; Richmond, Michael G.
The reactivity of the labile cluster Os3(CO)10(MeCN)2 (1) with the monofunctionalized heterocyclic ligands 6-R-2,2 -bipyridine (where R = Et, Ph) has been investigated. The alkyl-substituted heterocycle 6-Et-2,2 -bipyridine reacts with 1 in refluxing CH2Cl2 to give an isomeric mixture of HOs3(CO)9(N2C12H11) due to cyclometalation of the side-chain ethyl group (2) and ortho metalation of the unsubstituted bipyridine ring (3). The solid-state structure of the latter cluster, HOs3(CO)9(N2C10H6-6-Et) (3), has unequivocally established the site of the C-H bond activation in the product. Treatment of 1 with the aryl-substituted ligand 6-Ph-2,2 -bipyridine proceeds similarly with ortho metalation at the ancillary phenyl group and the C-6 ortho site of the unsubstituted bipyridine ring, as verified by 1H NMR spectroscopy. The X-ray diffraction structure of the thermodynamically more stable bipyridine-metalated cluster HOs3(CO)9(N2C10H6-6-Ph) (5) has been determined. The course of these reactions is discussed with respect to our recent study involving the reaction of cluster 1 with the ligand 6-Me-2,2 -bipyridine. Graphical Abstract The reaction between the labile cluster Os3(CO)10(MeCN)2 (1) and the monofunctionalized heterocyclic ligand 6-Et-2,2 -bipyridine proceeds readily at room temperature to furnish an isomeric mixture of the cyclometalated and ortho-metalated hydride-bridged clusters HOs3(CO)9(N2C12H11) (2 and 3). Treatment of 1 with 6-Ph-2,2 -bipyridine also yields two distinct hydride-containing clusters that result from independent ortho-metalation paths involving the 6-phenyl substituent and unsubstituted bipyridine group. The bipyridine-derived ortho metalation attendant in the new clusters HOs3(CO)9(N2C10H6-6-Et) (3) and HOs3(CO)9(N2C10H6-6-Ph) (5) has been established by X-ray crystallography.
Knowledge Regarding Basic Facts of Stroke Among Final Year MBBS Students and House Officers: A Cross-Sectional Survey of 708 Respondents from Pakistan
Khubaib, Mohammad U; Waqas, Ahmed; Jan, Mohsin M; Sohail, Sana
Introduction: Stroke is the leading cause of neurological disability in the world. In Pakistan, house officers (HOs) are usually the first contact for a stroke patient in the emergency department. Sometimes they need to make quick decisions regarding diagnosis and management without specialist supervision. Thousands of current final year MBBS (Bachelor of Medicine, Bachelor of Surgery) students will be performing the duties of HOs soon. This study documents the knowledge and confidence levels of final year students and HOs in Pakistan regarding basic facts related to initial diagnosis and management of stroke. Materials and Methods: A questionnaire was developed using two standard textbooks of medicine and current stroke guidelines of the American Heart Association. The pre-tested self-administered questionnaire was distributed among 800 final year MBBS students and HOs in 14 medical colleges and hospitals in four different cities. The response rate was 88.5%. Data analysis was done using SPSS V.21. The CMH Lahore Medical College Ethics Review Committee approved this project. Results: Respondents included medical students (n=496) and HOs (N= 212); most were female (n = 452, 63.9%). Of these, 31.4% had managed or assisted in the management of a patient with a stroke and had a higher confidence level in its diagnosis (p< 0.001) and management (p <0.001). Having a family member with stroke was associated with higher confidence in the diagnosis of stroke (p < 0.05) but not with confidence in its management (p = 0.41). Most correctly defined stroke (60.6%), identified the CT scan as the initial diagnostic modality (88.1%), knew the dosage of aspirin (64.9%), knew the time limit for thrombolysis (67.4%), and were familiar with the risk of deep vein thrombosis in immobilized stroke patients (85.4%). Less than half (44.5%) chose tissue plasminogen activator (t-PA) as the preferred initial intervention for acute ischemic stroke. Conclusion: This multicenter survey shows
Application of Omics Technologies in Occupational and Environmental Health Research; Current Status and Projections
Vlaanderen, Jelle; Moore, Lee E.; Smith, Martyn T.; Lan, Qing; Zhang, Luoping; Skibola, Christine F.; Rothman, Nathaniel; Vermeulen, Roel
OMICS technologies are relatively new biomarker discovery tools that can be applied to study large sets of biological molecules. Their application in human observational studies (HOS) has become feasible in recent years due to a spectacular increase in the sensitivity, resolution and throughput of OMICS based assays. Although, the number of OMIC techniques is ever expanding, the five most developed OMICS technologies are genotyping, transcriptomics, epigenomics, proteomics and metabolomics. These techniques have been applied in HOS to various extents. However, their application in Occupational Environmental Health (OEH) research has been limited. Here, we will discuss the opportunities these new techniques provide for OEH research. In addition we will address difficulties and limitations to the interpretation of the data that is generated by OMICS technologies. To illustrate the current status of the application of OMICS in OEH research, we will provide examples of studies that used OMICS technologies to investigate human health effects of two well known toxicants, benzene and arsenic. PMID:19933307
Electron heating due to microwave photoexcitation in the high mobility GaAs/AlGaAs two dimensional electron system
NASA Astrophysics Data System (ADS)
Ramanayaka, A. N.; Mani, R. G.; Wegscheider, W.
We extract the electron temperature in the microwave photo-excited high mobility GaAs/AlGaAs two dimensional electron system (2DES) by studying the influence of microwave radiation on the amplitude of Shubnikov-de Haas oscillations (SdHOs) in a regime where the cyclotron frequency, ωc, and the microwave angular frequency, ω, satisfy 2ω ≤ ωc ≤ 3.5ω The results indicate that increasing the incident microwave power has a weak effect on the amplitude of the SdHOs and therefore the electron temperature, in comparison to the influence of modest temperature changes on the dark-specimen SdH effect. The results indicate negligible electron heating under modest microwave photo-excitation, in good agreement with theoretical predictions.
Characterization of tissue plasminogen activator binding proteins isolated from endothelial cells and other cell types
Beebe, D.P.; Wood, L.L.; Moos, M. )
Human tissue plasminogen activator (t-PA) was shown to bind specifically to human osteosarcoma cells (HOS), and human epidermoid carcinoma cells (A-431 cells). Crosslinking studies with DTSSP demonstrated high molecular weight complexes (130,000) between {sup 125}I-t-PA and cell membrane protein on human umbilical vein endothelial cells (HUVEC), HOS, and A-431 cells. A 48-65,000 molecular weight complex was demonstrated after crosslinking t-PA peptide (res. 7-20) to cells. Ligand blotting of cell lysates which had been passed over a t-PA affinity column revealed binding of t-PA to 54,000 and 95,000 molecular weight proteins. Several t-PA binding proteins were identified in immunopurified cell lysates, including tubulin beta chain, plasminogen activator inhibitor type 1 and single chain urokinase.
Electron heating due to microwave photoexcitation in the high mobility GaAs/AlGaAs two dimensional electron system
Ramanayaka, A. N.; Mani, R. G.; Wegscheider, W.
We extract the electron temperature in the microwave photo-excited high mobility GaAs/AlGaAs two dimensional electron system (2DES) by studying the influence of microwave radiation on the amplitude of Shubnikov-de Haas oscillations (SdHOs) in a regime where the cyclotron frequency, ω{sub c}, and the microwave angular frequency, ω, satisfy 2ω ≤ ω{sub c} ≤ 3.5ω The results indicate that increasing the incident microwave power has a weak effect on the amplitude of the SdHOs and therefore the electron temperature, in comparison to the influence of modest temperature changes on the dark-specimen SdH effect. The results indicate negligible electron heating under modest microwave photo-excitation, in good agreement with theoretical predictions.
Morphologic characterization of osteosarcoma growth on the chick chorioallantoic membrane
Background The chick chorio-allantoic membrane (CAM) assay is a commonly used method for studying angiogenic or anti-angiogenic activities in vivo. The ease of access allows direct monitoring of tumour growth by biomicroscopy and the possibility to screen many samples in an inexpensive way. The CAM model provides a powerful tool to study effects of molecules, which interfere with physiological angiogenesis, or experimental tumours derived from cancer cell lines. We therefore screened eight osteosarcoma cell lines for their ability to form vascularized tumours on the CAM. Findings We implanted 3-5 million cells of human osteosarcoma lines (HOS, MG63, MNNG-HOS, OST, SAOS, SJSA1, U2OS, ZK58) on the CAM at day 10 of embryonic development. Tumour growth was monitored by in vivo biomicroscopy at different time points and tumours were fixed in paraformaldehyde seven days after cell grafting. The tissue was observed, photographed and selected cases were further analyzed using standard histology. From the eight cell lines the MNNG-HOS, U2OS and SAOS were able to form solid tumours when grafted on the CAM. The MNNG-HOS tumours showed the most reliable and consistent growth and were able to penetrate the chorionic epithelium, grow in the CAM stroma and induce a strong angiogenic response. Conclusions Our results show that the CAM assay is a useful tool for studying osteosarcoma growth. The model provides an excellent alternative to current rodent models and could serve as a preclinical screening assay for anticancer molecules. It might increase the speed and efficacy of the development of new drugs for the treatment of osteosarcoma. PMID:20202196
Overexpression of urokinase receptor increases matrix invasion without altering cell migration in a human osteosarcoma cell line.
Karikó, K; Kuo, A; Boyd, D; Okada, S S; Cines, D B; Barnathan, E S
Proteolysis triggered by receptor-bound urokinase-type plasminogen activator (uPA) involves a cascade of species-specific molecular interactions. To study the role of the uPA receptor (uPAR) in such interactions, a human osteosarcoma cell line (HOS), which normally expresses low levels of uPAR, was transfected with human uPAR complementary DNA. One of several stably transformed clonal cells lines, designated 2A2, was characterized and compared to the parental HOS, revealing the following: (a) stable incorporation of uPAR complementary DNA into the genome demonstrated by Southern blot analysis; (b) a 10-fold increase in steady state mRNA levels of uPAR assessed by Northern blot analysis; (c) a 2-fold increase in the surface expression of glycosylphosphatidylinositol anchored uPAR protein determined by enzyme-linked immunosorbent assay and by the specific binding of radiolabeled single chain uPA; (d) a 2-fold increase in internalization and degradation of radiolabeled uPA/PAI-1 complexes; and (e) a 2-fold increase in receptor-bound uPA-mediated plasmin generation measured by the cleavage of a chromogenic substrate and degradation of 125I-labeled laminin. The involvement of uPAR in cellular processes was determined by comparing 2A2 and HOS cells in in vitro migration and invasion assays. The migration of 2A2 cells were slower on fibronectin-coated surfaces in a linear under-agarose assay, but both cell lines migrated at the same rate on uncoated polycarbonate filters in Boyden chamber assays. In the invasion experiments, 4 times more 2A2 than HOS cells penetrated through the barrier of reconstituted basement membrane Matrigel. These data suggest that uPAR does not potentiate random cell migration but facilitates matrix degradation and subsequent cell invasion. PMID:8391387
Recent Applications of the Volterra Theory to Aeroelastic Phenomena
NASA Technical Reports Server (NTRS)
Silva, Walter A.; Haji, Muhammad R; Prazenica, Richard J.
The identification of nonlinear aeroelastic systems based on the Volterra theory of nonlinear systems is presented. Recent applications of the theory to problems in experimental aeroelasticity are reviewed. These results include the identification of aerodynamic impulse responses, the application of higher-order spectra (HOS) to wind-tunnel flutter data, and the identification of nonlinear aeroelastic phenomena from flight flutter test data of the Active Aeroelastic Wing (AAW) aircraft.
Oncogenic roles of carbonic anhydrase 8 in human osteosarcoma cells.
Wang, Tze-Kai; Lin, Yu-Ming; Lo, Che-Min; Tang, Chih-Hsin; Teng, Chieh-Lin Jerry; Chao, Wei-Ting; Wu, Min Huan; Liu, Chin-San; Hsieh, Mingli
Carbonic anhydrase 8 (CA8), a member of the carbonic anhydrase family, is one of the three isozymes that do not catalyze the reversible hydration of carbon dioxide due to the lack of one important histidine. In the present study, we observed increased expression of CA8 in more aggressive types of human osteosarcoma (OS) cells and found that CA8 expression is correlated with disease stages, such that more intense expression occurs in the disease late stage. We also demonstrated that overexpression of CA8 in human OS (HOS) cells significantly increased cell proliferation both in vitro and in vivo. Downregulated CA8 sensitized cells to apoptotic stress induced by staurosporine and cisplatin, suggesting a specific role of CA8 to protect cells from stresses. In addition, downregulation of CA8 in HOS cells reduced cell invasion and colony formation ability in soft agar and further decreased matrix metalloproteinase 9 and focal adhesion kinase expression, indicating that CA8 might facilitate cancer cell invasion via the activation of FAK-MMP9 signaling. Interestingly, HOS cells with CA8 knockdown showed a significant decrease in glycolytic activity and cell death under glucose withdrawal, further indicating that CA8 may be involved in regulating aerobic glycolysis and enhancing cell viability. Knockdown of CA8 significantly decreased phosphorylated Akt expression suggesting that the oncogenic role of CA8 may be mediated by the regulation of Akt activation through p-Akt induction. Importantly, the inhibition of glycolysis by 2-deoxyglucose sensitized CA8 HOS-CA8-myc cells to cisplatin treatment under low glucose condition, highlighting a new therapeutic option for OS cancer. PMID:26711783
The role of propionates in substrate binding to heme oxygenase from Neisseria meningitidis; A NMR study†
Peng, Dungeng; Ma, Li-Hua; Smith, Kevin M.; Zhang, Xuhong; Sato, Michihiko; La Mar, Gerd N.
Heme oxygenase, HO, cleaves hemin into biliverdin, iron and CO. For mammalian HOs, both native hemin propionates are required for substrate binding and activity. The HO from the pathogenic bacterium Neisseria meningitidis, NmHO, possesses a crystallographically undetected C-terminal fragment that by solution 1H NMR is found to fold and interact with the active site. One of the substrate propionates has been proposed to form a salt bridge to the C-terminus rather than to the conventional buried cationic side chain in other HOs. Moreover, the C-terminal dipeptide Arg208His209 cleaves spontaneously over ~24 hours at a rate dependent on substituent size. 2D 1H NMR of NmHO azide complexes with hemins with selectively deleted or rearranged propionates all bind to NmHO with a structurally conserved active site as reflected in optical spectra and NMR NOESY cross peak and hyperfine shift patterns. In contrast to mammalian HOs, NmHO requires only a single propionate interacting with the buried terminus of Lys16 to exhibit full activity and tolerates the existence of a propionate at the exposed 8-position. The structure of the C-terminus is qualitatively retained upon deletion of the 7-propionate but a dramatic change in the 7-propionate carboxylate 13C chemical shift upon C-terminal cleavage confirms its role in the interaction with the C-terminus. The stronger hydrophobic contacts between pyrroles A and B with NmHO contribute more substantially to the substrate binding free energy than in mammalian HOs, "liberating" one propionate to stabilize the C-terminus. The functional implications of the C-terminus in product release are discussed. PMID:22913621
Comparability of higher order structure in proteins: chemometric analysis of second-derivative amide I Fourier transform infrared spectra.
Stockdale, Gregory; Murphy, Brian M; D'Antonio, Jennifer; Manning, Mark Cornell; Al-Azzam, Wasfi
Comparing higher order structure (HOS) in therapeutic proteins is a significant challenge. Previously, we showed that changes in solution conditions produced detectable changes in the second-derivative amide I Fourier transform infrared (FTIR) spectra for a variety of model proteins. Those comparisons utilized vector-based approaches, such as spectral overlap and spectral correlation coefficients to quantify differences between spectra. In this study, chemometric analyses of the same data were performed, to classify samples into different groups based on the solution conditions received. The solution conditions were composed of various combinations of temperature, pH, and salt types. At first, principal component analysis (PCA) was used to visually demonstrate that FTIR spectra respond to changes in solution conditions, which, presumably indicates variations in HOS. This observed when samples from the same solution condition form clusters within a PCA score plot. The second approach, called soft independent modeling of class analogy (SIMCA), was conducted to account for the within-class experimental error for the lysozyme spectra. The DModX values, indicative of the distance of each spectra to their respective class models, was found to be a more sensitive quantitative indicator of changes in HOS, when compared with the modified area of overlap algorithm. The SIMCA approach provides a metric to determine whether new observations do, or do not belong to a particular class or group. Thus, SIMCA is the recommended approach when multiple samples from each condition are available. PMID:25382804
Higher-Order Statistics for the Detection of Small Objects in a Noisy Background Application on Sonar Imaging
NASA Astrophysics Data System (ADS)
Maussang, F.; Chanussot, J.; Hétet, A.; Amate, M.
An original algorithm for the detection of small objects in a noisy background is proposed. Its application to underwater objects detection by sonar imaging is addressed. This new method is based on the use of higher-order statistics (HOS) that are locally estimated on the images. The proposed algorithm is divided into two steps. In a first step, HOS (skewness and kurtosis) are estimated locally using a square sliding computation window. Small deterministic objects have different statistical properties from the background they are thus highlighted. The influence of the signal-to-noise ratio (SNR) on the results is studied in the case of Gaussian noise. Mathematical expressions of the estimators and of the expected performances are derived and are experimentally confirmed. In a second step, the results are focused by a matched filter using a theoretical model. This enables the precise localization of the regions of interest. The proposed method generalizes to other statistical distributions and we derive the theoretical expressions of the HOS estimators in the case of a Weibull distribution (both when only noise is present or when a small deterministic object is present within the filtering window). This enables the application of the proposed technique to the processing of synthetic aperture sonar data containing underwater mines whose echoes have to be detected and located. Results on real data sets are presented and quantitatively evaluated using receiver operating characteristic (ROC) curves.
Quantifying wave-breaking dissipation using nonlinear phase-resolved wave-field simulations with a phenomenological-based wave breaking model
NASA Astrophysics Data System (ADS)
Qi, Yusheng; Yue, Dick
We use direct nonlinear phase-resolved simulations based on a High-Order Spectral (HOS) method (Dommermuth & Yue 1987) to understand and quantify wave-breaking dissipation in the evolution of general irregular short-crested wave-fields. We achieve this by incorporating a robust phenomenological-based wave breaking model in HOS simulations to account for energy dissipation. This model can automatically simulate the onset of wave breaking, and the simulated wave-breaking dissipation strength differentiates corresponding to different wave breaking type (such as spilling or plunging breaking waves). The efficacy of this model is confirmed by direct comparisons against measurements for the energy loss in 2D and 3D breaking events. By comparing simulated wave-fields with and without the dissipation model in HOS, we obtain the dissipation field, which provides the times, locations and intensity of wave breaking events. From the dissipation field we further calculate the distribution of total length of breaking wave front per unit surface area per unit increment of breaking velocity (Phillips 1985), and obtain qualitative agreement with Phillips theoretical power-law.
OST1 kinase modulates freezing tolerance by enhancing ICE1 stability in Arabidopsis.
Ding, Yanglin; Li, Hui; Zhang, Xiaoyan; Xie, Qi; Gong, Zhizhong; Yang, Shuhua
Cold stress is a major environmental factor that limits plant growth and development. The C-repeat-binding factor (CBF)-dependent cold signaling pathway is extensively studied in Arabidopsis; however, the specific protein kinases involved in this pathway remain elusive. Here we report that OST1 (open stomata 1), a well-known Ser/Thr protein kinase in ABA signaling, acts upstream of CBFs to positively regulate freezing tolerance. The ost1 mutants show freezing hypersensitivity, whereas transgenic plants overexpressing OST1 exhibit enhanced freezing tolerance. The OST1 kinase is activated by cold stress. Moreover, OST1 interacts with both the transcription factor ICE1 and the E3 ligase HOS1 in the CBF pathway. Cold-activated OST1 phosphorylates ICE1 and enhances its stability and transcriptional activity. Meanwhile, OST1 interferes with the interaction between HOS1 and ICE1, thus suppressing HOS1-mediated ICE1 degradation under cold stress. Our results thus uncover the unexpected roles of OST1 in modulating CBF-dependent cold signaling in Arabidopsis. PMID:25669882
Improvement of liquid and frozen-thawed semen quality of Nili-Ravi buffalo bulls (Bubalus bubalis) through supplementation of fat.
Adeel, M; Ijaz, A; Aleem, M; Rehman, H; Yousaf, M S; Jabbar, M A
The aim of the study was to investigate the effects of dietary fat on quality of liquid and frozen-thawed semen of Nili-Ravi buffalo bulls. Adult bulls (n=21) were fed a balanced ration (Con; n=7) or the same ration either containing sunflower oil (SF-O; n=7) or whole sunflower seeds (SF-S; n=7) for 63 days. Body weight and body condition score of each bull was recorded on days 0, 30 and 60 of the experiment. Semen was collected on days 39, 46, 53 and 60, frozen by a fast method and stored at -196 degrees C for 24h. Sperm motility was assessed using a bright field microscope. Plasma membrane integrity of fresh and frozen-thawed spermatozoa was assessed using a hypo-osmotic swelling (HOS) assay. The concentration of spermatozoa and volume of semen was not different among groups on various days of collection. Sunflower-enriched diets did not affect the motility and number of HOS-positive spermatozoa in the fresh semen. Motility and HOS of post-thawed spermatozoa were higher (p<0.05) in bulls fed the sunflower-enriched diets. Similarly, diets did not affect the body condition score and body weight of bulls. In conclusion, feeding of sunflower oil or sunflower seed as fat sources can improve the quality of buffalo bull spermatozoa. PMID:19246083
Hematopoiesis toxicity induced by 4-methylumbelliferon determined in an invertebrate model organism.
Fang, Yan; He, Yue; Wang, Hua; Yan, Siqi; Xing, Rui; Yin, Weimin; Sima, Yanghu; Xu, Shiqing
Umbelliferone has potential value as it has an inhibitory effect on tumor cells; however, its impact on an animal's circulatory system and hematopoietic function has not been reported. In this study, 4-methylumbelliferon (4-MU), an umbelliferone derivative, was used as a model drug, and its potential toxicity on hemocytes and hematopoietic organs (HOs) was investigated using an invertebrate animal model, the silkworm, Bombyx mori. The results showed that the level of reactive oxygen species in HOs increased when larvae (third day of the fifth instar) were orally exposed to 4 mM 4-MU for 8 min, followed by the induction of improved antioxidative metabolism of coenzymes in hemolymph. Exposure to 4-MU also significantly upregulated the expression levels of several genes in the hemolymph and fat body (a detoxification tissue similar to the liver in mammals) including antimicrobial peptide gene cecropinA and moricin, and a phagocytosis-related gene, tetraspanin E, suggesting an increased antioxidant level and antimicrobial ability of the circulatory system. However, the percentage of dead hemocytes increased and hematopoiesis significantly decreased in HOs, indicating the toxic effect of 4-MU on hemocytes and hematopoiesis, despite it inducing enhanced antioxidant and antimicrobial activity in the circulatory system. PMID:26327572
Emotion classification in Parkinson's disease by higher-order spectra and power spectrum features using EEG signals: a comparative study.
Yuvaraj, R; Murugappan, M; Ibrahim, Norlinah Mohamed; Omar, Mohd Iqbal; Sundaraj, Kenneth; Mohamad, Khairiyah; Palaniappan, R; Satiyan, M
Deficits in the ability to process emotions characterize several neuropsychiatric disorders and are traits of Parkinson's disease (PD), and there is need for a method of quantifying emotion, which is currently performed by clinical diagnosis. Electroencephalogram (EEG) signals, being an activity of central nervous system (CNS), can reflect the underlying true emotional state of a person. This study applied machine-learning algorithms to categorize EEG emotional states in PD patients that would classify six basic emotions (happiness and sadness, fear, anger, surprise and disgust) in comparison with healthy controls (HC). Emotional EEG data were recorded from 20 PD patients and 20 healthy age-, education level- and sex-matched controls using multimodal (audio-visual) stimuli. The use of nonlinear features motivated by the higher-order spectra (HOS) has been reported to be a promising approach to classify the emotional states. In this work, we made the comparative study of the performance of k-nearest neighbor (kNN) and support vector machine (SVM) classifiers using the features derived from HOS and from the power spectrum. Analysis of variance (ANOVA) showed that power spectrum and HOS based features were statistically significant among the six emotional states (p < 0.0001). Classification results shows that using the selected HOS based features instead of power spectrum based features provided comparatively better accuracy for all the six classes with an overall accuracy of 70.10% ± 2.83% and 77.29% ± 1.73% for PD patients and HC in beta (13-30 Hz) band using SVM classifier. Besides, PD patients achieved less accuracy in the processing of negative emotions (sadness, fear, anger and disgust) than in processing of positive emotions (happiness, surprise) compared with HC. These results demonstrate the effectiveness of applying machine learning techniques to the classification of emotional states in PD patients in a user independent manner using EEG signals. The
Pre-operative Thresholds for Achieving Meaningful Clinical Improvement after Arthroscopic Treatment of Femoroacetabular Impingement
Nwachukwu, Benedict U.; Fields, Kara G.; Nawabi, Danyal H.; Kelly, Bryan T.; Ranawat, Anil S.
Objectives: Knowledge of the thresholds and determinants for successful femoroacetabular impingement (FAI) treatment is evolving. The primary purpose of this study was to define pre-operative outcome score thresholds that can be used to predict patients most likely to achieve meaningful clinically important difference (MCID) after arthroscopic FAI treatment. Secondarily determinants of achieving MCID were evaluated. Methods: A prospective institutional hip arthroscopy registry was reviewed to identify patients with FAI treated with arthroscopic labral surgery, acetabular rim trimming, and femoral osteochondroplasty. The modified Harris Hip Score (mHHS), the Hip Outcome Score (HOS) and the international Hip Outcome Tool (iHOT-33) tools were administered at baseline and at one year post-operatively. MCID was calculated using a distribution-based method. A receiver operating characteristic (ROC) analysis was used to calculate cohort-based threshold values predictive of achieving MCID. Area under the curve (AUC) was used to define predictive ability (strength of association) with AUC >0.7 considered acceptably predictive. Univariate and multivariable analyses were used to analyze demographic, radiographic and intra-operative factors associated with achieving MCID. Results: There were 374 patients (mean + SD age, 32.9 + 10.5) and 56.4% were female. The MCID for mHHS, HOS activities of daily living (HOS-ADL), HOS Sports, and iHOT-33 was 8.2, 8.4,14.5, and 12.0 respectively. ROC analysis (threshold, % achieving MCID, strength of association) for these tools in our population was: mHHS (61.6, 78%, 0.68), HOS-ADL (83.8, 68%, 0.84), HOS-Sports (63.9, 64%, 0.74), and iHOT-33 (54.3, 82%, 0.65). Likelihood for achieving MCID declined above and increased below these thresholds. In univariate analysis female sex, femoral version, lower acetabular outerbridge score and increasing CT sagittal center edge angle (CEA) were predictive of achieving MCID. In multivariable analysis
Classification of emotional states from electrocardiogram signals: a non-linear approach based on hurst
Background Identifying the emotional state is helpful in applications involving patients with autism and other intellectual disabilities; computer-based training, human computer interaction etc. Electrocardiogram (ECG) signals, being an activity of the autonomous nervous system (ANS), reflect the underlying true emotional state of a person. However, the performance of various methods developed so far lacks accuracy, and more robust methods need to be developed to identify the emotional pattern associated with ECG signals. Methods Emotional ECG data was obtained from sixty participants by inducing the six basic emotional states (happiness, sadness, fear, disgust, surprise and neutral) using audio-visual stimuli. The non-linear feature 'Hurst' was computed using Rescaled Range Statistics (RRS) and Finite Variance Scaling (FVS) methods. New Hurst features were proposed by combining the existing RRS and FVS methods with Higher Order Statistics (HOS). The features were then classified using four classifiers – Bayesian Classifier, Regression Tree, K- nearest neighbor and Fuzzy K-nearest neighbor. Seventy percent of the features were used for training and thirty percent for testing the algorithm. Results Analysis of Variance (ANOVA) conveyed that Hurst and the proposed features were statistically significant (p < 0.001). Hurst computed using RRS and FVS methods showed similar classification accuracy. The features obtained by combining FVS and HOS performed better with a maximum accuracy of 92.87% and 76.45% for classifying the six emotional states using random and subject independent validation respectively. Conclusions The results indicate that the combination of non-linear analysis and HOS tend to capture the finer emotional changes that can be seen in healthy ECG data. This work can be further fine tuned to develop a real time system. PMID:23680041
Strength and pain are related to different measures of functional ability in patients with end-stage hip osteoarthritis
Zeni, Joseph; Abujaber, Sumayah; Pozzi, Federico; Raisis, Leo
Objective Performance tests and self-reported questionnaires capture different domains of function in patients with lower extremity osteoarthritis, but the impairments related to each of these domains have not been elucidated. The purpose of this study was to determine how strength and joint pain influence performance-based tests and self-reported questionnaires of functional ability of individuals with end-stage hip osteoarthritis. Methods Fifty-six patients scheduled to undergo unilateral total hip arthroplasty were included in this cross sectional analysis. Subjects completed the Hip Outcome Survey (HOS) and pain in the affected hip was quantified on a scale from 0–10. Performance-based tests included the Six Minute Walk (6MW), Timed Up and Go (TUG), and Stair Climbing Test (SCT). Isometric strength of the hip abductors and knee extensors was collected. Hierarchical linear regressions were created to determine the contributions of pain, knee extensor strength, and hip strength for each outcome measure (TUG, 6MW, SCT, HOS). Height and BMI were entered first, followed by pain, then knee extensor strength, then hip abduction strength. Results Greater pain significantly predicted lower HOS scores, but hip strength did not significantly improve the model. Hip pain was also significantly related to TUG, SCT and 6MW values, but hip and knee strength explained additional variance for these performance-based measures. Conclusions Self-report and performance-based measures capture different aspects of disability and are influenced by different underlying impairments. Both types of outcome measures should be used in studies that evaluate functional changes in patients with hip osteoarthritis. PMID:24664893
Mesenchymal stem cells increase proliferation but do not change quiescent state of osteosarcoma cells: Potential implications according to the tumor resection status
Avril, Pierre; Le Nail, Louis-Romée; Brennan, Meadhbh Á.; Rosset, Philippe; De Pinieux, Gonzague; Layrolle, Pierre; Heymann, Dominique; Perrot, Pierre; Trichet, Valérie
Conventional therapy of primary bone tumors includes surgical excision with wide resection, which leads to physical and aesthetic defects. For reconstruction of bone and joints, allografts can be supplemented with mesenchymal stem cells (MSCs). Similarly, adipose tissue transfer (ATT) is supplemented with adipose-derived stem cells (ADSCs) to improve the efficient grafting in the correction of soft tissue defects. MSC-like cells may also be used in tumor-targeted cell therapy. However, MSC may have adverse effects on sarcoma development. In the present study, human ADSCs, MSCs and pre-osteoclasts were co-injected with human MNNG-HOS osteosarcoma cells in immunodeficient mice. ADSCs and MSCs, but not the osteoclast precursors, accelerated the local proliferation of MNNG-HOS osteosarcoma cells. However, the osteolysis and the metastasis process were not exacerbated by ADSCs, MSCs, or pre-osteoclasts. In vitro proliferation of MNNG-HOS and Saos-2 osteosarcoma cells was increased up to 2-fold in the presence of ADSC-conditioned medium. In contrast, ADSC-conditioned medium did not change the dormant, quiescent state of osteosarcoma cells cultured in oncospheres. Due to the enhancing effect of ADSCs/MSCs on in vivo/in vitro proliferation of osteosarcoma cells, MSCs may not be good candidates for osteosarcoma-targeted cell therapy. Although conditioned medium of ADSCs accelerated the cell cycle of proliferating osteosarcoma cells, it did not change the quiescent state of dormant osteosarcoma cells, indicating that ADSC-secreted factors may not be involved in the risk of local recurrence. PMID:26998421
Effect of incubation on freezability of cholesterol-loaded cyclodextrin treated buffalo (Bubalus bubalis) spermatozoa
Lone, S. A.; Prasad, J. K.; Ghosh, S. K.; Das, G. K.; Balamurugan, B.; Katiyar, R.; Verma, M. R.
Aim: The aim of this study was to investigate the effect of incubation on freezability of cholesterol loaded cyclodextrin (CLC) treated buffalo spermatozoa. Materials and Methods: Semen samples with mass motility of 3+ and greater, collected from Murrah buffalo bulls were utilized. Immediately after collection, four equal groups of semen sample were made. Group I was kept as control and diluted with Tris upto concentration of 60×106 sperm/ml, where as Groups II, III, and IV were treated with CLC at 3 mg/120× 106 spermatozoa, incubated at 37°C for action of CLC for 10, 15 and 20 min, respectively, and diluted with tris upto concentration of 60×106 sperm/ml. All groups were subjected to equilibration and freezing. The evaluation of semen samples from all groups was carried out at fresh, pre-freeze and post-thaw stage for progressive motility, viability and hypo-osmotic swelling response (HOS response). Results: At the pre-freeze stage, significantly (p<0.05) higher percentage of progressive motility and viability was observed in treatment groups as compared to control with no significant difference among treatment groups. HOS response was significantly (p<0.05) higher in treatment groups as compared to control at pre-freeze stage. At post-thaw stage, significantly (p<0.05) higher percentage of progressive motility, viability and HOS response was recorded in Group II as compared to control and other treatment groups (III and IV). Group II retained significant post-thaw motility and viability at various post-thaw incubation periods. Conclusion: Incubation period of 10 min for CLC treated buffalo spermatozoa yielded significantly higher results in terms of freezability as compared to incubation for 15 and 20 min. PMID:27051205
Effects of the crude extract of Polygala tenuifolia Willd on human sperm in vitro.
Qiu, Yi; Wang, Lei-guang; Jia, Yi-fang; Yang, Dan-tong; Zhang, Mei-hua; Zhang, Yan-ping; Zhang, Li-hong; Gai, Ling
The aim of the present study is to analyze sperm membrane changes and the spermicidal effect in treatment with the crude extract from Polygala tenuifolia Willd (PTW) in vitro. The root of PTW was extracted in distilled water. Normal human spermatozoa were used to assess the spermicidal activity (Sander-Cramer assay) of the extract from the PTW root. The hypo-osmotic swelling (HOS) test and the eosin Y (EY) staining were used to detect the integrity of sperm membrane and vitality. The sperm chromatin dispersion (SCD) test was performed to determine sperm DNA integrity. N-9 was used as a reference standard and semen added to physiological saline was used as the control. Semen samples were donated by 42 healthy fertile men. The crude extract from the root of PTW could immobilize and kill 100% spermatozoa within 20 s in vitro at the concentrations of 20.0 and 10.0 mg/ml; at the concentration of 5.0 mg/ml, spermatozoa were immobilized in (39.5±3.2) s. In the groups of the crude extract from the root of PTW and N-9 solution, the rate of the normal HOS (tails swollen) and the white head (unstained) was 0%, and the rate of the abnormal HOS (tails unswollen) and red head (stained) was 100%. Sperm DNA fragmentation showed no change in exposure to the crude extract from the root of PTW and N-9 solution. The sperm revival test did not show any spermatozoa that recovered their motilities. The rapid spermicidal activity of the crude extract from the root of PTW in vitro may occur by the disruption of the sperm membrane integrity. PMID:21634037
Changes in soil bacterial community triggered by drought-induced gap succession preceded changes in soil C stocks and quality.
Yuste, Jorge Curiel; Barba, Josep; Fernandez-Gonzalez, Antonio José; Fernandez-Lopez, Manuel; Mattana, Stefania; Martinez-Vilalta, Jordi; Nolis, Pau; Lloret, Francisco
The aim of this study was to understand how drought-induced tree mortality and subsequent secondary succession would affect soil bacterial taxonomic composition as well as soil organic matter (SOM) quantity and quality in a mixed Mediterranean forest where the Scots pine (Pinus sylvestris) population, affected by climatic drought-induced die-off, is being replaced by Holm-oaks (HO; Quercus ilex). We apply a high throughput DNA pyrosequencing technique and (13)C solid-state Nuclear Magnetic Resonance (CP-MAS (13)C NMR) to soils within areas of influence (defined as an surface with 2-m radius around the trunk) of different trees: healthy and affected (defoliated) pines, pines that died a decade ago and healthy HOs. Soil respiration was also measured in the same spots during a spring campaign using a static close-chamber method (soda lime). A decade after death, and before aerial colonization by the more competitive HOs have even taken place, we could not find changes in soil C pools (quantity and/or quality) associated with tree mortality and secondary succession. Unlike C pools, bacterial diversity and community structure were strongly determined by tree mortality. Convergence between the most abundant taxa of soil bacterial communities under dead pines and colonizer trees (HOs) further suggests that physical gap colonization was occurring below-ground before above-ground colonization was taken place. Significantly higher soil respiration rates under dead trees, together with higher bacterial diversity and anomalously high representation of bacteria commonly associated with copiotrophic environments (r-strategic bacteria) further gives indications of how drought-induced tree mortality and secondary succession were influencing the structure of microbial communities and the metabolic activity of soils. PMID:23301169
Piperine inhibits proliferation of human osteosarcoma cells via G2/M phase arrest and metastasis by suppressing MMP-2/-9 expression.
Zhang, Jian; Zhu, Xiaobing; Li, Hengyuan; Li, Binghao; Sun, Lingling; Xie, Tao; Zhu, Ting; Zhou, Hong; Ye, Zhaoming
The piperidine alkaloid piperine, a major ingredient in black pepper, inhibits the growth and metastasis of cancer cells both in vivo and in vitro, although its mechanism of action is unclear. Furthermore, its anticancer activity against osteosarcoma cells has not been reported. In this study, we show that piperine inhibited the growth of HOS and U2OS cells in dose- and time-dependent manners but had a weaker effect on the growth of normal hFOB cells. Piperine inhibited osteosarcoma cell proliferation by causing G2/M phase cell cycle arrest associated with decreased expression of cyclin B1 and increased phosphorylation of Cyclin-dependent kinase-1(CDK1) and checkpoint kinase 2 (Chk2). In addition, piperine treatment inhibited phosphorylation of Akt and activated phosphorylation of c-Jun N-terminal kinase (c-JNK) and p38 mitogen-activated protein kinase (MAPK) in HOS and U2OS cells. Piperine induced colony formation in these two cell types. We proved that piperine could suppress the metastasis of osteosarcoma cells using scratch migration assays and Transwell chamber tests. Moreover, gelatin zymography showed that piperine inhibited the activity of matrix metalloproteinase (MMP)-2/-9 and increased the expression of tissue inhibitor of metalloproteinase (TIMP)-1/-2. Taken together, our results indicate that piperine inhibits proliferation, by inducing G2/M cell cycle arrest, and the migration and invasion of HOS and U2OS cells, via increased expression of TIMP-1/-2 and down-regulation of MMP-2/-9. These findings support further study of piperine as a promising therapeutic agent in the treatment of osteosarcoma. PMID:25479727
Orthopedic manifestations in patients with mucopolysaccharidosis type II (Hunter syndrome) enrolled in the Hunter Outcome Survey
Link, Bianca; de Camargo Pinto, Louise Lapagesse; Giugliani, Roberto; Wraith, James Edmond; Guffon, Nathalie; Eich, Elke; Beck, Michael
Mucopolysaccharidosis type II (MPS II or Hunter syndrome) is a rare, inherited disorder caused by deficiency of the lysosomal enzyme iduronate-2-sulfatase. As a result of this deficiency, glycosaminoglycans accumulate in lysosomes in many tissues, leading to progressive multisystemic disease. The cardiopulmonary and neurological problems associated with MPS II have received considerable attention. Orthopedic manifestations are common but not as well characterized. This study aimed to characterize the prevalence and severity of orthopedic manifestations of MPS II and to determine the relationship of these signs and symptoms with cardiovascular, pulmonary and central nervous system involvement. Orthopedic manifestations of MPS II were studied using cross-sectional data from the Hunter Outcome Survey (HOS). The HOS is a global, physician-led, multicenter observational database that collects information on the natural history of MPS II and the long-term safety and effectiveness of enzyme replacement therapy. As of January 2009, the HOS contained baseline data on joint range of motion in 124 males with MPS II. In total, 79% of patients had skeletal manifestations (median onset, 3.5 years) and 25% had abnormal gait (median onset, 5.4 years). Joint range of motion was restricted for all joints assessed (elbow, shoulder, hip, knee and ankle). Extension was the most severely affected movement: the exception to this was the shoulder. Surgery for orthopedic problems was rare. The presence of orthopedic manifestations was associated with the presence of central nervous system and pulmonary involvement, but not so clearly with cardiovascular involvement. Orthopedic interventions should be considered on an individual-patient basis. Although some orthopedic manifestations associated with MPS II may be managed routinely, a good knowledge of other concurrent organ system involvement is essential. A multidisciplinary approach is required. PMID:21808707
Shubnikov-de Haas Effect measured on single crystals of CeOs4 Sb 12 and NdOs4 Sb 12 along the high symmetry directions
NASA Astrophysics Data System (ADS)
Ho, P.-C.; Singleton, J.; Maple, M. B.; Shrekenhamer, D. B.; X., Lee; Thale, A.; Yanagisawa, T.
The filled skutterudite compounds CeOs 4 Sb 12 , PrOs 4 Sb 12 , and NdOs 4 Sb 12 are respectively a 1~K antiferromagnetic (AFM) Kondo insulator, a 1.85~K unconventional superconductor, and a 1~K mean-field type ferromagnet (FM), suggesting that superconductivity in PrOs 4 Sb 12 may result from proximity to AFM and FM quantum-critical points. Fermi-surface measurements of NdOs 4 Sb 12 and CeOs 4 Sb 12 could therefore give insights into the pairing mechanism. We have used a MHz skin-depth technique to observe Shubnikov-de Haas oscillations (SdHos) in single crystals of these materials at fields of up to 60~T. In CeOs 4 Sb 12 for H [001], a previously-unobserved semimetal-to-metal transition was detected at ~ 25 ~T; above this, a series of SdHos with a frequency of 1700~T and mCR ~ 3.6me emerge. For H [011] in NdOs 4 Sb 12 , a single series of SdHos, frequency ~ 874 ~T, was found. These may correspond to the β band in PrOs 4 Sb 12 , but with a much smaller mCR ~ 1.5me . Research at CSU-Fresno is supported by RC CCSA # 7669 and the start-up fund; at NHMFL by DOE, NSF, and FL.; at UCSD by NSF # 0802478 and US DOE DE FG02-04ER46105; at Hokkaido U by MEXT, Japan.
Correlation between seminal lead and cadmium and seminal parameters in idiopathic oligoasthenozoospermic males
Taha, Emad A.; Sayed, Sohair K.; Ghandour, Nagwa M; Mahran, Ali M.; Saleh, Medhat A.; Amin, Magdy M.
Introduction The exact causes of the decline in semen quality are not yet known, environmental factors have been considered to play an important role. Lead (Pb) and Cadmium (Cd) are two of the well-known reproductive toxicants to which humans are exposed occupationally and environmentally and can lead to negative effects on the testicular functions. The aim of this study was to evaluate lead and cadmium levels in seminal plasma of men with idiopathic oligoasthenozoospermia in comparison to fertile healthy controls and to correlate these levels with conventional semen parameters, sperm hypo-osmotic swelling (HOS) percentage, sperm DNA fragmentation percentage, and semen reactive oxygen species (ROS) levels. Material and Methods Thirty infertile male patients with idiopathic oligo and/or asthenozoospermia and thirty healthy fertile men, which was the control group, were included in the study. Lead and cadmium levels in seminal plasma, semen parameters, sperm HOS, sperm DNA fragmentation percentage and semen ROS assay were measured in all subjects. Results There was a significant increase in seminal lead and cadmium levels among infertile males in comparison to controls. There were significant negative correlations between seminal lead and cadmium levels on one hand and certain semen parameters especially progressive sperm motility and vitality (HOS). Importantly, significant positive correlations were noted between seminal lead and cadmium levels on one hand and sperm DNA fragmentation percentage and semen ROS level in infertile men and controls on the other hand. Conclusions Thus, men with idiopathic male infertility had higher levels of lead and cadmium in their semen which correlated with impairment of sperm motility and vitality percentages and more importantly with higher sperm DNA fragmentation% and semen ROS level. PMID:24579002
Cathepsin H indirectly regulates morphogenetic protein-4 (BMP-4) in various human cell lines
Rojnik, Matija; Jevnikar, Zala; Mirkovic, Bojana; Janes, Damjan; Zidar, Nace; Kikelj, Danijel; Kos, Janko
Background Cathepsin H is a cysteine protease considered to play a major role in tumor progression, however, its precise function in tumorigenesis is unclear. Cathepsin H was recently proposed to be involved in processing of bone morphogenetic protein 4 (BMP-4) in mice. In order to clarify whether cathepsin H also regulates BMP-4 in humans, its impact on BMP-4 expression, processing and degradation was investigated in prostate cancer (PC-3), osteosarcoma (HOS) and pro-monocytic (U937) human cell lines. Materials and methods BMP-4 expression was founded to be regulated by cathepsin H using PCR array technology and confirmed by real time PCR. Immunoassays including Western blot and confocal microscopy were used to evaluate the influence of cathepsin H on BMP-4 processing. Results In contrast to HOS, the expression of BMP-4 mRNA in U937 and PC3 cells was significantly decreased by cathepsin H. The different regulation of BMP-4 synthesis could be associated with the absence of the mature 28 kDa cathepsin H form in HOS cells, where only the intermediate 30 kDa form was observed. No co-localization of BMP-4 and cathepsin H was observed in human cell lines and the multistep processing of BMP-4 was not altered in the presence of specific cathepsin H inhibitor. Isolated cathepsin H does not cleave mature recombinant BMP-4, neither with its amino- nor its endopeptidase activity. Conclusions Our results exclude direct proteolytic processing of BMP-4 by cathepsin H, however, they provide support for its involvement in the regulation of BMP-4 expression. PMID:22933963
Evaluation of muscle force classification using shape analysis of the sEMG probability density function: a simulation study.
Ayachi, F S; Boudaoud, S; Marque, C
In this work, we propose to classify, by simulation, the shape variability (or non-Gaussianity) of the surface electromyogram (sEMG) amplitude probability density function (PDF), according to contraction level, using high-order statistics (HOS) and a recent functional formalism, the core shape modeling (CSM). According to recent studies, based on simulated and/or experimental conditions, the sEMG PDF shape seems to be modified by many factors as: contraction level, fatigue state, muscle anatomy, used instrumentation, and also motor control parameters. For sensitivity evaluation against these several sources (physiological, instrumental, and neural control) of variability, a large-scale simulation (25 muscle anatomies, ten parameter configurations, three electrode arrangements) is performed, by using a recent sEMG-force model and parallel computing, to classify sEMG data from three contraction levels (20, 50, and 80% MVC). A shape clustering algorithm is then launched using five combinations of HOS parameters, the CSM method and compared to amplitude clustering with classical indicators [average rectified value (ARV) and root mean square (RMS)]. From the results screening, it appears that the CSM method obtains, using Laplacian electrode arrangement, the highest classification scores, after ARV and RMS approaches, and followed by one HOS combination. However, when some critical confounding parameters are changed, these scores decrease. These simulation results demonstrate that the shape screening of the sEMG amplitude PDF is a complex task which needs both efficient shape analysis methods and specific signal recording protocol to be properly used for tracking neural drive and muscle activation strategies with varying force contraction in complement to classical amplitude estimators. PMID:24961179
Changes in soil bacterial community triggered by drought-induced gap succession preceded changes in soil C stocks and quality
Yuste, Jorge Curiel; Barba, Josep; Fernandez-Gonzalez, Antonio José; Fernandez-Lopez, Manuel; Mattana, Stefania; Martinez-Vilalta, Jordi; Nolis, Pau; Lloret, Francisco
The aim of this study was to understand how drought-induced tree mortality and subsequent secondary succession would affect soil bacterial taxonomic composition as well as soil organic matter (SOM) quantity and quality in a mixed Mediterranean forest where the Scots pine (Pinus sylvestris) population, affected by climatic drought-induced die-off, is being replaced by Holm-oaks (HO; Quercus ilex). We apply a high throughput DNA pyrosequencing technique and 13C solid-state Nuclear Magnetic Resonance (CP-MAS 13C NMR) to soils within areas of influence (defined as an surface with 2-m radius around the trunk) of different trees: healthy and affected (defoliated) pines, pines that died a decade ago and healthy HOs. Soil respiration was also measured in the same spots during a spring campaign using a static close-chamber method (soda lime). A decade after death, and before aerial colonization by the more competitive HOs have even taken place, we could not find changes in soil C pools (quantity and/or quality) associated with tree mortality and secondary succession. Unlike C pools, bacterial diversity and community structure were strongly determined by tree mortality. Convergence between the most abundant taxa of soil bacterial communities under dead pines and colonizer trees (HOs) further suggests that physical gap colonization was occurring below-ground before above-ground colonization was taken place. Significantly higher soil respiration rates under dead trees, together with higher bacterial diversity and anomalously high representation of bacteria commonly associated with copiotrophic environments (r-strategic bacteria) further gives indications of how drought-induced tree mortality and secondary succession were influencing the structure of microbial communities and the metabolic activity of soils. PMID:23301169
Microfracture in the hip: a matched-control study with average 3-year follow-up
Lodhia, Parth; Gui, Chengcheng; Chandrasekaran, Sivashankar; Suarez-Ahedo, Carlos; Vemula, S. Pavan; Domb, Benjamin G.
There is a paucity of literature regarding microfracture surgery in the hip. The purpose of this study was to compare outcomes in patients undergoing hip arthroscopy predominantly for labral tears with focal full thickness chondral damage on the acetabulum or femoral head treated with microfracture and a matched control group that did not have focal full thickness chondral damage. A prospective matched-control study was performed examining four patient-reported outcome (PRO) scores: modified Harris Hip Score (mHHS), non-arthritic hip score, Hip Outcome Score—Activities of Daily Living (HOS-ADL), and Hip Outcome Score—Sports Specific Subscale (HOS-SSS) at minimum 2 years post-operatively between 35 patients undergoing microfracture for chondral defects during hip arthroscopy and 70 patients in a control group that did not have chondral defects. The patients were matched based on gender, age within 7 years, Workman's compensation claim, labral treatment and acetabular crossover percentage less than or greater than 20. There was no significant difference (P > 0.05) in PRO scores preoperatively between the groups. Both groups demonstrated significant improvement (P < 0.05) in all post-operative PRO scores at all time points. There was no statistically significant difference (P > 0.05) in post-operative PRO scores between the microfracture and control groups, except for HOS-ADL and the visual analog scale (VAS) score, both of which were superior in the control group (P < 0.05). Patient satisfaction was 6.9 for the microfracture group and 7.7 for the control group (P > 0.05). Arthroscopic microfracture of the hip during treatment of labral tears results in favorable outcomes that are similar to the results arthroscopic treatment of labral tears in patients without full thickness chondral damage. PMID:27011867
Alpha-CaMKII plays a critical role in determining the aggressive behavior of human osteosarcoma.
Daft, Paul G; Yuan, Kaiyu; Warram, Jason M; Klein, Michael J; Siegal, Gene P; Zayzafoon, Majd
Osteosarcoma is among the most frequently occurring primary bone tumors, primarily affecting adolescents and young adults. Despite improvements in osteosarcoma treatment, more specific molecular targets are needed as potential therapeutic options. One target of interest is α-Ca(2+)/calmodulin-dependent protein kinase II (α-CaMKII), a ubiquitous mediator of Ca(2+)-linked signaling, which has been shown to regulate tumor cell proliferation and differentiation. Here, we investigate the role of α-CaMKII in the growth and tumorigenicity of human osteosarcoma. We show that α-CaMKII is highly expressed in primary osteosarcoma tissue derived from 114 patients, and is expressed in varying levels in different human osteosarcoma (OS) cell lines [MG-63, N-methyl-N'-nitro-N-nitrosoguanidine (MNNG)/HOS, and 143B). To examine whether α-CaMKII regulates osteosarcoma tumorigenic properties, we genetically inhibited α-CaMKII in two osteosarcoma cell lines using two different α-CaMKII shRNAs delivered by lentiviral vectors and overexpressed α-CaMKII by retrovirus. The genetic deletion of α-CaMKII by short hairpin RNA (shRNA) in MG-63 and 143B cells resulted in decreased proliferation (50% and 41%), migration (22% and 25%), and invasion (95% and 90%), respectively. The overexpression of α-CaMKII in HOS cells resulted in increased proliferation (240%), migration (640%), and invasion (10,000%). Furthermore, α-CaMKII deletion in MG-63 cells significantly reduced tumor burden in vivo (65%), whereas α-CaMKII overexpression resulted in tumor formation in a previously nontumor forming osteosarcoma cell line (HOS). Our results suggest that α-CaMKII plays a critical role in determining the aggressive phenotype of osteosarcoma, and its inhibition could be an attractive therapeutic target to combat this devastating adolescent disease. PMID:23364534
Effect of management system and season on semen freezability in Jakhrana bucks
Kumar, Narendra; Rai, B.; Bhat, Showkat A.; Kharche, S. D.; Gangwar, Chetna; Jindal, S. K.; Chandra, Subhash
Aim: The objective of the study was to determine the effect of the management system (intensive and semi-intensive) and season (autumn and winter) on semen freezability in Jakhrana bucks. Materials and Methods: A total of 24 Jakhrana bucks of same body weight and age (BW=30 kg, age=1 year) were randomly allotted into two groups, viz., Group I (intensive system, 12 bucks) and Group II (semi-intensive system, 12 bucks). These two groups were statistically tested for their homogeneity with respect to age and BW. Semen was collected twice weekly using an artificial vagina during two seasons: autumn (September-November) and winter (December-February). A total of 240 semen samples (120 from each group and season) were evaluated for post-thaw motility (PTM), viability, abnormality, functional membrane integrity (hypo-osmotic swelling [HOS]) response and acrosomal integrity. Results: The mean values of PTM and acrosomal integrity of spermatozoa were significantly (p<0.01) higher in Group II as compared to Group I. The mean values of viability and abnormality were also differed significant (p<0.05) between groups. However, the mean values of HOS response were found non-significant (p>0.05) between groups. The season showed a significant effect on all parameters except viability and HOS response. The PTM and acrosomal integrity of spermatozoa were significantly (p<0.01) higher in winter as compared to autumn season. Abnormality of spermatozoa was significantly (p<0.05) lower in winter season. Conclusions: This study indicates that both management system and season influence semen freezability. The semen collected from bucks reared under the semi-intensive system and winter season showed better semen freezability characteristics. PMID:27051208
Higher-Order Spectral Analysis of a Nonlinear Pitch and Plunge Apparatus
NASA Technical Reports Server (NTRS)
Silva, Walter A.; Strganac, Thomas W.; Hajj, Muhammad R.
Simulated aeroelastic responses of a nonlinear pitch and plunge apparatus are analyzed using various statistical signal processing techniques including higher-order spectral methods. A MATLAB version of the Nonlinear Aeroelastic Testbed Apparatus (NATA) at the Texas A&M University is used to generate various aeroelastic response data including limit cycle oscillations (LCO). Traditional and higher-order spectral (HOS) methods are applied to the simulated aeroelastic responses. Higher-order spectral methods are used to identify critical signatures that indicate the transition from linear to nonlinear (LCO) aeroelastic behavior.
Recreational Athletes Return to Sport at a Comparable Rate to Elite Athletes Following Hip Arthroscopy for Femoroacetabular Impingement
Weber, Alexander E.; Kuhns, Benjamin; Cvetanovich, Gregory; Levy, David; Nho, Shane Jay
Objectives: The objective of the current study was to evaluate patient reported outcomes and return to sport in a cohort of distinctly recreational and amateur level athletes following hip arthroscopy for femoroacetabular impingement (FAI). Methods: Following IRB approval, clinical data was retrospectively retrieved for 66 consecutive FAI patients (26 men, 40 women) who had undergone hip arthroscopy and identified themselves as recreational or amateur athletes on intake forms. Two-year patient-reported outcomes (PRO) included a sport-specific questionnaire, modified Harris Hip Score (mHHS), and Hip Outcome Scores with Activities of Daily Living (HOS-ADL) and Sports-Specific (HOS-SS) subscales were analyzed. Results: The mean age and BMI of all subjects was 26.8 ± 7.6 years and 23.9 ± 3.2 kg/m2, respectively. Athletes had withdrawn from sport for an average of 9.5 ± 6.7 months prior to surgery and on average required 9.7 ± 5.1 months to return to sport. After two years, all mean PRO scores had improved significantly (Figure 1), and 57 patients (92%) had returned to play and continued participation. Patients who had withdrawn from sport for greater than 8 months before surgery returned to sport significantly more slowly than those who had withdrawn for less than 8 months (p=0.01). Greater withdrawal from sport prior to surgery also correlated with lower postoperative improvements in HOS-ADL and HOS-SS scores. Bivariate analysis revealed that increasing body-mass index (BMI) was associated with lower improvements in PROs. Conclusion: Recreational athletes, following hip arthroscopy for FAI, return to play at a high rate. Increasing BMI and preoperative withdrawal from sport both significantly prolong return to play and diminish two-year PROs. Most return-to-play studies following hip arthroscopy for FAI have focused on professional athletes, with limited generalizability to the average sports medicine surgeon practice. This is the first study of its kind to focus
A human operator simulator model of the NASA Terminal Configured Vehicle (TCV)
NASA Technical Reports Server (NTRS)
Glenn, F. A., III; Doane, S. M.
A generic operator model called HOS was used to simulate the behavior and performance of a pilot flying a transport airplane during instrument approach and landing operations in order to demonstrate the applicability of the model to problems associated with interfacing a crew with a flight system. The model which was installed and operated on NASA Langley's central computing system is described. Preliminary results of its application to an investigation of an innovative display system under development in Langley's terminal configured vehicle program are considered.
Shubnikov-de Haas oscillations in a two-dimensional electron gas under subterahertz radiation
NASA Astrophysics Data System (ADS)
Shi, Q.; Martin, P. D.; Hatke, A. T.; Zudov, M. A.; Watson, J. D.; Gardner, G. C.; Manfra, M. J.; Pfeiffer, L. N.; West, K. W.
We report on magnetotransport measurements in a two-dimensional (2D) electron gas subject to subterahertz radiation in the regime where Shubnikov-de Haas oscillations (SdHOs) and microwave-induced resistance oscillations (MIROs) coexist over a wide magnetic field range, spanning several harmonics of the cyclotron resonance. Surprisingly, we find that the SdHO amplitude is modified by the radiation in a nontrivial way, owing to the oscillatory correction which has the same period and phase as MIROs. This finding challenges our current understanding of microwave photoresistance in 2D electron gas, calling for future investigations.
High-fat meals rich in EPA plus DHA compared with DHA only have differential effects on postprandial lipemia and plasma 8-isoprostane F2α concentrations relative to a control high–oleic acid meal: a randomized controlled trial1234
Purcell, Robert; Latham, Sally H; Botham, Kathleen M; Hall, Wendy L; Wheeler-Jones, Caroline PD
Background: Eicosapentaenoic acid (EPA) plus docosahexaenoic acid (DHA) supplementation has beneficial cardiovascular effects, but postprandial influences of these individual fatty acids are unclear. Objectives: The primary objective was to determine the vascular effects of EPA + DHA compared with DHA only during postprandial lipemia relative to control high–oleic acid meals; the secondary objective was to characterize the effects of linoleic acid–enriched high-fat meals relative to the control meal. Design: We conducted a randomized, controlled, double-blind crossover trial of 4 high-fat (75-g) meals containing 1) high–oleic acid sunflower oil (HOS; control), 2) HOS + fish oil (FO; 5 g EPA and DHA), 3) HOS + algal oil (AO; 5 g DHA), and 4) high–linoleic acid sunflower oil (HLS) in 16 healthy men (aged 35–70 y) with higher than optimal fasting triacylglycerol concentrations (mean ± SD triacylglycerol, 1.9 ± 0.5 mmol/L). Results: Elevations in triacylglycerol concentration relative to baseline were slightly reduced after FO and HLS compared with the HOS control (P < 0.05). The characteristic decrease from baseline in plasma nonesterified fatty acids after a mixed meal was inhibited after AO (Δ 0–3 h, P < 0.05). HLS increased the augmentation index compared with the other test meals (P < 0.05), although the digital volume pulse–reflection index was not significantly different. Plasma 8-isoprostane F2α analysis revealed opposing effects of FO (increased) and AO (reduced) compared with the control (P < 0.05). No differences in nitric oxide metabolites were observed. Conclusions: These data show differential postprandial 8-isoprostane F2α responses to high-fat meals containing EPA + DHA–rich fish oil compared with DHA-rich AO, but these differences were not associated with consistent effects on postprandial vascular function or lipemia. More detailed analyses of polyunsaturated fatty acid–derived lipid mediators are required to determine possible
'HeART of Stroke (HoS)', a community-based Arts for Health group intervention to support self-confidence and psychological well-being following a stroke: protocol for a randomised controlled feasibility study
Ellis-Hill, Caroline; Gracey, Fergus; Thomas, Sarah; Lamont-Robinson, Catherine; Thomas, Peter W; Marques, Elsa M R; Grant, Mary; Nunn, Samantha; Cant, Robin P I; Galvin, Kathleen T; Reynolds, Frances; Jenkinson, Damian F
Introduction Over 152 000 people in the UK have strokes annually and a third experience residual disability. Low mood also affects a third of stroke survivors; yet psychological support is poor. While Arts for Health interventions have been shown to improve well-being in people with mild-to-moderate depression post-stroke, their role in helping people regain sense of self, well-being and confidence has yet to be evaluated. The main aim of this study is to explore the feasibility of conducting a pragmatic multicentre randomised controlled trial to assess the effectiveness and cost-effectiveness of an Arts for Health group intervention ('HeART of Stroke' (HoS)) for stroke survivors. HoS is a 10-session artist-facilitated group intervention held in the community over 14 weeks. It offers a non-judgemental, supportive environment for people to explore sense of self, potentially enhancing well-being and confidence. Methods and analysis Sixty-four people, up to 2 years post-stroke, recruited via secondary care research staff or community stroke/rehabilitation teams in two UK centres will be randomised to either HoS plus usual care or usual care only. Self-reported outcomes, measured at baseline and approximately 5 months postrandomisation, will include stroke-related, well-being, mood, self-esteem, quality of life and process measures. Analyses will focus on estimating key feasibility parameters (eg, rates of recruitment, retention, intervention attendance). We will develop outcome and resource use data collection methods to inform an effectiveness and cost-effectiveness analysis in the future trial. Interviews, with a sample of participants, will explore the acceptability of the intervention and study processes, as well as experiences of the HoS group. Ethics and dissemination National Health Service (NHS), Research and Development and University ethical approvals have been obtained. Two peer-reviewed journal publications are planned plus one service user led
Higher-order spectra for identification of nonlinear modal coupling
NASA Astrophysics Data System (ADS)
Hickey, Daryl; Worden, Keith; Platten, Michael F.; Wright, Jan R.; Cooper, Jonathan E.
Over the past four decades considerable work has been done in the area of power spectrum estimation. The information contained within the power spectrum relates to a signal's autocorrelation or 'second-order statistics'. The power spectrum provides a complete statistical description of a Gaussian process; however, a problem with this information is that it is phase blind. This problem is addressed if one turns to a system's frequency response function (FRF). The FRF graphs the magnitude and phase of the frequency response of a system; in order to do this it requires information regarding the frequency content of the input and output signals. Situations arise in science and engineering whereby signal analysts are required to look beyond second-order statistics and analyse a signal's higher-order statistics (HOS). HOS or spectra give information on a signal's deviation from Gaussianity and consequently are a good indicator function for the presence of nonlinearity within a system. One of the main problems in nonlinear system identification is that of high modal density. Many modelling schemes involve making some expansion of the nonlinear restoring force in terms of polynomial or other basis terms. If more than one degree-of-freedom is involved this becomes a multivariate problem and the number of candidate terms in the expansion grows explosively with the order of nonlinearity and the number of degrees-of-freedom. This paper attempts to use HOS to detect and qualify nonlinear behaviour for a number of symmetrical and asymmetrical systems over a range of degrees-of-freedom. In doing so the paper also attempts to show that HOS are a more sensitive tool than the FRF in detecting nonlinearity. Furthermore, the object of this paper is to try and identify which modes couple in a nonlinear manner in order to reduce the number of candidate coupling terms, for a model, as much as possible. The bispectrum method has previously been applied to simple low-DOF systems with high
Changes in Levels of Seminal Nitric Oxide Synthase, Macrophage Migration Inhibitory Factor, Sperm DNA Integrity and Caspase-3 in Fertile Men after Scrotal Heat Stress
Shi, Zhi-Da; Wang, Lei-Guang; Qiu, Yi
Background This study observes changes in levels of seminal nitric oxide (NO), nitric oxide synthase (NOS), macrophage migration inhibitory factor (MIF), sperm DNA integrity, chromatin condensation and Caspase-3in adult healthy men after scrotal heat stress (SHS). Methods Exposure of the scrotum of 25 healthy male volunteers locally at 40–43°C SHS belt warming 40 min each day for successive 2 d per week. The course of SHS was continuously 3 months. Routine semen analysis, hypo-osmotic swelling (HOS) test, Aniline blue (AB) staining, HOS/AB and terminal deoxynucleotidyl transferase-mediated d UDP nick-end labeling (TUNEL) were carried out before, during and after SHS. Seminal NO and NOS contents were determined by nitrate reduction method. The activated Caspase-3 levels of spermatozoa and MIF in seminal plasma were measured by the enzyme-linked immunosorbent assay (ELISA) method. Statistical significance between mean values was determined using statistical ANOVA tests. Results The mean parameters of sperm concentration, motile and progressive motile sperm and normal morphological sperm were significantly decreased in groups during SHS 1, 2 and 3 months compared with those in groups of pre-SHS (P<0.001). Statistically significant differences of sperm DNA fragmentation, normal sperm membrane, and Caspase-3 activity as well as the level of NO, NOS and MIF in semen were observed between the groups before SHS and after SHS 3 months and the groups during SHS 1, 2 and 3 months (P<0.001). After three months of the SHS, various parameters recovered to the level before SHS. WBC in semen showed a positively significant correlation with the levels of NO, NOS, MIF and Caspase-3 activity. The percentage of abnormal sperm by using the test of HOS showed a positively significant correlation with that of HOS/AB. Conclusions The continuously constant SHS can impact the semen quality and sperm DNA and chromatin, which may be contributed to the high level of NO, NOS, MIF and Caspase
Hypoosmotic swelling test for quality control of sperm prepared for assisted reproduction.
Mladenović, I; Mićić, S; Genbaćev, O; Papić, N
This study evaluates the clinical usefulness of one additional parameter for assessment of human sperm cell function in vitro--the hypoosmotic swelling test. The hypoosmotic swelling test evaluated the functional integrity of the sperm plasma membrane. The investigation included a comparison of the hypoosmotic swelling test in samples containing motile and immotile spermatozoa and their correlation with the intrauterine insemination outcome. Motile spermatozoa expressed better membrane characteristics, without any importance of the hypoosmotic conditions. Positive correlation exists between HOS results and the outcome of IUI. This test can be a useful addition to the standard battery of semen analyzing tests. PMID:7625880
Wettability and cellular response of UV light irradiated anodized titanium surface
Park, Kyou-Hwa; Koak, Jai-Young; Kim, Seong-Kyun
PURPOSE The object of this study was to investigate the effect of UV irradiation (by a general commercial UV sterilizer) on anodized titanium surface. Surface characteristics and cellular responses were compared between anodized titanium discs and UV irradiated anodized titanium discs. MATERIALS AND METHODS Titanium discs were anodized and divided into the following groups: Group 1, anodized (control), and Group 2, anodized and UV irradiated for 24 hours. The surface characteristics including contact angle, roughness, phase of oxide layer, and chemical elemental composition were inspected. The osteoblast-like human osteogenic sarcoma (HOS) cells were cultured on control and test group discs. Initial cellular attachment, MTS-based cell proliferation assay, and ALP synthesis level were compared between the two groups for the evaluation of cellular response. RESULTS After UV irradiation, the contact angle decreased significantly (P<.001). The surface roughness and phase of oxide layer did not show definite changes, but carbon showed a considerable decrease after UV irradiation. Initial cell attachment was increased in test group (P=.004). Cells cultured on test group samples proliferated more actively (P=.009 at day 2, 5, and 7) and the ALP synthesis also increased in cells cultured on the test group (P=.016 at day 3, P=.009 at day 7 and 14). CONCLUSION UV irradiation induced enhanced wettability, and increased initial cellular responses of HOS cells on anodized titanium surface. PMID:21814613
Swarm intelligence for detecting interesting events in crowded environments.
Kaltsa, Vagia; Briassouli, Alexia; Kompatsiaris, Ioannis; Hadjileontiadis, Leontios J; Strintzis, Michael Gerasimos
This paper focuses on detecting and localizing anomalous events in videos of crowded scenes, i.e., divergences from a dominant pattern. Both motion and appearance information are considered, so as to robustly distinguish different kinds of anomalies, for a wide range of scenarios. A newly introduced concept based on swarm theory, histograms of oriented swarms (HOS), is applied to capture the dynamics of crowded environments. HOS, together with the well-known histograms of oriented gradients, are combined to build a descriptor that effectively characterizes each scene. These appearance and motion features are only extracted within spatiotemporal volumes of moving pixels to ensure robustness to local noise, increase accuracy in the detection of local, nondominant anomalies, and achieve a lower computational cost. Experiments on benchmark data sets containing various situations with human crowds, as well as on traffic data, led to results that surpassed the current state of the art (SoA), confirming the method's efficacy and generality. Finally, the experiments show that our approach achieves significantly higher accuracy, especially for pixel-level event detection compared to SoA methods, at a low computational cost. PMID:25769154
Platelets promote osteosarcoma cell growth through activation of the platelet-derived growth factor receptor-Akt signaling axis
Takagi, Satoshi; Takemoto, Ai; Takami, Miho; Oh-hara, Tomoko; Fujita, Naoya
The interactions of tumor cells with platelets contribute to the progression of tumor malignancy, and the expression levels of platelet aggregation-inducing factors positively correlate with the metastatic potential of osteosarcoma cells. However, it is unclear how tumor-platelet interaction contributes to the proliferation of osteosarcomas. We report here that osteosarcoma-platelet interactions induce the release of platelet-derived growth factor (PDGF) from platelets, which promotes the proliferation of osteosarcomas. Co-culture of platelets with MG63 or HOS osteosarcoma cells, which could induce platelet aggregation, enhanced the proliferation of each cell line in vitro. Analysis of phospho-antibody arrays revealed that co-culture of MG63 cells with platelets induced the phosphorylation of platelet derived growth factor receptor (PDGFR) and Akt. The addition of supernatants of osteosarcoma-platelet reactants also increased the growth of MG63 and HOS cells as well as the level of phosphorylated-PDGFR and -Akt. Sunitinib or LY294002, but not erlotinib, significantly inhibited the platelet-induced proliferation of osteosarcoma cells, indicating that PDGF released from platelets plays an important role in the proliferation of osteosarcomas by activating the PDGFR and then Akt. Our results suggest that inhibitors that specifically target osteosarcoma-platelet interactions may eradicate osteosarcomas. PMID:24974736
Holt-Oram syndrome: a clinical genetic study.
Newbury-Ecob, R A; Leanage, R; Raeburn, J A; Young, I D
A clinical and genetic study of the Holt-Oram syndrome (HOS) has been carried out in the United Kingdom involving 55 cases designated Holt-Oram syndrome, together with their parents and sibs. Data from the clinical assessment of both familial and isolated cases were used to define the HOS phenotype and to outline the spectrum of abnormalities, especially factors affecting severity. Skeletal defects affected the upper limbs exclusively and were bilateral and asymmetrical. They ranged from minor signs such as clinodactyly, limited supination, and sloping shoulders to severe reduction deformities of the upper arm (4.5%). The radial ray was predominantly affected than the right. All affected cases showed evidence of upper limb involvement. Cardiac defects were seen in 95% of familial cases and included both atrial septal defect (ASD, 34%) and ventricular septal defect (VSD, 25%); 39% had only ECG changes. Cardiac involvement ranged from asymptomatic conduction disturbances to multiple structural defects requiring surgery in infancy. Sudden death could be caused by heart block. Inheritance was autosomal dominant with 100% penetrance and no evidence of reduced fitness. Increasing severity occurred in succeeding generations consistent with anticipation. Images PMID:8730285
Effects of dietary n-3 fatty acids and vitamin C on semen characteristics, lipid composition of sperm and blood metabolites in fat-tailed Moghani rams.
Jafaroghli, M; Abdi-Benemar, H; Zamiri, M J; Khalili, B; Farshad, A; Shadparvar, A A
Sixteen fertile rams were randomly allotted to four groups and fed either of the four diets for 14 weeks: (1) control diet (COD) without fish oil (FO) and vitamin C (VC), (2) diet containing 2.5% FO (FOD), (3) diet containing 300 mg/kg DM VC (VCD), and (4) diet containing 2.5% FO and 300 mg/kg DM VC (FCD). Semen was collected at 14-d intervals from 1 April to 10 July (out of the physiologic breeding season in Iran). Semen volume and percentages of motile and progressively motile sperm were increased by FO and VC feeding. A significant interaction was also found between FOD and VCD on motility and progressive motility percentage (P<0.05). HOS-test and percentage of sperm with normal acrosome improved significantly by FO and VC. Rams fed FCD had better HOS-test and higher proportion of sperm with normal acrosome than rams in other groups (82.4 and 93.6%, respectively). Diets containing FO and FO and VC increased the proportion of docosahexaenoic acid in sperm (P<0.05). The activity of lactate dehydrogenase in the seminal fluid was significantly affected by VC and the interaction between FO and VC (P<0.05). Blood metabolites, except glucose, were affected positively by FO. The results showed that dietary supplementation with FO and VC improved seminal quality and may have beneficial effects on fertility in Moghani rams. PMID:24745668
Proliferation and harvest of human mesenchymal stem cells using new thermoresponsive nanocomposite gels.
Kotobuki, Noriko; Murata, Kazutaka; Haraguchi, Kazutoshi
For tissue engineering and regenerative medicine, stem cells should be effectively cultured in vitro. New thermoresponsive nanocomposite gels (MD-NC gels), consisting of inorganic clay (hectorite) and copolymers composed of hydrophobic 2-methoxyethyl acrylate (MEA) and hydrophilic N,N-dimethylacrylamide (DMAA) units, could be applied in cell culture and cell harvesting without trypsinization, specifically using mesenchymal stem cells (MSCs). The composition of the MD-NC gel (the ratio of the two monomer types and the clay content) was found to determine its swelling properties in the culture medium, thermosensitivity, protein adsorption, and cell attachment and proliferation. Various kinds of human cells, including MSCs, osteoblast (HOS) cells, fibroblast (NHDF) cells, and epithelial cells could be effectively cultured on MD-NC gels. In particular, on an MD10-NC2 gel with relatively low DMAA and clay content, the cells could be harvested by decreasing the temperature, either as a cell sheet (MSCs or NHDF cells) or as a population of suspension cells (HOS cells). Further, it was found that the MD10-NC2 gel is suitable for stem cell differentiation. Because of their thermosensitivity, controllable modulus, and surface properties, MD-NC gels are promising cell culture substrates useful for tissue engineering and regenerative medicine. PMID:22926940
Biological performance of titania containing phosphate-based glasses for bone tissue engineering applications.
Abou Neel, Ensanya Ali; Chrzanowski, Wojciech; Knowles, Jonathan Campbell
The interplay between glass chemistry, structure, degradation kinetics, and biological activity provides flexibility for the development of scaffolds with highly specific cellular response. The aim of this study was therefore to investigate the role of titania inclusion into the phosphate-based glass on its ability to stimulate osteoblast-like human osteosarcoma (HOS) cells to adhere, proliferate and differentiate. In depth morphological and biochemical characterisation was performed on HOS cells cultured on the surface of glass discs. Cell proliferation was also studied in the presence of the glass extract. Cell differentiation, through osteoblast phenotype genes, alkaline phosphatase (ALP) activity and osteocalcin production, was carried out using normal or osteogenic media. Both Thermanox® and titania free glass were used as controls. The data demonstrated that titania inclusion provides desired cytocompatible surface that supported initial cell attachment, sustained viability, and increased cell proliferation similar or significantly higher than Thermanox®. The modified glasses regulated osteoblastic cell differentiation as detected by osteoblast phenotype gene transcription and upregulated ALP and osteocalcin expression. Using osteogenic media had no significant effect on ALP activity and osteocalcin expression. Therefore, titania modified phosphate glasses may have future use as bone tissue engineering scaffolds. PMID:24411382
Sperm characteristics following freezing in extenders supplemented with whole egg yolk and different concentrations of low-density lipoproteins in the collared peccary (Pecari tajacu).
Souza, Ana Liza Paz; Lima, Gabriela Liberalino; Peixoto, Gislayne Christianne Xavier; de Souza Castelo, Thibério; Oliveira, Maria Glaucia Carlos; de Paula, Valéria Veras; Silva, Alexandre Rodrigues
The aim of the current study was to compare sperm quality characteristics of the collared peccary (Pecari tajacu) following freezing in extenders supplemented with whole egg yolk and different concentrations of low-density lipoproteins (LDL). Semen from 11 adult males was obtained by electroejaculation and evaluated for sperm motility, vigor, morphology as well as membrane integrity analyzed by the hypo-osmotic swelling (HOS) test and a fluorescent staining. Moreover, the semen was diluted in a Tris-based extender containing 20% egg yolk (control group) or 5, 10 or 20% LDL (treatment groups). The semen samples were frozen in liquid nitrogen and thawed in a water bath for 60s at 37°C. The treatments did not affect (p>0.05) sperm vigor, morphology or membrane integrity analyzed by the HOS test. However, post-thaw sperm motility was significantly higher (p<0.05) in the extender supplemented with 20% LDL (36.4 ± 5.3%) compared with the egg yolk extender and extender supplemented with 10% LDL. Furthermore, the percentage of membrane-intact frozen-thawed spermatozoa analyzed by the fluorescent staining was significantly higher (p<0.05) in the extender supplemented with 20% LDL (27.4 ± 6.5%) than in the other groups. In conclusion, 20% LDL can be used to substitute the whole egg yolk as a cryoprotective additive for freezing semen of the collared peccary. PMID:26679163
The role of thermal shock in cyclic oxidation
NASA Technical Reports Server (NTRS)
Lowell, C. E.; Deadmore, D. L.
The effect of thermal shock on the spalling of oxides from the surfaces of several commercial alloys was determined. The average cooling rate was varied from approximately 240 C/sec to less than 1.0 C/sec during cyclic oxidation tests in air. The tests consisted of one hundred cycles of one hour at the maximum temperature (1100 or 1200 C). The alloys were HOS-875, TD-Ni, TD-NiCrAl, IN-601, IN-702, and B-1900 plus Hf. Thermal shock resulted in deformation of the metal which in turn resulted, in most cases, in changing the oxide failure mode from compressive to tensile. Tensile failures were characterized by cracking of the oxide and little loss, while compressive failures were characterized by explosive loss of platelets of oxide. The thermally shocked oxides spalled less than the slow cooled samples with the exception of TD-NiCrAl. This material failed in a brittle manner rather than by plastic deformation. The HOS-875 and the TD-Ni did not spall during either type of cooling. Thus, the effect of thermal shock on spalling is determined, in large part, by the mechanical properties of the metal.
Hypo-osmotic test in cat spermatozoa.
Comercio, E A; Monachesi, N E; Loza, M E; Gambarotta, M; Wanke, M M
The hypo-osmotic (HOS) test has been used in other species as an indicator of the fertilising capacity of spermatozoa. The aims of this study were to assess the response of domestic cat spermatozoa to the hypo-osmotic test, to determine the type of solution, concentration and time of incubation needed to obtain a maximum percentage of swelling, to correlate the selected combination with the percentages of progressive motility and to evaluate whether dilution of the ejaculate alters the results. Incubation for 30 and 45 min in solutions of fructose and of citrate of 50 and 100 mOsmol kg⁻¹ was evaluated. The highest percentage of swelling was obtained using the 50 mOsmol kg⁻¹ solution, and no significant differences were observed between the times of exposure to the solutions. A positive correlation was observed between the percentage of individual progressive motility and the percentage of sperm swelling in a 50 mOsmol kg⁻¹ fructose solution, with no significant differences being observed between raw and diluted semen samples. The results of this study suggest that the HOS test could be useful for evaluating membrane function in domestic cat spermatozoa, both in raw semen and in samples diluted in the EZ Mixin® commercial extender, and thus could be incorporated into routine semen evaluation protocols. PMID:22928866
TPR-MET oncogenic rearrangement: Detection by polymerase chain reaction amplification of the transcript and expression in human tumor cells lines
Soman, N.R.; Wogan, G.N. ); Rhim, J.S. )
Activation of the MET protooncogene by a rearrangement involving the fusion of TPR and MET specific gene sequences has been observed in a human osteosarcoma cell line (HOS) treated in vitro with N-methyl-N{prime}-nitro-N-nitrosoguanidine (MNNG). No information has been available about the possible occurrence of this rearrangement in human tumors. To facilitate rapid screening of human cell lines and tumor samples for this specific gene rearrangement; the authors developed a sensitive detection method based on polymerase chain reaction (PCR) amplification of TPR-MET mRNA. cDNA was generated from cellular transcripts by using one of the PCR primers, which was then used as a template for PCR amplification of a 205-base-pair region carrying the breakpoint. An end-labeled internal probe was hybridized in solution to an aliquot of the PCR product for detecting amplification. Cells could be directly screened by the assay without prior isolation of RNA. A 205-base-pair DNA fragment characteristic of the TRP-MET rearrangement was detected in cell lines previously known to contain this altered sequence. The rearrangement was also detected at very low levels in the parental (nontransformed) cell line, HOS TE-85. A preliminary survey of cell lines derived from a variety of human tumors indicates that TPR-MET rearrangement occurred and was expressed at very low frequencies by cells from 7 of 14 tumors of nonhematopoietic origin.
Coherent quantum squeezing due to the phase space noncommutativity
NASA Astrophysics Data System (ADS)
Bernardini, Alex E.; Mizrahi, Salomon S.
The effects of general noncommutativity of operators on producing deformed coherent squeezed states is examined in phase space. A two-dimensional noncommutative (NC) quantum system supported by a deformed mathematical structure, similar to that of Hadamard billiard, is obtained and the components behaviour is monitored in time. It is assumed that the independent degrees of freedom are two free 1D harmonic oscillators (HOs), so the system Hamiltonian does not contain interaction terms. Through the NC deformation parameterized by a Seiberg-Witten transform on the original canonical variables, one gets the standard commutation relations for the new ones, such that the obtained, new, Hamiltonian represents two interacting 1D HOs. By admitting that one HO is inverted relatively to the other, we show that their effective interaction induces a squeezing dynamics for initial coherent states imaged in the phase space. A suitable pattern of logarithmic spirals is obtained and some relevant properties are discussed in terms of Wigner functions, which are essential to put in evidence the effects of the noncommutativity.
Diallyl trisulfide induces osteosarcoma cell apoptosis through reactive oxygen species-mediated downregulation of the PI3K/Akt pathway.
Wang, Hongliang; Sun, Na; Li, Xin; Li, Ka; Tian, Jiguang; Li, Jianmin
Diallyl trisulfide (DATS) is a natural organosulfur compound isolated from garlic, and has been reported to possess anticancer activities. However, the cancer growth inhibitory effects and molecular mechanisms in human osteosarcoma cells have not been well studied. The present study demonstrated that DATS significantly reduced cell viability in a dose- and time-dependent manner in MG63 and MNNG/HOS cells. DATS-induced G0/G1 phase arrest was found to correlate with a decrease in cyclin D1 in concomitance with an increase in p21 and p27. DATS induced a marked increase in reactive oxygen species (ROS) levels and collapse of mitochondrial membrane potential (Δψm) in the osteosarcoma cells. DATS induced apoptosis in the MG63 and MNNG/HOS cells via inhibition of the PI3K/Akt signaling pathway and through the mitochondrial apoptotic pathway. The efficiency of DATS basically approached the efficacy of LY294002, a specific PI3K inhibitor. However, N-acetylcysteine (NAC), a general ROS scavenger, completely blocked the DATS-induced ROS increase, inhibition of the PI3K/Akt pathway and cell apoptosis. Overall, DATS has the potential to be developed as a new anticancer drug. The mechanisms of action involve the ROS-mediated downregulation of the PI3K/Akt pathway. PMID:27035545
Response of cyanobacteria to low atmosphere pressure
NASA Astrophysics Data System (ADS)
Qin, Lifeng; Ai, Weidang; Guo, Shuangsheng; Tang, Yongkang; Yu, Qingni; Shen, Yunze; Ren, Jin
Maintaining a low pressure environment would reduce the technological complexity and constructed cost of future lunar base. To estimate the effect of hypobaric of controlled ecological life support system in lunar base on terrestrial life, cyanobacteria was used as the model to exam the response of growth, morphology, physiology to it. The decrease of atmosphere pressure from 100 KPa to 50 KPa reducing the growth rates of Microcystis aeruginosa, Merismopedia.sp, Anabaena sp. PCC 7120, Anabaena Hos-aquae, the chlorophyll a content in Microcystis aeruginosa, Merismopedia.sp, Anabaena Hos-aquae, the carotenoid content in Microcystis aeruginosa, Merismopedia.sp and Anabaena sp. PCC 7120, the phycocyanin content in Microcystis aeruginosa. This study explored the biological characteristics of the cyanobacteria under low pressure condition, which aimed at understanding the response of the earth's life to environment for the future moon base, the results enrich the research contents of the lunar biology and may be referred for the research of other terrestrial life, such as human, plant, microbe and animal living in life support system of lunar base.
Flight Test of the F/A-18 Active Aeroelastic Wing Airplane
NASA Technical Reports Server (NTRS)
Voracek, David
A viewgraph presentation of flight tests performed on the F/A active aeroelastic wing airplane is shown. The topics include: 1) F/A-18 AAW Airplane; 2) F/A-18 AAW Control Surfaces; 3) Flight Test Background; 4) Roll Control Effectiveness Regions; 5) AAW Design Test Points; 6) AAW Phase I Test Maneuvers; 7) OBES Pitch Doublets; 8) OBES Roll Doublets; 9) AAW Aileron Flexibility; 10) Phase I - Lessons Learned; 11) Control Law Development and Verification & Validation Testing; 12) AAW Phase II RFCS Envelopes; 13) AAW 1-g Phase II Flight Test; 14) Region I - Subsonic 1-g Rolls; 15) Region I - Subsonic 1-g 360 Roll; 16) Region II - Supersonic 1-g Rolls; 17) Region II - Supersonic 1-g 360 Roll; 18) Region III - Subsonic 1-g Rolls; 19) Roll Axis HOS/LOS Comparison Region II - Supersonic (open-loop); 20) Roll Axis HOS/LOS Comparison Region II - Supersonic (closed-loop); 21) AAW Phase II Elevated-g Flight Test; 22) Region I - Subsonic 4-g RPO; and 23) Phase II - Lessons Learned
Application of higher-order spectra for automated grading of diabetic maculopathy.
Mookiah, Muthu Rama Krishnan; Acharya, U Rajendra; Chandran, Vinod; Martis, Roshan Joy; Tan, Jen Hong; Koh, Joel E W; Chua, Chua Kuang; Tong, Louis; Laude, Augustinus
Diabetic macular edema (DME) is one of the most common causes of visual loss among diabetes mellitus patients. Early detection and successive treatment may improve the visual acuity. DME is mainly graded into non-clinically significant macular edema (NCSME) and clinically significant macular edema according to the location of hard exudates in the macula region. DME can be identified by manual examination of fundus images. It is laborious and resource intensive. Hence, in this work, automated grading of DME is proposed using higher-order spectra (HOS) of Radon transform projections of the fundus images. We have used third-order cumulants and bispectrum magnitude, in this work, as features, and compared their performance. They can capture subtle changes in the fundus image. Spectral regression discriminant analysis (SRDA) reduces feature dimension, and minimum redundancy maximum relevance method is used to rank the significant SRDA components. Ranked features are fed to various supervised classifiers, viz. Naive Bayes, AdaBoost and support vector machine, to discriminate No DME, NCSME and clinically significant macular edema classes. The performance of our system is evaluated using the publicly available MESSIDOR dataset (300 images) and also verified with a local dataset (300 images). Our results show that HOS cumulants and bispectrum magnitude obtained an average accuracy of 95.56 and 94.39% for MESSIDOR dataset and 95.93 and 93.33% for local dataset, respectively. PMID:25894464
The Cancer-Related Transcription Factor Runx2 Modulates Cell Proliferation in Human Osteosarcoma Cell Lines
Lucero, Claudia M.J.; Vega, Oscar A.; Osorio, Mariana M.; Tapia, Julio C.; Antonelli, Marcelo; Stein, Gary S.; Van Wijnen, Andre J.; Galindo, Mario A.
Runx2 regulates osteogenic differentiation and bone formation, but also suppresses pre-osteoblast proliferation by affecting cell cycle progression in the G1 phase. The growth suppressive potential of Runx2 is normally inactivated in part by protein destabilization, which permits cell cycle progression beyond the G1/S phase transition, and Runx2 is again up-regulated after mitosis. Runx2 expression also correlates with metastasis and poor chemotherapy response in osteosarcoma. Here we show that six human osteosarcoma cell lines (SaOS, MG63, U2OS, HOS, G292, and 143B) have different growth rates, which is consistent with differences in the lengths of the cell cycle. Runx2 protein levels are cell cycle-regulated with respect to the G1/S phase transition in U2OS, HOS, G292, and 143B cells. In contrast, Runx2 protein levels are constitutively expressed during the cell cycle in SaOS and MG63 cells. Forced expression of Runx2 suppresses growth in all cell lines indicating that accumulation of Runx2 in excess of its pre-established levels in a given cell type triggers one or more anti-proliferative pathways in osteosarcoma cells. Thus, regulatory mechanisms controlling Runx2 expression in osteosarcoma cells must balance Runx2 protein levels to promote its putative oncogenic functions, while avoiding suppression of bone tumor growth. PMID:22949168
Pontiff, Mattie; Ithurburn, Matthew P.; Ellis, Thomas; Cenkus, Kathleen
ABSTRACT Background Generalized joint laxity is more prevalent in women than men and may lead to poorer post-operative outcomes in select orthopedic populations. There are no studies examining peri-operative function in patients with generalized joint laxity (GJL) and femoroacetabular impingement (FAI). Purpose The purpose of this study was to determine the difference in perceived function and quality of life as measured by the Hip Outcome Score ADL subscale (HOS-ADL), International Hip Outcomes Tool (iHOT-33) and the Short Form 12-Item Health Survey (SF-12) in women with and without GJL prior to and six months after undergoing hip arthroscopy for FAI. Study Design Cohort Study Methods Peri-operative data were collected from women with FAI from November 2011-September 2014. Lax subjects were women with laxity scores ≥4/9 on the Beighton and Horan Joint Mobility Index; Nonlax subjects were women with laxity scores <4/9. Functional outcomes were evaluated using the HOS-ADL, iHOT-33, PCS-12, and the MCS-12 pre-operatively and at 6 months post-operatively. Change scores (post-score – pre-score) were calculated for each outcome measure and compared between groups, along with pre-operative and post-operative means, using Mann-Whitney U tests. Results 166 women met the inclusion criteria: Nonlax (n = 131), Lax (n = 35). There were no statistically significant differences between groups in pre-operative functional outcomes (all p > .05). Additionally, there were no statistically significant differences between groups in post-operative means or change scores, respectively, for HOS-ADL (p = .696, .358), iHOT-33 (p = .550, .705), PCS-12 (p = .713, .191), and MCS-12 (p = .751, .082). Laxity score was not associated with any post-operative functional outcome score or change score (all p > .05). Conclusion Women with and without generalized joint laxity do not appear to report differences in hip function in the 6-month peri-operative period before and after hip
Gold(I) complexes of 9-deazahypoxanthine as selective antitumor and anti-inflammatory agents.
Vančo, Ján; Gáliková, Jana; Hošek, Jan; Dvořák, Zdeněk; Paráková, Lenka; Trávníček, Zdeněk
The gold(I) mixed-ligand complexes involving O-substituted derivatives of 9-deazahypoxanthine (HLn) and triphenylphosphine (PPh3) with the general formula [Au(Ln)(PPh3)] (1-5) were prepared and thoroughly characterized by elemental analysis, FT-IR and multinuclear NMR spectroscopy, ESI+ mass spectrometry, single crystal X-ray (HL5 and complex 2) and TG/DTA analyses. Complexes 1-5 were evaluated for their in vitro antitumor activity against nine human cancer lines, i.e. MCF7 (breast carcinoma), HOS (osteosarcoma), A549 (adenocarcinoma), G361 (melanoma), HeLa (cervical cancer), A2780 (ovarian carcinoma), A2780R (ovarian carcinoma resistant to cisplatin), 22Rv1 (prostate cancer) and THP-1 (monocytic leukaemia), for their in vitro anti-inflammatory activity using a model of LPS-activated macrophages, and for their in vivo antiedematous activity by λ-carrageenan-induced hind paw edema model on rats. The results showed that the complexes 1-5 exhibit selective in vitro cytotoxicity against MCF7, HOS, 22Rv1, A2780 and A2780R, with submicromolar IC50 values for 2 against the MCF7 (0.6 µM) and HOS (0.9 µM). The results of in vitro cytotoxicity screening on primary culture of human hepatocytes (HEP220) revealed up to 30-times lower toxicity of compounds against healthy cells as compared with cancer cells. Additionally, the complexes 1-5 significantly influence the secretion and expression of pro-inflammatory cytokines TNF-α and IL-1β by a similar manner as a commercially used anti-arthritic drug Auranofin. The tested complexes also significantly influence the rate and overall volume of the edema, caused by the intraplantar application of λ-carrageenan polysaccharide to rats. Based on these promising results, the presented compounds could qualify to become feasible candidates for advanced testing as potential antitumor and anti-inflammatory drug-like compounds. PMID:25333949
Gold(I) Complexes of 9-Deazahypoxanthine as Selective Antitumor and Anti-Inflammatory Agents
Vančo, Ján; Gáliková, Jana; Hošek, Jan; Dvořák, Zdeněk; Paráková, Lenka; Trávníček, Zdeněk
The gold(I) mixed-ligand complexes involving O-substituted derivatives of 9-deazahypoxanthine (HLn) and triphenylphosphine (PPh3) with the general formula [Au(Ln)(PPh3)] (1–5) were prepared and thoroughly characterized by elemental analysis, FT-IR and multinuclear NMR spectroscopy, ESI+ mass spectrometry, single crystal X-ray (HL5 and complex 2) and TG/DTA analyses. Complexes 1–5 were evaluated for their in vitro antitumor activity against nine human cancer lines, i.e. MCF7 (breast carcinoma), HOS (osteosarcoma), A549 (adenocarcinoma), G361 (melanoma), HeLa (cervical cancer), A2780 (ovarian carcinoma), A2780R (ovarian carcinoma resistant to cisplatin), 22Rv1 (prostate cancer) and THP-1 (monocytic leukaemia), for their in vitro anti-inflammatory activity using a model of LPS-activated macrophages, and for their in vivo antiedematous activity by λ-carrageenan-induced hind paw edema model on rats. The results showed that the complexes 1–5 exhibit selective in vitro cytotoxicity against MCF7, HOS, 22Rv1, A2780 and A2780R, with submicromolar IC50 values for 2 against the MCF7 (0.6 µM) and HOS (0.9 µM). The results of in vitro cytotoxicity screening on primary culture of human hepatocytes (HEP220) revealed up to 30-times lower toxicity of compounds against healthy cells as compared with cancer cells. Additionally, the complexes 1–5 significantly influence the secretion and expression of pro-inflammatory cytokines TNF-α and IL-1β by a similar manner as a commercially used anti-arthritic drug Auranofin. The tested complexes also significantly influence the rate and overall volume of the edema, caused by the intraplantar application of λ-carrageenan polysaccharide to rats. Based on these promising results, the presented compounds could qualify to become feasible candidates for advanced testing as potential antitumor and anti-inflammatory drug-like compounds. PMID:25333949
Crystallographic and Spectroscopic Insights into Heme Degradation by Mycobacterium tuberculosis MhuD
Mycobacterium heme utilization degrader (MhuD) is a heme-degrading protein from Mycobacterium tuberculosis responsible for extracting the essential nutrient iron from host-derived heme. MhuD has been previously shown to produce unique organic products compared to those of canonical heme oxygenases (HOs) as well as those of the IsdG/I heme-degrading enzymes from Staphylococcus aureus. Here, we report the X-ray crystal structure of cyanide-inhibited MhuD (MhuD–heme–CN) as well as detailed 1H nuclear magnetic resonance (NMR), UV/vis absorption, and magnetic circular dichroism (MCD) spectroscopic characterization of this species. There is no evidence for an ordered network of water molecules on the distal side of the heme substrate in the X-ray crystal structure, as was previously reported for canonical HOs. The degree of heme ruffling in the crystal structure of MhuD is greater than that observed for HO and less than that observed for IsdI. As a consequence, the Fe 3dxz-, 3dyz-, and 3dxy-based MOs are very close in energy, and the room-temperature 1H NMR spectrum of MhuD–heme–CN is consistent with population of both a 2Eg electronic state with a (dxy)2(dxz,dyz)3 electron configuration, similar to the ground state of canonical HOs, and a 2B2g state with a (dxz,dyz)4(dxy)1 electron configuration, similar to the ground state of cyanide-inhibited IsdI. Variable temperature, variable field MCD saturation magnetization data establishes that MhuD–heme–CN has a 2B2g electronic ground state with a low-lying 2Eg excited state. Our crystallographic and spectroscopic data suggest that there are both structural and electronic contributions to the α-meso regioselectivity of MhuD-catalyzed heme cleavage. The structural distortion of the heme substrate observed in the X-ray crystal structure of MhuD–heme–CN is likely to favor cleavage at the α- and γ-meso carbons, whereas the spin density distribution may favor selective oxygenation of the α-meso carbon. PMID
Human T-cell leukemia virus type 1 infection leads to arrest in the G1 phase of the cell cycle.
Liu, Meihong; Yang, Liangpeng; Zhang, Ling; Liu, Baoying; Merling, Randall; Xia, Zheng; Giam, Chou-Zen
Infection by the human T-cell leukemia virus type 1 (HTLV-1) is thought to cause dysregulated T-cell proliferation, which in turn leads to adult T-cell leukemia/lymphoma. Early cellular changes after HTLV-1 infection have been difficult to study due to the poorly infectious nature of HTLV-1 and the need for cell-to-cell contact for HTLV-1 transmission. Using a series of reporter systems, we show that HeLa cells cease proliferation within one or two division cycles after infection by HTLV-1 or transduction of the HTLV-1 tax gene. HTLV-1-infected HeLa cells, like their tax-transduced counterparts, expressed high levels of p21(CIP1/WAF1) and p27(KIP1), developed mitotic abnormalities, and became arrested in G(1) in senescence. In contrast, cells of a human osteosarcoma lineage (HOS) continued to divide after HTLV-1 infection or Tax expression, albeit at a reduced growth rate and with mitotic aberrations. Unique to HOS cells is the dramatic reduction of p21(CIP1/WAF1) and p27(KIP1) expression, which is in part associated with the constitutive activation of the phosphatidylinositol-3-kinase (PI3K)-protein kinase B (Akt) pathway. The loss of p21(CIP1/WAF1) and p27(KIP1) in HOS cells apparently allows HTLV-1- and Tax-induced G(1) arrest to be bypassed. Finally, HTLV-1 infection and Tax expression also cause human SupT1 T cells to arrest in the G(1) phase of the cell cycle. These results suggest that productive HTLV-1 infection ordinarily leads to Tax-mediated G(1) arrest. However, T cells containing somatic mutations that inactivate p21(CIP1/WAF1) and p27(KIP1) may continue to proliferate after HTLV-1 infection and Tax expression. These infected cells can expand clonally, accumulate additional chromosomal abnormalities, and progress to cancer. PMID:18596104
The use of the history of science as a motivational tool in middle school science
NASA Astrophysics Data System (ADS)
Chapel, Fredrick Milton
In science classrooms, the content of science is usually taught as sets of facts to be learned as they are, with little understanding as to how those facts were discovered, verified, and challenged by the scientific community (American Association for the Advancement of Science, 1990). Additionally, laboratory activities are not used to challenge those established facts, but only verify them (Roth & Roychoudhury, 1993). The result is science devoid of inherent interest for students, creating a downward spiral of decreasing engagement and achievement, a conclusion supported by recent national and international assessment measures (Schmidt, McKnight, Cogan, Jakwerth, & Houang, 1999). The challenge is to create classroom environments in middle school science that support and motivate students to become and remain engaged in productive learning activities. The objective of this study was to determine the effectiveness of using the history of science (HOS) as a motivational tool in middle school science education. The behaviors and work samples of students were observed and analyzed in relation to three research questions: (1) What patterns of motivated student behavior emerge during a unit of instruction? (2) What evidence of student conceptual change can be observed during a unit of instruction? (3) What is the role of the HOS in the science classroom in regard to student motivation? This naturalistic, ethnographic case study examined four eighth-grade science classrooms in an inner-city charter middle school. The data collected included transcripts of classroom dialogue, student work samples (notes, essays, projects), daily anecdotal notes of the teacher, and observations of the classroom by educational consultants and school personnel. Data analysis was based upon specialized forms of discourse and task analysis with ethnomethodological and phenomenological sensitivity as to how students make meaning of what they are learning. The findings identified patterns of
Hospital planning for weapons of mass destruction incidents.
Perry, Ronald W; Lindell, Michael K
As terrorist attacks increase in fre-quency, hospital disaster plans need to be scrutinized to ensure that they take into account issues unique to weapons of mass destruction. This paper reports a review of the literature addressing hospital experiences with such incidents and the planning lessons thus learned. Construction of hos-pital disaster plans is examined as an ongoing process guided by the disaster planning committee. A review is completed of six special elements of weapons of mass destruction incidents that should be addressed in hospital disaster plans. The paper closes with a dis-cussion of the importance of train-ing and exercises in maintaining and improving the disaster plan. PMID:17970447
10,000-Hour Cyclic Oxidation Behavior at 982 C (1800 F) of 68 High-Temperature Co-, Fe-, and Ni-Base Alloys
NASA Technical Reports Server (NTRS)
Barrett, Charles A.
Sixty-eight high temperature Co-, Fe-, and Ni-base alloys were tested for 10-one thousand hour cycles in static air at 982 C (1800 F). The oxidation behavior of the test samples was evaluated by specific weight change/time data, x-ray diffraction of the post-test samples, and their final appearance. The gravimetric and appearance data were combined into a single modified oxidation parameter, KB4 to rank the cyclic oxidation resistance from excellent to catastrophic. The alloys showing the 'best' resistance with no significant oxidation attack were the alumina/aluminate spinel forming Ni-base turbine alloys: U-700, NASA-VIA and B-1900; the Fe-base ferritic alloys with Al: TRW-Valve, HOS-875, NASA-18T, Thermenol and 18SR; and the Ni-base superalloy IN-702.
EEG signal analysis: a survey.
Subha, D Puthankattil; Joseph, Paul K; Acharya U, Rajendra; Lim, Choo Min
The EEG (Electroencephalogram) signal indicates the electrical activity of the brain. They are highly random in nature and may contain useful information about the brain state. However, it is very difficult to get useful information from these signals directly in the time domain just by observing them. They are basically non-linear and nonstationary in nature. Hence, important features can be extracted for the diagnosis of different diseases using advanced signal processing techniques. In this paper the effect of different events on the EEG signal, and different signal processing methods used to extract the hidden information from the signal are discussed in detail. Linear, Frequency domain, time - frequency and non-linear techniques like correlation dimension (CD), largest Lyapunov exponent (LLE), Hurst exponent (H), different entropies, fractal dimension(FD), Higher Order Spectra (HOS), phase space plots and recurrence plots are discussed in detail using a typical normal EEG signal. PMID:20433058
Inhibition of Plastocyanin to P700+ Electron Transfer in Chlamydomonas reinhardtii by Hyperosmotic Stress1
Cruz, Jeffrey A.; Salbilla, Brian A.; Kanazawa, Atsuko; Kramer, David M.
Oxygen electrode and fluorescence studies demonstrate that linear electron transport in the freshwater alga Chlamydomonas reinhardtii can be completely abolished by abrupt hyperosmotic shock. We show that the most likely primary site of inhibition of electron transfer by hyperosmotic shock is a blockage of electron transfer between plastocyanin (PC) or cytochrome c6 and P700. The effects on this reaction were reversible upon dilution of the osmolytes and the stability of plastocyanin or photosystem (PS) I was unaffected. Electron micrographs of osmotically shocked cells showed a significant decrease in the thylakoid lumen volume. Comparison of estimated lumenal width with the x-ray structures of plastocyanin and PS I suggest that lumenal space contracts during HOS so as to hinder the movement of docking to PS I of plastocyanin or cytochrome c6. PMID:11706196
Establishment of a cell line producing bone morphogenetic protein from a human osteosarcoma.
Takaoka, K; Yoshikawa, H; Masuhara, K; Sugamoto, K; Tsuda, T; Aoki, Y; Ono, K; Sakamoto, Y
A human osteosarcoma cell line was established from a biopsy specimen from a 13-year-old girl. The osteosarcoma tissue was maintained in athymic nude mice (Balb C nu/nu) by serial transplantation for three years. The tumor was excised from a host mouse and digested with collagenase. The isolated cells were cultured by 98 passages in 14 months, and clones of osteosarcoma cells were obtained by limiting dilution. A clone named human osteosarcoma cell 6 (H-OS-6) that showed the osteoblastic phenotypes of productions of bone morphogenetic protein (BMP) and alkaline phosphatase and a response to human parathyroid hormone (h-PTH 1-34) was selected. The morphology of its chromosomes indicated its human origin. This human osteosarcoma cell line is unique in producing BMP under in vitro conditions. PMID:2545399
The rovibrational nature of closed-shell third-row triatomics: HOX and HXO, X = Si+, P, S+, and Cl
NASA Astrophysics Data System (ADS)
Kitchens, Mason J. R.; Fortenberry, Ryan C.
Atmospheric and even interstellar chemistry are dominated by the presence of small molecules. This work examines closed-shell, triatomic, hydroxide molecules containing a single third-row atom: HOSi+, HSiO+, HPO, HOS+, HSO+, HOCl, and HClO as well as their deuterated isotopologues and those with 37Cl. The use of proven composite energy-defined quartic force fields is further benchmarked where experimental data is known in order to provide an estimate for the accuracy of other previously unknown values for these and even previously unexamined species. Rotational constants within 1 MHz and vibrational frequencies within 1 cm-1 of experiment are produced in some cases with typical comparison around 50 MHz and 10 cm-1, respectively. As such the remaining previously unreported geometries, spectroscopic constants, vibrational frequencies, and dipole moments will enhance the spectroscopic analysis of any environment, terrestrially or astronomically, where the properties of these molecules may serve better to inform the chemistry of these regions.
Biology of MET: a double life between normal tissue repair and tumor progression
MNNG HOS transforming gene (MET) is a class IV receptor tyrosine kinase, expressed on the surface of epithelial cells. The interaction with the hepatocyte grow factor (HGF) induces MET dimerization and the activation of multiple intracellular pathways leading to cell proliferation, anti-apoptosis, morphogenic differentiation, motility, invasion, and angiogenesis. Knock out mice have demonstrated that MET is necessary for normal embryogenesis including the formation of striate muscles, liver and trophoblastic structures. The overexpression of MET and HGF are common in solid tumors and contribute to determine their growth. Indeed, MET has been cloned as a transforming gene from a chemically induced human osteosarcoma cell line and therefore is considered a proto-oncogene. Germline MET mutations are characteristic of hereditary papillary kidney cancers and MET amplification is observed in tumors including lung and gastric adenocarcinomas. The inhibition of MET signaling is the target for specific drugs that are raising exciting expectation for medical treatment of cancer. PMID:25992381
Diabetes and alcohol: Double jeopardy with regard to oxidative toxicity and sexual dysfunction in adult male Wistar rats.
Himabindu, B; Madhu, P; Reddy, P Sreenivasula
The aim of this study was to test whether diabetic rats exposed to alcohol demonstrate a higher degree of reproductive toxicity and suffer with elevated oxidative toxicity when compared with alcohol exposed control rats. Diabetes was induced by injecting single dose of streptozotocin and alcohol was administered through orogastric tube once daily for a period of 55 days. Daily sperm production, epididymal sperm count, motile, viable and HOS-tail coiled sperms, serum testosterone levels and testicular 3β- and 17β-hydroxysteroid dehydrogenase activity levels were significantly decreased in diabetic rats. Significant reduction in testicular and epididymal superoxide dismutase and catalase activity levels, and elevation in lipid peroxidation products were observed in diabetic rats. Similar reproductive and oxidative toxicity was observed in alcohol treated control rats. Further, alcohol exposed diabetic rats showed additional deterioration in reproductive endpoints and noteworthy elevation in oxidative toxicity suggesting that treatment with alcohol further deteriorates sexual dysfunction in STZ-induced diabetic rats. PMID:25541261
Estrogen binding, receptor mRNA, and biologic response in osteoblast-like osteosarcoma cells
Komm, B.S.; Terpening, C.M.; Benz, D.J.; Graeme, K.A.; Gallegos, A.; Korc, M.; Greene, G.L.; O'Malley, B.W.; Haussler, M.R.
High specific activity estradiol labeled with iodine-125 was used to detect approximately 200 saturable, high-affinity (dissociation constant approximately equal to 1.0 nM) nuclear binding sites in rat (ROS 17/2.8) and human (HOS TE85) clonal osteoblast-like osteosarcoma cells. Of the steroids tested, only testosterone exhibited significant cross-reactivity with estrogen binding. RNA blot analysis with a complementary DNA probe to the human estrogen receptor revealed putative receptor transcripts of 6 to 6.2 kilobases in both rat and human osteosarcoma cells. Type I procollagen and transforming growth factor-beta messenger RNA levels were enhanced in cultured human osteoblast-like cells treated with 1 nM estradiol. Thus, estrogen can act directly on osteoblasts by a receptor-mediated mechanism and thereby modulate the extracellular matrix and other proteins involved in the maintenance of skeletal mineralization and remodeling.
Towards non-cooperative standoff biometrics using extremely low frequency (ELF) processing
NASA Astrophysics Data System (ADS)
Byrd, Kenneth A.; Szu, Harold; Wardlaw, Michael J.
Earth's Field Nuclear Magnetic Resonance (⊕-NMR) is a novel detection technique that exploits the uniformity of the earth's magnetic field to measure precessing magnetizations of isotope-having objects such as human beings. The ability to provide standoff medical surveillance will increase the Biomedical Wellness (BMW) of not only aging American citizens but also persons we seek to track from healthy to diseased states. Taking advantage of the uniform field in the earth, spatio-temporal integration (3 orders due to time and 4 orders due to space) and Higher Order Statistics (HOS), namely, change detection via kurtosis (ΔK), SNR and the ability to make non-locality measurements may be increased to at least the level of the typical hospital NMR devices. Moreover, we present theoretical calculations of magnitude and consider time dynamics of the resonant/relaxing magnetic fields as a function of 3D human motion and distance.
Fine-structure map of the histidine transport genes in Salmonella typhimurium.
Ames, G F; Noel, K D; Taber, H; Spudich, E N; Nikaido, K; Afong, J
Afine-structure genetic map of the histidine transport region of the Salmonella typhimurium chromosome was constructed. Twenty-five deletion mutants were isolated and used for dividing the hisJ and hisP genes into 8 and 13 regions respectively. A total of 308 mutations, spontaneous and mutagen induced, have been placed in these regions by deletion mapping. The histidine transport operon is presumed to be constituted of genes dhuA, hisJ, and hisP, and the regulation of the hosP and hisJ genes by dhuA is discussed. The orientation of this operon relative to purF has been established by three-point crosses as being: purF duhA hisJ hisP. PMID:321422
Fine-structure map of the histidine transport genes in Salmonella typhimurium.
Ames, G F; Noel, K D; Taber, H; Spudich, E N; Nikaido, K; Afong, J
Afine-structure genetic map of the histidine transport region of the Salmonella typhimurium chromosome was constructed. Twenty-five deletion mutants were isolated and used for dividing the hisJ and hisP genes into 8 and 13 regions respectively. A total of 308 mutations, spontaneous and mutagen induced, have been placed in these regions by deletion mapping. The histidine transport operon is presumed to be constituted of genes dhuA, hisJ, and hisP, and the regulation of the hosP and hisJ genes by dhuA is discussed. The orientation of this operon relative to purF has been established by three-point crosses as being: purF duhA hisJ hisP. PMID:321422
Design of biomaterials for intracellular delivery of carbon monoxide.
Inaba, Hiroshi; Fujita, Kenta; Ueno, Takafumi
Carbon monoxide (CO) is recognized as one of the most important gas signaling molecules involved in governing various therapeutic responses. Intracellular generation of CO is spatiotemporally controlled by catalytic reactions of heme oxygenases (HOs). Thus, the ability to control intracellular CO delivery with modulation of the CO-release rate in specific amounts and locations is expected to improve our fundamental understanding of the functions of CO and the development of clinical applications. For this purpose, CO-releasing molecules (CORMs) have been developed and investigated in vitro and in vivo. Most CORMs are based on transition metal carbonyl complexes. Recently, various biomaterials consisting of metal carbonyls with biomacromolecular scaffolds have been reported to improve the properties of bare metal carbonyls. In this mini-review, current progress in CO delivery, recent strategies for the development of CORMs, and future directions in this field are discussed. PMID:26252321
ActiveSeismoPick3D - automatic first arrival determination for large active seismic arrays
NASA Astrophysics Data System (ADS)
Paffrath, Marcel; Küperkoch, Ludger; Wehling-Benatelli, Sebastian; Friederich, Wolfgang
We developed a tool for automatic determination of first arrivals in active seismic data based on an approach, that utilises higher order statistics (HOS) and the Akaike information criterion (AIC), commonly used in seismology, but not in active seismics. Automatic picking is highly desirable in active seismics as the number of data provided by large seismic arrays rapidly exceeds of what an analyst can evaluate in a reasonable amount of time. To bring the functionality of automatic phase picking into the context of active data, the software package ActiveSeismoPick3D was developed in Python. It uses a modified algorithm for the determination of first arrivals which searches for the HOS maximum in unfiltered data. Additionally, it offers tools for manual quality control and postprocessing, e.g. various visualisation and repicking functionalities. For flexibility, the tool also includes methods for the preparation of geometry information of large seismic arrays and improved interfaces to the Fast Marching Tomography Package (FMTOMO), which can be used for the prediction of travel times and inversion for subsurface properties. Output files are generated in the VTK format, allowing the 3D visualization of e.g. the inversion results. As a test case, a data set consisting of 9216 traces from 64 shots was gathered, recorded at 144 receivers deployed in a regular 2D array of a size of 100 x 100 m. ActiveSeismoPick3D automatically checks the determined first arrivals by a dynamic signal to noise ratio threshold. From the data a 3D model of the subsurface was generated using the export functionality of the package and FMTOMO.
Designing Hydrologic Observatories as a Community Resource
NASA Astrophysics Data System (ADS)
Hooper, R. P.; Duncan, J. M.
CUAHSI convened a workshop in August 2004 to explore what makes a successful hydrologic observatory. Because of their high cost, only a small number of observatories will be operated, at least initially. (CUAHSI has recommended a pilot network of 5 observatories to develop operational experience and an eventual network of approximately 15 sites.) Because hydrologic scientists can work "in their backyard" (unlike oceanographers or astronomers), hydrologic observatories must offer significant advantages over current methods of field work to successfully attract researchers. Twenty-four teams of scientists submitted "prospectuses" of potential locations for hydrologic observatories for consideration by network attendees. These documents (available at http://www.cuahsi.org) were marketing documents to the workshop participants, who voted for a hypothetical network of 5 observatories from the 24 proposed sites. This network formed the basis for a day of discussions on necessary attributes of core data and how to form a network of observatories from a collection of sites that are designed and implemented individually. Key findings included: 1) Core data must be balanced among disciplines. Although the hydrologic cycle is an organizing principle for the design of HOs, physical data cannot dominate the core data; chemical and biological data, although more expensive to collect, must be given equal footing. 2) New data collection must strategically leverage existing data. Resources are always limited, so that a successful HO must carefully target gaps in existing data, as determined by an explicitly stated conceptual model, and fill them rather than designing an independent study. 3) Site logistics must support remote researchers. Significant resources will be necessary for on-site staff to handle housing, transportation, permitting and other needs. 4) Network-level hypotheses are required early in the implementation of HOs. A network will only emerge around hypotheses
Molecular perspective of antibody aggregates and their adsorption on Protein A resin.
Yu, Deqiang; Song, Yuanli; Huang, Richard Y-C; Swanson, Ryan K; Tan, Zhijun; Schutsky, Elizabeth; Lewandowski, Angela; Chen, Guodong; Li, Zheng Jian
Antibody aggregate is a common issue in therapeutic antibodies, which may compromise product efficacy and cause adverse effects. Antibody aggregate level is normally controlled in bioprocessing by polishing steps after Protein A capture. This paper studied the Higher Order Structures (HOS) of antibody aggregates (dimer H1 and H2) and their adsorption on Protein A resin and thus elucidated the mechanism using Protein A capture for enhanced aggregate removal. The HOS of antibody aggregates and their complex with Protein A were characterized using HDX-MS combined with SEC-MALS, Protein Conformational Array (PCA), and molecular modeling. The aggregate size and Protein A binding ratio suggested that H2 has much more compact structure than H1. HDX-MS and PCA further revealed that H1 was formed by single Fab-Fab interaction while H2 formed by Fab-Fab and likely Fc-Fc interaction. On Protein A resin, both the molar binding ratio and the correlation between protein size and ligand distance support that each monomer can only bind one Protein A ligand, while each dimer can bind two ligands, thus resulting in stronger resin binding. Furthermore, dimer H2 binds stronger than dimer H1 due to its compact structure. By integrating biophysical analysis and molecular modeling with process development, this study revealed the antibody aggregate structures and the mechanism of aggregate removal using Protein A chromatography. It also provided a general strategy for in-depth product and process understanding in antibody and other biologics development. PMID:27344283
A high-fat meal enriched with eicosapentaenoic acid reduces postprandial arterial stiffness measured by digital volume pulse analysis in healthy men.
Hall, Wendy L; Sanders, Katie A; Sanders, Thomas A B; Chowienczyk, Philip J
Diets rich in eicosapentaenoic acid [EPA; 20:5(n-3)] are associated with decreased arterial stiffness, but postprandial effects on vascular function are unknown. We investigated whether an EPA-enriched high-fat meal could improve postprandial vascular function. Seventeen healthy men ingested 2 test meals (51 g fat), 1 wk apart, in random order: 5 g EPA plus high-oleic sunflower oil (HOS) vs. HOS only. A second high-fat meal (44 g fat), the same on both study days, was provided 4 h later. Blood pressure and arterial function were measured using digital volume pulse (DVP) to derive a stiffness index (DVP-SI) and reflection index in fasting subjects at 3 and 6 h following the test meal. Blood samples were taken following the test meal for plasma 8-isoprostane F2alpha, nitric oxide (NO) metabolites (NOx), glucose, insulin, triacylglycerol, and fatty acid analysis. The plasma EPA concentration (mean +/- SD) reached a peak of 2.10 +/- 0.99 mmol/L following the EPA meal (5 h) and did not rise above 0.27 +/- 0.16 mmol/L 1 h following the placebo meal. DeltaDVP-SI did not differ between the 2 test meals at 3 h but was greater at 6 h following EPA (6 h -0.65 +/- 0.65 m/s) compared with placebo (6 h -0.33 +/- 1.26 m/s). Plasma 8-isoprostane F2alpha concentrations increased by 48% at 6 h compared with baseline following the EPA meal and plasma NOx decreased following both meals, with no differences between the meals in the changes. Changes in other variables measured also did not differ after subjects consumed the 2 meals. In conclusion, adding EPA to a high-fat meal results in acute changes in vascular tone, independent of changes in oxidative stress. PMID:18203893
Increased electrocatalyzed performance through hairpin oligonucleotide aptamer-functionalized gold nanorods labels and graphene-streptavidin nanomatrix: Highly selective and sensitive electrochemical biosensor of carcinoembryonic antigen.
Wen, Wei; Huang, Jing-Yi; Bao, Ting; Zhou, Jun; Xia, Hong-Xing; Zhang, Xiu-Hua; Wang, Sheng-Fu; Zhao, Yuan-Di
We report a triplex signal amplification strategy for sensitive biosensing of cancer biomarker by taking advantage of hairpin-shaped oligonucleotide-functionalized gold nanorods (HO-GNRs), graphene and the avidin-biotin reation. The strategy expands electrochemical detection of carcinoembryonic antigen (CEA) by using an aptamer as biosensor's recognition element and HO-GNRs as signal enhancer. To construct this biosensor, the GNR was used as a carrier of horseradish peroxidase (HRP) and HO aptamer with a biotin at the 3'-end and a thiol at the 5'-end, which amplified the electrochemical response because of a large molar ratio of HRP to HO. In the presence of target CEA, the binding reactions of CEA with the loop portions of the HOs caused HOs' loop-stem structure opened and exposed the biotins, and then HRP-GNRs-HO conjugates were captured on graphene and streptavidin modified electrodes via the reaction between the exposed biotins and preimmobilized streptavidins. The accumulation of HRP effectively catalyzed the hydrogen peroxide-mediated oxidation of o-phenylenediamine to generate an electrochemical reduction current for CEA detection. Under optimal conditions, the electrochemical biosensor exhibited a wide dynamic range of 5pgmL(-1) and 50ngmL(-1) toward CEA standards with a low detection limit of 1.5pgmL(-1) (signal-to-noise ratio of 3). The proposed biosensor accurately detected CEA concentration in 8 human serum samples from patients with lung diseases, showing excellent correlations with standard chemiluminescence immunoassay. Furthermore, these results of target DNA detection made it abundantly clear that the proposed strategy can also be extended for detection of other relative biomarkers using different functional DNA structures, which shows great prospects in single-nucleotide polymorphisms analysis, biomedical sensing and application for accurate clinical diseases diagnostic. PMID:27111123
The Semen pH Affects Sperm Motility and Capacitation
Hong, Zhiwei; Xie, Min; Chen, Shengrong; Yao, Bing
As the chemical environment of semen can have a profound effect on sperm quality, we examined the effect of pH on the motility, viability and capacitation of human sperm. The sperm in this study was collected from healthy males to avoid interference from other factors. The spermatozoa cultured in sperm nutrition solution at pH 5.2, 6.2, 7.2 and 8.2 were analyzed for sperm total motility, progressive motility (PR), hypo-osmotic swelling (HOS) rate, and sperm penetration. Our results showed that these parameters were similar in pH 7.2 and 8.2 sperm nutrition solutions, but decreased in pH 5.2 and 6.2 solutions. The HOS rate exhibited positive correlation with the sperm total motility and PR. In addition, the sperm Na+/K+-ATPase activity at different pHs was measured, and the enzyme activity was significantly lower in pH 5.2 and 6.2 media, comparing with that in pH 8.2 and pH 7.2 solutions. Using flow cytometry (FCM) and laser confocal scanning microscopy (LCSM) analysis, the intracellular Ca2+ concentrations of sperm cultured in sperm capacitation solution at pH 5.2, 6.2, 7.2 and 8.2 were determined. Compared with that at pH 7.2, the mean fluorescence intensity of sperm in pH 5.2 and 6.2 media decreased significantly, while that of pH 8.2 group showed no difference. Our results suggested that the declined Na+/K+-ATPase activity at acidic pHs result in decreased sperm movement and capacitation, which could be one of the mechanisms of male infertility. PMID:26173069
Evaluation of the Spermicidal and Contraceptive Activity of Platycodin D, a Saponin from Platycodon grandiflorum
Lu, Zongliang; Wang, Leiguang; Zhou, Rui; Qiu, Yi; Yang, Liuna; Zhang, Chanyu; Cai, Min; Mi, Mantian; Xu, Hongxia
Background The extract of Platycodon grandiflorum has been reported to have effective spermicidal activity. This study was designed to evaluate the spermicidal and contraceptive activity, as well as the safety, of Platycodin D (PD), a major saponin in Platycodon grandiflorum. Methods Using the computer-aided sperm analysis (CASA) test criteria, the sperm-immobilizing activity of PD was studied using highly motile human sperm. The sperm viability was assessed by fluorescent staining using SYBR-14 (living sperm) and propidium iodide (dead sperm). The sperm membrane integrity was assessed by evaluating the hypo-osmotic swelling (HOS) and examinations by transmission electron microscopy (TEM) and scanning electron microscopy (SEM). The in vivo contraceptive efficacy was evaluated in rats using post-intrauterine PD application. The comet assay was employed to determine whether PD caused DNA damage in the sperm. Vaginal biopsies were also performed to determine whether the PD gel induced vaginal inflammation. Results A dose-dependent effect of PD on the sperm motility and viability was observed. The maximum spermicidal effect was observed with a 0.25 mM concentration of PD. More than 70% of the PD-treated sperm lost their HOS responsiveness at a concentration of 0.20 mM PD, indicating that PD caused injury to the sperm plasma membrane. TEM and SEM revealed significant damage to both the head and tail membranes of the sperm. PD decreased the fertility to zero in rats, was non-DNA damaging and was not harmful to the vaginal tissue in the rats. Conclusion PD has significant spermicidal activity that should be explored in further studies. PMID:24303079
Ultramicroscopic and biochemical changes in ram spermatozoa cryopreserved with trehalose-based hypertonic extenders.
Aisen, E; Quintana, M; Medina, V; Morello, H; Venturino, A
The ability of a range of extenders to cryopreserve ram spermatozoa was tested. The extenders were modified by the inclusion of citrate, Tris buffer, trehalose, and EDTA. Ejaculates from three Pampinta rams were evaluated and pooled at 30 degrees C. The semen was diluted to contain 1 x 10(9) cells/mL, cooled to 5 degrees C, loaded into 0.25-mL straws, frozen and stored in liquid nitrogen. Evaluation was based on the hypoosmotic swelling test (HOS test), electron microscopy, and biochemical parameters such as lipid peroxidation and reduced and total glutathione levels, all measured after thawing. The HOS test indicated that the percentage of intact plasma membranes after freezing and thawing was significantly higher for the hypertonic extender containing trehalose (T), compared with an extender containing trehalose+EDTA (TE) or an isotonic Tris extender (B) (p < 0.05). Membrane evaluation by ultramicroscopy also indicated better sperm cryopreservation in extender T compared with the others, and there was a significant reduction in the number of damaged membranes (27%, p < 0.0002). The level of reduced glutathione was significantly higher after sperm cryopreservation in either hypertonic diluent (T and TE) with respect to the isotonic extender B, immediately after thawing (12%) and after a 3-h post-thawing thermotolerance test at 37 degrees C (17%, p = 0.007). Total glutathione levels did not show statistical differences among the extenders. After 3h post-thawing incubation at 37 degrees C, lipid peroxide levels in spermatozoa were statistically lower for T than TE (35%) or isotonic extender B (44%) (p = 0.002). Taken together these results indicate a reduction in the oxidative stress provoked by freezing and thawing when semen is cryopreserved in extender T. The antioxidant properties of extender T may be related to its effectiveness in membrane cryopreservation. PMID:15925576
Clusterin inhibition using OGX-011 synergistically enhances zoledronic acid activity in osteosarcoma
Lamoureux, Francois; Baud'huin, Marc; Ory, Benjamin; Guiho, Romain; Zoubeidi, Amina; Gleave, Martin; Heymann, Dominique; Rédini, Françoise
Purpose Despite recent improvements in therapeutic management of osteosarcoma, ongoing challenges in improving the response to chemotherapy warrants new strategies still needed to improve overall patient survival. Among new therapeutic approaches, zoledronic acid (ZOL) represents a promising adjuvant molecule to chemotherapy to limit the osteolytic component of bone tumors. However, ZOL triggers the elevation of heat shock proteins (Hsp), including Hsp27 and clusterin (CLU), which could enhance tumor cell survival and treatment resistance. We hypothesized that targeting CLU using siRNA or the antisense drug, OGX-011, will suppress treatment-induced CLU induction and enhance ZOL-induced cell death in osteosarcoma (OS) cells. Methods The combined effects of OGX-011 and ZOL were investigated in vitro on cell growth, viability, apoptosis and cell cycle repartition of ZOL-sensitive or -resistant human OS cell lines (SaOS2, U2OS, MG63 and MNNG/HOS). Results In OS cell lines, ZOL increased levels of HSPs, especially CLU, in a dose- and time-dependent manner by mechanism including increased HSF1 transcription activity. The OS resistant cells to ZOL exhibited higher CLU expression level than the sensitive cells. Moreover, CLU overexpression protects OS sensitive cells to ZOL-induced cell death by modulating the MDR1 and farnesyl diphosphate synthase expression. OGX-011 suppressed treatment-induced increases in CLU and synergistically enhanced the activity of ZOL on cell growth and apoptosis. These biologic events were accompanied by decreased expression of HSPs, MDR1 and HSF1 transcriptional activity. In vivo, OGX-011, administered 3 times a week (IP, 20mg/kg), potentiated the effect of ZOL (s.c; 50μg/kg), significantly inhibiting tumor growth by 50% and prolonging survival in MNNG/HOS xenograft model compared to ZOL alone. Conclusion These results indicate that ZOL-mediated induction of CLU can be attenuated by OGX-011, with synergistic effects on delaying progression of
The tumor suppressive role of RASSF1A in osteosarcoma through the Wnt signaling pathway.
Wang, Wei-Guo; Chen, Shi-Jie; He, Jin-Shen; Li, Jing-Song; Zang, Xiao-Fang
Ras-association domain family 1 isoform A (RASSF1A) is a tumor suppressor gene and its expression is lost in numerous types of cancer cells, including primary osteosarcoma cells. However, its functional significance in osteosarcoma has not been well defined. The messenger RNA (mRNA) expression of RASSF1A in osteosarcoma tissues and corresponding non-tumoral tissues was measured by real-time PCR. Overexpression of RASSF1A was established by an adenoviral vector expressing RASSF1A. Cell migration and invasion were analyzed in transwells. Apoptosis and cell cycle were analyzed using flow cytometry. Wnt/β-catenin activity was measured by TCF reporter dual-luciferase assay. Cell viability was measured by MTT assay. Protein expression was detected by Western blot. RASSF1A mRNA expression was significantly lower in osteosarcoma tissues than that in the corresponding non-tumoral tissues. The lowered RASSF1A expression correlated with the clinical severity of osteosarcoma. rAd-RASSF1A injection significantly inhibited the growth of xenograft MNNG/HOS tumors in mice. Overexpression of RASSF1A resulted in significant inhibition of the proliferation, migration, and invasion; induced apoptosis; and arrested cell cycle at G0/G1 phase in both the MNNG/HOS and SaOS2 cells. Overexpression of RASSF1A inhibited the Wnt/β-catenin activity, decreased phosphorylation of Akt/glycogen synthase kinase-3-β (GSK3-β), and increased phosphorylation of mammalian sterile 20-like kinase 1 (MST1). Overexpression of RASSF1A downregulated the cyclin D1, c-Myc, and matrix metalloproteinase-7 (MMP-7) protein levels. RASSF1A functions as a tumor suppressor in osteosarcoma and exerts anti-cancer roles through regulating Akt/GSK-3-Wnt/β-catenin signaling. PMID:26750098
In vitro biocompatibility of 45S5 Bioglass-derived glass-ceramic scaffolds coated with poly(3-hydroxybutyrate).
Bretcanu, Oana; Misra, Superb; Roy, Ipsita; Renghini, Chiara; Fiori, Fabrizio; Boccaccini, Aldo R; Salih, Vehid
The aim of this work was to study the in vitro biocompatibility of glass-ceramic scaffolds based on 45S5 Bioglass, using a human osteosarcoma cell line (HOS-TE85). The highly porous scaffolds were produced by the foam replication technique. Two different types of scaffolds with different porosities were analysed. They were coated with a biodegradable polymer, poly(3-hydroxybutyrate) (P(3HB)). The scaffold bioactivity was evaluated by soaking in a simulated body fluid (SBF) for different durations. Compression strength tests were performed before and after immersion in SBF. These experiments showed that the scaffolds are highly bioactive, as after a few days of immersion in SBF a hydroxyapatite-like layer was formed on the scaffold's surface. It was also observed that P(3HB)-coated samples exhibited higher values of compression strength than uncoated samples. Biocompatibility assessment was carried out by qualitative evaluation of cell morphology after different culture periods, using scanning electron microscopy, while cell proliferation was determined by using the AlamarBlue assay. Alkaline phosphatase (ALP) and osteocalcin (OC) assays were used as quantitative in vitro indicators of osteoblast function. Two different types of medium were used for ALP and OC tests: normal supplemented medium and osteogenic medium. HOS cells were seeded and cultured onto the scaffolds for up to 2 weeks. The AlamarBlue assay showed that cells were able to proliferate and grow on the scaffold surface. After 7 days in culture, the P(3HB)-coated samples had a higher number of cells on their surfaces than the uncoated samples. Regarding ALP- and OC-specific activity, no significant differences were found between samples with different pore sizes. All scaffolds containing osteogenic medium seemed to have a slightly higher level of ALP and OC concentration. These experiments confirmed that Bioglass/P(3HB) scaffolds have potential as osteoconductive tissue engineering substrates for
Heme oxygenase metabolites inhibit tubuloglomerular feedback (TGF).
Ren, YiLin; D'Ambrosio, Martin A; Wang, Hong; Liu, Ruisheng; Garvin, Jeffrey L; Carretero, Oscar A
Tubuloglomerular feedback (TGF) is the mechanism by which the macula densa (MD) senses increases in luminal NaCl concentration and sends a signal to constrict the afferent arteriole (Af-Art). The kidney expresses constitutively heme oxygenase-2 (HO-2) and low levels of HO-1. HOs release carbon monoxide (CO), biliverdin, and free iron. We hypothesized that renal HOs inhibit TGF via release of CO and biliverdin. Rabbit Af-Arts and attached MD were simultaneously microperfused in vitro. The TGF response was determined by measuring Af-Art diameter before and after increasing NaCl in the MD perfusate. When HO activity was inhibited by adding stannous mesoporphyrin (SnMP) to the MD perfusate, the TGF response increased from 2.1+/-0.2 to 4.1+/-0.4 microm (P=0.003, control vs. SnMP, n=7). When a CO-releasing molecule, (CORM-3; 50 microM), was added to the MD perfusate, the TGF response decreased by 41%, from 3.6+/-0.3 to 2.1+/-0.2 microm (P<0.001, control vs. CORM-3, n=12). When CORM-3 at 100 microM was added to the perfusate, it completely blocked the TGF response, from 4.2+/-0.4 to -0.2+/-0.3 microm (P<0.001, control vs. CORM-3, n=6). When biliverdin was added to the perfusate, the TGF response decreased by 79%, from 3.4+/-0.3 to 0.7+/-0.4 microm (P=0.001, control vs. biliverdin, n=6). The effects of SnMP and CORM-3 were not blocked by inhibition of nitric oxide synthase. We concluded that renal HO inhibits TGF probably via release of CO and biliverdin. HO regulation of TGF is a novel mechanism that could lead to a better understanding of the control of renal microcirculation and function. PMID:18715939
Simple NMR methods for evaluating higher order structures of monoclonal antibody therapeutics with quinary structure.
Chen, Kang; Long, Dianna S; Lute, Scott C; Levy, Michaella J; Brorson, Kurt A; Keire, David A
Monoclonal antibody (mAb) drugs constitute the largest class of protein therapeutics currently on the market. Correctly folded protein higher order structure (HOS), including quinary structure, is crucial for mAb drug quality. The quinary structure is defined as the association of quaternary structures (e.g., oligomerized mAb). Here, several commonly available analytical methods, i.e., size-exclusion-chromatography (SEC) FPLC, multi-angle light scattering (MALS), circular dichroism (CD), NMR and multivariate analysis, were combined and modified to yield a complete profile of HOS and comparable metrics. Rituximab and infliximab were chosen for method evaluation because both IgG1 molecules are known to be homologous in sequence, superimposable in Fab crystal structure and identical in Fc structure. However, herein the two are identified to be significantly different in quinary structure in addition to minor secondary structure differences. All data collectively showed rituximab was mostly monomeric while infliximab was in mono-oligomer equilibrium driven by its Fab fragment. The quinary structure differences were qualitatively inferred from the less used but more reproducible dilution-injection-SEC-FPLC curve method. Quantitative principal component analysis (PCA) was performed on NMR spectra of either the intact or the in-situ enzymatic-digested mAb samples. The cleavage reactions happened directly in NMR tubes without further separation, which greatly enhanced NMR spectra quality and resulted in larger inter- and intra-lot variations based on PCA. The new in-situ enzymatic digestion method holds potential in identifying structural differences on larger therapeutic molecules using NMR. PMID:27344629
Highly and Broad-Spectrum In Vitro Antitumor Active cis-Dichloridoplatinum(II) Complexes with 7-Azaindoles
Štarha, Pavel; Dvořák, Zdeněk; Trávníček, Zdeněk
The cis-[PtCl2(naza)2] complexes (1–3) containing monosubstituted 7-azaindole halogeno-derivatives (naza), showed significantly higher activity than cisplatin towards ovarian carcinoma A2780, its cisplatin-resistant variant A2780R, osteosarcoma HOS, breast carcinoma MCF7 and cervix carcinoma HeLa cell lines, with the IC50 values of 3.8, 3.5, 4.5, 2.7, and 9.2 μM, respectively, obtained for the most active complex 3. As for 4 and 5 having disubstituted 7-azaindoles in their molecule, the significant cytotoxicity was detected only for 4 against A2780 (IC50 = 4.8 μM), A2780R (IC50 = 3.8 μM) and HOS (IC50 = 4.3 μM), while 5 was evaluated as having only moderate antiproliferative effect against the mentioned cancer cell lines with IC50 = 33.4, 24.7 and 46.7 μM, respectively. All the studied complexes 1–5 effectively avoided the acquired resistance of ovarian carcinoma cell line. On the other hand, the complexes did not reveal any inhibition activity on the purified 20S proteasome from the A2780 cells. The representative complexes 3 and 5 showed low ability to be hydrolysed, but their stability was markedly lowered in the presence of physiological sulphur-containing biomolecule glutathione (GSH), as proved by the 1H NMR spectroscopy and mass spectrometry studies. A rate of interaction of the studied complexes with GSH was affected by an addition of another mechanistically relevant biomolecule guanosine monophosphate. The differences in interactions of 3 and 5 with GSH correlate well with their different cytotoxicity profiles. PMID:26309251
Highly and Broad-Spectrum In Vitro Antitumor Active cis-Dichloridoplatinum(II) Complexes with 7-Azaindoles.
Štarha, Pavel; Dvořák, Zdeněk; Trávníček, Zdeněk
The cis-[PtCl2(naza)2] complexes (1-3) containing monosubstituted 7-azaindole halogeno-derivatives (naza), showed significantly higher activity than cisplatin towards ovarian carcinoma A2780, its cisplatin-resistant variant A2780R, osteosarcoma HOS, breast carcinoma MCF7 and cervix carcinoma HeLa cell lines, with the IC50 values of 3.8, 3.5, 4.5, 2.7, and 9.2 μM, respectively, obtained for the most active complex 3. As for 4 and 5 having disubstituted 7-azaindoles in their molecule, the significant cytotoxicity was detected only for 4 against A2780 (IC50 = 4.8 μM), A2780R (IC50 = 3.8 μM) and HOS (IC50 = 4.3 μM), while 5 was evaluated as having only moderate antiproliferative effect against the mentioned cancer cell lines with IC50 = 33.4, 24.7 and 46.7 μM, respectively. All the studied complexes 1-5 effectively avoided the acquired resistance of ovarian carcinoma cell line. On the other hand, the complexes did not reveal any inhibition activity on the purified 20S proteasome from the A2780 cells. The representative complexes 3 and 5 showed low ability to be hydrolysed, but their stability was markedly lowered in the presence of physiological sulphur-containing biomolecule glutathione (GSH), as proved by the 1H NMR spectroscopy and mass spectrometry studies. A rate of interaction of the studied complexes with GSH was affected by an addition of another mechanistically relevant biomolecule guanosine monophosphate. The differences in interactions of 3 and 5 with GSH correlate well with their different cytotoxicity profiles. PMID:26309251
In vitro cytotoxicity, DNA cleavage and SOD-mimic activity of copper(II) mixed-ligand quinolinonato complexes.
Buchtík, Roman; Trávníček, Zdeněk; Vančo, Ján
Six mixed-ligand copper(II) complexes with the composition [Cu(qui)(L)]BF(4)·xH(2)O (1-6), where Hqui=2-phenyl-3-hydroxy-4(1H)-quinolinone, L=2,2'-bipyridine (bpy) (1), 1,10-phenanthroline (phen) (2), bis(2-pyridyl)amine (ambpy) (3), 5-methyl-1,10-phenanthroline (mphen) (4), 5-nitro-1,10-phenanthroline (nphen) (5) and bathophenanthroline (bphen) (6), were prepared, fully characterized and studied for their in vitro cytotoxicity on human osteosarcoma (HOS) and human breast adenocarcinoma (MCF7) cancer cell lines. The overall promising results of the cytotoxicity were found for all the complexes, while the best results were achieved for complex 6, with IC(50)=2.6 ± 0.8 μM (HOS), and 1.3 ± 0.5 μM (MCF7). The interactions of the Cu(II) complexes 1-6 with calf thymus DNA were investigated by the UV-visible spectral titration. An agarose-gel electrophoretic method of oxidative damage determination to circular plasmid pUC19 was used to assess the ability of the complexes to act as chemical nucleases. A high effectiveness of DNA cleavage was observed for 2, 4 and 5. In vitro antioxidative activity of the complexes was studied by the superoxide dismutase-mimic (SOD-mimic) method. The best result was afforded by complex 1 with IC(50)=4.7 ± 1.0 μM, which corresponds to 10.2% of the native Cu,Zn-SOD enzyme activity. The ability of the tested complexes to interact with sulfur-containing biomolecules (cysteine and reduced glutathione) at physiological levels was proved by electrospray-ionization mass spectrometry (ESI-MS). PMID:23022693
How to modify 7-azaindole to form cytotoxic Pt(II) complexes: highly in vitro anticancer effective cisplatin derivatives involving halogeno-substituted 7-azaindole.
Štarha, Pavel; Trávníček, Zdeněk; Popa, Alexandr; Popa, Igor; Muchová, Tereza; Brabec, Viktor
The platinum(II) dichlorido and oxalato complexes of the general formula cis-[PtCl(2)(nHaza)(2)] (1-3) [Pt(ox)(nHaza)(2)] (4-6) involving 7-azaindole halogeno-derivatives (nHaza) were prepared and thoroughly characterized. A single-crystal X-ray analysis of cis-[PtCl(2)(3ClHaza)(2)]·DMF (1·DMF; 3ClHaza symbolizes 3-chloro-7-azaindole) revealed a distorted square-planar arrangement with both the 3ClHaza molecules coordinated through their N7 atoms in a cis fashion. In vitro cytotoxicity of the complexes was evaluated by an MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay against the HOS (osteosarcoma), MCF7 (breast adenocarcinoma) and LNCaP (prostate adenocarcinoma) human cancer cell lines. The dichlorido complexes 1-3 (IC(50)=3.8, 3.9, and 2.5 μM, respectively) showed significantly higher in vitro anticancer effect against HOS as compared with cisplatin, whose IC(50)=37.7 μM. The biological effect of cisplatin against MCF7 (IC(50)=24.5 μM) and LNCaP (IC(50)=3.8 μM) was also exceeded by 1-3 (except for 2 against LNCaP), but the difference can be classified as significant only in the case of 1 (IC(50)=3.4 μM) and 3 (IC(50)=2.0 μM) against MCF7. The molecular pharmacological studies (RNA synthesis by T7 RNA polymerase in vitro) proved that 1-3 bind to DNA in a similar manner as cisplatin, since the RNA synthesis products of 1-3 and cisplatin showed a similar sequence profile of major bands. PMID:22922312
Investigating the role of a backbone to substrate hydrogen bond in OMP decarboxylase using a site-specific amide to ester substitution
Desai, Bijoy J.; Goto, Yuki; Cembran, Alessandro; Fedorov, Alexander A.; Almo, Steven C.; Gao, Jiali; Suga, Hiroaki; Gerlt, John A.
Hydrogen bonds between backbone amide groups of enzymes and their substrates are often observed, but their importance in substrate binding and/or catalysis is not easy to investigate experimentally. We describe the generation and kinetic characterization of a backbone amide to ester substitution in the orotidine 5′-monophosphate (OMP) decarboxylase from Methanobacter thermoautotrophicum (MtOMPDC) to determine the importance of a backbone amide–substrate hydrogen bond. The MtOMPDC-catalyzed reaction is characterized by a rate enhancement (∼1017) that is among the largest for enzyme-catalyzed reactions. The reaction proceeds through a vinyl anion intermediate that may be stabilized by hydrogen bonding interaction between the backbone amide of a conserved active site serine residue (Ser-127) and oxygen (O4) of the pyrimidine moiety and/or electrostatic interactions with the conserved general acidic lysine (Lys-72). In vitro translation in conjunction with amber suppression using an orthogonal amber tRNA charged with l-glycerate (HOS) was used to generate the ester backbone substitution (S127HOS). With 5-fluoro OMP (FOMP) as substrate, the amide to ester substitution increased the value of Km by ∼1.5-fold and decreased the value of kcat by ∼50-fold. We conclude that (i) the hydrogen bond between the backbone amide of Ser-127 and O4 of the pyrimidine moiety contributes a modest factor (∼102) to the 1017 rate enhancement and (ii) the stabilization of the anionic intermediate is accomplished by electrostatic interactions, including its proximity of Lys-72. These conclusions are in good agreement with predictions obtained from hybrid quantum mechanical/molecular mechanical calculations. PMID:25275007
Imatinib Mesylate Exerts Anti-Proliferative Effects on Osteosarcoma Cells and Inhibits the Tumour Growth in Immunocompetent Murine Models
Ory, Benjamin; Charrier, Céline; Brion, Régis; Blanchard, Frederic; Redini, Françoise; Heymann, Dominique
Osteosarcoma is the most common primary malignant bone tumour characterized by osteoid production and/or osteolytic lesions of bone. A lack of response to chemotherapeutic treatments shows the importance of exploring new therapeutic methods. Imatinib mesylate (Gleevec, Novartis Pharma), a tyrosine kinase inhibitor, was originally developed for the treatment of chronic myeloid leukemia. Several studies revealed that imatinib mesylate inhibits osteoclast differentiation through the M-CSFR pathway and activates osteoblast differentiation through PDGFR pathway, two key cells involved in the vicious cycle controlling the tumour development. The present study investigated the in vitro effects of imatinib mesylate on the proliferation, apoptosis, cell cycle, and migration ability of five osteosarcoma cell lines (human: MG-63, HOS; rat: OSRGA; mice: MOS-J, POS-1). Imatinib mesylate was also assessed as a curative and preventive treatment in two syngenic osteosarcoma models: MOS-J (mixed osteoblastic/osteolytic osteosarcoma) and POS-1 (undifferentiated osteosarcoma). Imatinib mesylate exhibited a dose-dependent anti-proliferative effect in all cell lines studied. The drug induced a G0/G1 cell cycle arrest in most cell lines, except for POS-1 and HOS cells that were blocked in the S phase. In addition, imatinib mesylate induced cell death and strongly inhibited osteosarcoma cell migration. In the MOS-J osteosarcoma model, oral administration of imatinib mesylate significantly inhibited the tumour development in both preventive and curative approaches. A phospho-receptor tyrosine kinase array kit revealed that PDGFRα, among 7 other receptors (PDFGFRβ, Axl, RYK, EGFR, EphA2 and 10, IGF1R), appears as one of the main molecular targets for imatinib mesylate. In the light of the present study and the literature, it would be particularly interesting to revisit therapeutic evaluation of imatinib mesylate in osteosarcoma according to the tyrosine-kinase receptor status of patients
Application of power spectrum, cepstrum, higher order spectrum and neural network analyses for induction motor fault diagnosis
NASA Astrophysics Data System (ADS)
Liang, B.; Iwnicki, S. D.; Zhao, Y.
The power spectrum is defined as the square of the magnitude of the Fourier transform (FT) of a signal. The advantage of FT analysis is that it allows the decomposition of a signal into individual periodic frequency components and establishes the relative intensity of each component. It is the most commonly used signal processing technique today. If the same principle is applied for the detection of periodicity components in a Fourier spectrum, the process is called the cepstrum analysis. Cepstrum analysis is a very useful tool for detection families of harmonics with uniform spacing or the families of sidebands commonly found in gearbox, bearing and engine vibration fault spectra. Higher order spectra (HOS) (also known as polyspectra) consist of higher order moment of spectra which are able to detect non-linear interactions between frequency components. For HOS, the most commonly used is the bispectrum. The bispectrum is the third-order frequency domain measure, which contains information that standard power spectral analysis techniques cannot provide. It is well known that neural networks can represent complex non-linear relationships, and therefore they are extremely useful for fault identification and classification. This paper presents an application of power spectrum, cepstrum, bispectrum and neural network for fault pattern extraction of induction motors. The potential for using the power spectrum, cepstrum, bispectrum and neural network as a means for differentiating between healthy and faulty induction motor operation is examined. A series of experiments is done and the advantages and disadvantages between them are discussed. It has been found that a combination of power spectrum, cepstrum and bispectrum plus neural network analyses could be a very useful tool for condition monitoring and fault diagnosis of induction motors.
Prostate tissue characterization/classification in 144 patient population using wavelet and higher order spectra features from transrectal ultrasound images.
Pareek, Gyan; Acharya, U Rajendra; Sree, S Vinitha; Swapna, G; Yantri, Ratna; Martis, Roshan Joy; Saba, Luca; Krishnamurthi, Ganapathy; Mallarini, Giorgio; El-Baz, Ayman; Al Ekish, Shadi; Beland, Michael; Suri, Jasjit S
In this work, we have proposed an on-line computer-aided diagnostic system called "UroImage" that classifies a Transrectal Ultrasound (TRUS) image into cancerous or non-cancerous with the help of non-linear Higher Order Spectra (HOS) features and Discrete Wavelet Transform (DWT) coefficients. The UroImage system consists of an on-line system where five significant features (one DWT-based feature and four HOS-based features) are extracted from the test image. These on-line features are transformed by the classifier parameters obtained using the training dataset to determine the class. We trained and tested six classifiers. The dataset used for evaluation had 144 TRUS images which were split into training and testing sets. Three-fold and ten-fold cross-validation protocols were adopted for training and estimating the accuracy of the classifiers. The ground truth used for training was obtained using the biopsy results. Among the six classifiers, using 10-fold cross-validation technique, Support Vector Machine and Fuzzy Sugeno classifiers presented the best classification accuracy of 97.9% with equally high values for sensitivity, specificity and positive predictive value. Our proposed automated system, which achieved more than 95% values for all the performance measures, can be an adjunct tool to provide an initial diagnosis for the identification of patients with prostate cancer. The technique, however, is limited by the limitations of 2D ultrasound guided biopsy, and we intend to improve our technique by using 3D TRUS images in the future. PMID:23745787
Entropy analysis of muscular near-infrared spectroscopy (NIRS) signals during exercise programme of type 2 diabetic patients: quantitative assessment of muscle metabolic pattern.
Molinari, Filippo; Acharya, U Rajendra; Martis, Roshan Joy; De Luca, Riccardo; Petraroli, Giuliana; Liboni, William
Diabetes mellitus (DM) is a metabolic disorder that is widely rampant throughout the world population these days. The uncontrolled DM may lead to complications of eye, heart, kidney and nerves. The most common type of diabetes is the type 2 diabetes or insulin-resistant DM. Near-infrared spectroscopy (NIRS) technology is widely used in non-invasive monitoring of physiological signals. Three types of NIRS signals are used in this work: (i) variation in the oxygenated haemoglobin (O2Hb) concentration, (ii) deoxygenated haemoglobin (HHb), and (iii) ratio of oxygenated over the sum of the oxygenated and deoxygenated haemoglobin which is defined as: tissue oxygenation index (TOI) to analyze the effect of exercise on diabetes subjects. The NIRS signal has the characteristics of non-linearity and non-stationarity. Hence, the very small changes in this time series can be efficiently extracted using higher order statistics (HOS) method. Hence, in this work, we have used sample and HOS entropies to analyze these NIRS signals. These computer aided techniques will assist the clinicians to diagnose and monitor the health accurately and easily without any inter or intra observer variability. Results showed that after a one-year of physical exercise programme, all diabetic subjects increased the sample entropy of the NIRS signals, thus revealing a better muscle performance and an improved recruitment by the central nervous system. Moreover, after one year of physical therapy, diabetic subjects showed a NIRS muscular metabolic pattern that was not distinguished from that of controls. We believe that sample and bispectral entropy analysis is need when the aim is to compare the inner structure of the NIRS signals during muscle contraction, particularly when dealing with neuromuscular impairments. PMID:24075080
Influence of substrate modification and C-terminal truncation on the active site structure of substrate-bound heme oxygenase from Neisseriae meningitidis; A 1H NMR study†
Peng, Dungeng; Satterlee, James D.; Ma, Li-Hua; Dallas, Jerry L.; Smith, Kevin M.; Zhang, Xuhong; Sato, Michihiko; La Mar, Gerd N.
Heme oxygenase, HO, from the pathogenic bacterium N. meningitidis, NmHO, which secures host iron, shares many properties with mammalian HOs, but also exhibits some key differences. The crystal structure appears more compact and the crystal-undetected C-terminus interacts with substrate in solution. The unique nature of substrate-protein, specifically pyrrole-I/II-helix-2, peripheral interactions in NmHO are probed by 2D 1H NMR to reveal unique structural features controlling substrate orientation. The thermodynamics of substrate orientational isomerism are mapped for substrates with individual vinyl → methyl → hydrogen substitutions and with enzyme C-terminal deletions. NmHO exhibits significantly stronger orientational preference, reflecting much stronger and selective pyrrole-I/II interactions with the protein matrix, than in mammalian HOs. Thus, replacing bulky vinyls with hydrogens results in a 180° rotation of substrate about the α,γ-meso axis in the active site. A "collapse" of the substrate pocket as substrate size decreases is reflected in movement of helix-2 toward the substrate as indicated by significant and selective increased NOESY cross peak intensity, increase in steric Fe-CN tilt reflected in the orientation of the major magnetic axis, and decrease in steric constraints controlling the rate of aromatic ring reorientation. The active site of NmHO appears "stressed" for native protohemin and its "collapse" upon replacing vinyls by hydrogen leads to a factor ~102 increase in substrate affinity. Interaction of the C-terminus with the active site destabilizes the crystallographic protohemin orientation by ~0.7 kcal/mol, which is consistent with optimizing the His207-Asp27 H-bond. Implications of the active site "stress" for product release are discussed. PMID:21870860
Teaching Elderly Adults to Use the Internet to Access Health Care Information: Before-After Study
Nolfi, David A
Background Much has been written about the Internet's potential to revolutionize health care delivery. As younger populations increasingly utilize Internet-based health care information, it will be essential to ensure that the elderly become adept at using this medium for health care purposes, especially those from minority, low income, and limited educational backgrounds. Objective This paper presents the results of a program designed to teach elderly adults to use the Internet to access health care information. The objective was to examine whether the training led to changes in participant's perceptions of their health, perceptions of their interactions with health care providers, health information–seeking behaviors, and self-care activities. Methods Participants attended a 5-week training course held in public libraries and senior community centers within the greater Pittsburgh and Allegheny County region. Classes within each seminar lasted 2 hours and consisted of lecture and hands-on training. Baseline surveys were administered prior to the course, 5-week follow-up surveys were administered immediately after the course, and final surveys were mailed 1 year later. Instruments included the Multidimensional Health Locus of Control (MHLC) Scale, which measures three domains of locus of control (internal, external, and chance); the Krantz Health Opinion Survey (HOS); and the Lau, Hartman, and Ware Health Value Survey. Two additional questionnaires included multiple choice and qualitative questions designed to measure participants' Internet utilization and levels of health care participation. The Health Participation Survey was administered with the baseline survey. The Internet Use Survey was administered at the 1-year mark and contained several items from the Health Participation Survey, which allowed comparison between baseline and 1-year responses. Results Of the 60 elderly adults who began the training course, 42 (mean age 72) completed the entire 5-week
Automated determination of P-phase arrival times at regional and local distances using higher order statistics
NASA Astrophysics Data System (ADS)
Küperkoch, L.; Meier, T.; Lee, J.; Friederich, W.; Working Group, EGELADOS
We present an algorithm for automatic P-phase arrival time determination for local and regional seismic events based on higher order statistics (HOS). Using skewness or kurtosis a characteristic function is determined to which a new iterative picking algorithm is applied. For P-phase identification we apply the Akaike Information Criterion to the characteristic function, while for a precise determination of the P-phase arrival time a pragmatic picking algorithm is applied to a recalculated characteristic function. In addition, an automatic quality estimate is obtained, based on the slope and the signal-to-noise ratio, both calculated from the characteristic function. To get rid of erroneous picks, a Jackknife procedure and an envelope function analysis is used. The algorithm is applied to a large data set with very heterogeneous qualities of P-onsets acquired by a temporary, regional seismic network of the EGELADOS-project in the southern Aegean. The reliability and robustness of the proposed algorithm is tested by comparing more than 3000 manually derived P readings, serving as reference picks, with the corresponding automatically estimated P-wave arrival times. We find an average deviation from the reference picks of 0.26 +/- 0.64s when using kurtosis and 0.38 +/- 0.75s when using skewness. If automatically as excellent classified picks are considered only, the average difference from the reference picks is 0.07 +/- 0.31s and 0.07 +/- 0.41s, respectively. However, substantially more P-arrival times are determined when using kurtosis, indicating that the characteristic function derived from kurtosis estimation is to be preferred. Since the characteristic function is calculated recursively, the algorithm is very fast and hence suited for earthquake early warning purposes. Furthermore, a comparative study with automatically derived P-readings using Allen's and Baer & Kradolfer's picking algorithms applied to the same data set demonstrates better quantitative and
Discovering miRNA Regulatory Networks in Holt–Oram Syndrome Using a Zebrafish Model
D'Aurizio, Romina; Russo, Francesco; Chiavacci, Elena; Baumgart, Mario; Groth, Marco; D'Onofrio, Mara; Arisi, Ivan; Rainaldi, Giuseppe; Pitto, Letizia; Pellegrini, Marco
MicroRNAs (miRNAs) are small non-coding RNAs that play an important role in the post-transcriptional regulation of gene expression. miRNAs are involved in the regulation of many biological processes such as differentiation, apoptosis, and cell proliferation. miRNAs are expressed in embryonic, postnatal, and adult hearts, and they have a key role in the regulation of gene expression during cardiovascular development and disease. Aberrant expression of miRNAs is associated with abnormal cardiac cell differentiation and dysfunction. Tbx5 is a member of the T-box gene family, which acts as transcription factor involved in the vertebrate heart development. Alteration of Tbx5 level affects the expression of hundreds of genes. Haploinsufficiency and gene duplication of Tbx5 are at the basis of the cardiac abnormalities associated with Holt–Oram syndrome (HOS). Recent data indicate that miRNAs might be an important part of the regulatory circuit through which Tbx5 controls heart development. Using high-throughput technologies, we characterized genome-widely the miRNA and mRNA expression profiles in WT- and Tbx5-depleted zebrafish embryos at two crucial developmental time points, 24 and 48 h post fertilization (hpf). We found that several miRNAs, which are potential effectors of Tbx5, are differentially expressed; some of them are already known to be involved in cardiac development and functions, such as miR-30, miR-34, miR-190, and miR-21. We performed an integrated analysis of miRNA expression data with gene expression profiles to refine computational target prediction approaches by means of the inversely correlation of miRNA–mRNA expressions, and we highlighted targets, which have roles in cardiac contractility, cardiomyocyte proliferation/apoptosis, and morphogenesis, crucial functions regulated by Tbx5. This approach allowed to discover complex regulatory circuits involving novel miRNAs and protein coding genes not considered before in the HOS such as miR-34a and
Oligocene-Early Miocene Planktonic Microbiostratigraphy and Paleoenvironments of the South Slovakian Basin (Lučenec Depression)
NASA Astrophysics Data System (ADS)
Ozdínová, Silvia; Soták, Ján
Oligocene and Lower Miocene sediments of the Lučenec Depression were studied to demonstrate the planktonic bioevents and climatic proxies from the Číž Formation (Rupelian) and Lučenec Formation (Chattian-Aquitanian) on the basis of quantitative analyses of foraminifers and calcareous nannofossils. The oldest nannofossil assemblages of the Číž Formation belonged to the NP23 Zone and were dominated by Reticulofenestra ornata known for preference of temperate eutrophic water conditions. An increase in bioproductivity was documented by abundant large-sized planktonic foraminifers (e.g. Turborotalia ampliapertura, Paragloborotalia nana, Subbotina gortanii) and epifaunal to shal-low-infaunal benthic species. The middle part of the Číž Formation reveals a lowstand progradation of deltaic sediments of the Rapovce Member. There, the planktonic foraminifers are impoverished in both size and diversity, containing mostly tenuitellid and cassigerinellid species, probably as a result of decreased salinity and increased anoxia in the Tard Clay. Contrary of this, the benthic foraminifers are rich, mainly the infaunal forms of uvigerinid species. They probably proliferated due to a high organic flux from riverine input. Open marine conditions were restored in the upper part of the Číž Formation above the lowest occurrence (LO) of Cyclicargolithus abisectus on the NP23-NP24 zone boundary. The transitional interval between the Číž and Lučenec formations (O5/O6 - NP24/25) was approximated by the HOs of Paragloborotalia opima and Sphenolithus distentus and the LOs of Globigerinoides primordius and Pontosphaera enormis. Benthic foraminifera of the Lučenec Formation indicate a high productivity and oxygen-deplected environ-ments. The Oligocene-Miocene boundary in the Lučenec Formation was appointed by the HOs of Helicospahera recta and Dictyococcites bisectus. Foraminiferal markers of this boundary were established from the HO of Globigerina ciperoensis and the LO of
A systems biology approach reveals common metastatic pathways in osteosarcoma
Background Osteosarcoma (OS) is the most common malignant bone tumor in children and adolescents. The survival rate of patients with metastatic disease remains very dismal. Nevertheless, metastasis is a complex process and a single-level analysis is not likely to identify its key biological determinants. In this study, we used a systems biology approach to identify common metastatic pathways that are jointly supported by both mRNA and protein expression data in two distinct human metastatic OS models. Results mRNA expression microarray and N-linked glycoproteomic analyses were performed on two commonly used isogenic pairs of human metastatic OS cell lines, namely HOS/143B and SaOS-2/LM7. Pathway analysis of the differentially regulated genes and glycoproteins separately revealed pathways associated to metastasis including cell cycle regulation, immune response, and epithelial-to-mesenchymal-transition. However, no common significant pathway was found at both genomic and proteomic levels between the two metastatic models, suggesting a very different biological nature of the cell lines. To address this issue, we used a topological significance analysis based on a "shortest-path" algorithm to identify topological nodes, which uncovered additional biological information with respect to the genomic and glycoproteomic profiles but remained hidden from the direct analyses. Pathway analysis of the significant topological nodes revealed a striking concordance between the models and identified significant common pathways, including "Cytoskeleton remodeling/TGF/WNT", "Cytoskeleton remodeling/Cytoskeleton remodeling", and "Cell adhesion/Chemokines and adhesion". Of these, the "Cytoskeleton remodeling/TGF/WNT" was the top ranked common pathway from the topological analysis of the genomic and proteomic profiles in the two metastatic models. The up-regulation of proteins in the "Cytoskeleton remodeling/TGF/WNT" pathway in the SaOS-2/LM7 and HOS/143B models
Metabolism of vitamin D3 in human osteoblasts: evidence for autocrine and paracrine activities of 1 alpha,25-dihydroxyvitamin D3.
Atkins, Gerald J; Anderson, Paul H; Findlay, David M; Welldon, Katie J; Vincent, Cristina; Zannettino, Andrew C W; O'Loughlin, Peter D; Morris, Howard A
Circulating 1 alpha,25-dihydroxyvitamin D(3) (1,25D) derives from renal conversion of 25-hydroxyvitamin D(3) (25D), by the 25D 1 alpha-hydroxylase (CYP27B1). Blood 25D levels, but not 1,25D levels, are the best indicator of vitamin D status and predict fracture risk in the elderly. We examined the extent to which osteoblasts can metabolize 25D. Well-characterized human primary osteoblasts and osteosarcoma (OS) cell lines were examined for the expression and regulation of genes associated with vitamin D metabolism, using real-time PCR. Primary osteoblasts and OS cell lines were found to express CYP27B1 mRNA and secreted detectable 1,25D in response to 25D. Of the OS cell lines tested, HOS expressed the most CYP27B1 mRNA and secreted the highest levels of 1,25D. All osteoblastic cells examined up-regulated expression of the catabolic regulator of 1,25D, the 25-hydroxyvitamin D-24-hydroxylase (CYP24), when incubated with either 1,25D or 25D. Exposure to physiological levels of 25D resulted in up-regulated transcription of the 1,25D responsive genes, osteocalcin (OCN), osteopontin (OPN) and RANKL. Specific knockdown of CYP27B1 in HOS cells using siRNA resulted in up to 80% reduction in both 1,25D secretion and the transcription of OCN and CYP24, strongly implying that the 25D effect in osteoblasts is preceded by conversion to 1,25D. Incubation with 25D, like 1,25D, inhibited primary osteoblast proliferation and promoted in vitro mineralization. Finally, we detected expression by osteoblasts of receptors for vitamin D binding protein (DBP), cubilin and megalin, suggesting that osteoblasts are able to internalize DBP-25D complexes in vivo. Together, our results suggest that autocrine, and perhaps paracrine, pathways of vitamin D(3) metabolism may regulate key osteoblast functions independently of circulating, kidney derived 1,25D. Our results are therefore consistent with the reported benefits of maintaining a healthy vitamin D status in the elderly to reduce the risk of
Discovering miRNA Regulatory Networks in Holt-Oram Syndrome Using a Zebrafish Model.
D'Aurizio, Romina; Russo, Francesco; Chiavacci, Elena; Baumgart, Mario; Groth, Marco; D'Onofrio, Mara; Arisi, Ivan; Rainaldi, Giuseppe; Pitto, Letizia; Pellegrini, Marco
MicroRNAs (miRNAs) are small non-coding RNAs that play an important role in the post-transcriptional regulation of gene expression. miRNAs are involved in the regulation of many biological processes such as differentiation, apoptosis, and cell proliferation. miRNAs are expressed in embryonic, postnatal, and adult hearts, and they have a key role in the regulation of gene expression during cardiovascular development and disease. Aberrant expression of miRNAs is associated with abnormal cardiac cell differentiation and dysfunction. Tbx5 is a member of the T-box gene family, which acts as transcription factor involved in the vertebrate heart development. Alteration of Tbx5 level affects the expression of hundreds of genes. Haploinsufficiency and gene duplication of Tbx5 are at the basis of the cardiac abnormalities associated with Holt-Oram syndrome (HOS). Recent data indicate that miRNAs might be an important part of the regulatory circuit through which Tbx5 controls heart development. Using high-throughput technologies, we characterized genome-widely the miRNA and mRNA expression profiles in WT- and Tbx5-depleted zebrafish embryos at two crucial developmental time points, 24 and 48 h post fertilization (hpf). We found that several miRNAs, which are potential effectors of Tbx5, are differentially expressed; some of them are already known to be involved in cardiac development and functions, such as miR-30, miR-34, miR-190, and miR-21. We performed an integrated analysis of miRNA expression data with gene expression profiles to refine computational target prediction approaches by means of the inversely correlation of miRNA-mRNA expressions, and we highlighted targets, which have roles in cardiac contractility, cardiomyocyte proliferation/apoptosis, and morphogenesis, crucial functions regulated by Tbx5. This approach allowed to discover complex regulatory circuits involving novel miRNAs and protein coding genes not considered before in the HOS such as miR-34a and mi
Intermediate clinical and radiological results of cervical TDR (Mobi-C®) with up to 2 years of follow-up
Beaurain, J.; Bernard, P.; Dufour, T.; Fuentes, J. M.; Hovorka, I.; Huppert, J.; Steib, J. P.; Vital, J. M.; Aubourg, L.
The interest in cervical total disc replacement (TDR) as an alternative to the so-far gold standard in the surgical treatment of degenerative disc disease (DDD), e.g anterior cervical discectomy and fusion (ACDF), is growing very rapidly. Many authors have established the fact that ACDF may result in progressive degeneration in adjacent segments. On the contrary, but still theoretically, preservation of motion with TDR at the surgically treated level may potentially reduce the occurrence of adjacent-level degeneration (ALD). The authors report the intermediate results of an undergoing multicentre prospective study of TDR with Mobi-C® prosthesis. The aim of the study was to assess the safety and efficacy of the device in the treatment of DDD and secondary to evaluate the radiological status of adjacent levels and the occurrence of ossifications, at 2-year follow-up (FU). 76 patients have performed their 2-year FU visit and have been analyzed clinically and radiologically. Clinical outcomes (NDI, VAS, SF-36) and ROM measurements were analyzed pre-operatively and at the different post-operative time-points. Complications and re-operations were also assessed. Occurrences of heterotopic ossifications (HOs) and of adjacent disc degeneration radiographic changes have been analyzed from 2-year FU X-rays. The mean NDI and VAS scores for arm and neck are reduced significantly at each post-operative time-point compared to pre-operative condition. Motion is preserved over the time at index levels (mean ROM = 9° at 2 years) and 85.5% of the segments are mobile at 2 years. HOs are responsible for the fusion of 6/76 levels at 2 years. However, presence of HO does not alter the clinical outcomes. The occurrence rate of radiological signs of ALD is very low at 2 years (9.1%). There has been no subsidence, no expulsion and no sub-luxation of the implant. Finally, after 2 years, 91% of the patients assume that they would undergo the procedure again. These intermediate
Acetabular Labral Tears in Patients with Sports Injury
Kang, Chan; Cha, Soo-Min
Background We wanted to investigate acetabular labral tears and their correlation with femoroacetabular impingement in patients with sports injury. Methods Among 111 patients who were diagnosed with the acetabular labral tears after arthroscopic treatment from January 2004 to December 2007, we selected 41 patients with sports injury. There were 12 cases of Taekwondo injury, 5 of golf injury, 4 of soccer injury, 3 of gymnastics injury, 2 of Hapkido injury, 2 of aerobics injury, 2 of rock-climbing injury, 2 of fitness training injury and 9 of other sports injuries. We checked the subtypes of acetabular labral tears and the accompanying femoroacetabular impingement. For the cases with accompanying femoroacetabular impingement, we investigated the subtypes according to the types of sports, gender and age. At last follow-up, we checked the Harris Hip Score (HHS), the Hip Outcome Score (HOS) sports scale and the percentage of patients who returned to their sports activity. Results The average age of symptomatic onset was 26 years (range, 12 to 65 years). The ratio of males to females was 29 : 12. An average duration of the hip pain was 17 months (range, 1 to 60 months). The degenerative type of acetabular labral tears was the most prevalent with 32 cases (78%), and there were 9 cases (22%) of the partial tear type. Thirty cases (73%) were accompanied by femoroacetabular impingement. The average age of the 23 cases (56%) of the cam-type was 23 years (range, 12 to 48 years), and it was more likely to occur in men (87%) and for people practicing martial arts such as Taekwondo or Hapkido. An average age of the 5 cases (12%) of the pincer-type was 26 (range, 16 to 43 years), it usually occurred in women (60%) and for non-martial arts such as golf and gymnastics. There were 2 cases of the mixed type (cam + pincer-type). At 27 months follow-up, the HHS was 61 to 92 points, the HOS sports scale increased 43 to 75%, and the rate of returning to sports was 71%. Conclusions In
A Ferric-Peroxo Intermediate in the Oxidation of Heme by IsdI.
Takayama, Shin-Ichi J; Loutet, Slade A; Mauk, A Grant; Murphy, Michael E P
The canonical heme oxygenases (HOs) catalyze heme oxidation via a heme-bound hydroperoxo intermediate that is stabilized by a water cluster at the active site of the enzyme. In contrast, the hydrophobic active site of IsdI, a heme-degrading enzyme from Staphylococcus aureus, lacks a water cluster and is expected to oxidize heme by an alternative mechanism. Reaction of the IsdI-heme complex with either H2O2 or m-chloroperoxybenzoic acid fails to produce a specific oxidized heme iron intermediate, suggesting that ferric-hydroperoxo or ferryl derivatives of IsdI are not involved in the catalytic mechanism of this enzyme. IsdI lacks a proton-donating group in the distal heme pocket, so the possible involvement of a ferric-peroxo intermediate has been evaluated. Density functional theory (DFT) calculations indicate that heme oxidation involving a ferric-peroxo intermediate is energetically accessible, whereas the energy barrier for a reaction involving a ferric-hydroperoxo intermediate is too great in the absence of a proton donor. We propose that IsdI catalyzes heme oxidation through nucleophilic attack by the heme-bound peroxo species. This proposal is consistent with our previous demonstration by nuclear magnetic resonance spectroscopy that heme ruffling increases the susceptibility of the meso-carbon of heme to nucleophilic attack. PMID:25853501
Interferon but not MxB inhibits foamy retroviruses.
Bähr, Ariane; Singer, Anna; Hain, Anika; Vasudevan, Ananda Ayyappan Jaguva; Schilling, Mirjam; Reh, Juliane; Riess, Maximilian; Panitz, Sylvia; Serrano, Vanessa; Schweizer, Matthias; König, Renate; Chanda, Sumit; Häussinger, Dieter; Kochs, Georg; Lindemann, Dirk; Münk, Carsten
Foamy viruses (FV) are retroviruses that are widely distributed in primate and non-primate animal species. We tested here FV with capsids of simian and non-simian origin for sensitivity to interferon-β (IFN-β). Our data show significant inhibition of FV by IFN-β early in infection of human HOS and THP-1 but not of HEK293T cells. The post-entry restriction of FV was not mediated by the interferon-induced MxB protein that was recently identified as a capsid-interacting restriction factor targeting Human immunodeficiency virus (HIV) before integration. Neither the ectopic expression of MxA or MxB in HEK293T cells nor the lack of MxB expression in CRISPR/CAS MxB THP-1 knockout cells impacted the infection of the tested FV. IFN-β treated THP-1 and THP-1 KO MxB cells showed the same extend of restriction to FV. Together, the data demonstrate that IFN-β inhibits FV early in infection and that MxB is not a restriction factor of FV. PMID:26609934
Concise Synthesis and Biological Evaluation of 2-Aroyl-5-Amino Benzo[b]thiophene Derivatives As a Novel Class of Potent Antimitotic Agents
Romagnoli, Romeo; Baraldi, Pier Giovanni; Lopez-Cara, Carlota; Preti, Delia; Tabrizi, Mojgan Aghazadeh; Balzarini, Jan; Bassetto, Marcella; Brancale, Andrea; Fu, Xian-Hua; Gao, Yang; Li, Jun; Zhang, Su-Zhan; Hamel, Ernest; Bortolozzi, Roberta; Basso, Giuseppe; Viola, Giampietro
The biological importance of microtubules make them an interesting target for the synthesis of antitumor agents. The 2-(3′,4′,5′-trimethoxybenzoyl)-5-aminobenzo[b]thiophene moiety was identified as a novel scaffold for the preparation of potent inhibitors of microtubule polymerization acting through the colchicine site of tubulin. The position of the methoxy group on the benzo[b]thiophene was important for maximal antiproliferative activity. Structure–activity relationship analysis established that the best activities were obtained with amino and methoxy groups placed at the C-5 and C-7 positions, respectively. Compounds 3c–e showed more potent inhibition of tubulin polymerization than combretastatin A-4 and strong binding to the colchicine site. These compounds also demonstrated substantial antiproliferative activity, with IC50 values ranging from 2.6 to 18 nM in a variety of cancer cell lines. Importantly, compound 3c (50 mg/kg), significantly inhibited the growth of the human osteosarcoma MNNG/HOS xenograft in nude mice. PMID:24164557
TRIM66 overexpresssion contributes to osteosarcoma carcinogenesis and indicates poor survival outcome
Chen, Yu; Guo, Yongfei; Yang, Haisong; Shi, Guodong; Xu, Guohua; Shi, Jiangang; Na, Yin; Chen, Deyu
TRIM66 belongs to the family of tripartite motif (TRIM)-containing proteins. Alterations in TRIM proteins have been implicated in several malignancies. This study was aimed at elucidating the expression and biological function of TRIM66 in osteosarcoma. Here, TRIM66 expression level was higher in osteosarcoma tissues than in normal tissues. High TRIM66 expression was correlated with high rate of local recurrence and lung metastasis, and short survival time. Then, we found that knockdown of TRIM66 in two osteosarcoma cell lines, MG63 and HOS, significantly inhibited cell proliferation and induced G1-phase arrest. Moreover, inhibition of TRIM66 in osteosarcoma cells significantly induced cell apoptosis, while remarkably inhibited cell migration, invasion as well as tumorigenicity in nude mice. Gene set enrichment analysis in Gene Expression Omnibus dataset revealed that apoptosis, epithelial-mesenchymal transition (EMT) and transforming growth factor-β (TGF-β) signaling pathway-related genes were enriched in TRIM66 higher expression patients, which was confirmed by western blot analysis in osteosarcoma cells with TRIM66 silenced. In conclusion, TRIM66 may act as an oncogene through suppressing apoptosis pathway and promoting TGF-β signaling in osteosarcoma carcinogenesis. TRIM66 may be a prognostic factor and potential therapeutic target in osteosarcoma. PMID:26247633
Protective Effect of Chitin Urocanate Nanofibers against Ultraviolet Radiation.
Ito, Ikuko; Yoneda, Toshikazu; Omura, Yoshihiko; Osaki, Tomohiro; Ifuku, Shinsuke; Saimoto, Hiroyuki; Azuma, Kazuo; Imagawa, Tomohiro; Tsuka, Takeshi; Murahata, Yusuke; Ito, Norihiko; Okamoto, Yoshiharu; Minami, Saburo
Urocanic acid is a major ultraviolet (UV)-absorbing chromophore. Chitins are highly crystalline structures that are found predominantly in crustacean shells. Alpha-chitin consists of microfibers that contain nanofibrils embedded in a protein matrix. Acid hydrolysis is a common method used to prepare chitin nanofibrils (NFs). We typically obtain NFs by hydrolyzing chitin with acetic acid. However, in the present study, we used urocanic acid to prepare urocanic acid chitin NFs (UNFs) and examined its protective effect against UVB radiation. Hos: HR-1 mice coated with UNFs were UVB irradiated (302 nm, 150 mJ/cm²), and these mice showed markedly lower UVB radiation-induced cutaneous erythema than the control. Additionally, sunburn cells were rarely detected in the epidermis of UNFs-coated mice after UVB irradiation. Although the difference was not as significant as UNFs, the number of sunburn cells in mice treated with acetic acid chitin nanofibrils (ANFs) tended to be lower than in control mice. These results demonstrate that ANFs have a protective effect against UVB and suggest that the anti-inflammatory and antioxidant effects of NFs influence the protective effect of ANFs against UVB radiation. The combination of NFs with other substances that possess UV-protective effects, such as urocanic acid, may provide an enhanced protective effect against UVB radiation. PMID:26703629
HDA2 and HDA3 are related proteins that interact with and are essential for the activity of the yeast histone deacetylase HDA1
Wu, Jiansheng; Carmen, Andrew A.; Kobayashi, Ryuji; Suka, Noriyuki; Grunstein, Michael
Histone deacetylase HDA1, the prototype for the class II mammalian deacetylases, is likely the catalytic subunit of the HDA1-containing complex that is involved in TUP1-specific repression and global deacetylation in yeast. Although the class I RPD3-like enzymatic complexes have been well characterized, little is known about the identity and interactions of the factors that associate to form the HDA1 complex. In this paper, we identify related HDA2 and HDA3 proteins that are found in the HDA1 complex and show that HDA1 interacts with itself and with the HDA2-HDA3 subcomplex to form a likely tetramer. These interactions are necessary for catalytic activity because mutations in any of the three components disrupt activity both in vitro and in vivo. In this respect the HDA1 complex differs from yeast RPD3, which has components such as SIN3 that are not essential for activity in vitro, and yeast HOS3, which has intrinsic in vitro activity as a homodimer in the absence of other subunits. PMID:11287668
Protective Effect of Chitin Urocanate Nanofibers against Ultraviolet Radiation
Ito, Ikuko; Yoneda, Toshikazu; Omura, Yoshihiko; Osaki, Tomohiro; Ifuku, Shinsuke; Saimoto, Hiroyuki; Azuma, Kazuo; Imagawa, Tomohiro; Tsuka, Takeshi; Murahata, Yusuke; Ito, Norihiko; Okamoto, Yoshiharu; Minami, Saburo
Urocanic acid is a major ultraviolet (UV)-absorbing chromophore. Chitins are highly crystalline structures that are found predominantly in crustacean shells. Alpha-chitin consists of microfibers that contain nanofibrils embedded in a protein matrix. Acid hydrolysis is a common method used to prepare chitin nanofibrils (NFs). We typically obtain NFs by hydrolyzing chitin with acetic acid. However, in the present study, we used urocanic acid to prepare urocanic acid chitin NFs (UNFs) and examined its protective effect against UVB radiation. Hos: HR-1 mice coated with UNFs were UVB irradiated (302 nm, 150 mJ/cm2), and these mice showed markedly lower UVB radiation-induced cutaneous erythema than the control. Additionally, sunburn cells were rarely detected in the epidermis of UNFs-coated mice after UVB irradiation. Although the difference was not as significant as UNFs, the number of sunburn cells in mice treated with acetic acid chitin nanofibrils (ANFs) tended to be lower than in control mice. These results demonstrate that ANFs have a protective effect against UVB and suggest that the anti-inflammatory and antioxidant effects of NFs influence the protective effect of ANFs against UVB radiation. The combination of NFs with other substances that possess UV-protective effects, such as urocanic acid, may provide an enhanced protective effect against UVB radiation. PMID:26703629
Detection of Neovascularization Based on Fractal and Texture Analysis with Interaction Effects in Diabetic Retinopathy
Lee, Jack; Zee, Benny Chung Ying; Li, Qing
Diabetic retinopathy is a major cause of blindness. Proliferative diabetic retinopathy is a result of severe vascular complication and is visible as neovascularization of the retina. Automatic detection of such new vessels would be useful for the severity grading of diabetic retinopathy, and it is an important part of screening process to identify those who may require immediate treatment for their diabetic retinopathy. We proposed a novel new vessels detection method including statistical texture analysis (STA), high order spectrum analysis (HOS), fractal analysis (FA), and most importantly we have shown that by incorporating their associated interactions the accuracy of new vessels detection can be greatly improved. To assess its performance, the sensitivity, specificity and accuracy (AUC) are obtained. They are 96.3%, 99.1% and 98.5% (99.3%), respectively. It is found that the proposed method can improve the accuracy of new vessels detection significantly over previous methods. The algorithm can be automated and is valuable to detect relatively severe cases of diabetic retinopathy among diabetes patients. PMID:24358105
An approach to predict Sudden Cardiac Death (SCD) using time domain and bispectrum features from HRV signal.
Houshyarifar, Vahid; Chehel Amirani, Mehdi
In this paper we present a method to predict Sudden Cardiac Arrest (SCA) with higher order spectral (HOS) and linear (Time) features extracted from heart rate variability (HRV) signal. Predicting the occurrence of SCA is important in order to avoid the probability of Sudden Cardiac Death (SCD). This work is a challenge to predict five minutes before SCA onset. The method consists of four steps: pre-processing, feature extraction, feature reduction, and classification. In the first step, the QRS complexes are detected from the electrocardiogram (ECG) signal and then the HRV signal is extracted. In second step, bispectrum features of HRV signal and time-domain features are obtained. Six features are extracted from bispectrum and two features from time-domain. In the next step, these features are reduced to one feature by the linear discriminant analysis (LDA) technique. Finally, KNN and support vector machine-based classifiers are used to classify the HRV signals. We used two database named, MIT/BIH Sudden Cardiac Death (SCD) Database and Physiobank Normal Sinus Rhythm (NSR). In this work we achieved prediction of SCD occurrence for six minutes before the SCA with the accuracy over 91%. PMID:27567781
Zoledronate blocks geranylgeranylation not farnesylation to suppress human osteosarcoma U2OS cells metastasis by EMT via Rho A activation and FAK-inhibited JNK and p38 pathways
Cheng, Hsin-Lin; Lin, Chiao-Wen; Yang, Jia-Sin; Hsieh, Ming-Ju; Yang, Shun-Fa; Lu, Ko-Hsiu
Zoledronate is a standard treatment for preventing skeletal complications of osteoporosis and some types of cancer associated with bone metastases, but we little know whether the effect of zoledronate on metastasis of osteosarcoma. Here, we investigated the inhibitory effects of zoledronate on cell viability, motility, migration and invasion of 4 osteosarcoma cell lines (Saos2, MG-63, HOS and U2OS) by affecting cell morphology, epithelial-mesenchymal transition (EMT) and cytoskeletal organization as well as induction of E-cadherin and reduction of N-cadherin with activation of transcription factors Slug and Twist, especially in U2OS cells. Zoledronate decreased JNK and p38 phosphorylation and upper streams of focal adhesion kinase (FAK) and Src to suppress the motility, invasiveness and migration of U2OS cells. In addition to zoledronate-inhibited Rho A and Cdc42 membrane translocation and GTPγS activities, the anti-metastatic effects in U2OS cells including inhibition of adhesion were reversed by geranylgeraniol, but not farnesol. In conclusion, Zoledronate blocks geranylgeranylation not farnesylation to suppress human osteosarcoma U2OS cell-matrix and cell-cell interactions, migration potential, the invasive activity, and the adhesive ability by EMT via Rho A activation and FAK-inhibited JNK and p38 pathways. PMID:26848867
Antileukemic Effect of Tualang Honey on Acute and Chronic Leukemia Cell Lines
Nik Man, Nik Muhd Khuzaimi; Hassan, Rosline; Ang, Cheng Yong; Abdullah, Abu Dzarr; Mohd Radzi, Muhammad Amiro Rasheeq; Sulaiman, Siti Amrah
Complementary medicine using natural product as antitumor is on the rise. Much research has been performed on Tualang Honey and it was shown to have therapeutic potential in wound healing, and antimicrobial activity and be antiproliferative against several cancer models such as human osteosarcoma (HOS), human breast (MCF-7 and MDA-MB-231), and cervical (HeLa) cancer cell lines. To date, there was limited study on antileukemic properties of Tualang (Koompassia excelsa) Honey. The aim of this study was to evaluate the antileukemic effect of Tualang Honey on acute and chronic leukemia cell lines. Leukemia cell lines (K562 and MV4-11) and human mononuclear cell isolated from peripheral blood were grown in RPM1 1640 culture medium. The cells were incubated with increasing concentrations of Tualang Honey. After incubation, the evaluation of viability and apoptosis was performed. The morphological changes of leukemia cells were the presence of cytoplasmic blebs followed by apoptotic bodies and round shape of cells. IC50 against K562 and MV4-11 was determined. Tualang Honey gave 53.9% and 50.6% apoptosis activity on K562 and MV4-11, respectively, while on human mononuclear cell it was 37.4%. Tualang Honey has the apoptosis-inducing ability for acute and chronic myeloid leukemia (K562 and MV4-11) cell lines. PMID:26613081
ICE1 Ser403 is necessary for protein stabilization and regulation of cold signaling and tolerance.
Miura, Kenji; Ohta, Masaru; Nakazawa, Machiko; Ono, Michiyuki; Hasegawa, Paul M
ICE1, a MYC-type transcription factor, has an important role in the induction of CBF3/DREB1A for regulation of cold signaling and tolerance. Here we reveal that serine 403 of ICE1 is involved in regulating the transactivation and stability of the ICE1 protein. Substitution of serine 403 by alanine enhanced the transactivational activity of ICE1 in Arabidopsis protoplasts. Over-expression of ICE1(S403A) conferred more freezing tolerance than ICE1(WT) in Arabidopsis, and the expression of cold-regulated genes such as CBF3/DREB1A, COR47 and KIN1 was enhanced in plants over-expressing ICE1(S403A). Furthermore, the ICE1(S403A) protein level was not changed after cold treatment, whereas the ICE1(WT) protein level was reduced. Interestingly, polyubiquitylation of the ICE1(S403A) protein in vivo was apparently blocked. These results demonstrate that serine 403 of ICE1 has roles in both transactivation and cold-induced degradation of ICE1 via the ubiquitin/26S proteasome pathway, suggesting that serine 403 is a key residue for the attenuation of cold-stress responses by HOS1-mediated degradation of ICE1. PMID:21447070
The low temperature response pathways for cold acclimation and vernalization are independent.
Bond, Donna M; Dennis, Elizabeth S; Finnegan, E Jean
Vernalization is the promotion of flowering in response to the prolonged cold of winter. To survive sub-zero winter temperatures, plants must first acclimate to low, non-freezing temperatures (cold acclimation). Induction of VERNALIZATION INSENSITIVE 3 (VIN3), the first gene in the vernalization pathway, is initiated within the same time frame as the induction of genes in the cold acclimation pathway raising the question of whether there are common elements in the signal transduction pathways that activate these two responses to cold. We show that none of the signalling components required for cold acclimation, including the 'master regulator'INDUCTION OF CBF EXPRESSION1 (ICE1) or HIGH EXPRESSION OF OSMOTICALLY RESPONSIVE GENE1 (HOS1), which has been described as a link between cold acclimation and vernalization, play a role in VIN3 induction. We also show that the hormone abscisic acid (ABA) does not modulate VIN3 induction, consistent with earlier reports that ABA signalling plays no role in the vernalization response. The cold acclimation pathway is activated at 12 °C, at which temperature there is no induction of VIN3 expression. Taken together, our data demonstrate that the responses to low temperatures leading to cold acclimation and vernalization are controlled by distinct signalling pathways. PMID:21631537
CBF-dependent signaling pathway: a key responder to low temperature stress in plants.
Zhou, M Q; Shen, C; Wu, L H; Tang, K X; Lin, J
Plants under low temperature (LT) stress exhibit a C-repeat binding factor (CBF)-dependent responsive pathway. The transcription factors in the CBF family, existing in multiple plant species, are the key regulators of the cold-responsive (COR) genes. CBF1 and CBF3 are regulated in a different way from CBF2, and CBF4 is the only known CBF gene definitely involved in abscisic acid (ABA)-dependent signaling pathways. RAP2.1 and RAP2.6 are the downstream regulators under CBFs. The upstream regulators of the CBF named inducer of CBF expression (ICE) acts as a positive regulator of CBFs. Meanwhile, these CBF signaling pathway components could associate with many other transcription activators and repressors in regulating gene expression when plants are under LT stress. HOS1 negatively regulates ICE1, which down regulates MYB15, an upstream repressor of CBFs. ZAT12 participates in the repression of CBFs, while ZAT10 and FRY2 negatively regulate the CBF-target genes. ADF5 was recently also found to repress CBFs. LOS2 works against ZAT10, and LOS4 positively regulates CBFs. SFR6 is involved in the modification of CBFs to activate the COR genes, and SIZ1-dependent sumoylation plays a positive role in the regulation of ICE1. The utilization of CBF-dependent signaling components has a broad perspective in the field of plant breeding for enhancing crop LT tolerance. PMID:20919819
The unified ICE-CBF pathway provides a transcriptional feedback control of freezing tolerance during cold acclimation in Arabidopsis.
Kim, Ye Seul; Lee, Minyoung; Lee, Jae-Hyung; Lee, Hyo-Jun; Park, Chung-Mo
During cold acclimation, C-repeat binding factors (CBFs) activate downstream targets, such as cold-regulated genes, leading to the acquisition of freezing tolerance in plants. Inducer of CBF expression 1 (ICE1) plays a key role by activating CBF3 expression in shaping the cold-induced transcriptome. While the ICE1-CBF3 regulon constitutes a major cold acclimation pathway, gene regulatory networks governing the CBF signaling are poorly understood. Here, we demonstrated that ICE1 and its paralog ICE2 induce CBF1, CBF2, and CBF3 by binding to the gene promoters. ICE2, like ICE1, was ubiquitinated by the high expression of osmotically responsive gene 1 (HOS1) E3 ubiquitin ligase. Whereas ICE2-defective ice2-2 mutant did not exhibit any discernible freezing-sensitive phenotypes, ice1-2 ice2-2/+ plant, which is defective in ICE1 and has a heterozygotic ice2 mutation, exhibited significantly reduced freezing tolerance. Accordingly, all three CBF genes were markedly down-regulated in the ice1-2 ice2-2/+ plant, indicating that ICE1 and ICE2 are functionally redundant with different implementations in inducing CBF genes. Together with the negative regulation of CBF3 by CBF2, we propose that the unified ICE-CBF pathway provides a transcriptional feedback of freezing tolerance to sustain plant development and survival during cold acclimation. PMID:26311645
Automated diagnosis of dry eye using infrared thermography images
NASA Astrophysics Data System (ADS)
Acharya, U. Rajendra; Tan, Jen Hong; Koh, Joel E. W.; Sudarshan, Vidya K.; Yeo, Sharon; Too, Cheah Loon; Chua, Chua Kuang; Ng, E. Y. K.; Tong, Louis
Dry Eye (DE) is a condition of either decreased tear production or increased tear film evaporation. Prolonged DE damages the cornea causing the corneal scarring, thinning and perforation. There is no single uniform diagnosis test available to date; combinations of diagnostic tests are to be performed to diagnose DE. The current diagnostic methods available are subjective, uncomfortable and invasive. Hence in this paper, we have developed an efficient, fast and non-invasive technique for the automated identification of normal and DE classes using infrared thermography images. The features are extracted from nonlinear method called Higher Order Spectra (HOS). Features are ranked using t-test ranking strategy. These ranked features are fed to various classifiers namely, K-Nearest Neighbor (KNN), Nave Bayesian Classifier (NBC), Decision Tree (DT), Probabilistic Neural Network (PNN), and Support Vector Machine (SVM) to select the best classifier using minimum number of features. Our proposed system is able to identify the DE and normal classes automatically with classification accuracy of 99.8%, sensitivity of 99.8%, and specificity if 99.8% for left eye using PNN and KNN classifiers. And we have reported classification accuracy of 99.8%, sensitivity of 99.9%, and specificity if 99.4% for right eye using SVM classifier with polynomial order 2 kernel.
Response of goat sperm to hypoosmotic steps modelled probit analysis.
Leboeuf, B; Le Vern, Y; Furstoss, V; Kerboeuf, D; Guillouet, P; Magistrini, M
Hypoosmotic swelling test (HOS) has been proposed by many authors to evaluate the functional integrity of the sperm membrane. Our approach in this experiment has consisted in exposing spermatozoa to a wide range of osmotic pressures then evaluating the reacted sperm cells by flow cytometry and finally modelling the sperm cell responses. Semen samples were diluted in skim milk or NPPC (native phosphocaseinate) extenders, and stored at 4 degrees C for 3 days. At D0 and D3 aliquots from each ejaculate (n=12) were submitted to seven hypoosmotic solutions varying from 230 to 10mOsm/kg. Sperm samples were analyzed using flow cytometry to determine two populations of spermatozoa identified by propidium iodide (PI): PI+ (including PI, red fluorescence) and PI- (excluding PI, no fluorescence). Spermatozoa PI+ were considered as spermatozoa with membrane damages. PI+ exhibited a high variation from 230 to 10mOsm/kg which was considered as a dose-response curve. Data were modelled using Mixed procedure and probit analysis to a sigmoid curve. Each model curve characterized the profile of response of the variable PI+ to the range of osmotic pressure from 230 to 10mOsm/kg. The estimated parameters modelling the sigmoid curves are discussed in order to evaluate the effect of extender (skim milk versus NPPC) and duration of preservation (D0 versus D3). Such modelling could help to differentiate storage method ejaculates within males or between male, contributing therefore to improve semen technology. PMID:15985346
Set3 contributes to heterochromatin integrity by promoting transcription of subunits of Clr4-Rik1-Cul4 histone methyltransferase complex in fission yeast
Yu, Yao; Zhou, Huan; Deng, Xiaolong; Wang, Wenchao; Lu, Hong
Heterochromatin formation in fission yeast depends on RNAi machinery and histone-modifying enzymes. One of the key histone-modifying complexes is Clr4-Rik1-Cul4 methyltransferase complex (CLRC), which mediates histone H3K9 methylation, a hallmark for heterochromatin. CLRC is composed of the Clr4 histone methyltransferase, Rik1, Raf1, Raf2 and Pcu4. However, transcriptional regulation of the CLRC subunits is not well understood. In this study, we identified Set3, a core subunit of the Set3/Hos2 histone deacetylase complex (Set3C), as a contributor to the integrity and silencing of heterochromatin at centromeres, telomeres and silent mating-type locus. This novel role of Set3 relies on its PHD finger, but is independent of deacetylase activity or structural integrity of Set3C. Set3 is not located to the centromeric region. Instead, Set3 is targeted to the promoters of clr4+ and rik1+, probably through its PHD finger. Set3 promotes transcription of clr4+ and rik1+. Consistently, the protein levels of Clr4 and Rik1 were reduced in the set3Δ mutant. The heterochromatin silencing defect in the set3Δ mutant could be rescued by overexpressing of clr4+ or rik1+. Our study suggests transcriptional activation of essential heterochromatin factors underlies the tight regulation of heterochromatin integrity. PMID:27538348
Theoretical survey of the reaction between osmium and acetaldehyde
NASA Astrophysics Data System (ADS)
Dai, Guo-Liang; Wang, Chuan-Feng
The mechanism of the reaction of osmium atom with acetaldehyde has been investigated with a DFT approach. All the stationary points are determined at the UB3LYP/ sdd/6-311++G** level of the theory. Both ground and excited state potential energy surfaces are investigated in detail. The present results show that the title reaction start with the formation of a CH3CHO-metal complex followed by C-C, aldehyde C-H, C-O, and methyl C-H activation. These reactions can lead to four different products (HOsCH3 + CO, OsCO + CH4, OsCOCH3 + H, and OsO + C2H4). The minimum energy reaction path is found to involve the spin inversion in the initial reaction step. This potential energy curve-crossing dramatically affects reaction exothermic. The present results may be helpful in understanding the mechanism of the title reaction and further experimental investigation of the reaction.
Capacitation and acrosome reaction induction on thawed Dama dama deer spermatozoa: glycine effect as cryopreservation diluent supplement.
Fernández, Silvina; Sestelo, Adrián; Rivolta, Miguel; Córdoba, Mariana
The aim of this research was to evaluate two different diluents for sperm cryopreservation and to study functional parameters in relation to the response to heparin, lysophosphatidylcholine and progesterone, in frozen-thawed semen of fallow deer (Dama dama) during the reproductive season (brama). In this way, fallow deer can be used as a biological model of endangered cervids. Semen was obtained by electroejaculation. Heparin, progesterone and lysophosphatidylcholine were used as capacitation and acrosome reaction inducers, respectively. Capacitation and acrosome reaction were evaluated by chlorotetracycline epifluorescence technique (CTC), membrane integrity by Hypo-osmotic swelling test (HOS) and viability and acrosome integrity by trypan blue stain/DIC. Data was analyzed by ANOVA and Tukey Test (P < 0.05). Semen was cryopreserved in different diluents and Fructose-Tris-Glycine extender was selected. Capacitation with heparin at different incubation times determined that the highest capacitation percentage was obtained at 45 minutes incubation. Progesterone (1 'M) and lysophosphatidylcholine in heparin capacitated sperm induced acrosome reaction (P < 0.05). This study contributes to improve cryopreservation methods and to increase the knowledge about capacitation and acrosome reaction in vitro in deer spermatozoa, allowing an advance in the development of reproductive biotechnologies. PMID:24320190
Simian immunodeficiency virus variants with differential T-cell and macrophage tropism use CCR5 and an unidentified cofactor expressed in CEMx174 cells for efficient entry.
Kirchhoff, F; Pöhlmann, S; Hamacher, M; Means, R E; Kraus, T; Uberla, K; Di Marzio, P
The recent identification of coreceptors that mediate efficient entry of human immunodeficiency virus type 1 (HIV-1) suggests new therapeutic and preventive strategies. We analyzed simian immunodeficiency virus (SIV) entry cofactors to investigate whether the macaque SIV model can be used as an experimental model to evaluate these strategies. Similar to primary HIV-1 isolates, a well-characterized molecular clone, SIVmac239, which replicates poorly but efficiently enters into rhesus alveolar macrophages and an envelope variant, SIVmac239/316Env, with an approximately 1,000-fold-higher replicative capacity in macrophages used the beta-chemokine receptor CCR5 for efficient entry. The transmembrane portion of 316Env allowed low-level entry into cells expressing CCR1, CCR2B, and CCR3. A single amino acid substitution in the V3 loop of SIVmac239/316Env, 321P-->S, impaired the ability to enter into the T-B hybrid cell line CEMx174 but had relatively little effect on entry into primary cells and HOS.CD4 cells expressing CCR5. Although CEMx174 cells do not express CCR5, most SIVmac variants entered this hybrid cell line efficiently but did not enter the parental T-cell line CEM. It seems likely that CEMx174 cells express an as-yet-unidentified, perhaps B-cell-derived cofactor which allows efficient entry of SIVmac. PMID:9261370
The role of thermal shock in cyclic oxidation
NASA Technical Reports Server (NTRS)
Lowell, C. E.; Deadmore, D. L.
The effect of thermal shock on the spalling of oxides from the surfaces of several commercial alloys was determined. The average cooling rate was varied from approximately 240 C per second to less than 1.0 C per second during cyclic oxidation tests in air. The tests consisted of one hundred cycles of one hour at the maximum temperature (1100 or 1200 C). The alloys were HOS-875, TD-Ni, TD-NiCrAl, IN-601, IN-702, and B-1900 plus Hf. All of these alloys exhibited partial spalling within the oxide rather than total oxide loss down to bare metal. Thermal shock resulted in deformation of the metal which in turn resulted, in most cases, in changing the oxide failure mode from compressive to tensile. Tensile failures were characterized by cracking of the oxide and little loss, while compressive failures were characterized by explosive loss of platelets of oxide. This behavior was confirmed by examination of mechanically stressed oxide scales. The thermally shocked oxides spalled less than the slow cooled samples with the exception of TD-NiCrAl. This material failed in a brittle manner rather than by plastic deformation.
First report of ivermectin resistance in field populations of Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) in Punjab districts of India.
Singh, Nirbhay Kumar; Singh, Harkirat; Jyoti; Prerna, Mranalini; Rath, Shitanshu S
The larval immersion test (LIT) was used on the progenies of Rhipicephalus (Boophilus) microplus collected from four districts of the Punjab state, India to test the resistance to ivermectin. The regression graphs of probit mortality of larval ticks plotted against log values of increasing concentrations of ivermectin were utilized for the determination of slope of mortality, lethal concentration for 50%(LC50), 95%(LC95), resistance ratios (RR50, RR95) and the resistance levels (RL). Values of the coefficient of determination (R(2)) for LIT ranged from 0.900 to 0.978, and the RR50 and RR95 values against ivermectin ranged from 2.97 to 8.85 and 2.42 to 8.47, respectively, indicating resistance status in all field isolates. On the basis of RR values, three field isolates (BAT, GUR, HOS) showed level II, whereas PTK isolate showed presence of level I resistance status against ivermectin. This appears to be the first report of ivermectin resistance in R. (B.) microplus from Punjab, India. PMID:26404476
Antileukemic Effect of Tualang Honey on Acute and Chronic Leukemia Cell Lines.
Nik Man, Nik Muhd Khuzaimi; Hassan, Rosline; Ang, Cheng Yong; Abdullah, Abu Dzarr; Mohd Radzi, Muhammad Amiro Rasheeq; Sulaiman, Siti Amrah
Complementary medicine using natural product as antitumor is on the rise. Much research has been performed on Tualang Honey and it was shown to have therapeutic potential in wound healing, and antimicrobial activity and be antiproliferative against several cancer models such as human osteosarcoma (HOS), human breast (MCF-7 and MDA-MB-231), and cervical (HeLa) cancer cell lines. To date, there was limited study on antileukemic properties of Tualang (Koompassia excelsa) Honey. The aim of this study was to evaluate the antileukemic effect of Tualang Honey on acute and chronic leukemia cell lines. Leukemia cell lines (K562 and MV4-11) and human mononuclear cell isolated from peripheral blood were grown in RPM1 1640 culture medium. The cells were incubated with increasing concentrations of Tualang Honey. After incubation, the evaluation of viability and apoptosis was performed. The morphological changes of leukemia cells were the presence of cytoplasmic blebs followed by apoptotic bodies and round shape of cells. IC50 against K562 and MV4-11 was determined. Tualang Honey gave 53.9% and 50.6% apoptosis activity on K562 and MV4-11, respectively, while on human mononuclear cell it was 37.4%. Tualang Honey has the apoptosis-inducing ability for acute and chronic myeloid leukemia (K562 and MV4-11) cell lines. PMID:26613081
Recovery of lead-induced suppressed reproduction in male rats by testosterone.
Reshma Anjum, M; Sreenivasula Reddy, P
The objective of the present study was to investigate the effects of testosterone in recuperation of lead-induced suppressed reproduction in adult male rats. Lead acetate was administered orally to adult male rats (95 ± 5 days) at dosage level of 0.05 and 0.15% for 55 days through drinking water and injected intraperitoneally with either testoviron depot at a dose of 4.16 mg kg(-1) body weight or vehicle alone on days 1, 7 and 14 respectively. At the end of treatment, control and treated males were cohabited with untreated normal-cycling females. After cohabitation for 5 days, all the male rats were killed and weights of reproductive organs were determined. Significant increase in the indices of testis, epididymis, seminal vesicles, vas deferens and prostate glands was observed in testosterone (T)-treated rats when compared to those of lead-exposed rats. Testosterone treatment significantly increased epididymal sperm count, motile spermatozoa, viable spermatozoa and HOS tail-coiled spermatozoa and also the activity levels of testicular 3β- and 17β-hydroxysteroid dehydrogenases when compared to those of lead-exposed males. From the results, it can be hypothesised that supplementation of testosterone mitigated lead-induced suppressed reproduction in male rats. PMID:24909355
Structure analysis and photochemistry of adsorbates on platinum and palladium surfaces. [1,2-dichloroethene
Grassian, V.H.
The vibrational spectra of benzene and toluene adsorbed on Pd(111) indicates at 180K these molecules weakly bond to the surface. The adsorption of benzene and toluene on Pt(111) is much stronger as indicated by large frequency shifts from gas phase values. Pyridine adsorption on both Pt(111) and Pd(111) was studied as a function of temperature. At room temperature pyridine decomposes on the surface to form an ..cap alpha..-pyridyl fragment (NC/sub 5/H/sub 4/) on Pt(111), whereas the molecule remains intact on Pd(111). The electron energy loss spectra of pyridine adsorbed on these surfaces is compared to the ir spectra of two osmium cluster compounds: Os/sub 3/(CO)/sub 11/(NC/sub 5/H/sub 5/), a pyridine complex, and HOs/sub 3/(CO)/sub 10/(NC/sub 5/H/sub 4/), a pyridyl complex. The stronger interaction of these molecules to the platinum surface is a consequence of the stronger bonding of the 5d orbitals as compared to the 4d orbitals. The uv photochemistry of 2-butene and 1,2-dichloroethene when adsorbed on Pt and Pd surfaces was also studied.
Platinum(II) oxalato complexes involving adenosine-based N-donor ligands: synthesis, characterization and cytotoxicity evaluation.
Starha, Pavel; Popa, Igor; Trávníček, Zdeněk
A one-step synthetic procedure using the reaction of potassium bis(oxalato)platinate(II) with the corresponding N6-benzyladenosine derivative (nL) provided the [Pt(ox)(nL)₂]∙1.5H₂O oxalato (ox) complexes 1-5, involving the nL molecules as monodentate coordinated N-donor ligands. The complexes were thoroughly characterized by elemental analysis, multinuclear (¹H, ¹³C, ¹⁵N, 1¹⁹⁵Pt) and two dimensional NMR, infrared and Raman spectroscopy, and mass spectrometry, proving their composition and purity as well as coordination of nL through the N7 atom of the purine moiety. Geometry of [Pt(ox)(4FL)₂] (5) was optimized at the B3LYP/LANLTZ/6-311G** level of theory. The complexes were screened for their in vitro cytotoxicity against two human cancer cell lines (HOS osteosarcoma and MCF7 breast adenocarcinoma), but they did not show any effect up to the concentration of 50.0 µM (compounds 1, 2) or 20.0 µM (compounds 3-5). PMID:24662093
Synthesis, characterization and in vitro antitumor activity of platinum(II) oxalato complexes involving 7-azaindole derivatives as coligands.
Štarha, Pavel; Trávníček, Zdeněk; Popa, Igor; Dvořák, Zdeněk
The platinum(II) oxalato complexes [Pt(ox)(naza)2] (1-3) were synthesized and characterized by elemental analysis (C, H, N), multinuclear NMR spectroscopy ((1)H, (13)C, (15)N, (195)Pt) and electrospray ionization mass spectrometry (ESI-MS); naza = 4-chloro-7-azaindole (4Claza; 1), 3-bromo-7-azaindole (3Braza; 2) or 4-bromo-7-azaindole (4Braza; 3). The prepared substances were screened for their in vitro antitumor activity on the osteosarcoma (HOS) and breast adenocarcinoma (MCF7) human cancer cell lines, where 2 showed moderate antitumor effect (IC50 = 27.5 μM, and 18.3 μM, respectively). The complex 2 was further tested on a panel of six others human cancer cell lines, including the malignant melanoma (G361), cervix carcinoma (HeLa), ovarian carcinoma (A2780), cisplatin-resistant ovarian carcinoma (A2780R), lung carcinoma (A549) and prostate adenocarcinoma (LNCaP). This substance was found to be moderate antitumor effective against G361 (IC50 = 17.3 μM), HeLa (IC50 = 31.8 μM) and A2780 (IC50 = 19.2 μM) cell lines. The complex 2 was also studied by NMR for its solution stability and by ESI-MS experiments for its ability to interact with biomolecules, such as cysteine, glutathione or guanosine 5'-monophosphate. PMID:25068781
Iron-salophen complexes involving azole-derived ligands: A new group of compounds with high-level and broad-spectrum in vitro antitumor activity.
Vančo, Ján; Šindelář, Zdeněk; Dvořák, Zdeněk; Trávníček, Zdeněk
A series of iron(II/III) salophen (salph) complexes involving monodentate azole-derived ligands, having the composition [Fe(II)(salph)(HL1)] (1) and [Fe(III)(salph)(L)] (2-6), where HL1=imidazole, L=1,2,4-triazol-1-ido (L2), benzo[d][1,2,3]triazol-1-ido (L3), 5-aminotetrazol-1-ido (L4), 5-phenyltetrazol-1-ido (L5), and 5-methyltetrazol-1-ido (L6) ligand, was prepared and characterized by elemental analyses, infrared, Mössbauer and X-ray photolelectron spectroscopy, magnetic data and electrospray-ionization mass spectrometry. X-ray structure of 1 revealed a distorted square-pyramidal geometry in the vicinity of the iron(II) atom. The complexes were evaluated for their in vitro antitumor activity against the panel of six human cancer cell lines (HOS, MCF7, A549, HeLa, A2780 and G-361) and were found to be highly cytotoxic, showing the best IC50 value of 58nM for [Fe(III)(salph)(L6)] (6) against the ovarian carcinoma A2780 cell line, being 200-times more effective than cisplatin. In vitro cytotoxicity of complexes 1-6 on primary culture of human hepatocytes and calf-thymus DNA (CT-DNA) binding studies using the fluorescence titration were also performed. PMID:25450023
Novel Schiff bases based on the quinolinone skeleton: syntheses, X-ray structures and fluorescent properties.
Trávníček, Zdeněk; Buchtík, Roman; Nemec, Ivan
A series of a new type of Schiff bases 1-7, derived from 2-phenyl-3-amino-4(1H)-quinolinone and R-salicyladehyde derivatives wherein R = 3-hydroxy (1), 3,4-dihydroxy (2), 3-methoxy (3), 3-carboxy (4), 3-allyl (5), 5-chloro (6), and 5-nitro (7), was synthesized and structurally characterized. Each of the molecules 1, 3 and 7 consists of three planar moieties (i.e., a quinolinone and two phenyl rings), which are mutually oriented differently depending on the appropriate substituent R and the extent of non-covalent contacts stabilizing the crystal structures. The compounds were studied for their fluorescence properties, where compound 6 yielded the strongest intensity both in the solid phase and in 100 μM ethanol solution with a quantum yield of φ = 3.6% as compared to quinine sulfate used as a standard. The in vitro cytotoxicity of these compounds was tested against the human osteosarcoma (HOS) and breast adenocarcinoma (MCF7) cell lines, revealing no activity up to the concentration of 50 µM. PMID:25255753
Hypoxia-induced resistance to cisplatin-mediated apoptosis in osteosarcoma cells is reversed by gambogic acid independently of HIF-1α.
Zhao, Wei; Xia, Shi-Qi; Zhuang, Jin-Peng; Zhang, Zhi-Peng; You, Chang-Cheng; Yan, Jing-Long; Xu, Gong-Ping
In vitro evidence of hypoxia-induced resistance to cisplatin (CDDP)-mediated apoptosis exists in human osteosarcoma (OS). Gambogic acid (GA) is a promising chemotherapeutic compound that could increase the chemotherapeutic effectiveness of CDDP in human OS cells by inducing cell cycle arrest and promoting apoptosis. This study examined whether GA could overcome OS cell resistance to CDDP. Hypoxia significantly reduced levels of CDDP-induced apoptosis in the OS cell lines MG63 and HOS. However, combined treatment with GA and CDDP revealed a strong synergistic action between these drugs, and higher protein levels of the apoptosis-related factor Fas, cleaved caspase-8 and cleaved caspase-3 and lower expression of hypoxia-inducible factor (HIF)-1α are detected in both cell lines. Meanwhile, drug resistance was not reversed by exposure to the HIF-1α inhibitor 2-methoxyestradiol. These findings strongly suggest that hypoxia-induced resistance to CDDP is reversed by GA in OS cells independently of HIF-1α. Furthermore, in vivo studies using xenograft mouse models revealed that combination therapy with CDDP and GA exerted increased antitumor effects by inducing apoptosis. Taken together, our results demonstrate that GA may be a new potent therapeutic agent useful for targeting human OS cells. PMID:27473145
Mechanosensitivity of human osteosarcoma cells and phospholipase C {beta}2 expression
Hoberg, M. . E-mail: email@example.com; Gratz, H.-H.; Noll, M.; Jones, D.B.
Bone adapts to mechanical load by osteosynthesis, suggesting that osteoblasts might respond to mechanical stimuli. We therefore investigated cell proliferation and phospholipase C (PLC) expression in osteoblasts. One Hertz uniaxial stretching at 4000 {mu}strains significantly increased the proliferation rates of human osteoblast-like osteosarcoma cell line MG-63 and primary human osteoblasts. However, U-2/OS, SaOS-2, OST, and MNNG/HOS cells showed no significant changes in proliferation rate. We investigated the expression pattern of different isoforms of PLC in these cell lines. We were able to detect PLC {beta}1, {beta}3, {gamma}1, {gamma}2, and {delta}1 in all cells, but PLC {beta}2 was only detectable in the mechanosensitive cells. We therefore investigated the possible role of PLC {beta}2 in mechanotransduction. Inducible antisense expression for 24 h inhibited the translation of PLC {beta}1 in U-2/OS cells by 35% and PLC {beta}2 in MG-63 by 29%. Fluid shear flow experiments with MG-63 lacking PLC {beta}2 revealed a significantly higher level of cells losing attachment to coverslips and a significantly lower number of cells increasing intracellular free calcium.
Long noncoding RNA MALAT1 as a potential therapeutic target in osteosarcoma.
Cai, Xianyi; Liu, Yunlu; Yang, Wen; Xia, Yun; Yang, Cao; Yang, Shuhua; Liu, Xianzhe
Recent studies have revealed that long noncoding RNA metastasis-associated lung adenocarcinoma transcript 1 (MALAT1) plays an important role in the development of several solid tumors. However, the function of MALAT1 in the tumorigenesis of osteosarcoma remains unknown. In the present study, levels of MALAT1 in human osteosarcoma cell lines and tissues were detected by quantitative real-time polymerase chain reaction (RT-PCR). The roles of MALAT1 in osteosarcoma were investigated by using in vitro and in vivo assays. We observed that MALAT1 expression was up-regulated in human osteosarcoma cell lines and tissues. In vitro knockdown of MALAT1 by siRNA significantly inhibited cell proliferation and migration, and induced cell cycle arrest and apoptosis in osteosarcoma cells. In addition, MALAT1 knockdown markedly suppressed the formation of tubular network structures and caused breakage of stress fibers in osteosarcoma cell lines U2OS and MNNG/HOS. Furthermore, MALAT1 knockdown delayed tumor growth in an osteosarcoma xenograft model. Specifically, we found that administration of MALAT1 siRNA decreased the protein levels of RhoA and its downstream effectors Rho-associated coiled-coil containing protein kinases (ROCKs). Taken together, these findings suggest that MALAT1 plays an oncogenic role in osteosarcoma and may be a promising therapeutic target for the treatment of osteosarcoma patients. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:932-941, 2016. PMID:26575981
A new 2D monolayer BiXene, M2C (M = Mo, Tc, Os).
Sun, Weiwei; Li, Yunguo; Wang, Baotian; Jiang, Xue; Katsnelson, Mikhail I; Korzhavyi, Pavel; Eriksson, Olle; Di Marco, Igor
The existence of BiXenes, a new family of 2D monolayers, is hereby predicted. Theoretically, BiXenes have 1H symmetry (P6[combining macron]m2) and can be formed from the 4d/5d binary carbides. As the name suggests, they are close relatives of MXenes, which instead have 1T symmetry (P3[combining macron]m1). The newly found BiXenes, as well as some new MXenes, are shown to have formation energies close to that of germanene, which suggests that these materials should be possible to be synthesised. Among them, we illustrate that 1H-Tc2C and 1T-Mo2C are dynamically stable at 0 K, while 1H-Mo2C, 1T-Tc2C, 1H-Os2C, and 1T-Rh2C are likely to be stabilised via strain or temperature. In addition, the nature of the chemical bonding is analysed, emphasizing that the covalency between the transition metal ions and carbon is much stronger in BiXenes than in MXenes. The emergence of BiXenes can not only open up a new era of conducting 2D monolayers, but also provide good candidates for carrier materials aimed at energy storage and spintronic devices that have already been unveiled in MXenes. PMID:27528499
Zoledronate blocks geranylgeranylation not farnesylation to suppress human osteosarcoma U2OS cells metastasis by EMT via Rho A activation and FAK-inhibited JNK and p38 pathways.
Cheng, Hsin-Lin; Lin, Chiao-Wen; Yang, Jia-Sin; Hsieh, Ming-Ju; Yang, Shun-Fa; Lu, Ko-Hsiu
Zoledronate is a standard treatment for preventing skeletal complications of osteoporosis and some types of cancer associated with bone metastases, but we little know whether the effect of zoledronate on metastasis of osteosarcoma. Here, we investigated the inhibitory effects of zoledronate on cell viability, motility, migration and invasion of 4 osteosarcoma cell lines (Saos2, MG-63, HOS and U2OS) by affecting cell morphology, epithelial-mesenchymal transition (EMT) and cytoskeletal organization as well as induction of E-cadherin and reduction of N-cadherin with activation of transcription factors Slug and Twist, especially in U2OS cells. Zoledronate decreased JNK and p38 phosphorylation and upper streams of focal adhesion kinase (FAK) and Src to suppress the motility, invasiveness and migration of U2OS cells. In addition to zoledronate-inhibited Rho A and Cdc42 membrane translocation and GTPγS activities, the anti-metastatic effects in U2OS cells including inhibition of adhesion were reversed by geranylgeraniol, but not farnesol. In conclusion, Zoledronate blocks geranylgeranylation not farnesylation to suppress human osteosarcoma U2OS cell-matrix and cell-cell interactions, migration potential, the invasive activity, and the adhesive ability by EMT via Rho A activation and FAK-inhibited JNK and p38 pathways. PMID:26848867
Wellness engineering for better quality of life of aging baby boomer
NASA Astrophysics Data System (ADS)
Szu, Harold
Current health care system serving 78M aging baby-boomers is no longer sustainable, as the cost about 1/5 GDP will reach 1/4 GDT when all is retired in decades, unless the system is changed. We design a high-tech safe net to enhance the timeliness of early correct treatment execution (otherwise, causing 1/4 mortality associated with an escalating legal fee waste). We follow the common sense that "a stitch in time saves nine," and adopt the military surveillance know-how in designing early warning health management system, comprising of smart sensor pairs for point-care surveillance. However, the grand plan of affordable smart sensors hardware for households requires an ODM & OEM teaming to conduct parallel designing and sequential marketing strategy. The military software strategy combating a treacherous adversary enemy match well with point cares surveillance overcoming real world microorganism variability. Moreover, such smart military software provides self-reference change detection, not by traditional cohort ensemble average, but by individual own higher order statistics (HOS) independent component analysis (ICA), which take the advantage of known initial condition for each individual and desirable over-sampling daily dynamics. The triggering of warning follows the military algorithms comprising of Receiver Operation Characteristics (ROC) and Automatic Target Recognition (ATR). To further reduce the unwanted false negative rate, a benchmarked is made against the traditional cohort-ensemble baseline average & the upper & lower bounds of variance as adopted by the gatekeepers - Medical Doctors (MD) and Nurses.
Application of higher order spectra for accurate delineation of atrial arrhythmia.
Prasad, Hari; Martis, Roshan Joy; Acharya, U Rajendra; Min, Lim Choo; Suri, Jasjit S
The electrocardiogram (ECG) is being commonly used as a diagnostic tool to distinguish different types of atrial tachyarrhythmias. The inherent complexity and mechanistic and clinical inter-relationships often brings about diagnostic difficulties to treating physicians and primary health care professionals creating frequent misdiagnoses and cross classifications using visual criteria. The current paper presents a methodology for ECG based pattern analysis for detection of atrial flutter, atrial fibrillation and normal sinus rhythm beats. ECG is an inherently non-linear and non-stationary signal; its variation may contain indicators of current disease, or warnings about impending cardiac diseases. Routinely used time domain and frequency domain methods will not be able to capture the hidden information present in the ECG beats. In the present study, we have used non-linear features of higher order spectra (HOS) to differentiate the normal, atrial fibrillation and atrial flutter ECG beats. The bispectrum features were subjected to independent component analysis (ICA) for data reduction. The ICA coefficients were subsequently subjected to K-nearest-neighbor (KNN), classification and regression tree (CART) and neural network (NN) classifiers to evaluate the best automated classifier. We have obtained an average accuracy of 97.65%, sensitivity and specificity of 98.75% and 99.53% respectively using ten-fold cross validation. Overall, the results show that application of higher order spectra statistics is useful for the classification of atrial tachyarrhythmias with reasonably high accuracies. Further validation of the proposed technique will yield acceptable results for clinical implementation. PMID:24109623
Alleviating anastrozole induced bone toxicity by selenium nanoparticles in SD rats.
Vekariya, Kiritkumar K; Kaur, Jasmine; Tikoo, Kulbhushan
Aromatase inhibitors like anastrozole play an undisputed key role in the treatment of breast cancer, but on the other hand, various side effects like osteoporosis and increased risk of bone fracture accompany the chronic administration of these drugs. Here we show for the first time that selenium nanoparticles, when given in conjugation to anastrozole, lower the bone toxicity caused by anastrozole and thus reduce the probable damage to the bone. Selenium nanoparticles at a dose of 5μg/ml significantly reduced the cell death caused by anastrozole (1μM) in HOS (human osteoblast) cells. In addition, our results also highlighted that in female SD rat model, SeNPs (0.25, 0.5, 1mg/kg/day) significantly prevented the decrease in bone density and increase in biochemical markers of bone resorption induced by anastrozole (0.2mg/kg/day) treatment. Histopathological examination of the femurs of SeNP treated group revealed ossification, mineralization, calcified cartilaginous deposits and a marginal osteoclastic activity, all of which indicate a marked restorative action, suggesting the protective action of the SeNPs. Interestingly, SeNPs (1mg/kg/day) also exhibited protective effect in ovariectomized rat model, by preventing osteoporosis, which signifies that bone loss due to estrogen deficiency can be effectively overcome by using SeNPs. PMID:23415680
Propagation of 3D nonlinear waves over complex bathymetry using a High-Order Spectral method
NASA Astrophysics Data System (ADS)
Gouin, Maïté; Ducrozet, Guillaume; Ferrant, Pierre
Scattering of regular and irregular surface gravity waves propagating over a region of arbitrary three-dimensional varying bathymetry is considered here. The three-dimensional High-Order Spectral method (HOS) with an extension to account for a variable bathymetry is used. The efficiency of the model has been proved to be conserved even with this extension. The method is first applied to a bathymetry consisting of an elliptical lens, as used in the Vincent and Briggs (1989) experiment. Incident waves passing across the lens are transformed and a strong convergence region is observed after the elliptical mound. The wave amplification depends on the incident wave. Numerical results for regular and irregular waves are analysed and compared with other methods and experimental data demonstrating the efficiency and practical applicability of the present approach. Then the method is used to model waves propagating over a real bathymetry: the canyons of Scripps/La Jolla in California. The implementation of this complex bathymetry in the model is presented, as well as the first results achieved. They will be compared to the ones obtained with another numerical model.
Reactions occurring during the sulfation of sodium chloride deposited on alumina substrates
NASA Technical Reports Server (NTRS)
Wu, C. S.; Birks, N.
The reaction between solid NaCl and air containing 1 pct SO2 has been studied between 500 and 700 C. The reaction product, Na2SO4, forms not only on the surface of the NaCl but also on surrounding areas of the substrate due to the volatility of the NaCl at these temperatures. At the higher temperatures, the vapor pressure of NaCl is so high that the majority of the reaction product is distributed on the substrate. Above 625 C, the reaction product is a liquid solution of NaCl and Na2SO4 that exists only so long as NaCl is supplied from the original crystal source. Eventually, the liquid solidifies by constitutional solidification as the NaCl is converted to Na2SO4. While it exists, the liquid NaCl-Na2SO4 solution is shown to be highly corrosive to Al2O3 and, on a scale of Al2O3 growing on alloy HOS 875, particularly attacks the grain boundaries of the scale at preferred sites where chromium and iron oxides and sulfides rapidly develop. This is proposed as one mechanism by which NaCl deposition contributes to the initiation of low temperature hot corrosion.
Surveying the Impact of Work Hours and Schedules on Commercial Motor Vehicle Driver Sleep
Hege, Adam; Perko, Michael; Johnson, Amber; Yu, Chong Ho; Sönmez, Sevil; Apostolopoulos, Yorghos
Background Given the long hours on the road involving multiple and interacting work stressors (i.e., delivery pressures, irregular shifts, ergonomic hazards), commercial drivers face a plethora of health and safety risks. Researchers goal was to determine whether and to what extent long-haul trucker work schedules influence sleep duration and quality. Methods Survey and biometric data collected from male long-haul truck drivers at a major truckstop in central North Carolina over a six month period. Results Daily hours worked (mean = 11 hours, 55 minutes) and frequency of working over government-mandated daily HOS regulations (23.8% "frequently or always") were statistically significant predictors of sleep duration. Miles driven per week (mean = 2,812.61), irregular daily hours worked (63.8%), and frequency of working over the daily hour limit (23.8% "frequently or always") were statistically significant predictors of sleep quality. Conclusion Implications of findings suggest a comprehensive review of the regulations and operational conditions for commercial motor vehicle drivers be undertaken. PMID:26106509
Improved Near-Infrared Up-Conversion Emission of Tm3+ Sensitized by Yb3+ and Ho3+ in LuF3 Nanocrystals.
Xu, Xuhui; Wu, Yumei; Bian, Wenjuan; Yu, Xue; Zhang, Buhao; Li, Qianyue; Qiu, Jianbei; Liu, BiTao
In the present work, mono-disperse and uniform orthorhombic lutetium fluoride (LuF3) nanocrystals with an average size of about 35 nm have been successfully synthesized by a simple ionothermal method without any template. The infrared (IR) to visible up-conversion (UC) photoluminescence of LuF3 doped with Yb3+, Tm3+, and Ho3+ under 980 nm excitation was systemically studied. The intensity of near infrared (NIR) to visible up-conversion emission of Tm3+ was improved efficiently by adding Yb3+ and Ho3+ in LuF3, especially for the broad NIR emission band located at 812 nm. Meanwhile, compared to the Yb3+ and Tm3+ co-doped LuF3, the ratio of red to green emission in the Yb3+, TmS+, and HoS+ co-doped LuF3 changed greatly, and a bright yellowish-green emission was observed under 980 nm laser excitation. It shows that Yb3+, Tm3+ and Ho3+ co-doped LuF3 nanocrystals provided a potential application in vitro and in vivo bio-imaging, color displays and optical storage. PMID:27451685
Set3 contributes to heterochromatin integrity by promoting transcription of subunits of Clr4-Rik1-Cul4 histone methyltransferase complex in fission yeast.
Yu, Yao; Zhou, Huan; Deng, Xiaolong; Wang, Wenchao; Lu, Hong
Heterochromatin formation in fission yeast depends on RNAi machinery and histone-modifying enzymes. One of the key histone-modifying complexes is Clr4-Rik1-Cul4 methyltransferase complex (CLRC), which mediates histone H3K9 methylation, a hallmark for heterochromatin. CLRC is composed of the Clr4 histone methyltransferase, Rik1, Raf1, Raf2 and Pcu4. However, transcriptional regulation of the CLRC subunits is not well understood. In this study, we identified Set3, a core subunit of the Set3/Hos2 histone deacetylase complex (Set3C), as a contributor to the integrity and silencing of heterochromatin at centromeres, telomeres and silent mating-type locus. This novel role of Set3 relies on its PHD finger, but is independent of deacetylase activity or structural integrity of Set3C. Set3 is not located to the centromeric region. Instead, Set3 is targeted to the promoters of clr4(+) and rik1(+), probably through its PHD finger. Set3 promotes transcription of clr4(+) and rik1(+). Consistently, the protein levels of Clr4 and Rik1 were reduced in the set3Δ mutant. The heterochromatin silencing defect in the set3Δ mutant could be rescued by overexpressing of clr4(+) or rik1(+). Our study suggests transcriptional activation of essential heterochromatin factors underlies the tight regulation of heterochromatin integrity. PMID:27538348
Relative efficacy of egg yolk and soya milk-based extenders for cryopreservation (−196°C) of buffalo semen
Chaudhari, D.V.; Dhami, A. J.; Hadiya, K. K.; Patel, J. A.
Aim: The aim was to compare commercially available soybean milk-based extenders, viz. Bioxcell® and Optixcell® (IMV, France) with standard Tris-citrate-fructose-egg yolk-glycerol (TFYG) extender for cryopreservation of buffalo semen. Materials and Methods: Semen was collected twice a week in artificial vagina from six sexually mature, 4-6 years old, healthy breeding bulls of Surti buffalo breed. In all 48 qualifying ejaculates (8 per bull) having initial motility >70% were split into three equal aliquots and were diluted (at 34°C keeping 100×106 sperm ml−1) in TFYG, Bioxcell and Optixcell extenders. The French mini straws filled from each aliquot were gradually cooled to 4-5°C, equilibrated at 4°C for 4 h and frozen in liquid nitrogen 2 vapor using programmable biofreezer. Just before freezing (post-equilibration) and 24 h after frozen storage, the samples were evaluated for various sperm quality parameters using standard protocols. Frozen semen straws were thawed in a water bath at 37°C for 30 s. The post-thaw incubation survival (37°C for 1 h) was assessed through motility rating at 0, 30 and 60 min of incubation. Results: The mean percentages of prefreeze sperms in TFYG, Bioxcell and Optixcell extenders in terms of progressive motility (69.48±0.37, 68.02±0.49, 70.94±0.38), viability (79.21±0.39, 77.38±0.48, 81.58±0.38), total abnormalities (7.90±0.14, 8.60±0.16, 7.08±0.15), intact acrosome (89.54± 0.18, 88.58±0.22, 90.52±0.21) and hypoosmotic swelling (HOS) reactivity (67.96±0.32, 65.65±0.42, 70.23±0.37) varied significantly (p<0.05) between extenders. Similar pattern of significant (p<0.05) variations between these extenders for post-thaw sperm progressive motility (47.71±0.79, 44.38±0.85, 49.90±0.90), viability (57.19±0.79, 53.85±0.84, 59.67±0.91), total abnormalities (12.33±0.17, 12.75±0.21, 11.27±0.18), intact acrosome (76.83±0.23, 75.90± 0.27, 78.50±0.25) and HOS reactivity (45.02±0.84, 42.31±0.82, 47.81±0.90) was
Assessment of semen quality in pure and crossbred Jersey bulls
Kumar, Umesh; Gawande, Ajay P.; Sahatpure, Sunil K.; Patil, Manoj S.; Lakde, Chetan K.; Bonde, Sachin W.; Borkar, Pradnyankur L.; Poharkar, Ajay J.; Ramteke, Baldeo R.
Aim: To compare the seminal attributes of neat, pre-freeze (at equilibration), and post-freeze (24 h after freezing) semen in pure and crossbred Jersey bulls. Materials and Methods: Total 36 ejaculates (3 ejaculates from each bull) were collected from 6 pure Jersey and 6 crossbred Jersey bulls and evaluated for various seminal attributes during neat, pre-freeze, and post-freeze semen. Results: The mean (±standard error [SE]) values of neat semen characteristics in pure and crossbred Jersey bulls were recorded such as volume (ml), color, consistency, mass activity (scale: 0-5), and sperm concentration (millions/ml). The extended semen was further investigated at pre-freeze and post-freeze stages and the mean (±SE) values recorded at neat, pre-freeze, and post-freeze semen were compared between pure and crossbred Jersey bulls; sperm motility (80.55±1.70%, 62.77±1.35%, 46.11±1.43% vs. 80.00±1.80%, 65.00±1.66%, 47.22±1.08%), live sperm count (83.63±1.08%, 71.72±1.09%, 58.67±1.02% vs. 80.00±1.08%, 67.91±1.20%, 51.63±0.97%), total abnormal sperm count (8.38±0.32%, 12.30±0.39%, 16.75±0.42% vs. 9.00±0.45%, 12.19±0.48%, 18.11±0.64%), hypo-osmotic swelling (HOS) reacted spermatozoa (71.88±0.77%, 62.05±0.80%, 47.27±1.05% vs. 72.77±1.02%, 62.11±0.89%, 45.94±1.33%), acrosome integrity (89.05±0.83%, 81.33±0.71%, 71.94±0.86% vs. 86.55±0.57%, 78.66±0.42%, 69.38±0.53%), and DNA integrity (99.88±0.07%, 100, 99.66±0.11% vs. 99.94±0.05%, 100, 99.44±0.18%,). The volume, color, consistency, sperm concentration, and initial motility in pure and crossbred Jersey bulls did not differ significantly (p>0.05). The mass activity was significantly (p<0.05) higher in pure Jersey as compare to crossbred Jersey bulls. Live sperm percentage and acrosome integrity was significantly (p<0.01) higher in pure Jersey bulls as compared to crossbred Jersey bulls. However, no statistical difference (p>0.05) was observed in abnormal sperm; HOS reacted spermatozoa and DNA
Experimental and Computational Studies of the Isomerization Reactions of Bidentate Phosphine Ligands in Triosmium Clusters
Xue, Zhang; Kandala, Srikanth; Yang, Li; Watson, William H.; Wang, Xiaoping; Hrovat, David A.
The diphosphine ligand 1,2-bis(diphenylphosphino)benzene (dppbz) reacts with the activated cluster 1,2-Os{sub 3}(CO){sub 10}(MeCN){sub 2} (1) at room temperature to furnish a mixture of the triosmium clusters 1,2-Os{sub 3}(CO){sub 10}(dppbz) (2) and 1,1-Os{sub 3}(CO){sub 10}(dppbz) (3), along with a trace amount of the hydride cluster HOs{sub 3}(CO){sub 9}[{mu}-1,2-PhP(C{sub 6}H{sub 4}-{eta}{sup 1})C{sub 6}H{sub 4}PPh{sub 2}] (4). The dppbz-bridged cluster 2 forms as the kinetically controlled product and irreversibly transforms to the corresponding chelated isomer 3 at ambient temperature. The disposition of the dppbz ligand in 2 and 3 has been established by X-ray crystallography and {sup 31}P NMR spectroscopy, and the kinetics for the conversion 2 {yields} 3 have been followed by UV-vis spectroscopy in toluene over the temperature range 318-343 K. The calculated activation parameters ({Delta}H{sub {+-}} = 21.6(3) kcal/mol; {Delta}S{sub {+-}} = -11(1) eu) and lack of CO inhibition support an intramolecular isomerization mechanism that involves the simultaneous migration of phosphine and CO groups about the cluster polyhedron. The reaction between 1 and the fluorinated diphosphine ligand 1,2-bis(diphenylphosphino)tetrafluorobenzene (dppbzF{sub 4}) was examined under similar reaction conditions and was found to afford the chelated cluster 1,1-Os{sub 3}(CO){sub 10}(dppbzF{sub 4}) (6) as the sole observable product. The absence of the expected bridged isomer 1,2-Os{sub 3}(CO){sub 10}(dppbzF{sub 4}) (5) suggests that the dppbzF{sub 4} ligand destabilizes 5, thus accounting for the rapid isomerization of 5 to 6. Near-UV irradiation of clusters 3 and 6 leads to CO loss and ortho metalation of an ancillary aryl group. The resulting hydride clusters 4 and HOs{sub 3}(CO){sub 9}[{mu}-1,2-PhP(C{sub 6}H{sub 4}-{eta}{sup 1})C{sub 6}F{sub 4}PPh{sub 2}] (7) have been isolated and fully characterized by spectroscopic and X-ray diffraction analyses. Both 4 and 7 react with added
Nonlinearity detection for condition monitoring utilizing higher-order spectral analysis diagnostics
NASA Astrophysics Data System (ADS)
Park, Hyeonsu
In this dissertation, we investigate the theory and application of higher-order spectral analysis techniques to condition monitoring in shipboard electrical power systems. Monitoring and early detection of faults in rotating machines, such as induction motors, are essential for both preventive maintenance and to avoid potentially severe damage. As machines degrade, they often tend to become more nonlinear. This increased nonlinearity results in the introduction of new frequencies which satisfy particular frequency selection rules; the exact selection rule depends on the order of the nonlinearity. In addition, the phases of the newly generated frequencies satisfy a similar phase selection rule. This results in a phase coherence, or phase coupling, between the "original" interacting frequencies and the "new" frequencies. This phase coupling is a true signature of nonlinearity. Since the classical auto-power spectrum contains no phase information, the phase coupling signature associated with nonlinear interactions is not available. However, various higher-order spectra (HOS) are capable of detecting such nonlinear-induced phase coupling. The efficacy of the various proposed HOS-based methodologies is investigated using real-world vibration time-series data from a faulted induction motor driving a dc generator. The fault is controlled by varying a resistor placed in one phase of the three-phase line to the induction motor. First, we propose a novel method using a bispectral change detection (BCD) for condition monitoring. Even though the bicoherence is dominant and powerful in the detection of phase coupling of nonlinearly interacting frequencies, it has some difficulties in its application to machine condition monitoring. Basically, the bicoherence may not be able to distinguish between intrinsic nonlinearities associated with healthy machines and fault-induced nonlinearities. Therefore, the ability to discriminate the fault-only nonlinearities from the intrinsic
Alleviating anastrozole induced bone toxicity by selenium nanoparticles in SD rats
Vekariya, Kiritkumar K.; Kaur, Jasmine; Tikoo, Kulbhushan
Aromatase inhibitors like anastrozole play an undisputed key role in the treatment of breast cancer, but on the other hand, various side effects like osteoporosis and increased risk of bone fracture accompany the chronic administration of these drugs. Here we show for the first time that selenium nanoparticles, when given in conjugation to anastrozole, lower the bone toxicity caused by anastrozole and thus reduce the probable damage to the bone. Selenium nanoparticles at a dose of 5 μg/ml significantly reduced the cell death caused by anastrozole (1 μM) in HOS (human osteoblast) cells. In addition, our results also highlighted that in female SD rat model, SeNPs (0.25, 0.5, 1 mg/kg/day) significantly prevented the decrease in bone density and increase in biochemical markers of bone resorption induced by anastrozole (0.2 mg/kg/day) treatment. Histopathological examination of the femurs of SeNP treated group revealed ossification, mineralization, calcified cartilaginous deposits and a marginal osteoclastic activity, all of which indicate a marked restorative action, suggesting the protective action of the SeNPs. Interestingly, SeNPs (1 mg/kg/day) also exhibited protective effect in ovariectomized rat model, by preventing osteoporosis, which signifies that bone loss due to estrogen deficiency can be effectively overcome by using SeNPs. - Highlights: ► SeNPs significantly reduce bone toxicity in anastrozole treated rats. ► SeNPs successfully prevented osteoporosis in ovariectomized rats. ► SeNP treatment lowered the levels of TRAP and increased the levels of ALKP.
Automated diagnosis of Age-related Macular Degeneration using greyscale features from digital fundus images.
Mookiah, Muthu Rama Krishnan; Acharya, U Rajendra; Koh, Joel E W; Chandran, Vinod; Chua, Chua Kuang; Tan, Jen Hong; Lim, Choo Min; Ng, E Y K; Noronha, Kevin; Tong, Louis; Laude, Augustinus
Age-related Macular Degeneration (AMD) is one of the major causes of vision loss and blindness in ageing population. Currently, there is no cure for AMD, however early detection and subsequent treatment may prevent the severe vision loss or slow the progression of the disease. AMD can be classified into two types: dry and wet AMDs. The people with macular degeneration are mostly affected by dry AMD. Early symptoms of AMD are formation of drusen and yellow pigmentation. These lesions are identified by manual inspection of fundus images by the ophthalmologists. It is a time consuming, tiresome process, and hence an automated diagnosis of AMD screening tool can aid clinicians in their diagnosis significantly. This study proposes an automated dry AMD detection system using various entropies (Shannon, Kapur, Renyi and Yager), Higher Order Spectra (HOS) bispectra features, Fractional Dimension (FD), and Gabor wavelet features extracted from greyscale fundus images. The features are ranked using t-test, Kullback-Lieber Divergence (KLD), Chernoff Bound and Bhattacharyya Distance (CBBD), Receiver Operating Characteristics (ROC) curve-based and Wilcoxon ranking methods in order to select optimum features and classified into normal and AMD classes using Naive Bayes (NB), k-Nearest Neighbour (k-NN), Probabilistic Neural Network (PNN), Decision Tree (DT) and Support Vector Machine (SVM) classifiers. The performance of the proposed system is evaluated using private (Kasturba Medical Hospital, Manipal, India), Automated Retinal Image Analysis (ARIA) and STructured Analysis of the Retina (STARE) datasets. The proposed system yielded the highest average classification accuracies of 90.19%, 95.07% and 95% with 42, 54 and 38 optimal ranked features using SVM classifier for private, ARIA and STARE datasets respectively. This automated AMD detection system can be used for mass fundus image screening and aid clinicians by making better use of their expertise on selected images that
The Hemagglutinin of Canine Distemper Virus Determines Tropism and Cytopathogenicity
von Messling, Veronika; Zimmer, Gert; Herrler, Georg; Haas, Ludwig; Cattaneo, Roberto
Canine distemper virus (CDV) and measles virus (MV) cause severe illnesses in their respective hosts. The viruses display a characteristic cytopathic effect by forming syncytia in susceptible cells. For CDV, the proficiency of syncytium formation varies among different strains and correlates with the degree of viral attenuation. In this study, we examined the determinants for the differential fusogenicity of the wild-type CDV isolate 5804Han89 (CDV5804), the small- and large-plaque-forming variants of the CDV vaccine strain Onderstepoort (CDVOS and CDVOL, respectively), and the MV vaccine strain Edmonston B (MVEdm). The cotransfection of different combinations of fusion (F) and hemagglutinin (H) genes in Vero cells indicated that the H protein is the main determinant of fusion efficiency. To verify the significance of this observation in the viral context, a reverse genetic system to generate recombinant CDVs was established. This system is based on a plasmid containing the full-length antigenomic sequence of CDVOS. The coding regions of the H proteins of all CDV strains and MVEdm were introduced into the CDV and MV genetic backgrounds, and recombinant viruses rCDV-H5804, rCDV-HOL, rCDV-HEdm, rMV-H5804, rMV-HOL, and rMV-HOS were recovered. Thus, the H proteins of the two morbilliviruses are interchangeable and fully functional in a heterologous complex. This is in contrast with the glycoproteins of other members of the family Paramyxoviridae, which do not function efficiently with heterologous partners. The fusogenicity, growth characteristics, and tropism of the recombinant viruses were examined and compared with those of the parental strains. All these characteristics were found to be predominantly mediated by the H protein regardless of the viral backbone used. PMID:11413309
The HSP90 inhibitor alvespimycin enhances the potency of telomerase inhibition by imetelstat in human osteosarcoma
Hu, Yafang; Bobb, Daniel; He, Jianping; Hill, D Ashley; Dome, Jeffrey S
The unsatisfactory outcomes for osteosarcoma necessitate novel therapeutic strategies. This study evaluated the effect of the telomerase inhibitor imetelstat in pre-clinical models of human osteosarcoma. Because the chaperone molecule HSP90 facilitates the assembly of telomerase protein, the ability of the HSP90 inhibitor alvespimycin to potentiate the effect of the telomerase inhibitor was assessed. The effect of single or combined treatment with imetelstat and alvespimycin on long-term growth was assessed in osteosarcoma cell lines (143B, HOS and MG-63) and xenografts derived from 143B cells. Results indicated that imetelstat as a single agent inhibited telomerase activity, induced telomere shortening, and inhibited growth in all 3 osteosarcoma cell lines, though the bulk cell cultures did not undergo growth arrest. Combined treatment with imetelstat and alvespimycin resulted in diminished telomerase activity and shorter telomeres compared to either agent alone as well as higher levels of γH2AX and cleaved caspase-3, indicative of increased DNA damage and apoptosis. With dual telomerase and HSP90 inhibition, complete growth arrest of bulk cell cultures was achieved. In xenograft models, all 3 treatment groups significantly inhibited tumor growth compared with the placebo-treated control group, with the greatest effect seen in the combined treatment group (imetelstat, p = 0.045, alvespimycin, p = 0.034; combined treatment, p = 0.004). In conclusion, HSP90 inhibition enhanced the effect of telomerase inhibition in pre-clinical models of osteosarcoma. Dual targeting of telomerase and HSP90 warrants further investigation as a therapeutic strategy. PMID:25920748
Extender osmolality and sugar supplementation exert a complex effect on the cryopreservation of Iberian red deer (Cervus elaphus hispanicus) epididymal spermatozoa.
Fernández-Santos, M R; Martínez-Pastor, F; García-Macías, V; Esteso, M C; Soler, A J; de Paz, P; Anel, L; Garde, J J
We have carried out two experiments to study the cryobiology of red deer epididymal spermatozoa and to improve freezing extenders: (1) effect of extender (Tris-citrate-fructose) osmolality (300-600 mOsm/kg), and (2) effect of sugar (0.4M) supplementation to the extender (no sugar, glucose, fructose, mannose, sucrose, maltose, threalose and raffinose). Sperm quality was assessed pre-freezing, post-thawing, and after 2h at 37 degrees C post-thawing: sperm motility index (SMI), acrosome integrity and membrane integrity (HOS test) were assessed subjectively; mitochondrial activity (JC-1) and membrane stability (merocyanine 540) were assessed by flow cytometry. In experiment 2, DNA status was assessed using acridine orange and flow cytometry. To find an optimal osmolality, we fitted the data to a quadratic curve. Four hundred Osmolal per kilogram rendered better results pre-freezing and post-thawing. However, post-thawing viability and most parameters after 2h incubation fitted a linear model. Osmolalities above 425 mOsm/kg were deleterious (P<0.05). In experiment 2, fructose had a positive effect respect to control after 2h of incubation at 37 degrees C post-thawing. Di and trisaccharides were deleterious. Trehalose showed impaired DNA status after 2h incubation. In conclusion, the osmolality of the extenders should be around 400 mOsm/kg, although the change from quadratic to lineal may indicate a complex effect that must be further studied. Monosaccharides may enhance red deer epididymal sperm cryopreservation, especially fructose, whereas di and trisaccharides may not be adequate. PMID:17140651
Reduced glycosylation of human cell lines increases susceptibility to CD4-independent infection by human immunodeficiency virus type 2 (LAV-2/B).
Talbot, S J; Weiss, R A; Schulz, T F
The human immunodeficiency virus type 2 (HIV-2) strain LAV-2/B is able to infect a variety of human cell lines via a CD4-independent pathway. We have used the glycosylation inhibitors tunicamycin, swainsonine, and deoxymannojirimycin to further characterize this putative alternative receptor for HIV-2 (LAV-2/B). These antibiotics resulted in an increase (5- to 30-fold) in the susceptibility of a variety of CD4- human cell lines to infection by LAV-2/B (RD, HeLa, HT29, Rsb, Heb7a, Hos, and Daudi). Several nonprimate cell lines (mink Mv-1-lu, rabbit SIRC, hamster a23, mouse NIH 3T3, cat CCC, and rat HSN) remained resistant to infection by LAV-2/B after treatment with glycosylation inhibitors, suggesting that they do not express the HIV-2 CD4-independent receptor. Two of these nonprimate cell lines are readily infected by HIV-2 when they express CD4 (Mv-1-lu and CCC). Treatment of human cells with neuraminidase had no effect on subsequent infection by LAV-2/B, suggesting that the increase in susceptibility to infection of deglycosylated cells is not due to a change in the electrostatic charge of the cell surface. Treatment of RD CD4- cells and HeLa CD4+ cells with a variety of proteases resulted in a 75 to 90% decrease in infection by LAV-2/B when compared with untreated cells. Taken together, all these data suggest that HIV-2 can utilize a membrane glycoprotein other than CD4 to attach and fuse with a variety of human cells. PMID:7745686
Genetic differentiation of Mexican Holstein cattle and its relationship with Canadian and U.S. Holsteins
García-Ruiz, Adriana; Ruiz-López, Felipe de J.; Van Tassell, Curtis P.; Montaldo, Hugo H.; Huson, Heather J.
The Mexican Holstein (HO) industry has imported Canadian and US (CAN + USA) HO germplasm for use in two different production systems, the conventional (Conv) and the low income (Lowi) system. The objective of this work was to study the genetic composition and differentiation of the Mexican HO cattle, considering the production system in which they perform and their relationship with the Canadian and US HO populations. The analysis included information from 149, 303, and 173 unrelated or with unknown pedigree HO animals from the Conv, Lowi, and CAN + USA populations, respectively. Canadian and US Jersey (JE) and Brown Swiss (BS) genotypes (162 and 86, respectively) were used to determine if Mexican HOs were hybridized with either of these breeds. After quality control filtering, a total of 6,617 out of 6,836 single nucleotide polymorphism markers were used. To describe the genetic diversity across the populations, principal component (PC), admixture composition, and linkage disequilibrium (LD; r2) analyses were performed. Through the PC analysis, HO × JE and HO × BS crossbreeding was detected in the Lowi system. The Conv system appeared to be in between Lowi and CAN + USA populations. Admixture analysis differentiated between the genetic composition of the Conv and Lowi systems, and five ancestry groups associated to sire's country of origin were identified. The minimum distance between markers to estimate a useful LD was found to be 54.5 kb for the Mexican HO populations. At this average distance, the persistence of phase across autosomes of Conv and Lowi systems was 0.94, for Conv and CAN + USA was 0.92 and for the Lowi and CAN + USA was 0.91. Results supported the flow of germplasm among populations being Conv a source for Lowi, and dependent on migration from CAN + USA. Mexican HO cattle in Conv and Lowi populations share common ancestry with CAN + USA but have different genetic signatures. PMID:25709615
The use of kernel local Fisher discriminant analysis for the channelization of the Hotelling model observer
NASA Astrophysics Data System (ADS)
Wen, Gezheng; Markey, Mia K.
It is resource-intensive to conduct human studies for task-based assessment of medical image quality and system optimization. Thus, numerical model observers have been developed as a surrogate for human observers. The Hotelling observer (HO) is the optimal linear observer for signal-detection tasks, but the high dimensionality of imaging data results in a heavy computational burden. Channelization is often used to approximate the HO through a dimensionality reduction step, but how to produce channelized images without losing significant image information remains a key challenge. Kernel local Fisher discriminant analysis (KLFDA) uses kernel techniques to perform supervised dimensionality reduction, which finds an embedding transformation that maximizes betweenclass separability and preserves within-class local structure in the low-dimensional manifold. It is powerful for classification tasks, especially when the distribution of a class is multimodal. Such multimodality could be observed in many practical clinical tasks. For example, primary and metastatic lesions may both appear in medical imaging studies, but the distributions of their typical characteristics (e.g., size) may be very different. In this study, we propose to use KLFDA as a novel channelization method. The dimension of the embedded manifold (i.e., the result of KLFDA) is a counterpart to the number of channels in the state-of-art linear channelization. We present a simulation study to demonstrate the potential usefulness of KLFDA for building the channelized HOs (CHOs) and generating reliable decision statistics for clinical tasks. We show that the performance of the CHO with KLFDA channels is comparable to that of the benchmark CHOs.
Rapid phosphatidic acid accumulation in response to low temperature stress in Arabidopsis is generated through diacylglycerol kinase.
Arisz, Steven A; van Wijk, Ringo; Roels, Wendy; Zhu, Jian-Kang; Haring, Michel A; Munnik, Teun
Phosphatidic acid (PtdOH) is emerging as an important signaling lipid in abiotic stress responses in plants. The effect of cold stress was monitored using (32)P-labeled seedlings and leaf discs of Arabidopsis thaliana. Low, non-freezing temperatures were found to trigger a very rapid (32)P-PtdOH increase, peaking within 2 and 5 min, respectively. In principle, PtdOH can be generated through three different pathways, i.e., (1) via de novo phospholipid biosynthesis (through acylation of lyso-PtdOH), (2) via phospholipase D hydrolysis of structural phospholipids, or (3) via phosphorylation of diacylglycerol (DAG) by DAG kinase (DGK). Using a differential (32)P-labeling protocol and a PLD-transphosphatidylation assay, evidence is provided that the rapid (32)P-PtdOH response was primarily generated through DGK. A simultaneous decrease in the levels of (32)P-PtdInsP, correlating in time, temperature dependency, and magnitude with the increase in (32)P-PtdOH, suggested that a PtdInsP-hydrolyzing PLC generated the DAG in this reaction. Testing T-DNA insertion lines available for the seven DGK genes, revealed no clear changes in (32)P-PtdOH responses, suggesting functional redundancy. Similarly, known cold-stress mutants were analyzed to investigate whether the PtdOH response acted downstream of the respective gene products. The hos1, los1, and fry1 mutants were found to exhibit normal PtdOH responses. Slight changes were found for ice1, snow1, and the overexpression line Super-ICE1, however, this was not cold-specific and likely due to pleiotropic effects. A tentative model illustrating direct cold effects on phospholipid metabolism is presented. PMID:23346092
Combined MET inhibition and topoisomerase I inhibition block cell growth of small cell lung cancer.
Rolle, Cleo E; Kanteti, Rajani; Surati, Mosmi; Nandi, Suvobroto; Dhanasingh, Immanuel; Yala, Soheil; Tretiakova, Maria; Arif, Qudsia; Hembrough, Todd; Brand, Toni M; Wheeler, Deric L; Husain, Aliya N; Vokes, Everett E; Bharti, Ajit; Salgia, Ravi
Small cell lung cancer (SCLC) is a devastating disease, and current therapies have not greatly improved the 5-year survival rates. Topoisomerase (Top) inhibition is a treatment modality for SCLC; however, the response is short lived. Consequently, our research has focused on improving SCLC therapeutics through the identification of novel targets. Previously, we identified MNNG HOS transforming gene (MET) to be overexpressed and functional in SCLC. Herein, we investigated the therapeutic potential of combinatorial targeting of MET using SU11274 and Top1 using 7-ethyl-10-hydroxycamptothecin (SN-38). MET and TOP1 gene copy numbers and protein expression were determined in 29 patients with limited (n = 11) and extensive (n = 18) disease. MET gene copy number was significantly increased (>6 copies) in extensive disease compared with limited disease (P = 0.015). Similar TOP1 gene copy numbers were detected in limited and extensive disease. Immunohistochemical staining revealed a significantly higher Top1 nuclear expression in extensive (0.93) versus limited (0.15) disease (P = 0.04). Interestingly, a significant positive correlation was detected between MET gene copy number and Top1 nuclear expression (r = 0.5). In vitro stimulation of H82 cells revealed hepatocyte growth factor (HGF)-induced nuclear colocalization of p-MET and Top1. Furthermore, activation of the HGF/MET axis enhanced Top1 activity, which was abrogated by SU11274. Combination of SN-38 with SU11274 dramatically decreased SCLC growth as compared with either drug alone. Collectively, these findings suggest that the combinatorial inhibition of MET and Top1 is a potentially efficacious treatment strategy for SCLC. PMID:24327519
ADRA2A is involved in neuro-endocrine regulation of bone resorption
Mlakar, Vid; Jurkovic Mlakar, Simona; Zupan, Janja; Komadina, Radko; Prezelj, Janez; Marc, Janja
Adrenergic stimulation is important for osteoclast differentiation and bone resorption. Previous research shows that this happens through β2-adrenergic receptor (AR), but there are conflicting evidence on presence and role of α2A-AR in bone. The aim of this study was to investigate the presence of α2A-AR and its involvement in neuro-endocrine signalling of bone remodelling in humans. Real-time polymerase chain reaction (PCR) and immunohistochemistry were used to investigate α2A-AR receptor presence and localization in bone cells. Functionality of rs553668 and rs1800544 single nucleotide polymorphism SNPs located in α2A-AR gene was analysed by qPCR expression on bone samples and luciferase reporter assay in human osteosarcoma HOS cells. Using real-time PCR, genetic association study between rs553668 A>G and rs1800544 C>G SNPs and major bone markers was performed on 661 Slovenian patients with osteoporosis. α2A-AR is expressed in osteoblasts and lining cells but not in osteocytes. SNP rs553668 has a significant influence on α2A-AR mRNA level in human bone samples through the stability of mRNA. α2A-AR gene locus associates with important bone remodelling markers (BMD, CTX, Cathepsin K and pOC). The results of this study are providing comprehensive new evidence that α2A-AR is involved in neuro-endocrine signalling of bone turnover and development of osteoporosis. As shown by our results the neurological signalling is mediated through osteoblasts and result in bone resorption. Genetic study showed association of SNPs in α2A-AR gene locus with bone remodelling markers, identifying the individuals with higher risk of development of osteoporosis. PMID:25818344
A bulk segregant gene expression analysis of a peach population reveals components of the underlying mechanism of the fruit cold response.
Pons, Clara; Martí, Cristina; Forment, Javier; Crisosto, Carlos H; Dandekar, Abhaya M; Granell, Antonio
Peach fruits subjected for long periods of cold storage are primed to develop chilling injury once fruits are shelf ripened at room temperature. Very little is known about the molecular changes occurring in fruits during cold exposure. To get some insight into this process a transcript profiling analyses was performed on fruits from a PopDG population segregating for chilling injury CI responses. A bulked segregant gene expression analysis based on groups of fruits showing extreme CI responses indicated that the transcriptome of peach fruits was modified already during cold storage consistently with eventual CI development. Most peach cold-responsive genes have orthologs in Arabidopsis that participate in cold acclimation and other stresses responses, while some of them showed expression patterns that differs in fruits according to their susceptibility to develop mealiness. Members of ICE1, CBF1/3 and HOS9 regulons seem to have a prominent role in differential cold responses between low and high sensitive fruits. In high sensitive fruits, an alternative cold response program is detected. This program is probably associated with dehydration/osmotic stress and regulated by ABA, auxins and ethylene. In addition, the observation that tolerant siblings showed a series of genes encoding for stress protective activities with higher expression both at harvest and during cold treatment, suggests that preprogrammed mechanisms could shape fruit ability to tolerate postharvest cold-induced stress. A number of genes differentially expressed were validated and extended to individual genotypes by medium-throughput RT-qPCR. Analyses presented here provide a global view of the responses of peach fruits to cold storage and highlights new peach genes that probably play important roles in the tolerance/sensitivity to cold storage. Our results provide a roadmap for further experiments and would help to develop new postharvest protocols and gene directed breeding strategies to better
SUB1 Plays a Negative Role during Starvation Induced Sporulation Program in Saccharomyces cerevisiae
Gupta, Ritu; Vijayraghavan, Usha
Saccharomyces cerevisiae Sub1 is involved in several cellular processes such as, transcription initiation, elongation, mRNA processing and DNA repair. It has also been reported to provide cellular resistance during conditions of oxidative DNA damage and osmotic stress. Here, we report a novel role of SUB1 during starvation stress-induced sporulation, which leads to meiosis and spore formation in diploid yeast cells. Deletion of SUB1 gene significantly increased sporulation efficiency as compared to the wild-type cells in S288c genetic background. Whereas, the sporulation functions of the sub1(Y66A) missense mutant were similar to Sub1. SUB1 transcript and protein levels are downregulated during sporulation, in highly synchronized and sporulation proficient wild-type SK1 cells. The changes in Sub1 levels during sporulation cascade correlate with the induction of middle sporulation gene expression. Deletion of SUB1 increased middle sporulation gene transcript levels with no effect on their induction kinetics. In wild-type cells, Sub1 associates with chromatin at these loci in a temporal pattern that correlates with their enhanced gene expression seen in sub1Δ cells. We show that SUB1 genetically interacts with HOS2, which led us to speculate that Sub1 might function with Set3 repressor complex during sporulation. Positive Cofactor 4, human homolog of Sub1, complemented the sub1Δ sporulation phenotype, suggesting conservation of function. Taken together, our results suggest that SUB1 acts as a negative regulator of sporulation. PMID:26147804
Further evidence against a direct genotoxic mode of action for arsenic-induced cancer
Klein, Catherine B.; Leszczynska, Joanna; Hickey, Christina; Rossman, Toby G.
Arsenic in drinking water, a mixture of arsenite and arsenate, is associated with increased skin and other cancers in Asia and Latin America, but not the United States. Arsenite alone in drinking water does not cause skin cancers in experimental animals; therefore, it is not a complete carcinogen in skin. We recently showed that low concentrations of arsenite enhanced the tumorigenicity of solar UV irradiation in hairless mice, suggesting arsenic cocarcinogenesis with sunlight in skin cancer and perhaps with different carcinogenic partners for lung and bladder tumors. Cocarcinogenic mechanisms could include blocking DNA repair, stimulating angiogenesis, altering DNA methylation patterns, dysregulating cell cycle control, induction of aneuploidy and blocking apoptosis. Arsenicals are documented clastogens but not strong mutagens, with weak mutagenic activity reported at highly toxic concentrations of inorganic arsenic. Previously, we showed that arsenite, but not monomethylarsonous acid (MMA[III]), induced delayed mutagenesis in HOS cells. Here, we report new data on the mutagenicity of the trivalent methylated arsenic metabolites MMA(III) and dimethylarsinous acid [DMA(III)] at the gpt locus in Chinese hamster G12 cells. Both methylated arsenicals seemed mutagenic with apparent sublinear dose responses. However, significant mutagenesis occurred only at highly toxic concentrations of MMA(III). Most mutants induced by MMA(III) and DMA(III) exhibited transgene deletions. Some non-deletion mutants exhibited altered DNA methylation. A critical discussion of cell survival leads us to conclude that clastogenesis occurs primarily at highly cytotoxic arsenic concentrations, casting further doubt as to whether a genotoxic mode of action (MOA) for arsenicals is supportable.
Eradication of osteosarcoma by fluorescence-guided surgery with tumor labeling by a killer-reporter adenovirus.
Yano, Shuya; Miwa, Shinji; Kishimoto, Hiroyuki; Urata, Yasuo; Tazawa, Hiroshi; Kagawa, Shunsuke; Bouvet, Michael; Fujiwara, Toshiyoshi; Hoffman, Robert M
In a previous study, we developed fluorescence-guided surgery (FGS) for osteosarcoma using an orthotopic model with 143B human osteosarcoma cells expressing red fluorescent protein (RFP) implanted into the intramedullary cavity of the tibia in nude mice. The FGS-treated mice had a significantly higher disease-free survival (DFS) rate than the bright-light surgery (BLS). However, although FGS significantly reduced the recurrence of the primary tumor, it did not reduce lung metastasis. In the present study, we utilized the OBP-401 telomerase-dependent killer-reporter adenovirus, carrying green fluorescent protein (GFP), to label human osteosarcoma in situ in orthotopic mouse models. OBP-401-illuminated human osteosarcoma cell lines, 143B and MNNG/HOS cells in vitro and in vivo. OBP-401 tumor illumination enabled effective FGS of the 143B-derived orthotopic mouse model of human osteosarcoma model as well as FGS eradication of residual cancer cells after BLS. OBP-401-assisted FGS significantly inhibited local recurrence and lung metastasis after surgery, thereby prolonging DFS and overall survival (OS), achieving a very important improvement of therapeutic outcomes over our previously reported FGS study. These therapeutic benefits of FGS were demonstrated using a clinically-viable methodology of direct labeling of human osteosarcoma in situ with the OBP-401 killer-reporter adenovirus in contrast with previous reports, which used genetically engineered labeled cells or antibody-based fluorescent labels for FGS. © 2015 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 34:836-844, 2016. PMID:26479501
Delayed germination of Arabidopsis seeds under chilling stress by overexpressing an abiotic stress inducible GhTPS11.
Wang, Cai-Li; Zhang, Shi-Cai; Qi, Sheng-Dong; Zheng, Cheng-Chao; Wu, Chang-Ai
Trehalose-6-phosphate synthase (TPS) plays an important role in metabolic regulation and stress responses in a variety of organisms. However information about cotton TPS is poor. Here a cotton TPS gene GhTPS11 was isolated and characterized. Expression analysis revealed that GhTPS11 was induced in 20-day old cotton seedlings by heat drought and high salt stresses as well as GA and ABA. Moreover GhTPS11 was induced by chilling stress and mannitol while was depressed by sucrose. Tissue expression analysis indicated that GhTPS11 expressed higher in leaves than in stems and roots of 20-day old cotton seedlings. The GhTPS11 overexpressing Arabidopsis seeds germinated slower than the wild-type (WT) under chilling stress. Trehalose-6-phosphate (T6P) and trehalose contents were evidently higher in GhTPS11 overexpressing lines 3, 5, and 22 than in WT under normal germination condition as well as chilling stress. Further analysis demonstrated that the expression of ICE1 CBF3 and RCI2A was induced lower whereas that of CBF1 and CBF2 was induced higher under chilling stress in the GhTPS11 overexpressing seeds than WT respectively. These results suggested that GhTPS11 encoded a stress-responsive TPS protein and functioned in chilling stress during seed germination. Perhaps the chilling stress sensitivity of transgenic Arabidopsis seeds was caused by the expression changes of at least some chilling-related genes such as ICE1 CBFs and RCI2A other than HOS1. So this article provided the useful information for GhTPS11 usage for crop molecular breeding. PMID:26325072
Study on the description method of upper limb's muscle force levels during simulated in-orbit operations
NASA Astrophysics Data System (ADS)
Zhao, Yan; Li, DongXu; Liu, ZhiZhen; Liu, Liang
The dexterous upper limb serves as the most important tool for astronauts to implement in-orbit experiments and operations. This study developed a simulated weightlessness experiment and invented new measuring equipment to quantitatively evaluate the muscle ability of the upper limb. Isometric maximum voluntary contractions (MVCs) and surface electromyography (sEMG) signals of right-handed pushing at the three positions were measured for eleven subjects. In order to enhance the comprehensiveness and accuracy of muscle force assessment, the study focused on signal processing techniques. We applied a combination method, which consists of time-, frequency-, and bi-frequency-domain analyses. Time- and frequency-domain analyses estimated the root mean square (RMS) and median frequency (MDF) of sEMG signals, respectively. Higher order spectra (HOS) of bi-frequency domain evaluated the maximum bispectrum amplitude ( B max), Gaussianity level (Sg) and linearity level (S l ) of sEMG signals. Results showed that B max, S l , and RMS values all increased as force increased. MDF and Sg values both declined as force increased. The research demonstrated that the combination method is superior to the conventional time- and frequency-domain analyses. The method not only described sEMG signal amplitude and power spectrum, but also deeper characterized phase coupling information and non-Gaussianity and non-linearity levels of sEMG, compared to two conventional analyses. The finding from the study can aid ergonomist to estimate astronaut muscle performance, so as to optimize in-orbit operation efficacy and minimize musculoskeletal injuries.
Preparation of bone-implants by coating hydroxyapatite nanoparticles on self-formed titanium dioxide thin-layers on titanium metal surfaces.
Wijesinghe, W P S L; Mantilaka, M M M G P G; Chathuranga Senarathna, K G; Herath, H M T U; Premachandra, T N; Ranasinghe, C S K; Rajapakse, R P V J; Rajapakse, R M G; Edirisinghe, Mohan; Mahalingam, S; Bandara, I M C C D; Singh, Sanjleena
Preparation of hydroxyapatite coated custom-made metallic bone-implants is very important for the replacement of injured bones of the body. Furthermore, these bone-implants are more stable under the corrosive environment of the body and biocompatible than bone-implants made up of pure metals and metal alloys. Herein, we describe a novel, simple and low-cost technique to prepare biocompatible hydroxyapatite coated titanium metal (TiM) implants through growth of self-formed TiO2 thin-layer (SFTL) on TiM via a heat treatment process. SFTL acts as a surface binder of HA nanoparticles in order to produce HA coated implants. Colloidal HA nanorods prepared by a novel surfactant-assisted synthesis method, have been coated on SFTL via atomized spray pyrolysis (ASP) technique. The corrosion behavior of the bare and surface-modified TiM (SMTiM) in a simulated body fluid (SBF) medium is also studied. The highest corrosion rate is found to be for the bare TiM plate, but the corrosion rate has been reduced with the heat-treatment of TiM due to the formation of SFTL. The lowest corrosion rate is recorded for the implant prepared by heat treatment of TiM at 700 °C. The HA-coating further assists in the passivation of the TiM in the SBF medium. Both SMTiM and HA coated SMTiM are noncytotoxic against osteoblast-like (HOS) cells and are in high-bioactivity. The overall production process of bone-implant described in this paper is in high economic value. PMID:27040209
A zinc(II) quinolinone complex (Et3NH)[Zn(qui)Cl2]: Synthesis, X-ray structure, spectral properties and in vitro cytotoxicity
NASA Astrophysics Data System (ADS)
Buchtík, Roman; Nemec, Ivan; Trávníček, Zdeněk
A new zinc(II) complex with 2-phenyl-3-hydroxy-4(1H)-quinolinone (Hqui) of the composition (Et3NH)[Zn(qui)Cl2] was prepared and characterized by elemental analysis, FT IR, 1D and 2D NMR, and fluorescence spectroscopy, mass spectrometry and single crystal X-ray analysis. The molecular structure is composed of the triethylammonium (Et3NH+) cations and tetrahedral [ZnII(qui)Cl2]- complex anions, in which the Zn(II) atoms are bidentate coordinated by one qui ligand through keto (OK) and phenolate (OP) oxygen atoms and by two chlorido ligands, thus forming the {O2Cl2} donor set, with Zn-OK = 1.9860(14) Å, Zn-OP 1.9961(14) Å and Zn-Cl = 2.2509(6) Å and 2.2266(6) Å. The complex cations are aligned into 1D supramolecular chains through the NH⋯Cl hydrogen bonding between the amine group of the quinolinone ligand and the chlorido ligand of the adjacent complex anion. The amine group from the Et3NH+ cations provides the NH⋯OP hydrogen bond with the phenolate oxygen atoms from the complex anion. Screening of in vitro cytotoxicity of the compound was studied on human osteosarcoma (HOS) and human breast adenocarcinoma (MCF7) cell lines, with IC50 > 50 μM. The fluorescence study showed that the complex exhibits a relatively high integral intensity (29%) as compared to the standard quinine sulfate, and 1.6-fold enhancement of emission with respect to free Hqui.
Spermicidal and contraceptive potential of desgalactotigonin: a prospective alternative of nonoxynol-9.
Chakraborty, Debanjana; Maity, Arindam; Jha, Tarun; Mondal, Nirup Bikash
Crude decoction of Chenopodium album seed showed spermicidal effect at MIC 2 mg/ml in earlier studies. Systematic isolation, characterization and evaluation revealed that the major metabolite Desgalactotigonin (DGT) is the most effective principle in both in vitro and in vivo studies. The in vitro studies comprises (a) rat and human sperm motility and immobilizing activity by Sander-Cramer assay; (b) sperm membrane integrity was observed by HOS test and electron microscopy; (c) microbial potential was examined in Lactobacillus broth culture, and (d) the hemolytic index was determined by using rat RBCs. The in vivo contraceptive efficacy was evaluated by intra uterine application of DGT in rat. Lipid peroxidation and induction of apoptosis by DGT on human spermatozoa were also studied. The minimum effective concentration (MEC) of DGT that induced instantaneous immobilization in vitro was 24.18 µM for rat and 58.03 µM for human spermatozoa. Microbial study indicated DGT to be friendly to Lactobacillus acidophilus. Implantation was prevented in DGT treated uterine horn while no hindrance occurred in the untreated contra lateral side. At the level of EC50, DGT induced apoptosis in human spermatozoa as determined by increased labeling with Annexin-V and decreased polarization of sperm mitochondria. Desgalactotigonin emerged 80 and 2×10(4) times more potent than the decoction and Nonoxynol-9 respectively. It possesses mechanism based detrimental action on both human and rat spermatozoa and spares lactobacilli and HeLa cells at MEC which proves its potential as a superior ingredient for the formulation of a contraceptive safer/compatible to vaginal microflora. PMID:25243914
Acid-, water- and high-temperature-stable ruthenium complexes for the total catalytic deoxygenation of glycerol to propane.
Taher, Deeb; Thibault, Michelle E; Di Mondo, Domenico; Jennings, Michael; Schlaf, Marcel
The ruthenium aqua complexes [Ru(H(2)O)(2)(bipy)(2)](OTf)(2), [cis-Ru(6,6'-Cl(2)-bipy)(2)(OH(2))(2)](OTf)(2), [Ru(H(2)O)(2)(phen)(2)](OTf)(2), [Ru(H(2)O)(3)(2,2':6',2''-terpy)](OTf)(2) and [Ru(H(2)O)(3)(Phterpy)](OTf)(2) (bipy = 2,2'-bipyridine; OTf(-) = triflate; phen = phenanthroline; terpy = terpyridine; Phterpy = 4'-phenyl-2,2':6',2''-terpyridine) are water- and acid-stable catalysts for the hydrogenation of aldehydes and ketones in sulfolane solution. In the presence of HOS(O)(2)CF(3) (triflic acid) as a dehydration co-catalyst they directly convert 1,2-hexanediol to n-hexanol and hexane. The terpyridine complexes are stable and active as catalysts at temperatures > or = 250 degrees C and in either aqueous sulfolane solution or pure water convert glycerol into n-propanol and ultimately propane as the final reaction product in up to quantitative yield. For the terpy complexes the active catalyst is postulated to be a carbonyl species [(4'-R-2,2':6',2''-terpy)Ru(CO)(H(2)O)(2)](OTf)(2) (R = H, Ph) formed by the decarbonylation of aldehydes (hexanal for 1,2-hexanediol and 3-hydroxypropanal for glycerol) generated in the reaction mixture through acid-catalyzed dehydration. The structure of the dimeric complex [{(4'-phenyl-2,2':6',2''-terpy)Ru(CO)}(2)(mu-OCH(3))(2)](OTf)(2) has been determined by single crystal X-ray crystallography (Space group P1 (a = 8.2532(17); b = 12.858(3); c = 14.363(3) A; alpha = 64.38(3); beta = 77.26(3); gamma = 87.12(3) degrees, R = 4.36 %). PMID:19693757
Escape from Mitotic Arrest: An Unexpected Connection Between Microtubule Dynamics and Epigenetic Regulation of Centromeric Chromatin in Schizosaccharomyces pombe.
George, Anuja A; Walworth, Nancy C
Accurate chromosome segregation is necessary to ensure genomic integrity. Segregation depends on the proper functioning of the centromere, kinetochore, and mitotic spindle microtubules and is monitored by the spindle assembly checkpoint (SAC). In the fission yeast Schizosaccharomyces pombe, defects in Dis1, a microtubule-associated protein that influences microtubule dynamics, lead to mitotic arrest as a result of an active SAC and consequent failure to grow at low temperature. In a mutant dis1 background (dis1-288), loss of function of Msc1, a fission yeast homolog of the KDM5 family of proteins, suppresses the growth defect and promotes normal mitosis. Genetic analysis implicates a histone deacetylase (HDAC)-linked pathway in suppression because HDAC mutants clr6-1, clr3∆, and sir2∆, though not hos2∆, also promote normal mitosis in the dis1-288 mutant. Suppression of the dis phenotype through loss of msc1 function requires the spindle checkpoint protein Mad2 and is limited by the presence of the heterochromatin-associated HP1 protein homolog Swi6. We speculate that alterations in histone acetylation promote a centromeric chromatin environment that compensates for compromised dis1 function by allowing for successful kinetochore-microtubule interactions that can satisfy the SAC. In cells arrested in mitosis by mutation of dis1, loss of function of epigenetic determinants such as Msc1 or specific HDACs can promote cell survival. Because the KDM5 family of proteins has been implicated in human cancers, an appreciation of the potential role of this family of proteins in chromosome segregation is warranted. PMID:26510788
Triboluminescent Materials for Smart Optical Damage Sensors for Space Applications
NASA Technical Reports Server (NTRS)
Aggarwal, Mohan D.; Penn, Benjamin G.; Miller, Jim
Triboluminescence is light that is produced by pressure, friction or mechanical shock. New composite materials are constantly being reengi neered in an effort to make lightweight spacecrafts for various NASA missions. For these materials there is interest in monitoring the con dition of the composite in real time to detect any delamination or cr acking due to damage, fatigue or external forces. Methods of periodic inspection of composite structures for mechanical damage such as ult rasonic testing are rather mature. However, there is a need to develop a new technique of damage detection for composites, which could dete ct cracking or delamination from any desired location within a materi al structure in real time. This could provide a valuable tool in the confident use of composite materials for various space applications. Recently, triboluminnescent materials have been proposed as smart sen sors of structural damage. To sense the damage, these materials can b e epoxy bonded or coated in a polymer matrix or embedded in a composi te host structure. When the damage or fracture takes place in the hos t structure, it will lead to the fracture of triboluminescent crystal s resulting in a light emission. This will warn, in real time, that a structural damage has occurred. The triboluminescent emission of the candidate phosphor has to be sufficiently bright, so that the light signal reaching from the point of fracture to the detector through a fiber optic cable is sufficiently strong to be detected. There are a large number of triboluminescent materials, but few satisfy the above criterion. Authors have synthesized a Eu based organic material know n as Europium tetrakis (dibenzoylmethide) triethylammonium .(EuD(sub 4)TEA), one of the bright triboluminescent materials, which is a pote ntial candidate for application as a damage sensor and could be made into a wireless sensor with the addition of microchip, antenna and el ectronics. Preliminary results on the synthesis and
Hyperfine structure in the X8, A[19.1]9 and B[21.68]8 states of HoF and X8 and A[15.6]9 state of HoCl
NASA Astrophysics Data System (ADS)
Dick, M. J.; Linton, C.; Adam, A. G.
High resolution spectra of holmium monofluoride, HoF and holmium monochloride, HoCl, prepared in a laser ablation source, have been obtained using laser induced fluorescence. Spectra of the A[19.1]9-X8 0-0 and 1-0 and B[21.68]8-X8 0-0 bands of HoF and the A[15.6]9-X8 0-0 band of HoCl all show resolved hyperfine structure. Analysis of the spectra yielded magnetic hyperfine parameters, h = 0.2240(5), 0.2210(6), 0.2177(6) and 0.2488(5) cm-1 for the X (v = 0), A (v = 0 and 1) and B (v = 0) states of HoF and 0.2355(32) and 0.2448(29) cm-1 for the X (v = 0) and A (v = 0) states of HoCl, respectively. The following quadrupole coupling constants were obtained for the above six states; eQq0(HoF) = -0.0874(67), -0.0586(44), -0.0579(56), -0.0840(64) cm-1 and eQq0(HoCl) = -0.082(11), -0.060(11) cm-1. Comparison with previously determined values for HoO and HoS show that the ground state magnetic hyperfine structure in HoF and HoCl is entirely due to the Ho 4f electron and is consistent with the ground state, X8, configuration of Ho+{4f10(5I8)6s2}X- (X = F, Cl). Calculations of the ground state magnetic, h(X8), and quadrupole, eQq0(X8) hyperfine parameters from atomic hyperfine parameters are found to be consistent with the observed values for both molecules.
BYL719, a new α-specific PI3K inhibitor: single administration and in combination with conventional chemotherapy for the treatment of osteosarcoma.
Gobin, Bérengère; Huin, Marc Baud'; Lamoureux, François; Ory, Benjamin; Charrier, Céline; Lanel, Rachel; Battaglia, Séverine; Redini, Françoise; Lezot, Frédéric; Blanchard, Frédéric; Heymann, Dominique
It has been established that disturbances in intracellular signaling pathways play a considerable part in the oncologic process. Phosphatidylinositol-3-kinase (PI3K) has become of key interest in cancer therapy because of its high mutation frequency and/or gain in function of its catalytic subunits in cancer cells. We investigated the therapeutic value of BYL719, a new specific PI3Kα inhibitor that blocks the ATP site, on osteosarcoma and bone cells. The in vitro effects of BYL719 on proliferation, apoptosis, and cell cycle were assessed in human and murine osteosarcoma cell. Its impact on bone cells was determined using human mesenchymal stem cells (hMSC) and human CD14+ osteoclast precursors. Two different murine preclinical models of osteosarcoma were used to analyze the in vivo biological activities of BYL719. BYL719 decreased cell proliferation by blocking cell cycle in G0/G1 phase with no outstanding effects on apoptosis cell death in HOS and MOS-J tumor cells. BYL719 inhibited cell migration and can thus be considered as a cytostatic drug for osteosarcoma. In murine preclinical models of osteosarcoma, BYL719 significantly decreased tumor progression and tumor ectopic bone formation as shown by a decrease of Ki67+ cells and tumor vascularization. To explore the maximum therapeutic potential of BYL719, the drug was studied in combination with conventional chemotherapeutic drugs, revealing promising efficacy with ifosfamide. BYL719 also exhibited dual activities on osteoblast and osteoclast differentiation. Overall, the present work shows that BYL719 is a promising drug in either a single or multidrug approach to curing bone sarcoma. PMID:24961790
Development of an Adaptive Multi-Method Algorithm for Automatic Picking of First Arrival Times: Application to Near Surface Seismic Data
NASA Astrophysics Data System (ADS)
Khalaf, A.; Camerlynck, C. M.; Schneider, A. C.; Florsch, N.
Accurate picking of first arrival times plays an important role in many seismic studies, particularly in seismic tomography and reservoirs or aquifers monitoring. Many techniques have been developed for picking first arrivals automatically or semi-automatically, but most of them were developed for seismological purposes which does not attain the accuracy objectives due to the complexity of near surface structures, and to usual low signal-to-noise ratio. We propose a new adaptive algorithm for near surface data based on three picking methods, combining multi-nested windows (MNW), Higher Order Statistics (HOS), and Akaike Information Criterion (AIC). They exploit the benefits of integrating many properties, which reveal the presence of first arrivals, to provide an efficient and robust first arrivals picking. This strategy mimics the human first-break picking, where at the beginning the global trend is defined. Then the exact first-breaks are searched in the vicinity of the now defined trend. In a multistage algorithm, three successive phases are launched, where each of them characterize a specific signal property. Within each phase, the potential picks and their error range are automatically estimated, and then used sequentially as leader in the following phase picking. The accuracy and robustness of the implemented algorithm are successfully validated on synthetic and real data which have special challenges for automatic pickers. The comparison of resulting P-wave arrival times with those picked manually, and other algorithms of automatic picking, demonstrated the reliable performance of the new scheme under different noisy conditions. All parameters of our multi-method algorithm are auto-adaptive thanks to the integration in series of each sub-algorithm results in the flow. Hence, it is nearly a parameter-free algorithm, which is straightforward to implement and demands low computational resources.
Development of antibody arrays for monoclonal antibody Higher Order Structure analysis
Wang, Xing; Li, Qing; Davies, Michael
Antibody arrays were developed to probe a monoclonal antibody's three-dimensional structure (3-D structure). Peptides with overlapping regions were designed to cover the whole mAb light chain and heavy chain, respectively, and used to generate polyclonal antibodies after the conjugation of the peptides to a carrier protein, KLH. It was shown that good peptide specificity was achieved from the antibodies generated. Using more than 30 different polyclonal antibodies to measure the surface epitope distribution, it was shown that the mAb antibody array can detect epitope exposure as low as 0.1% of defined mAb populations. This ELISA-based analysis of mAb epitope exposure can be considered as a measurement of "conformational impurity" in biologics development, similar to the analysis of other product-related impurities such as different forms of glycosylation, deamidation, and oxidation. This analysis of "conformational impurity" could provide valuable information on the mAb conformational comparability for biosimilar mAbs as well as novel mAbs, especially in the area of protein immunogenicity. Furthermore, stability studies indicated that there are several conformational "hot spots" in many mAbs tested, especially in the hinge region. This antibody array technology can be used for novel mAb Higher Order Structure (HOS) analysis during process and formulation development. Another important area of application is for biosimilar mAb development where the innovator molecule and biosimilar molecule could be compared based on their systemic "fingerprint" from the 30 plus antibodies. PMID:23970865
The heme oxygenase/carbon monoxide system is involved in the auxin-induced cucumber adventitious rooting process.
Xuan, Wei; Zhu, Fu-Yuan; Xu, Sheng; Huang, Ben-Kai; Ling, Teng-Fang; Qi, Ji-Yan; Ye, Mao-Bing; Shen, Wen-Biao
Indole acetic acid (IAA) is an important regulator of adventitious rooting via the activation of complex signaling cascades. In animals, carbon monoxide (CO), mainly generated by heme oxygenases (HOs), is a significant modulator of inflammatory reactions, affecting cell proliferation and the production of growth factors. In this report, we show that treatment with the auxin transport inhibitor naphthylphthalamic acid prevented auxin-mediated induction of adventitious rooting and also decreased the activity of HO and its by-product CO content. The application of IAA, HO-1 activator/CO donor hematin, or CO aqueous solution was able to alleviate the IAA depletion-induced inhibition of adventitious root formation. Meanwhile, IAA or hematin treatment rapidly activated HO activity or HO-1 protein expression, and CO content was also enhanced. The application of the HO-1-specific inhibitor zinc protoporphyrin IX (ZnPPIX) could inhibit the above IAA and hematin responses. CO aqueous solution treatment was able to ameliorate the ZnPPIX-induced inhibition of adventitious rooting. Molecular evidence further showed that ZnPPIX mimicked the effects of naphthylphthalamic acid on the inhibition of adventitious rooting, the down-regulation of one DnaJ-like gene (CSDNAJ-1), and two calcium-dependent protein kinase genes (CSCDPK1 and CSCDPK5). Application of CO aqueous solution not only dose-dependently blocked IAA depletion-induced inhibition of adventitious rooting but also enhanced endogenous CO content and up-regulated CSDNAJ-1 and CSCDPK1/5 transcripts. Together, we provided pharmacological, physiological, and molecular evidence that auxin rapidly activates HO activity and that the product of HO action, CO, then triggers the signal transduction events that lead to the auxin responses of adventitious root formation in cucumber (Cucumis sativus). PMID:18689445
The Heme Oxygenase/Carbon Monoxide System Is Involved in the Auxin-Induced Cucumber Adventitious Rooting Process1
Xuan, Wei; Zhu, Fu-Yuan; Xu, Sheng; Huang, Ben-Kai; Ling, Teng-Fang; Qi, Ji-Yan; Ye, Mao-Bing; Shen, Wen-Biao
Indole acetic acid (IAA) is an important regulator of adventitious rooting via the activation of complex signaling cascades. In animals, carbon monoxide (CO), mainly generated by heme oxygenases (HOs), is a significant modulator of inflammatory reactions, affecting cell proliferation and the production of growth factors. In this report, we show that treatment with the auxin transport inhibitor naphthylphthalamic acid prevented auxin-mediated induction of adventitious rooting and also decreased the activity of HO and its by-product CO content. The application of IAA, HO-1 activator/CO donor hematin, or CO aqueous solution was able to alleviate the IAA depletion-induced inhibition of adventitious root formation. Meanwhile, IAA or hematin treatment rapidly activated HO activity or HO-1 protein expression, and CO content was also enhanced. The application of the HO-1-specific inhibitor zinc protoporphyrin IX (ZnPPIX) could inhibit the above IAA and hematin responses. CO aqueous solution treatment was able to ameliorate the ZnPPIX-induced inhibition of adventitious rooting. Molecular evidence further showed that ZnPPIX mimicked the effects of naphthylphthalamic acid on the inhibition of adventitious rooting, the down-regulation of one DnaJ-like gene (CSDNAJ-1), and two calcium-dependent protein kinase genes (CSCDPK1 and CSCDPK5). Application of CO aqueous solution not only dose-dependently blocked IAA depletion-induced inhibition of adventitious rooting but also enhanced endogenous CO content and up-regulated CSDNAJ-1 and CSCDPK1/5 transcripts. Together, we provided pharmacological, physiological, and molecular evidence that auxin rapidly activates HO activity and that the product of HO action, CO, then triggers the signal transduction events that lead to the auxin responses of adventitious root formation in cucumber (Cucumis sativus). PMID:18689445
Electron energy loss and thermal desorption spectroscopy of pyridine adsorbed on Pt(111)
Grassian, V.H.; Muetterties, E.L.
The chemisorption behavior of pyridine (NC/sub 5/H/sub 5/) on a Pt(111) surface has been examined by using thermal desorption and electron energy loss spectroscopy as a function of adsorption temperature. The vibrational spectrum of pyridine adsorbed to room temperature on this surface shows intense loss peaks in the specular direction from vibrational modes which can be characterized as in-plane stretching and bending modes. This vibrational spectrum has been interpreted as the formation of an ..cap alpha..-pyridyl species (NC/sub 5/H/sub 4/) on the surface. The pyridyl moiety is bonded to the platinum surface through the nitrogen and one of the ..cap alpha..-carbon atoms with the pyridyl plane perpendicular to the metal surface. When pyridine is adsorbed at low temperature (120 K), it bonds to the surface through both the nitrogen atom and the ..pi.. and ..pi../sup */ orbitals of the pyridine ring. As the crystal is warmed to 260 K, at saturation coverage, approximately 50% of the molecules desorb as molecular pyridine. The remaining pyridine molecules partially decompose on the surface to form an ..cap alpha..-pyridyl fragment. The electron energy loss spectra of pyridine adsorbed at both low and room temperature is compared to the infrared spectra of two osmium cluster compounds: Os/sub 3/(CO)/sub 11/(NC/sub 5/H/sub 5/), a pyridine complex, and HOs/sub 3/(CO)/sup 18/NC/sub 5/H/sub 4/), a pyridyl complex.
Evaluation of in vitro cytotoxicity of one-dimensional chain [Fe(salen)(L)](n) complexes against human cancer cell lines.
Dvořák, Zdeněk; Starha, Pavel; Sindelář, Zdeněk; Trávníček, Zdeněk
The 1d-polymeric iron(III) complexes [Fe(salen)(μ-L)](n) (1-6), involving a deprotonated form of the N-donor heterocyclic compounds (L) imidazole (complex 1), 1,2,4-triazole (2), benztriazole (3), 5-methyltetrazole (4), 5-aminotetrazole (5) and 5-phenyltetrazole (6), were studied for their in vitro cytotoxic activity against human cancer cell lines including lung carcinoma (A549), cervix epithelial carcinoma (HeLa), osteosarcoma (HOS), malignant melanoma (G361), breast adenocarcinoma (MCF7), ovarian carcinoma (A2780) and cisplatin-resistant ovarian carcinoma (A2780cis). Cytotoxicity in vitro (IC50=0.39-0.48 μM) was achieved for 2-6 against A2780 (IC50 of cisplatin equals 11.5 μM) as well as for 5 and 6 against all the tested cells, with IC50=2.5-37.7 μM. The Uv-Vis spectroscopic study showed that the complexes are unstable in organic solvents (e.g., dimethyl sulfoxide, dimethylformamide) containing even trace amounts of water (and thus also in the medium, i.e., 0.1% DMF, v/v, used in the MTT assay), where they partially or completely decompose to the mixtures involving, besides [Fe(salen)(μ-L)](n) itself, also the starting compounds [{Fe(salen)}2(μ-O)] and appropriate organic compound (HL). In efforts to find how the resulting cytotoxicity of the most active compounds 5 and 6 is influenced by this fact, the in vitro cytotoxicity testing of mixtures of reactants [{Fe(salen)}2(μ-O)] and HL, as well as the respective reactants, was also performed. It has been found that the cytotoxicity of 5 and 6 against all the tested cell lines is probably caused by a combined effect of the individual components presented within the corresponding mixture in the medium used. PMID:22274660
Gold(I)-triphenylphosphine complexes with hypoxanthine-derived ligands: in vitro evaluations of anticancer and anti-inflammatory activities.
Křikavová, Radka; Hošek, Jan; Vančo, Ján; Hutyra, Jakub; Dvořák, Zdeněk; Trávníček, Zdeněk
A series of gold(I) complexes involving triphenylphosphine (PPh3) and one N-donor ligand derived from deprotonated mono- or disubstituted hypoxanthine (HLn) of the general composition [Au(Ln)(PPh3)] (1-9) is reported. The complexes were thoroughly characterized, including multinuclear high resolution NMR spectroscopy as well as single crystal X-ray analysis (for complexes 1 and 3). The complexes were screened for their in vitro cytotoxicity against human cancer cell lines MCF7 (breast carcinoma), HOS (osteosarcoma) and THP-1 (monocytic leukaemia), which identified the complexes 4-6 as the most promising representatives, who antiproliferative activity was further tested against A549 (lung adenocarcinoma), G-361 (melanoma), HeLa (cervical cancer), A2780 (ovarian carcinoma), A2780R (ovarian carcinoma resistant to cisplatin), 22Rv1 (prostate cancer) cell lines. Complexes 4-6 showed a significantly higher in vitro anticancer effect against the employed cancer cells, except for G-361, as compared with the commercially used anticancer drug cisplatin, with IC50 ≈ 1-30 µM. Anti-inflammatory activity was evaluated in vitro by the assessment of the ability of the complexes to modulate secretion of the pro-inflammatory cytokines, i.e. tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), in the lipopolysaccharide-activated macrophage-like THP-1 cell model. The results of this study identified the complexes as auspicious anti-inflammatory agents with similar or better activity as compared with the clinically applied gold-based antiarthritic drug Auranofin. In an effort to explore the possible mechanisms responsible for the biological effect, the products of interactions of selected complexes with sulfur-containing biomolecules (L-cysteine and reduced glutathione) were studied by means of the mass-spectrometry study. PMID:25226034
Gold(I)-Triphenylphosphine Complexes with Hypoxanthine-Derived Ligands: In Vitro Evaluations of Anticancer and Anti-Inflammatory Activities
Křikavová, Radka; Hošek, Jan; Vančo, Ján; Hutyra, Jakub; Dvořák, Zdeněk; Trávníček, Zdeněk
A series of gold(I) complexes involving triphenylphosphine (PPh3) and one N-donor ligand derived from deprotonated mono- or disubstituted hypoxanthine (HLn) of the general composition [Au(Ln)(PPh3)] (1–9) is reported. The complexes were thoroughly characterized, including multinuclear high resolution NMR spectroscopy as well as single crystal X-ray analysis (for complexes 1 and 3). The complexes were screened for their in vitro cytotoxicity against human cancer cell lines MCF7 (breast carcinoma), HOS (osteosarcoma) and THP-1 (monocytic leukaemia), which identified the complexes 4–6 as the most promising representatives, who antiproliferative activity was further tested against A549 (lung adenocarcinoma), G-361 (melanoma), HeLa (cervical cancer), A2780 (ovarian carcinoma), A2780R (ovarian carcinoma resistant to cisplatin), 22Rv1 (prostate cancer) cell lines. Complexes 4–6 showed a significantly higher in vitro anticancer effect against the employed cancer cells, except for G-361, as compared with the commercially used anticancer drug cisplatin, with IC50 ≈ 1–30 µM. Anti-inflammatory activity was evaluated in vitro by the assessment of the ability of the complexes to modulate secretion of the pro-inflammatory cytokines, i.e. tumour necrosis factor-α (TNF-α) and interleukin-1β (IL-1β), in the lipopolysaccharide-activated macrophage-like THP-1 cell model. The results of this study identified the complexes as auspicious anti-inflammatory agents with similar or better activity as compared with the clinically applied gold-based antiarthritic drug Auranofin. In an effort to explore the possible mechanisms responsible for the biological effect, the products of interactions of selected complexes with sulfur-containing biomolecules (L-cysteine and reduced glutathione) were studied by means of the mass-spectrometry study. PMID:25226034
A Comparative Study of Serum and Follicular Fluid Leptin Concentrations among Explained Infertile, Unexplained Infertile and Fertile Women
Kamyabi, Zahra; Gholamalizade, Tayebe
Background The relationship between metabolism and reproduction has been always considered as an important topic in female endocrinology. It seems that leptin is one of the involved factors in infertility. Leptin, in addition to regulating body weight plays an important role in regulation of endocrine, reproductive and immune systems. The aim of this stduy is to compare serum and follicular fluid leptin concentrations in order to find the role of leptin level in infertility. Materials and Methods This case-control study was performed from September 2010 to March 2013. A total of 90 women referred to the Infertility Center of Afzalipour Hos- pital, Kerman, Iran, and divided into three equal groups (n=30/per group) of explained infertile (including 4 subgroups), unexplained infertile and normal fertile (control group). The three groups were matched in regard to demographic features [age: 20-40 years and body mass index (BMI): 20-25]. In order to determine leptin level, blood sample and fol- licular fluid were taken one hour prior and at the time of follicular puncture, respectively. Serum and follicular fluid leptin levels were measured using enzyme-linked immune sorbent assay (ELISA). Data were analyzed using descriptive-analytic tests, like Mann- Whitney and Kruskal Wallis tests, through Statistical Package for the Social Sciences (SPSS) version 16. Results In explained infertile and fertile groups, as opposed to unexplained infertile group, mean leptin level was lower in follicular fluid than in serum. Mean follicular fluid leptin concentration in women with unexplained infertility was higher com- pared to the other two groups. Women with unexplained infertility had lower level of serum leptin in comparison to the other two groups. Follicular fluid leptin level in all subgroups of explained infertile group was lower as compared to unexplained and fertile women. Conclusion The results suggested that high leptin level of follicular fluid is one of the main factors
Generalized structure of higher order nonclassicality
NASA Astrophysics Data System (ADS)
Verma, Amit; Pathak, Anirban
A generalized notion of higher order nonclassicality (in terms of higher order moments) is introduced. Under this generalized framework of higher order nonclassicality, conditions of higher order squeezing and higher order subpoissonian photon statistics are derived. A simpler form of the Hong-Mandel higher order squeezing criterion is derived under this framework by using an operator ordering theorem introduced by us in [A. Pathak, J. Phys. A 33 (2000) 5607]. It is also generalized for multi-photon Bose operators of Brandt and Greenberg. Similarly, condition for higher order subpoissonian photon statistics is derived by normal ordering of higher powers of number operator. Further, with the help of simple density matrices, it is shown that the higher order antibunching (HOA) and higher order subpoissonian photon statistics (HOSPS) are not the manifestation of the same phenomenon and consequently it is incorrect to use the condition of HOA as a test of HOSPS. It is also shown that the HOA and HOSPS may exist even in absence of the corresponding lower order phenomenon. Binomial state, nonlinear first order excited squeezed state (NLESS) and nonlinear vacuum squeezed state (NLVSS) are used as examples of quantum state and it is shown that these states may show higher order nonclassical characteristics. It is observed that the Binomial state which is always antibunched, is not always higher order squeezed and NLVSS which shows higher order squeezing does not show HOSPS and HOA. The opposite is observed in NLESS and consequently it is established that the HOSPS and HOS are two independent signatures of higher order nonclassicality.
A Survey on Oocyte Donation: Turkish Fertile and Infertile Women's Opinions
Akyuz, Aygul; Sever, Nese; Karasahin, Emre; Guvenc, Gulten; Cek, Suzan
Background There are various treatment options for infertility, and new techniques are also being developed as it is an important healthcare problem affecting approximately 15-20% of married couples. The purpose of this study was to evaluate the level of infor- mation of fertile and infertile Turkish women on oocyte donation in order to understand their awareness of the legal, ethical, social and religious issues regarding this technique and to compare these two groups in terms of these variables. Materials and Methods This cross-sectional study included infertile women being treated at the assisted reproductive technologies (ART) program of a university hos- pital and women who had presented at the gynecology outpatients department of the same university for routine check-ups and who had no previous history of infertility. After consulting with specialists in the field and searching the related literature, a data collection form having 22 questions for infertile women and 18 questions for fertile women was prepared. Results The women were asked whether they would use the oocytes of another woman if necessary. The results showed that 67.6% of the fertile women said they would never want to use this method, while 63.9% of the infertile women stated they may accept to use this method under certain conditions (two distinct answers appeared in the answers, some women stated they would prefer donated oocytes from close relatives, while others stated they would prefer oocytes from total strangers), such as from a close relative or from someone they do not know at all. Conclusion Infertile women mentioned that they could use illegal routes if necessary to have a child at much higher rates than stated by fertile women. This shows that desire to have a child is a strong source of motivation in Turkey. PMID:25379158
Transformation of human osteoblast cells to the tumorigenic phenotype by depleted uranium-uranyl chloride.
Miller, A C; Blakely, W F; Livengood, D; Whittaker, T; Xu, J; Ejnik, J W; Hamilton, M M; Parlette, E; John, T S; Gerstenberg, H M; Hsu, H
Depleted uranium (DU) is a dense heavy metal used primarily in military applications. Although the health effects of occupational uranium exposure are well known, limited data exist regarding the long-term health effects of internalized DU in humans. We established an in vitro cellular model to study DU exposure. Microdosimetric assessment, determined using a Monte Carlo computer simulation based on measured intracellular and extracellular uranium levels, showed that few (0.0014%) cell nuclei were hit by alpha particles. We report the ability of DU-uranyl chloride to transform immortalized human osteoblastic cells (HOS) to the tumorigenic phenotype. DU-uranyl chloride-transformants are characterized by anchorage-independent growth, tumor formation in nude mice, expression of high levels of the k-ras oncogene, reduced production of the Rb tumor-suppressor protein, and elevated levels of sister chromatid exchanges per cell. DU-uranyl chloride treatment resulted in a 9.6 (+/- 2.8)-fold increase in transformation frequency compared to untreated cells. In comparison, nickel sulfate resulted in a 7.1 (+/- 2.1)-fold increase in transformation frequency. This is the first report showing that a DU compound caused human cell transformation to the neoplastic phenotype. Although additional studies are needed to determine if protracted DU exposure produces tumors in vivo, the implication from these in vitro results is that the risk of cancer induction from internalized DU exposure may be comparable to other biologically reactive and carcinogenic heavy-metal compounds (e.g., nickel). Images Figure 1 Figure 2 Figure 3 PMID:9681973
Long non-coding RNA tumor suppressor candidate 7 functions as a tumor suppressor and inhibits proliferation in osteosarcoma.
Cong, Menglin; Li, Jianmin; Jing, Rui; Li, Zhenzhong
Osteosarcoma is the most common malignant tumor of bone. Recent studies have proven long non-coding RNAs (lncRNAs) play important roles in the tumorigenesis and progression of cancer. However, few lncRNAs have been investigated in osteosarcoma. Here, we reported a novel lncRNA, tumor suppressor candidate 7 (TUSC7), was significantly downregulated in osteosarcoma tissues compared with paired non-tumor tissues and low expression of TUSC7 indicated poor survival (HR = 0.313, 95 % confidence interval (CI) 0.092-0.867) of osteosarcoma patients. Further analysis revealed that loss copy number of TUSC7 was correlated with low expression of TUSC7, and additionally, loss of TUSC7 copy number also indicated poor prognosis (HR = 3.994, 95 % CI 1.147-13.91) of osteosarcoma patients. Two osteosarcoma cell lines, HOS and MG63, were utilized to investigate biological function of TUSC7. Cell counting kit 8 (CCK-8) assay revealed that after silence of TUSC7, cell proliferation ability increased and the colony formation ability also increased. Further results showed that cell cycle was not affected by treatment of si-TUSC7, while the percentage of apoptotic cells decreased. Western blot showed that after silence of TUSC7, the proapoptotic Bcl2 expression was downregulated. Finally, we established xenograft tumor models in nude mice with MG63 cells. Compared with negative control group, silence of TUSC7 significantly promoted tumor growth in vivo. Thus, we demonstrated that TUSC7 could be a potential tumor suppressor in osteosarcoma. PMID:26781978
Spermicidal and Contraceptive Potential of Desgalactotigonin: A Prospective Alternative of Nonoxynol-9
Chakraborty, Debanjana; Maity, Arindam; Jha, Tarun; Mondal, Nirup Bikash
Crude decoction of Chenopodium album seed showed spermicidal effect at MIC 2 mg/ml in earlier studies. Systematic isolation, characterization and evaluation revealed that the major metabolite Desgalactotigonin (DGT) is the most effective principle in both in vitro and in vivo studies. The in vitro studies comprises (a) rat and human sperm motility and immobilizing activity by Sander-Cramer assay; (b) sperm membrane integrity was observed by HOS test and electron microscopy; (c) microbial potential was examined in Lactobacillus broth culture, and (d) the hemolytic index was determined by using rat RBCs. The in vivo contraceptive efficacy was evaluated by intra uterine application of DGT in rat. Lipid peroxidation and induction of apoptosis by DGT on human spermatozoa were also studied. The minimum effective concentration (MEC) of DGT that induced instantaneous immobilization in vitro was 24.18 µM for rat and 58.03 µM for human spermatozoa. Microbial study indicated DGT to be friendly to Lactobacillus acidophilus. Implantation was prevented in DGT treated uterine horn while no hindrance occurred in the untreated contra lateral side. At the level of EC50, DGT induced apoptosis in human spermatozoa as determined by increased labeling with Annexin-V and decreased polarization of sperm mitochondria. Desgalactotigonin emerged 80 and 2×104 times more potent than the decoction and Nonoxynol-9 respectively. It possesses mechanism based detrimental action on both human and rat spermatozoa and spares lactobacilli and HeLa cells at MEC which proves its potential as a superior ingredient for the formulation of a contraceptive safer/compatible to vaginal microflora. PMID:25243914
The post-thaw irradiation of avian spermatozoa with He-Ne laser differently affects chicken, pheasant and turkey sperm quality.
Iaffaldano, N; Paventi, G; Pizzuto, R; Passarella, S; Cerolini, S; Zaniboni, L; Marzoni, M; Castillo, A; Rosato, M P
The effects of post-thaw Helium-Neon (He-Ne) laser irradiation on mobility and functional integrity of frozen/thawed chicken, pheasant and turkey spermatozoa were investigated. Cytochrome C oxidase (COX) activity was also determined as a measure of the effect of irradiation on mitochondrial bioenergetics. Semen samples from each species were collected, processed and frozen according to the pellet procedure. After thawing, each semen sample was divided into two subsamples: the first one was the control; the second one was irradiated with a single mode continuous He-Ne laser wave (wavelength 632.8 nm; 6 mW; 3.96 J/cm(2)). Then the samples were assessed for sperm mobility (Accudenz(®) swim-down test), viability (SYBR-14/PI staining), osmotic-resistance (HOS test) and COX activity. The irradiation was effective P<0.05 increasing sperm motility in the turkey semen (0.228 ± 0.01 compared with 0.294 ± 0.02). The irradiation also caused an increase (P<0.05) of the COX activity in pheasant (+135 ± 4%) and turkey (+116 ± 4%) sperm, without affecting viability and osmotic-resistance. The COX was positively correlated (P<0.05) with the viability of chicken sperm, however no significant interactions were found between mobility and COX activity in the three avian species. Due to the difference in energetic metabolism among avian species used in this study, the He-Ne laser irradiation has a differential action on bio-stimulation of turkey, chicken and pheasant spermatozoa. The present results are the first to elucidate the possibility for restoration of motility of cryopreserved avian spermatozoa by bio-stimulation provided via He-Ne laser irradiation. PMID:24125852
Rapid phosphatidic acid accumulation in response to low temperature stress in Arabidopsis is generated through diacylglycerol kinase
Arisz, Steven A.; van Wijk, Ringo; Roels, Wendy; Zhu, Jian-Kang; Haring, Michel A.; Munnik, Teun
Phosphatidic acid (PtdOH) is emerging as an important signaling lipid in abiotic stress responses in plants. The effect of cold stress was monitored using 32P-labeled seedlings and leaf discs of Arabidopsis thaliana. Low, non-freezing temperatures were found to trigger a very rapid 32P-PtdOH increase, peaking within 2 and 5 min, respectively. In principle, PtdOH can be generated through three different pathways, i.e., (1) via de novo phospholipid biosynthesis (through acylation of lyso-PtdOH), (2) via phospholipase D hydrolysis of structural phospholipids, or (3) via phosphorylation of diacylglycerol (DAG) by DAG kinase (DGK). Using a differential 32P-labeling protocol and a PLD-transphosphatidylation assay, evidence is provided that the rapid 32P-PtdOH response was primarily generated through DGK. A simultaneous decrease in the levels of 32P-PtdInsP, correlating in time, temperature dependency, and magnitude with the increase in 32P-PtdOH, suggested that a PtdInsP-hydrolyzing PLC generated the DAG in this reaction. Testing T-DNA insertion lines available for the seven DGK genes, revealed no clear changes in 32P-PtdOH responses, suggesting functional redundancy. Similarly, known cold-stress mutants were analyzed to investigate whether the PtdOH response acted downstream of the respective gene products. The hos1, los1, and fry1 mutants were found to exhibit normal PtdOH responses. Slight changes were found for ice1, snow1, and the overexpression line Super-ICE1, however, this was not cold-specific and likely due to pleiotropic effects. A tentative model illustrating direct cold effects on phospholipid metabolism is presented. PMID:23346092
A Histone Deacetylase Adjusts Transcription Kinetics at Coding Sequences during Candida albicans Morphogenesis
Hnisz, Denes; Bardet, Anaïs F.; Nobile, Clarissa J.; Petryshyn, Andriy; Glaser, Walter; Schöck, Ulrike; Stark, Alexander; Kuchler, Karl
Despite their classical role as transcriptional repressors, several histone deacetylases, including the baker's yeast Set3/Hos2 complex (Set3C), facilitate gene expression. In the dimorphic human pathogen Candida albicans, the homologue of the Set3C inhibits the yeast-to-filament transition, but the precise molecular details of this function have remained elusive. Here, we use a combination of ChIP–Seq and RNA–Seq to show that the Set3C acts as a transcriptional co-factor of metabolic and morphogenesis-related genes in C. albicans. Binding of the Set3C correlates with gene expression during fungal morphogenesis; yet, surprisingly, deletion of SET3 leaves the steady-state expression level of most genes unchanged, both during exponential yeast-phase growth and during the yeast-filament transition. Fine temporal resolution of transcription in cells undergoing this transition revealed that the Set3C modulates transient expression changes of key morphogenesis-related genes. These include a transcription factor cluster comprising of NRG1, EFG1, BRG1, and TEC1, which form a regulatory circuit controlling hyphal differentiation. Set3C appears to restrict the factors by modulating their transcription kinetics, and the hyperfilamentous phenotype of SET3-deficient cells can be reverted by mutating the circuit factors. These results indicate that the chromatin status at coding regions represents a dynamic platform influencing transcription kinetics. Moreover, we suggest that transcription at the coding sequence can be transiently decoupled from potentially conflicting promoter information in dynamic environments. PMID:23236295
Cyclophilin A-Dependent Restriction of Human Immunodeficiency Virus Type 1 Capsid Mutants for Infection of Nondividing Cells ▿
Qi, Mingli; Yang, Ruifeng; Aiken, Christopher
Among retroviruses, lentiviruses are unusual in their ability to efficiently infect both dividing and nondividing cells, such as activated T cells and macrophages, respectively. Recent studies implicate the viral capsid protein (CA) as a key determinant of cell-cycle-independent infection by human immunodeficiency virus type 1 (HIV-1). We investigated the effects of the host cell protein cyclophilin A (CypA), which binds to HIV-1 CA, on HIV-1 infection of nondividing cells. The HIV-1 CA mutants A92E, T54A, and R132K were impaired for infection of aphidicolin-arrested HeLa cells, but not HOS cells. The mutants synthesized normal quantities of two-long-terminal-repeat circles in arrested HeLa cells, indicating that the mutant preintegration complexes can enter the nuclei of both dividing and nondividing cells. The impaired infectivity of the CA mutants on both dividing and nondividing HeLa cells was relieved by either pharmacological or genetic disruption of the CypA-CA interaction or by RNA interference-mediated depletion of CypA expression in target cells. A second-site suppressor of the CypA-restricted phenotype also restored the ability of CypA-restricted HIV-1 mutants to infect growth-arrested HeLa cells. These results indicate that CypA-restricted mutants are specifically impaired at a step between nuclear import and integration in nondividing HeLa cells. This study reveals a novel target cell-specific restriction of HIV-1 CA mutants in nondividing cells that is dependent on CypA-CA interactions. PMID:18829762
The HSP90 inhibitor alvespimycin enhances the potency of telomerase inhibition by imetelstat in human osteosarcoma.
Hu, Yafang; Bobb, Daniel; He, Jianping; Hill, D Ashley; Dome, Jeffrey S
The unsatisfactory outcomes for osteosarcoma necessitate novel therapeutic strategies. This study evaluated the effect of the telomerase inhibitor imetelstat in pre-clinical models of human osteosarcoma. Because the chaperone molecule HSP90 facilitates the assembly of telomerase protein, the ability of the HSP90 inhibitor alvespimycin to potentiate the effect of the telomerase inhibitor was assessed. The effect of single or combined treatment with imetelstat and alvespimycin on long-term growth was assessed in osteosarcoma cell lines (143B, HOS and MG-63) and xenografts derived from 143B cells. Results indicated that imetelstat as a single agent inhibited telomerase activity, induced telomere shortening, and inhibited growth in all 3 osteosarcoma cell lines, though the bulk cell cultures did not undergo growth arrest. Combined treatment with imetelstat and alvespimycin resulted in diminished telomerase activity and shorter telomeres compared to either agent alone as well as higher levels of γH2AX and cleaved caspase-3, indicative of increased DNA damage and apoptosis. With dual telomerase and HSP90 inhibition, complete growth arrest of bulk cell cultures was achieved. In xenograft models, all 3 treatment groups significantly inhibited tumor growth compared with the placebo-treated control group, with the greatest effect seen in the combined treatment group (imetelstat, p = 0.045, alvespimycin, p = 0.034; combined treatment, p = 0.004). In conclusion, HSP90 inhibition enhanced the effect of telomerase inhibition in pre-clinical models of osteosarcoma. Dual targeting of telomerase and HSP90 warrants further investigation as a therapeutic strategy. PMID:25920748
Modelization of highly nonlinear waves in coastal regions
NASA Astrophysics Data System (ADS)
Gouin, Maïté; Ducrozet, Guillaume; Ferrant, Pierre
The proposed work deals with the development of a highly non-linear model for water wave propagation in coastal regions. The accurate modelization of surface gravity waves is of major interest in ocean engineering, especially in the field of marine renewable energy. These marine structures are intended to be settled in coastal regions where the effect of variable bathymetry may be significant on local wave conditions. This study presents a numerical model for the wave propagation with complex bathymetry. It is based on High-Order Spectral (HOS) method, initially limited to the propagation of non-linear wave fields over flat bottom. Such a model has been developed and validated at the LHEEA Lab. (Ecole Centrale Nantes) over the past few years and the current developments will enlarge its application range. This new numerical model will keep the interesting numerical properties of the original pseudo-spectral approach (convergence, efficiency with the use of FFTs, …) and enable the possibility to propagate highly non-linear wave fields over long time and large distance. Different validations will be provided in addition to the presentation of the method. At first, Bragg reflection will be studied with the proposed approach. If the Bragg condition is satisfied, the reflected wave generated by a sinusoidal bottom patch should be amplified as a result of resonant quadratic interactions between incident wave and bottom. Comparisons will be provided with experiments and reference solutions. Then, the method will be used to consider the transformation of a non-linear monochromatic wave as it propagates up and over a submerged bar. As the waves travel up the front slope of the bar, it steepens and high harmonics are generated due to non-linear interactions. Comparisons with experimental data will be provided. The different test cases will assess the accuracy and efficiency of the method proposed.
Absent Left Main Coronary Artery and Separate Ostia of Left Coronary System in a Patient with Holt-Oram Syndrome and Sinus Node Dysfunction
Aung, Thein Tun; Roberto, Edward Samuel; Wase, Abdul
Patient: Male, 41 Final Diagnosis: Sick Sinus Syndrome and absent left main coronary artery • separate ostia of left anterior descending and circumflex arteries in Holt-Oram Syndrome Symptoms: Conduction disturbance • seizure-like activity • upper extremity malformations Medication: — Clinical Procedure: Electro physiology study • coronary catheterization • pacemaker Specialty: Cardiology Objective: Congenital defects/diseases Background: Holt-Oram syndrome (HOS) is a rare but significant syndrome consisting of structural heart defects, conduction abnormalities, and upper extremity anomalies. It was first described in the British Heart Journal in 1960 by Mary Holt and Samuel Oram as a report of atrial septal defect, conduction disturbances, and hand malformations occurring in family members. Patients can present with heart blocks or symptoms of underlying congenital heart defects. Case Report: A 41-year-old man with Holt-Oram syndrome presented with seizure-like activity and was found to have an underlying conduction disturbance. Physical exam showed bilateral atrophic upper extremities with anatomic disfiguration, and weakness of the intrinsic hand muscles. Cardiovascular exam revealed a slow heart rate with irregular rhythm. EKG showed sinus arrest with junctional escape rhythm. Cardiac catheterization revealed coronary anomalies, including absent left main coronary artery and separate ostia of the left anterior ascending and left circumflex coronary artery. Coronary arteries were patent. Following electrophysiology study, sick sinus syndrome and AV block were diagnosed, and the patient received implantation of a permanent pacemaker. Conclusions: This patient presented with a seizure-like episode attributed to hypoxia during asystole from an underlying cardiac conduction defect associated with Holt-Oram syndrome. Arrhythmias and heart blocks are seen in these patients, and conduction defects are highly associated with congenital heart defects. Holt
Diversity of reaction characteristics of glucan branching enzymes and the fine structure of α-glucan from various sources.
Sawada, Takayuki; Nakamura, Yasunori; Ohdan, Takashi; Saitoh, Asami; Francisco, Perigio B; Suzuki, Eiji; Fujita, Naoko; Shimonaga, Takahiro; Fujiwara, Shoko; Tsuzuki, Mikio; Colleoni, Christophe; Ball, Steven
To investigate the functional properties of 10 α-glucan branching enzymes (BEs) from various sources, we determined the chain-length distribution of BE enzymatic products and their phosphorylase-limit dextrins (Φ-LD). All BEs could be classified into either of the three rice BE isozymes: OsBEI, OsBEIIa, or OsBEIIb. Escherichia coli BE (EcoBE) had the same enzymatic properties as OsBEI, while Synechococcus elongatus BE (ScoBE) and Chlorella kessleri BE (ChlBE) had BEIIb-type properties. Human BE (HosBE), yeast BE (SacBE), and two Porphyridium purpureum BEs (PopBE1 and PopBE2) exhibited the OsBEIIa-type properties. Analysis of chain-length profile of Φ-LD of the BE reaction products revealed that EcoBE, ScoBE, PopBE1, and PopBE2 preferred A-chains as acceptors, while OsBEIIb used B-chains more frequently than A-chains. Both EcoBE and ScoBE specifically formed the branch linkages at the third glucose residue from the reducing end of the acceptor chain. The present results provide evidence for the first time that great variation exists as to the preference of BEs for their acceptor chain, either A-chain or B-chain. In addition, EcoBE and ScoBE recognize the location of branching points in their acceptor chain during their branching reaction. Nevertheless, no correlation exists between the primary structure of BE proteins and their enzymatic characteristics. PMID:25107532
Consensus by Democracy. Using Meta-Analyses of Microarray and Genomic Data to Model the Cold Acclimation Signaling Pathway in Arabidopsis1[W
Benedict, Catherine; Geisler, Matt; Trygg, Johan; Huner, Norman; Hurry, Vaughan
The whole-genome response of Arabidopsis (Arabidopsis thaliana) exposed to different types and durations of abiotic stress has now been described by a wealth of publicly available microarray data. When combined with studies of how gene expression is affected in mutant and transgenic Arabidopsis with altered ability to transduce the low temperature signal, these data can be used to test the interactions between various low temperature-associated transcription factors and their regulons. We quantized a collection of Affymetrix microarray data so that each gene in a particular regulon could vote on whether a cis-element found in its promoter conferred induction (+1), repression (−1), or no transcriptional change (0) during cold stress. By statistically comparing these election results with the voting behavior of all genes on the same gene chip, we verified the bioactivity of novel cis-elements and defined whether they were inductive or repressive. Using in silico mutagenesis we identified functional binding consensus variants for the transcription factors studied. Our results suggest that the previously identified ICEr1 (induction of CBF expression region 1) consensus does not correlate with cold gene induction, while the ICEr3/ICEr4 consensuses identified using our algorithms are present in regulons of genes that were induced coordinate with observed ICE1 transcript accumulation and temporally preceding genes containing the dehydration response element. Statistical analysis of overlap and cis-element enrichment in the ICE1, CBF2, ZAT12, HOS9, and PHYA regulons enabled us to construct a regulatory network supported by multiple lines of evidence that can be used for future hypothesis testing. PMID:16896234
Consensus by democracy. Using meta-analyses of microarray and genomic data to model the cold acclimation signaling pathway in Arabidopsis.
Benedict, Catherine; Geisler, Matt; Trygg, Johan; Huner, Norman; Hurry, Vaughan
The whole-genome response of Arabidopsis (Arabidopsis thaliana) exposed to different types and durations of abiotic stress has now been described by a wealth of publicly available microarray data. When combined with studies of how gene expression is affected in mutant and transgenic Arabidopsis with altered ability to transduce the low temperature signal, these data can be used to test the interactions between various low temperature-associated transcription factors and their regulons. We quantized a collection of Affymetrix microarray data so that each gene in a particular regulon could vote on whether a cis-element found in its promoter conferred induction (+1), repression (-1), or no transcriptional change (0) during cold stress. By statistically comparing these election results with the voting behavior of all genes on the same gene chip, we verified the bioactivity of novel cis-elements and defined whether they were inductive or repressive. Using in silico mutagenesis we identified functional binding consensus variants for the transcription factors studied. Our results suggest that the previously identified ICEr1 (induction of CBF expression region 1) consensus does not correlate with cold gene induction, while the ICEr3/ICEr4 consensuses identified using our algorithms are present in regulons of genes that were induced coordinate with observed ICE1 transcript accumulation and temporally preceding genes containing the dehydration response element. Statistical analysis of overlap and cis-element enrichment in the ICE1, CBF2, ZAT12, HOS9, and PHYA regulons enabled us to construct a regulatory network supported by multiple lines of evidence that can be used for future hypothesis testing. PMID:16896234
An integrated index for automated detection of infarcted myocardium from cross-sectional echocardiograms using texton-based features (Part 1).
Sudarshan, Vidya K; Acharya, U Rajendra; Ng, E Y K; Tan, Ru San; Chou, Siaw Meng; Ghista, Dhanjoo N
Cross-sectional view echocardiography is an efficient non-invasive diagnostic tool for characterizing Myocardial Infarction (MI) and stages of expansion leading to heart failure. An automated computer-aided technique of cross-sectional echocardiography feature assessment can aid clinicians in early and more reliable detection of MI patients before subsequent catastrophic post-MI medical conditions. Therefore, this paper proposes a novel Myocardial Infarction Index (MII) to discriminate infarcted and normal myocardium using features extracted from apical cross-sectional views of echocardiograms. The cross-sectional view of normal and MI echocardiography images are represented as textons using Maximum Responses (MR8) filter banks. Fractal Dimension (FD), Higher-Order Statistics (HOS), Hu's moments, Gabor Transform features, Fuzzy Entropy (FEnt), Energy, Local binary Pattern (LBP), Renyi's Entropy (REnt), Shannon's Entropy (ShEnt), and Kapur's Entropy (KEnt) features are extracted from textons. These features are ranked using t-test and fuzzy Max-Relevancy and Min-Redundancy (mRMR) ranking methods. Then, combinations of highly ranked features are used in the formulation and development of an integrated MII. This calculated novel MII is used to accurately and quickly detect infarcted myocardium by using one numerical value. Also, the highly ranked features are subjected to classification using different classifiers for the characterization of normal and MI LV ultrasound images using a minimum number of features. Our current technique is able to characterize MI with an average accuracy of 94.37%, sensitivity of 91.25% and specificity of 97.50% with 8 apical four chambers view features extracted from only single frame per patient making this a more reliable and accurate classification. PMID:26898671
Blockade of X4-Tropic HIV-1 Cellular Entry by GSK812397, a Potent Noncompetitive CXCR4 Receptor Antagonist▿
Jenkinson, Stephen; Thomson, Michael; McCoy, David; Edelstein, Mark; Danehower, Susan; Lawrence, Wendell; Wheelan, Pat; Spaltenstein, Andrew; Gudmundsson, Kristjan
GSK812397 is a potent entry inhibitor of X4-tropic strains of HIV-1, as demonstrated in multiple in vitro cellular assays (e.g., in peripheral blood mononuclear cells [PBMCs] and a viral human osteosarcoma [HOS] assay, mean 50% inhibitory concentrations [IC50s] ± standard errors of the means were 4.60 ± 1.23 nM and 1.50 ± 0.21 nM, respectively). The primary in vitro potency of GSK812397 was not significantly altered by the addition of serum proteins (2.55 [±0.12]-fold shift in the presence of human serum albumin and α-acid glycoprotein in the PBMC assay). Pharmacological characterization of GSK812397 in cell-based functional assays revealed it to be a noncompetitive antagonist of the CXCR4 receptor, with GSK812397 producing a concentration-dependent decrease in both an SDF-1-mediated chemotaxis and intracellular calcium release (IC50s were 0.34 ± 0.01 nM and 2.41 ± 0.50 nM, respectively). With respect to the antiviral activity of GSK812397, it was effective against a broad range of X4- and X4R5-utilizing clinical isolates. The potency and efficacy of GSK812397 were dependent on the individual isolate, with complete inhibition of infection observed with 24 of 30 isolates. GSK812397 did not show any detectable in vitro cytotoxicity and was highly selective for CXCR4, as determined using a wide range of receptors, enzymes, and transporters. Moreover, GSK812397 demonstrated acceptable pharmacokinetic properties and bioavailability across species. The data demonstrate that GSK812397 has antiviral activity against a broad range of X4-utilizing strains of HIV-1 via a noncompetitive antagonism of the CXCR4 receptor. PMID:19949058
Plumbagin exhibits an anti-proliferative effect in human osteosarcoma cells by downregulating FHL2 and interfering with Wnt/β-catenin signalling
Xue, Yuan-Liang; Meng, Xiang-Qi; Ma, Long-Jun; Yuan, Zhen
Plumbagin, a naphthoquinone constituent of Plumbago zeylanica L. (Plumbaginaceae) is widely used in traditional Chinese medicine as an antifungal, antibacterial and anti-inflammatory agent. Plumbagin is known to exhibit proapoptotic, antiangiogenic and antimetastatic effects in cancer cells. The transcriptional co-factor four and a half LIM domains 2 (FHL2) is a multifunctional adaptor protein that is involved in the regulation of gene expression, signal transduction and cell proliferation and differentiation, and also acts as a tumor suppressor or oncoprotein depending on the tissue microenvironment. The present study investigated the effect of plumbagin on FHL2 expression, Wnt/β-catenin signalling and its anti-proliferative activity in various human osteosarcoma cell lines, including SaOS2, MG63, HOS and U2OS. The cells were exposed to plumbagin and the expression of FHL2 was evaluated using western blot analysis. Furthermore, the anti-proliferative effect of plumbagin was evaluated using a 3-(4,5 dimethyl-thiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. In addition, since FHL2 is involved in Wnt/β-catenin signaling, the effect of plumbagin on β-catenin and its primary target genes, including v-myc avian myelocytomatosis viral oncogene homolog (c-Myc) and WNT1 inducible signaling pathway protein-1 (WISP-1), was evaluated using western blot analysis. It was observed that plumbagin suppressed the expression of FHL2 and exhibited significant anti-proliferative activity in osteosarcoma cells. It also attenuated Wnt/β-catenin signalling by downregulating β-catenin and its target genes, including c-Myc and WISP-1. In conclusion, plumbagin demonstrated anti-proliferative activity in osteosarcoma cells by downregulating FHL2 and interfering with Wnt/β-catenin signalling. PMID:27446400
Isocyanides inhibit human heme oxygenases at the verdoheme stage.
Evans, John P; Kandel, Sylvie; Ortiz de Montellano, Paul R
Heme oxygenases (HO) catalyze the oxidative cleavage of heme to generate biliverdin, CO, and free iron. In humans, heme oxygenase-1 (hHO-1) is overexpressed in tumor tissues, where it helps to protect cancer cells from anticancer agents, while HOs in fungal pathogens, such as Candida albicans, function as the primary means of iron acquisition. Thus, HO can be considered a potential therapeutic target for certain diseases. In this study, we have examined the equilibrium binding of three isocyanides, isopropyl, n-butyl, and benzyl, to the two major human HO isoforms (hHO-1 and hHO-2), Candida albicans HO (CaHmx1), and human cytochrome P450 CYP3A4 using electronic absorption spectroscopy. Isocyanides coordinate to both ferric and ferrous HO-bound heme, with tighter binding by the more hydrophobic isocyanides and 200-300-fold tighter binding to the ferrous form. Benzyl isocyanide was the strongest ligand to ferrous heme in all the enzymes. Because the dissociation constants (KD) of the ligands for ferrous heme-hHO-1 were below the limit of accuracy for equilibrium titrations, stopped-flow kinetic experiments were used to measure the binding parameters of the isocyanides to ferrous hHO-1. Steady-state activity assays showed that benzyl isocyanide was the most potent uncompetitive inhibitor with respect to heme with a KI = 0.15 microM for hHO-1. Importantly, single turnover assays revealed that the reaction was completely stopped by coordination of the isocyanide to the verdoheme intermediate rather than to the ferric heme complex. Much tighter binding of the inhibitor to the verdoheme intermediate differentiates it from inhibition of, for example, CYP3A4 and offers a possible route to more selective inhibitor design. PMID:19694439
Recent progresses of neural network unsupervised learning: I. Independent component analyses generalizing PCA
NASA Astrophysics Data System (ADS)
Szu, Harold H.
The early vision principle of redundancy reduction of 108 sensor excitations is understandable from computer vision viewpoint toward sparse edge maps. It is only recently derived using a truly unsupervised learning paradigm of artificial neural networks (ANN). In fact, the biological vision, Hubel- Wiesel edge maps, is reproduced seeking the underlying independent components analyses (ICA) among 102 image samples by maximizing the ANN output entropy (partial)H(V)/(partial)[W] equals (partial)[W]/(partial)t. When a pair of newborn eyes or ears meet the bustling and hustling world without supervision, they seek ICA by comparing 2 sensory measurements (x1(t), x2(t))T equalsV X(t). Assuming a linear and instantaneous mixture model of the external world X(t) equals [A] S(t), where both the mixing matrix ([A] equalsV [a1, a2] of ICA vectors and the source percentages (s1(t), s2(t))T equalsV S(t) are unknown, we seek the independent sources
approximately equals [I] where the approximated sign indicates that higher order statistics (HOS) may not be trivial. Without a teacher, the ANN weight matrix [W] equalsV [w1, w2] adjusts the outputs V(t) equals tanh([W]X(t)) approximately equals [W]X(t) until no desired outputs except the (Gaussian) 'garbage' (neither YES '1' nor NO '-1' but at linear may-be range 'origin 0') defined by Gaussian covariance HOS theories since 1991 (advanced by Jutten & Herault, Comon, Oja, Bell-Sejnowski, Amari-Cichocki, Cardoso), the LYAPONOV function L(v1,...,vn, w1,...wn,) equals E(v1,...,vn) - H(w1,...wn) is constructed as the HELMHOTZ free energy to prove both convergences of supervised energy E and unsupervised entropy H learning. Consequently, rather
Early Paleozoic magmatism and metallogeny in Northeast China: a record from the Tongshan porphyry Cu deposit
NASA Astrophysics Data System (ADS)
Hu, Xinlu; Yao, Shuzhen; Ding, Zhenju; He, Mouchun
The Tongshan Cu deposit is located in the northern segment of the Great Xing'an Range and represents one of the few early Paleozoic porphyry Cu deposits in northeastern China. The granitic rocks in the Tongshan Cu deposit include concealed granodiorite and exposed tonalite, which yield LA-ICP-MS zircon U-Pb ages of 478 ± 3 Ma and 214 ± 3 Ma, respectively. The granodiorite has relatively high SiO2 (60.5-63.5 wt%) and Sr (596-786 ppm) contents, low Yb (1.21-1.53 ppm) and Y (9.81-13.0 ppm) contents, and initial 87Sr/86Sr ratios (0.7038-0.7040), suggesting adakitic affinity. Combined with its positive ɛNd(t) values (3.5-5.4), low Mg# values (41-50), and low contents of Cr (18.6-29.0 ppm) and Ni (7.3-9.1 ppm), we propose an origin by partial melting of a juvenile mafic lower crust in a post-collisional setting after the amalgamation of the Erguna and Xing'an Blocks. The tonalite is characterized by high SiO2 (63.1-65.9 wt%) and Al2O3 (16.0-16.3 wt%) contents, low (87Sr/86Sr) i ratios (0.7041-0.7042), positive ɛNd(t) values (2.6-3.0), along with LILE and LREE enrichments and Nb-Ta-Ti depletions, suggesting an origin by partial melting of juvenile mafic lower crust, coupled with fractional crystallization, in a post-orogenic setting after the collision between the Xing'an and Songnen Blocks. The δD values of ore-forming fluids range from -100 to -93 ‰, and the δ18O values calculated from hydrothermal quartz are between -3 and 10 ‰. The δ34S values of sulfides vary from -2.6 to -1.1 ‰. Field observations, as well as the geochronological and H-O-S isotopic data, suggest that the Cu mineralization at Tongshan was genetically linked with the granodiorite.
MicroRNA-200b acts as a tumor suppressor in osteosarcoma via targeting ZEB1
Li, Yusheng; Zeng, Chao; Tu, Min; Jiang, Wei; Dai, Zixun; Hu, Yuling; Deng, Zhenhan; Xiao, Wenfeng
Osteosarcoma is the most common type of cancer that develops in bone, mainly arising from the metaphysis of the long bones. MicroRNA (miR)-200b has been found to generally act as a tumor suppressor in multiple types of human cancers. However, the detailed role of miR-200b in osteosarcoma still remains to be fully understood. This study aimed to investigate the exact role of miR-200b in the progression of osteosarcoma and the underlying mechanism. Real-time reverse transcription-polymerase chain reaction data showed that miR-200b was significantly downregulated in osteosarcoma tissues compared to their matched adjacent nontumor tissues. Low miR-200b level was associated with the advanced clinical stage and positive distant metastasis. Besides, it was also downregulated in osteosarcoma cell lines (U2OS, Saos2, HOS, and MG63) compared to normal osteoblast cell line NHOst. In vitro study showed that restoration of miR-200b led to a significant decrease in proliferation, migration, and invasion of osteosarcoma cells. Moreover, ZEB1 was identified as a target gene of miR-200b, and its expression levels were negatively mediated by miR-200b in osteosarcoma cells. In addition, ZEB1 was significantly upregulated in osteosarcoma cells compared to the normal osteoblast cell line NHOst, and inhibition of ZEB1 expression also suppressed the proliferation, migration, and invasion in osteosarcoma cells. Finally, we showed that ZEB1 was frequently upregulated in osteosarcoma tissues compared to their matched adjacent normal tissues, and its expression was reversely correlated to the miR-200b levels in osteosarcoma tissues. Based on these findings, our study suggests that miR-200b inhibits the proliferation, migration, and invasion of osteosarcoma cells, probably via the inhibition of ZEB1 expression. Therefore, miR-200b/ZEB1 may become a potential target for the treatment of osteosarcoma. PMID:27307751
Carbon monoxide in biology and microbiology: surprising roles for the "Detroit perfume".
Davidge, Kelly S; Motterlini, Roberto; Mann, Brian E; Wilson, Jayne Louise; Poole, Robert K
Carbon monoxide (CO) is a colorless, odorless gas with a reputation for being an anthropogenic poison; there is extensive documentation of the modes of human exposure, toxicokinetics, and health effects. However, CO is also generated endogenously by heme oxygenases (HOs) in mammals and microbes, and its extraordinary biological activities are now recognized and increasingly utilized in medicine and physiology. This review introduces recent advances in CO biology and chemistry and illustrates the exciting possibilities that exist for a deeper understanding of its biological consequences. However, the microbiological literature is scant and is currently restricted to: 1) CO-metabolizing bacteria, CO oxidation by CO dehydrogenase (CODH) and the CO-sensing mechanisms that enable CO oxidation; 2) the use of CO as a heme ligand in microbial biochemistry; and 3) very limited information on how microbes respond to CO toxicity. We demonstrate how our horizons in CO biology have been extended by intense research activity in recent years in mammalian and human physiology and biochemistry. CO is one of several "new" small gas molecules that are increasingly recognized for their profound and often beneficial biological activities, the others being nitric oxide (NO) and hydrogen sulfide (H2S). The chemistry of CO and other heme ligands (oxygen, NO, H2S and cyanide) and the implications for biological interactions are briefly presented. An important advance in recent years has been the development of CO-releasing molecules (CO-RMs) for aiding experimental administration of CO as an alternative to the use of CO gas. The chemical principles of CO-RM design and mechanisms of CO release from CO-RMs (dissociation, association, reduction and oxidation, photolysis, and acidification) are reviewed and we present a survey of the most commonly used CO-RMs. Amongst the most important new applications of CO in mammalian physiology and medicine are its vasoactive properties and the
Multi-scale cell/surface interaction on modified titanium aluminum vanadium surfaces
NASA Astrophysics Data System (ADS)
Chen, Jianbo
This dissertation presents a series of experimental studies of the effects of multi-scale cell/surface interactions on modified Ti-6Al-4V surfaces. These include laser-grooved surfaces; porous structures and RGD-coated laser-grooved surfaces. A nano-second DPSS UV lasers with a Gaussian pulse energy profile was used to introduce the desired micro-groove geometries onto Ti-6Al-4V surfaces. This was done without inducing micro-cracks or significant changes in surface chemistry within the heat affected zones. The desired 8-12 mum groove depths and widths were achieved by the control of pulse frequency, scan speed, and the lens focal length that controls spot size. The interactions between human osteosarcoma (HOS) cells and laser-grooved Ti-6Al-4V surfaces were investigated after 48 hours of cell culture. The cell behavior, including cell spreading, alignment and adhesion, was elucidated using scanning electronic microscopy (SEM), immuno-fluorescence staining and enzymatic detachment. Contact guidance was shown to increase as grooved spacing decreased. For the range of micro-groove geometries studied, micro-grooves with groove spacings of 20 mum provided the best combination of cell orientation and adhesion. Short-term adhesion experiments (15 mins to 1 day) also revealed that there is a positive correlation between cell orientation and cell adhesion. Contact guidance on the micro-grooved surfaces is shown to be enhanced by nano- and micro-scale asperities that provide sites for the attachment of lamellopodia during cell locomotion and spreading. Contact guidance is also promoted by the geometrical confinement provided by laser grooves. An experimental study of initial cell spreading and ingrowth into Ti-6Al-4V porous structures was also carried out on porous structures with different pore sizes and geometries. A combination of SEM, the tetrazolium salt (MTT) colorimetric assay and enzymatic detachment were used to study cell spreading and adhesion. The extent of cell
N6-benzyladenosine derivatives as novel N-donor ligands of platinum(II) dichlorido complexes.
Starha, Pavel; Popa, Igor; Trávníček, Zdeněk; Vančo, Ján
The platinum(II) complexes trans-[PtCl₂(Ln)₂]∙xSolv 1-13 (Solv = H₂O or CH3OH), involving N6-benzyladenosine-based N-donor ligands, were synthesized; L(n) stands for N6-(2-methoxybenzyl)adenosine (L₁, involved in complex 1), N6-(4-methoxy-benzyl)adenosine (L₂, 2), N6-(2-chlorobenzyl)adenosine (L₃, 3), N6-(4-chlorobenzyl)-adenosine (L₄, 4), N6-(2-hydroxybenzyl)adenosine (L₅, 5), N6-(3-hydroxybenzyl)-adenosine (L₆, 6), N6-(2-hydroxy-3-methoxybenzyl)adenosine (L₇, 7), N6-(4-fluoro-benzyl)adenosine (L₈, 8), N6-(4-methylbenzyl)adenosine (L₉, 9), 2-chloro-N6-(3-hydroxy-benzyl)adenosine (L₁₀, 10), 2-chloro-N6-(4-hydroxybenzyl)adenosine (L₁₁, 11), 2-chloro-N6-(2-hydroxy-3-methoxybenzyl)adenosine (L₁₂, 12) and 2-chloro-N6-(2-hydroxy-5-methylbenzyl)adenosine (L₁₃, 13). The compounds were characterized by elemental analysis, mass spectrometry, IR and multinuclear (¹H-, ¹³C-, ¹⁹⁵Pt- and ¹⁵N-) and two-dimensional NMR spectroscopy, which proved the N7-coordination mode of the appropriate N6-benzyladenosine derivative and trans-geometry of the title complexes. The complexes 1-13 were found to be non-toxic in vitro against two selected human cancer cell lines (HOS and MCF7; with IC₅₀ > 50.0 µM). However, they were found (by ESI-MS study) to be able to interact with the physiological levels of the sulfur-containing biogenic biomolecule L-methionine by a relatively simple 1:1 exchange mechanism (one L(n) molecule was replaced by one L-methionine molecule), thus forming a mixed-nitrogen/sulfur-ligand dichlorido-platinum(II) coordination species. PMID:23771060
Structure and Function of CC-Chemokine Receptor 5 Homologues Derived from Representative Primate Species and Subspecies of the Taxonomic Suborders Prosimii and Anthropoidea
Kunstman, Kevin J.; Puffer, Bridget; Korber, Bette T.; Kuiken, Carla; Smith, Una R.; Kunstman, Jennifer; Stanton, Jennifer; Agy, Michael; Shibata, Riri; Yoder, Anne D.; Pillai, Satish; Doms, Robert W.; Marx, Preston; Wolinsky, Steven M.
A chemokine receptor from the seven-transmembrane-domain G-protein-coupled receptor superfamily is an essential coreceptor for the cellular entry of human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency virus (SIV) strains. To investigate nonhuman primate CC-chemokine receptor 5 (CCR5) homologue structure and function, we amplified CCR5 DNA sequences from peripheral blood cells obtained from 24 representative species and subspecies of the primate suborders Prosimii (family Lemuridae) and Anthropoidea (families Cebidae, Callitrichidae, Cercopithecidae, Hylobatidae, and Pongidae) by PCR with primers flanking the coding region of the gene. Full-length CCR5 was inserted into pCDNA3.1, and multiple clones were sequenced to permit discrimination of both alleles. Compared to the human CCR5 sequence, the CCR5 sequences of the Lemuridae, Cebidae, and Cercopithecidae shared 87, 91 to 92, and 96 to 99% amino acid sequence homology, respectively. Amino acid substitutions tended to cluster in the amino and carboxy termini, the first transmembrane domain, and the second extracellular loop, with a pattern of species-specific changes that characterized CCR5 homologues from primates within a given family. At variance with humans, all primate species examined from the suborder Anthropoidea had amino acid substitutions at positions 13 (N to D) and 129 (V to I); the former change is critical for CD4-independent binding of SIV to CCR5. Within the Cebidae, Cercopithecidae, and Pongidae (including humans), CCR5 nucleotide similarities were 95.2 to 97.4, 98.0 to 99.5, and 98.3 to 99.3%, respectively. Despite this low genetic diversity, the phylogeny of the selected primate CCR5 homologue sequences agrees with present primate systematics, apart from some intermingling of species of the Cebidae and Cercopithecidae. Constructed HOS.CD4 cell lines expressing the entire CCR5 homologue protein from each of the Anthropoidea species and subspecies were tested for their ability
Comparative effects of n-3, n-6 and n-9 unsaturated fatty acid-rich diet consumption on lupus nephritis, autoantibody production and CD4+ T cell-related gene responses in the autoimmune NZBWF1 mouse.
Pestka, James J; Vines, Laura L; Bates, Melissa A; He, Kaiyu; Langohr, Ingeborg
Mortality from systemic lupus erythematosus (SLE), a prototypical autoimmune disease, correlates with the onset and severity of kidney glomerulonephritis. There are both preclinical and clinical evidence that SLE patients may benefit from consumption of n-3 polyunsaturated fatty acids (PUFA) found in fish oil, but the mechanisms remain unclear. Here we employed the NZBWF1 SLE mouse model to compare the effects of dietary lipids on the onset and severity of autoimmune glomerulonephritis after consuming: 1) n-3 PUFA-rich diet containing docosahexaenoic acid-enriched fish oil (DFO), 2) n-6 PUFA-rich Western-type diet containing corn oil (CRN) or 3) n-9 monounsaturated fatty acid (MUFA)-rich Mediterranean-type diet containing high oleic safflower oil (HOS). Elevated plasma autoantibodies, proteinuria and glomerulonephritis were evident in mice fed either the n-6 PUFA or n-9 MUFA diets, however, all three endpoints were markedly attenuated in mice that consumed the n-3 PUFA diet until 34 wk of age. A focused PCR array was used to relate these findings to the expression of 84 genes associated with CD4+ T cell function in the spleen and kidney both prior to and after the onset of the autoimmune nephritis. n-3 PUFA suppression of autoimmunity in NZBWF1 mice was found to co-occur with a generalized downregulation of CD4+ T cell-related genes in kidney and/or spleen at wk 34. These genes were associated with the inflammatory response, antigen presentation, T cell activation, B cell activation/differentiation and leukocyte recruitment. Quantitative RT-PCR of representative affected genes confirmed that n-3 PUFA consumption was associated with reduced expression of CD80, CTLA-4, IL-10, IL-18, CCL-5, CXCR3, IL-6, TNF-α and osteopontin mRNAs in kidney and/or spleens as compared to mice fed n-6 PUFA or n-9 MUFA diets. Remarkably, many of the genes identified in this study are currently under consideration as biomarkers and/or biotherapeutic targets for SLE and other autoimmune
Nitric oxide and iron modulate heme oxygenase activity as a long distance signaling response to salt stress in sunflower seedling cotyledons.
Singh, Neha; Bhatla, Satish C
Nitric oxide is a significant component of iron signaling in plants. Heme is one of the iron sensors in plants. Free heme is highly toxic and can cause cell damage as it catalyzes the formation of reactive oxygen species (ROS). Its catabolism is carried out by heme oxygenase (HOs; EC 188.8.131.52) which uses heme both as a prosthetic group and as a substrate. Two significant events, which accompany adaptation to salt stress in sunflower seedlings, are accumulation of ROS and enhanced production of nitric oxide (NO) in roots and cotyledons. Present investigations on the immunolocalization of heme oxygenase distribution in sunflower seedling cotyledons by confocal laser scanning microscopic (CLSM) imaging provide new information on the differential spatial distribution of the inducible form of HO (HO-1) as a long distance in response to NaCl stress. The enzyme is abundantly distributed in the specialized cells around the secretory canals (SCs) in seedling cotyledons. Abundance of tyrosine nitrated proteins has also been observed in the specialized cells around the secretory canals in cotyledons derived from salt stressed seedlings. The spatial distribution of tyrosine nitrated proteins and HO-1 expression further correlates with the abundance of mitochondria in these cells. Present findings, thus, highlight a link among distribution of HO-1 expression, abundance of tyrosine nitrated proteins and mitochondria in specialized cells around the secretory canal as a long distance mechanism of salt stress tolerance in sunflower seedlings. Enhanced spatial distribution of HO-1 in response to NaCl stress in seedling cotyledons is in congruence with the observed increase in specific activity of HO-1 in NaCl stressed conditions. The enzyme activity is further enhanced by hemin (HO-1 inducer) both in the absence or presence of NaCl stress and inhibited by zinc protoporphyrin. Western blot analysis of cotyledon homogenates using anti-HO-1 polyclonal antibody shows one major band (29
Selection and stability of quantitative stratigraphic age models: Plio-Pleistocene glaciomarine sediments in the ANDRILL 1B drillcore, McMurdo Ice Shelf
NASA Astrophysics Data System (ADS)
Cody, Rosemary; Levy, Richard; Crampton, James; Naish, Timothy; Wilson, Gary; Harwood, David
Interpretation of glacimarine sedimentary records from Antarctic shelf drillholes has been hampered by the ambiguous age of strata where erosional unconformities and coarse diamictite deposits truncate or omit the magnetostratigraphic and biostratigraphic units used for correlation. However, new quantitative biostratigraphic techniques enable the correlation of sparse, incomplete, and often reworked Plio-Pleistocene records of Ross Sea fossil diatom flora with the more extensively documented but potentially diachronous offshore history of species' first and last appearance datums (FADs and LADs). The approach uses comprehensive regional databases of fossil records and computer-automated search algorithms to find the multidimensional line of correlation (LOC) that best fits local observations, and to identify regionally isochronous biostratigraphic markers. Different model configurations can be used to produce LOCs that represent alternative working hypotheses regarding reworking and other sources of misfit in the biostratigraphic record, and that together provide an envelope of uncertainty for age interpretation. An integrated, quantitative chronostratigraphic model for the ANDRILL-1B drillcore was developed iteratively: an initial age model was constructed solely from preliminary on-ice observations of fossil diatom highest and lowest occurrences (HOs and LOs) and their correlation with a database of other local event records from 24 DVDP, CIROS, and IODP drillcore sections. The model was subsequently updated as off-ice work yielded additional biostratigraphic marker events and revised event horizons, Ar/Ar ages for volcanic material, better-constrained magnetostratigraphic interpretation, and refinements to computational/analytical methodology. The current quantitative biostratigraphic age model for the AND-1B hole integrates the local ranges of 29 diatom taxa, five dated volcanic samples, and independently constrained ages of five paleomagnetic reversals. During
A formylpeptide receptor, FPRL1, acts as an efficient coreceptor for primary isolates of human immunodeficiency virus
Shimizu, Nobuaki; Tanaka, Atsushi; Mori, Takahisa; Ohtsuki, Takahiro; Hoque, Aliful; Jinno-Oue, Atsushi; Apichartpiyakul, Chatchawann; Kusagawa, Shigeru; Takebe, Yutaka; Hoshino, Hiroo
different conditions. FPRL1 has been reported to be abundantly expressed in the lung, spleen, testis, and neutrophils. We detected mRNA expression of FPRL1 in 293T (embryonal kidney cell line), C8166 (T cell line), HOS (osteosarcoma cell line), Molt4#8 (T cell line), U251MG (astrocytoma cell line), U87/CD4 (CD4-transduced glioma cell line), and peripheral blood lymphocytes. Roles of FPRL1 in HIV-1 infection in vivo should be further investigated. PMID:18577234
Atmospheric and land surface measurements in a prototype hydrologic observatory
NASA Astrophysics Data System (ADS)
Scanlon, B.; Krajewski, W.; Famiglietti, J.; Duffy, C.
different microclimate, vegetation, land use, and soils regions to assess the relative importance of these parameters as controls on fluxes and to provide baseline data for predicting potential impacts of future climate and land use change on these fluxes. Critical parameters for hydrological modeling will be included in the measurement design. Although this prototype HO focuses on a porous media system in a humid environment, many of the design aspects are generic and should apply to HOs in other settings (fractured media, arid settings). Another goal of the program is to locate ground-based measurement stations to provide ground referencing for satellite systems. An example application is the use of satellite data (LandSat TM, AVHRR, and MODIS) to estimate ET with comparison to ground-based systems (Eddy Covariance/Bowen Ratio) and scintillometers. The use of Advanced Microwave Scanning Radiometer (AMSR-E) for monitoring soil moisture will also be evaluated using ground-based water content monitoring and field campaigns. Precipitation gauge clusters will be optimally located to evaluate NEXRAD and satellite precipitation data. Understanding the correlation between satellite and ground-based data is essential because spaceborne observations provide spatially and temporally distributed measurements and a framework for scaling local measurements to regional areas. The HO networks will provide valuable information on many aspects of the water cycle to advance our understanding on controls on spatial and temporal variability in fluxes across interfaces.
[Current malaria situation in the Republic of Uzbekistan].
Razakov, Sh A; Shakhgunova, G Sh
Malaria was once one of the most common diseases in Uzbekistan. There were massive epidemics with high mortality rates, wherein 140,000 to 700,000 cases of malaria were recorded. Following large-scale malaria control measures, the disease was eradicated in Uzbekistan in 1961 and the epidemiological situation is still favorable. The natural and climatic conditions that prevail in the Republic of Uzbekistan mean that the country is very susceptible to malaria. There are large water areas varying in type and origin, which provide a habitat for a number of epidemiologically dangerous species of malaria-transmitting mosquitoes in a single area. These are Anopheles maculipennis, An. pulcherrimus and An. superpictus. The prevailing temperatures promote rapid growth of vector mosquitoes and parasites and the malaria transmission season is over 5 months long. Seven malaria-transmitting mosquito species have been recently recorded in the Republic. DDT resistance has been so far noted in Anopheles maculipennis, An. hyrcanus and An. bifurcatus. An. superpictus is sensitive to all insecticides used in clinical practice (organophosphorus and organochlorine compounds, HOS, carbamates, pyrethroids). The most dangerous areas for transmitting malaria by importation are the flood plains of the country's main rivers, such as Syrdarya, Amudarya, Chirchik, Surkhana, etc., and rice-growing areas (an area of about 150,000 ha was under rice cultivation in 1999). The Republic is still very subjected to large-scale importations of malaria particularly in the towns and areas along the border with Tajikistan. There has been recently an increase in the incidence of infections imported into the Republic: 27 cases in 1995, 51 in 1996, 52 in 1997, 74 in 1998, and 78 in 1999. Eight regions of Uzbekistan border Tajikistan, their population is over 5.6 million people. In addition, close family ties between the populations of the frontier towns and regions further increase the risk for malaria to be

subset: commoncrawl