[Influence of LPS and Toll-like receptor 4 antagonist on progesterone receptor, interleukin-1β, and cyclooxygenase-2 in decidual cells].
To observe the expression of progesterone receptor (PR), interleukin-1β (IL-1β), and cyclooxygenase-2 (COX-2) induced by lipopolysaccharide (LPS) or Toll-like receptor 4 antagonist (TLR4 mAb) in decidual cells in vitro, and then to explore the effect of LPS and its antagonist on PR of decidual cells and the relation between PR and inflammatory cytokines. We isolated and cultured human decidua of early abortion in the sterile state. When the cells passaged to the 4th generation, the cells were randomly divided into 6 pore plates: A control group was added the culture medium alone; experimental group I was added 100 ng/mL of LPS; experimental group II was add 1 μg/mL of TLR4 mAb; experimental group III was added 3 μg/ mL of TLR4 mAb; experimental group IV was added 1 μg/mL of TLR4 mAb pretreatment for 24 h, and then 100 ng/mL LPS; and experimental group V was added 3 μg/mL of TLR4 mAb pretreatment for 24 h, and then 100 ng/mL LPS for 24 h culture. Subsequently, HE staining and immunofluorescence were used to observe the morphology and identify the purity of decidual cells in the 6 groups. The levels of mRNA expression of PR, IL-1β, and COX-2 were detected by reverse transcription PCR (RT-PCR). LPS reduced the mRNA expression of PR (P<0.05), increased the mRNA expression of IL-1β and COX-2 (P<0.05). TLR4 mAb increased the mRNA expression of PR (P<0.05) and reduced the mRNA expression of IL-1β (P<0.05) after LPS-stimulated decidual cells. High concentrations of TLR4 mAb reduced the mRNA expression of COX-2 (P<0.05) after LPS stimulated decidual cells. The mRNA expression of PR is reduced, and the mRNA expressions of IL-1β and COX-2 are increased after LPS-stimulated decidual cells in vitro. TLR4 mAb antagonize the role of LPS on PR, IL-1β, and COX-2.


Distribution and location of Daxx in cervical epithelial cells with high risk human papillomavirus positive.
To provide the basis for further exploring the effect and its mechanism of Death domain associated protein (Daxx) on the progress of cervical carcinoma induced by human papillomavirus (HPV), the distribution and location of Daxx in cervical carcinoma with high risk HPV(HR-HPV) positive was analyzed. The samples of normal cervical epithelial cells, cervical intraepithelial neoplasia grade I (CINI), CINII CINIII and cervical cancers were collected. Immunohistochemistry assay was used to analyze the distributions and locations of Daxx in the cervical tissue. Indirect immunoinfluorescence test was utilized to observe the locations of Daxx in Caski cells with HPV16 positive. Under the light microscopy, the brown signals of Daxx distributed in the nuclei of normal cervical epithelial cells; Daxx mainly distributed in nuclear membrane and there were a small amount of Daxx in the nuclei in CINI. Daxx intensively distributed in the cytoplasm and cell membrane in CINII, CINIII and cervical cancer. Under fluorescent microscopy, the distribution and location of Daxx in Caski cells was similarly to that in cervical cells of CINII, CINIII and cervical cancer. In the progress of the cervical cancer, Daxx gradually translocates from nucleus into nuclear membrane, cytoplasm and cell membrane. Daxx locates in the cytoplasm and cell membrane in CINII, CINIII and cervical cancer. The virtual slide(s) for this article can be found here:http://www.diagnosticpathology.diagnomx.eu/vs/4671548951113870.


Peptide QSAR on substance P analogues, enkephalins and bradykinins containing L- and D-amino acids.
Peptide QSARs are constructed for substance P analogues, enkephalins (two examples) and bradykinins containing both L- and D-amino acids. As descriptors in the QSARs, the previously developed descriptors z1 (hydrophobicity), z2 (bulk) and z3 (electronic effect) are used together with a qualitative variable coding for variation in chirality. Two parametrizations of the peptide sequences are tested. In the first no chiral description is used at all, and in the second chirality is described by the qualitative variable. It is concluded that for the current series of peptides, the biological response to variation in amino acid sequence and chirality can be modelled.


Comparison of the pharmacological properties of GK11 and MK801, two NMDA receptor antagonists: towards an explanation for the lack of intrinsic neurotoxicity of GK11.
Over-stimulation of NMDA receptors (NMDARs) is involved in many neurodegenerative disorders. Thus, developing safe NMDAR antagonists is of high therapeutic interest. GK11 is a high affinity uncompetitive NMDAR antagonist with low intrinsic neurotoxicity, shown to be promising for treating CNS trauma. In the present study, we investigated the molecular basis of its interaction with NMDARs and compared this with the reference molecule MK801. We show, on primary cultures of hippocampal neurons, that GK11 exhibits neuroprotection properties similar to those of MK801, but in contrast with MK801, GK11 is not toxic to neurons. Using patch-clamp techniques, we also show that on NR1a/NR2B receptors, GK11 totally blocks the NMDA-mediated currents but has a six-fold lower IC(50) than MK801. On NR1a/NR2A receptors, it displays similar affinity but fails to totally prevent the currents. As NR2A is preferentially localized at synapses and NR2B at extrasynaptic sites, we investigated, using calcium imaging and patch-clamp approaches, the effects of GK11 on either synaptic or extrasynaptic NMDA-mediated responses. Here we demonstrate that in contrast with MK801, GK11 better preserve the synaptic NMDA-mediated currents. Our study supports that the selectivity of GK11 for NR2B containing receptors accounts contributes, at least partially, for its safer pharmacological profile.


[Effects of species richness on colony biomass and tests of sampling- and complementary effect].
With eight species of unicellular algae cultured in random combinations to construct the assemblages with different degrees of richness, the increase pattern of colony biomass with the change of species richness was investigated, and the sampling effect was detected by using multiple variance analysis (MANOVA) method. Three analysis methods, i. e., over-yielding analysis, relative yield total (RYT) index, and subset approach, were applied to explore the complementary effect, and additive partitioning method was applied to separate the sampling effect and complementary effect quantitatively. The results showed that the colony biomass presented a saturated rise with the increase of species richness, and a strong complementary effect was observed in phytoplankton colonies. At exponential growth phase, some specific algal species affected the colony biomass significantly, but after reaching the stable phase, the biomass was not affected by the presence of given phytoplankton. The sampling effect was generally negative at stable phase, while the complementary effect and net biodiversity effect were always positive.


Dental injuries in younger emergency department patients.
Dental trauma is a relatively common occurrence in childhood, with an impact exceeding that of periodontal diseases. It places a significant burden on the public health system because of its high frequency, impact on quality of life and consumption of resources. To identify changing trends of dental injury in patients between 0-18 years of age in Canada and provide a detailed assessment of dental injury patterns in patients attending a large pediatric tertiary care hospital. This retrospective study was carried out in two parts. Firstly, data from patients who presented to the emergency departments of the 15 Canadian hospitals involved in the Injury Reporting and Prevention Program (CHIRPP) from 1st January 1990 to 31st December 2013 was collected. Secondly, at the Hospital for Sick Children, charts of patients identified from the aforementioned database from the years 2008, 2012 and 2015 were accessed for additional clinical data. Based on CHIRPP data there appears to be an increasing incidence in dental injuries presenting to emergency departments nationally. Participation in sports and cycling contributed to the majority of dental injuries. The use of safety equipment such as helmets with a face-grill and mouth-guard was low. Our study demonstrates a rising incidence of dental injuries in young patients. Most dental injuries occurred during sports and playground related activities. The low rate of use of safety equipment and playground safety measures suggests that Canadian Standards Association standards for playgrounds could be more effective if strengthened by mandatory legislation.


Global mapping of cancers: The Cancer Genome Atlas and beyond.
Cancer genomes have been explored from the early 2000s through massive exome sequencing efforts, leading to the publication of The Cancer Genome Atlas in 2013. Sequencing techniques have been developed alongside this project and have allowed scientists to bypass the limitation of costs for whole-genome sequencing (WGS) of single specimens by developing more accurate and extensive cancer sequencing projects, such as deep sequencing of whole genomes and transcriptomic analysis. The Pan-Cancer Analysis of Whole Genomes recently published WGS data from more than 2600 human cancers together with almost 1200 related transcriptomes. The application of WGS on a large database allowed, for the first time in history, a global analysis of features such as molecular signatures, large structural variations and noncoding regions of the genome, as well as the evaluation of RNA alterations in the absence of underlying DNA mutations. The vast amount of data generated still needs to be thoroughly deciphered, and the advent of machine-learning approaches will be the next step towards the generation of personalized approaches for cancer medicine. The present manuscript wants to give a broad perspective on some of the biological evidence derived from the largest sequencing attempts on human cancers so far, discussing advantages and limitations of this approach and its power in the era of machine learning.


The Role of Platelets in Cardiovascular Disease: Molecular Mechanisms.
The role of platelets in atherosclerotic process and subsequently in the pathophysiology of cardiovascular disease is essential as platelets in addition to their contribution to thrombosis and hemostasis modulating inflammatory reactions and immune response. Platelets after adhesion on the injured vascular endothelium and activation release a wide range of molecules stored in platelets granules such as chemokines, proinflammatory molecules and other biological response modulators accelerating interaction among platelets, endothelial cells and leukocytes. These interactions establish a localized inflammatory response that promotes the atherosclerotic process. Moreover, activated platelets give rise to microparticles another active participant within the blood stream. The purpose of this review is to present the role of platelets in the above mechanisms giving an emphasis on the nature of the platelet derived- molecules and their contribution to the atherosclerotic process.


Loss of Emi1-dependent anaphase-promoting complex/cyclosome inhibition deregulates E2F target expression and elicits DNA damage-induced senescence.
Expression of the anaphase-promoting complex/cyclosome (APC/C) inhibitor Emi1 is required for the accumulation of APC/C substrates crucial for DNA synthesis and mitotic entry. We show that in vivo Emi1 expression correlates with the proliferative status of the cellular compartment and that cells lacking Emi1 undergo cellular senescence. Emi1 depletion leads to strong decreases in E2F target mRNA and APC/C substrate protein abundances. However, cyclin E mRNA and cyclin E protein levels and associated kinase activities are increased. Cells lacking Emi1 undergo DNA damage, likely explained by replication stress upon deregulated cyclin E- and A-associated kinase activities. Inhibition of ATM kinase prevents induction of senescence, implying that senescence is a consequence of DNA damage. Surprisingly, no senescence or no extensive amount of senescence is evident upon depletion of the Emi1-stabilizing factor Evi5 or Pin1, respectively. Our data suggest that maintenance of a protein stabilization/mRNA expression positive-feedback circuit fueled by Emi1 is required for accurate cell cycle progression, maintenance of DNA integrity, and prevention of cellular senescence.


Change in Student Pharmacists' Perceptions and Confidence of Pediatric Related Topics After Participation in Pediatric Pharmacy-Focused Skills Laboratory.
The objective of this study was to assess the impact of a pediatric pharmacy-focused skills laboratory on student pharmacist confidence and comfort with pediatric topics. This study compared student responses on a questionnaire completed both pre- and post-lab. The lab activities included pre-readings, a pre-lab lecture, and a 2-hour laboratory session. The questionnaire assessed perceptions related to counseling pediatric patients and confidence in communicating with children and caregivers. Wilcoxon rank sum tests compared the differences in the preand post-lab questionnaire results. The McNemar test was used to test for conversion. A total of 187 of 221 pharmacy students completed the pre-lab questionnaire (85% response rate) and 116 completed the post-lab questionnaire (52% response rate). Significantly higher confidence levels were reported in the post-lab questionnaire for 5 of the 7 questionnaire items. Specifically, concerning level of confidence in measuring oral solutions, 17.5% of students' pre-lab, compared with 42.6% post-lab felt completely confident in being able to determine the appropriate measuring devices for liquid medications. No statistically significant differences were noted regarding comfort level or knowledge. Although an increase in confidence was seen in several areas, some students still lacked confidence after the lab session, suggesting that more pediatric-focused activities may be needed in the curriculum to reinforce pediatric concepts.


New Jersey-Wide Survey of Spotted Fever Group Rickettsia (Proteobacteria: Rickettsiaceae) in Dermacentor variabilis and Amblyomma americanum (Acari: Ixodida: Ixodidae).
For the last decade, the New Jersey (NJ) Department of Health has reported between 42 and 144 new cases each year of "spotted fever group rickettsiosis" (SFGR), a statistic that reflects uncertainty regarding which rickettsial agents (Proteobacteria: Rickettsiaceae: Rickettsia) are infecting NJ residents. To identify the Rickettsia circulating in NJ ticks, we used a combination of conventional and real time PCR approaches to screen 560 Dermacentor variabilis Say and 245 Amblyomma americanum L. obtained from a 1-day state-wide surveillance in May 2018 and an additional 394 D. variabilis collected across NJ in 2013-2018. We found zero D. variabilis infected with Rickettsia rickettsii, the agent of Rocky Mountain spotted fever and, on average, 1.3% infected with presumed nonpathogenic Rickettsia montanensis. We also found zero A. americanum infected with R. rickettsii, and 20% infected with Rickettsia amblyommatis, a prevalence somewhat lower than in more southern states. Overall, we conclude that it is unlikely that R. rickettsii vectored by D. variabilis is a primary cause of SFGR cases in NJ and discuss our findings in the context of known facts and current limitations. We conclude that understanding the causes of SFGR east of the Mississippi will require collaboration among medical doctors, public health authorities, and medical entomologists to follow up presumptive human cases of SFGR with detailed histories of exposure, species-specific molecular assays, and active surveillance of putative vectors and the pathogens they may carry.


Advances in Nanostructure-mediated Hyperthermia in Tumor Therapies.
Hyperthermia has been proposed as a promising treatment modality for its advantageous profiles such as mini/non-invasiveness, good tolerability and cost-effectiveness. Quick development of nanotechnology in recent years has greatly broadened the application area of hyperthermia and endowed it with attractive new functions. This review aims to present an overview of different nanostructures mediated hyperthermia in terms of external stimuli source. We performed to review for the development and current status of nanostructure-mediated hyperthermia, by searching MEDLINE, EMBASE, and Cochrane Library database for identification of relevant articles. In the present study, the systemic results of hyperthermia mediated by nanostructures were researched, and five different kinds of external sources were found and listed in this review. The brief mechanism and commonly explored nanostructures were introduced and then combined therapies of nanostructure-mediated hyperthermia stimulated by different external sources were investigated. Finally, challenges with current nanostructures mediated hyperthermia were discussed in order to give advice to the future development of nanostructure-mediated hyperthermia. Despite all the achievements the new technology of nanostructure-mediated hyperthermia have made in pre-clinical animal experiments, there are still much to be pursued in the further development to be biocompatible, effective and precise.


Malaria Pathogenesis.
In the mosquito-human life cycle, the six species of malaria parasites infecting humans (Plasmodium falciparum, Plasmodium vivax, Plasmodium ovale wallickeri, Plasmodium ovale curtisi, Plasmodium malariae, and Plasmodium knowlesi) undergo 10 or more morphological states, replicate from single to 10,000+ cells, and vary in total population from one to many more than 106 organisms. In the human host, only a small number of these morphological stages lead to clinical disease and the vast majority of all malaria-infected patients in the world produce few (if any) symptoms in the human. Human clinical disease (e.g., fever, anemia, coma) is the result of the parasite preprogrammed biology in concert with the human pathophysiological response. Caveats and corollaries that add variation to this host-parasite interaction include parasite genetic diversity of key proteins, coinfections, comorbidities, delays in treatment, human polymorphisms, and environmental determinants.


Specific effects of microRNAs on the plant transcriptome.
Most plant microRNAs (miRNAs) have perfect or near-perfect complementarity with their targets. This is consistent with their primary mode of action being cleavage of target mRNAs, similar to that induced by perfectly complementary small interfering RNAs (siRNAs). However, there are natural targets with up to five mismatches. Furthermore, artificial siRNAs can have substantial effects on so-called off-targets, to which they have only limited complementarity. By analyzing the transcriptome of plants overexpressing different miRNAs, we have deduced a set of empirical parameters for target recognition. Compared to artificial siRNAs, authentic plant miRNAs appear to have much higher specificity, which may reflect their coevolution with the remainder of the transcriptome. We also demonstrate that miR172, previously thought to act primarily by translational repression, can efficiently guide mRNA cleavage, although the effects on steady-state levels of target transcripts are obscured by strong feedback regulation. This finding unifies the view of plant miRNA action.


[Retroviruses-derived sequences in the human genome. Human endogenous retroviruses (HERVs)].
Retroviruses-derived elements in the human genome constitute 90% of non-coding mobile sequences. Reverse transcriptase (RT) plays an essential role in their transposition as do long terminal repeats (LTRs), which contain promotors, enhancers, and regulatory sequences. Some retroelements (pseudogens and retrogenes, e.g. SINE) are non-autonomic and do not possess their own RT. These elements are dependent on autonomic elements (retroposons, e.g. LINE, retrotransposons, exo- and endogenous retroviruses). The genome of retroviruses is composed of gag, pol, and env genes flanked by long terminal repeats. Endogenous retroviruses are probably the remnants of ancient germ cell infection by exogenous retroviruses and are transmissible to the next generation in a Mendelian way. Most of them are defective (because of mutation accumulation), but some are still active and their expression is regulated by different factors (UV radiation, inflammatory cytokines, steroid hormones, and exogenous virus products). Retroelements as well as their gene products exert influence on the organism's functions. They influence the plasticity and evolution of genomes, are a source of promotors and regulatory sequences, but they also supply additional signals of transcription initiation, mRNA splicing, and STOP codons. One of the positive aspects of human endogenous retroviruses (HERVs) is the participation of their products in normal syncytiotrophoblast formation. They also block exogenous retrovirus replication by receptor interference or antisense mRNA. Their presence is considered to be connected with a number of autoimmunological diseases (multiple sclerosis, insulin-dependent diabetes mellitus, systemic lupus erythematosus), cancer, or even psychiatric disorders (schizophrenia). There are also other problems connected with the potential role of ERVs in genomic therapy (with retroviruses vectors) and transplantology (xenotransplantation).


Community dental clinics in British Columbia, Canada: examining the potential as health equity interventions.
Community dental clinics (CDCs) have emerged to provide oral healthcare for those with low incomes. In British Columbia, the establishment of community clinics has been quite rapid in recent years. However, the expansion has occurred with very little assessment of their impact or effectiveness. While oral health inequities are well recognised, there is limited documentation on healthcare interventions to reduce oral health inequities. This study examines CDCs as health equity interventions from the perspectives of individuals establishing and operating the clinics. The study included interviews with 17 participants--4 dentists, 6 dental hygienists and 7 clinic managers--from 10 clinics operating in British Columbia, Canada in 2008-2009. A thematic analysis of the interview transcripts, explored through a health equity framework, found many ways in which the CDCs exemplify health equity interventions. Although their design and mandates are varied, they potentially enable access to dental treatment, but without necessarily ensuring sustainable outcomes. Moreover, the scalability of CDCs is questionable because frequently they are limited to emergency care and are less responsive to the gradients of needs for populations facing multiple barriers to care. Many of them operate on a charitable base with input from volunteer dentists; however, this foundation probably eases the pressure on dentists and dental hygienists rather than offering a safety net to underserved populations.


Safety and efficacy of systematic lesion preparation with a novel generation scoring balloon in complex percutaneous interventions: results from a prospective registry.
Coronary lesions predilatation with semicompliant (SC) or non-compliant balloons (NC) may be insufficient to obtain an optimal stent expansion, which can lead to in-stent restenosis or thrombosis. Moreover, increasing evidence supporting an optimal lesion preparation is mandatory when drug coated balloons (DCB) are used. To this extent, more "aggressive tools" such as cutting/scoring balloons, atherectomy or lithotripsy may play an important role and improve outcomes. We enrolled 78 consecutive patients from March 2020 to October 2020 with calcific/fibrotic or ostially-located lesions, which were prepared using scoring balloons, in addition to SC/NC balloons and other plaque modification strategies. The final treatment consisted in either stent or DCB usage. The primary endpoint was the rate of clinically-driven target lesion revascularization. Secondary endpoints entailed the procedural success and the individual rates of major adverse cardiac events (MACE) at 12 months. Most of the patients had left main (LM) or ostial lesions, 65% of them being moderate/severely calcified, with further debulking strategies being required in 15 (19.2%) patients (rotational atherectomy, 3.8% or coronary intravascular lithotripsy, 15.3%). A high-rate of DCB usage was reported. Angiographic and procedural success was obtained in 77 and 76 patients, respectively. We encountered one vessel perforation, which was sealed with a covered stent, without consequence. During follo- up, we observed only 6 MACE, 6 target lesion revascularizations (TLR) and 2 cardiovascular deaths. Among patients with high complexity and calcific lesions, an optimal lesion preparation using a dedicated scoring balloon was associated with low clinical events at mid-term follow-up and may be considered to improve immediate procedural success rate.


Outcomes in a community sex offender treatment program: a comparison between polygraphed and matched non-polygraphed offenders.
This study compared a group of 104 adult male sex offenders who received community cognitive-behavioral treatment, correctional supervision, and periodic polygraph compliance exams with a matched group of 104 sex offenders who received the same type of treatment and supervision services but no polygraph exams. Polygraph exams focused on whether participants were following their conditions of community supervision and treatment and had avoided committing new sexual offenses. The two groups were exact pair-wise matched on three variables: (1) Static-99 risk score (Hanson & Thornton 2000, Law and Human Behavior, 24, 119-136), (2) status as a completer of prison sex offender treatment, and (3) date placed in the community. At fixed 5-year follow-up periods, the number of individuals in the polygraph group charged with committing a new non-sexual violent offense was significantly lower than in the no polygraph group (2.9% versus 11.5%). However, there were no significant between-group differences for the number of individuals charged for new sexual (5.8% versus 6.7%), any sexual or violent (8.7% versus 16.3%), or any criminal offense (39.4% versus 34.6%). The results are discussed in terms of their clinical and research implications.


Simultaneous detection of genetic and copy number alterations in BRCA1/2 genes.
Germline mutations in BRCA1 and BRCA2 genes (BRCA1/2) predispose to hereditary breast and ovarian cancer syndrome (HBOC), and their dysregulation increases the risk of cancers. The detection of pathogenic BRCA1/2 variants is essential for the diagnosis and prevention of HBOC, and for offering treatment decisions for patients. Therefore, there is a growing demand for the development of accurate, rapid assay systems that simultaneously detect pathogenic variants and copy number alterations. Here, we tested Thermo Fisher Scientific's newly developed Oncomine®BRCA1/2 Panel. We showed that all mutations in standard reference DNA were detected with high accuracy, and that values of allelic fractions were detected with high concordance (R2 = 0.9986). The Oncomine®BRCA1/2 Panel detected 21 pathogenic germline variants in 147 patients with breast and/or ovarian cancer, of which 20 were detected by the previously-launched Ion AmpliSeq™ BRCA1/2 Panel, except for one frameshift mutation. The Oncomine®BRCA1/2 Panel precisely captured one additional frameshift mutation, which is difficult to detect because of the homopolymer site. Large genomic deletion was identified in one sample, which was previously detected by multiplex ligation-dependent probe amplification. Oncomine®BRCA1/2 Panel could accurately detect pathogenic variant and copy number alteration, and be an alternative assay to investigate BRCA1/2 germline and somatic mutations.


Narrow Band Imaging versus White Light for the Detection of Sessile Serrated Colorectal Lesions: A Randomized Clinical Trial.
Colorectal cancer (CRC) is a leading cause of cancer. The detection of pre-malignant lesions by colonoscopy is associated with reduced CRC incidence and mortality. Narrow band imaging has shown promising but conflicting results for the detection of serrated lesions. We performed a randomized clinical trial to compare the mean detection of serrated lesions and hyperplastic polyps ≥10 mm with NBI or high-definition white light (HD-WL) withdrawal. We also compared all sessile serrated lesions (SSLs), adenoma, and polyp prevalence and rates. Overall, 782 patients were randomized (WL group 392 patients; NBI group 390 patients). The average number of serrated lesions and hyperplastic polyps ≥10 mm detected per colonoscopy (primary endpoint) was similar between the HD-WL and NBI group (0.118 vs. 0.156, p = 0.44). Likewise, the adenoma detection rate (55.2% vs. 53.2%, p = 0.58) and SSL detection rate (6.8% vs. 7.5%, p = 0.502) were not different between the two study groups. Withdrawal time was higher in the NBI group (10.88 vs. 9.47 min, p = 0.004), with a statistically nonsignificant higher total procedure time (20.97 vs. 19.30 min, p = 0.052). The routine utilization of narrow band imaging does not improve the detection of serrated class lesions or any pre-malignant lesion and increases the withdrawal time.


Localization of hydrogen-bonds within modules in barnase.
Proteins in eukaryotes are composed of structural units, each encoded by discrete exons. The protein module is one such structural unit; it has been defined as the least extended or the most compact contiguous segment in a globular domain. To elucidate roles of modules in protein evolution and folding, we examined roles of hydrogen bonds and hydrophobic cores, as related to the stability of these modules. For this purpose we studied barnase, a bacterial RNase from Bacillus amylolique-faciens. Barnase is decomposed into at least six modules, M1-M6; the module boundaries are identified at amino acid residues 24, 52, 73, 88, and 98. Hydrogen bonds are localized mainly within each of the modules, with only a few between them, thereby indicating that their locations are designed to primarily stabilize each individual module. To obtain support for this notion, an analysis was made of hypothetical modules defined as segments starting at a center of one module and ending at the center of the following one. We found that the hydrogen bonds did not localize in each hypothetical module and that many formed between the hypothetical modules. The native conformations of modules of barnase may be specified predominantly by interactions within the modules.


[Catalase activity in the agent of tularemia].
The catalase activity was studied in the extracts of 27 strains of Francisella tularensis belonging to three geographical races. The most active enzyme was possessed by nonarctic strains, and the least--by holarctic. Japanese variant of the holarctic race was separated in an individual group since it possessed a higher catalase activity in comparison with the typical strains. The Central Asian race occupied an intermediate position by this sign and was placed between the Japanese variant and the nonarctic race. Attenuation of the strains of all the three races of F. tularensis led to a marked reduction of the catalase activity.


[ESTABLISHMENT OF INTERVERTEBRAL DISC DEGENERATION OF RABBITS BY USING MINIMALLY INVASIVE ACUPUNCTURE AND ROTARY-CUTTING].
To investigate the feasibility of establishing intervertebral disc degeneration (IDD) model by using minimally invasive acupuncture and rotary-cutting. Forty New Zealand white rabbits [male or female, (2.9 ± 0.3) kg in weight] were randomly divided into control group (n = 20) and experimental group (n = 20). No treatment was done in the control group; percutaneous puncture was performed on L4,5 and L5,6 intervertebral disc by using 18G needle under C-arm X-ray monitoring for rotary-cutting of nucleus pulposus to promote degeneration of the disc in the experimental group. At 4, 8, 12, and 16 weeks after operation, general observation and MRI observation were done, and intervertebral disc degeneration was accessed based on Pfirrmann grade; the specimens were harvested for Masson staining and Safranine O staining. The nucleus pulposus showed dark colors and reduced elasticity in the experimental group when compared with the control group. T2-weighted MRI images indicated that the disc signal intensity of control group had no obvious change at early stage, and weakened slightly at late stage; disc signal intensity of the experimental group decreased with time. According to Pfirrmann grade for disc degeneration, disc degeneration degree was significantly aggravated with time in 2 groups (P < 0.05); degeneration was significantly more severe in the experimental group than the control group at the other time points (P < 0.05) except 4 weeks (P > 0.05). Masson staining results showed that irregular arrangement of annulus with integrate structure was observed in the control group with time; the annulus of the experimental group arranged in disorder, or even disc fibrous circle rupture appeared with time. Safranin O staining showed that the nucleus pulposus cells reduced significantly in the experimental group, but did not in the control group. Minimally invasive acupuncture and rotary-cutting could successfully establish the IDD model in rabbits.


Countermeasures against pulmonary threat agents.
Inhaled toxicants are used for diverse purposes, ranging from industrial applications such as agriculture, sanitation, and fumigation, to crowd control and chemical warfare, and acute exposure can induce lasting respiratory complications. The intentional release of chemical warfare agents (CWAs) during World War I caused life-long damage for survivors, and CWA use is outlawed by international treaties. However, in the past two decades chemical warfare use has surged in the Middle East and Eastern Europe with a shift toward lung toxicants. The potential use of industrial and agricultural chemicals in rogue activities are a major concern, as they are often stored and transported near populated areas, where intentional or accidental release can cause severe injuries and fatalities. Despite laws and regulatory agencies that regulate use, storage, transport, emissions, and disposal, inhalational exposures continue to cause lasting lung injury. Industrial irritants (e.g., ammonia) aggravate the upper respiratory tract, causing pneumonitis, bronchoconstriction, and dyspnea. Irritant gases (e.g., acrolein, chloropicrin) affect epithelial barrier integrity and cause tissue damage, through reactive intermediates or by direct adduction of cysteine-rich proteins. Symptoms from CWAs (e.g., chlorine gas, phosgene, sulfur mustard) progress from airway obstruction and pulmonary edema to acute lung injury (ALI) and acute respiratory distress syndrome (ARDS) that results in respiratory depression days later. Emergency treatment is limited to supportive care using bronchodilators to control airway constriction, and rescue with mechanical ventilation to improve gas-exchange. Complications from acute exposure can promote obstructive lung disease and/or pulmonary fibrosis which require long-term clinical care. Significance Statement Inhaled chemical threats are of growing concern in both civilian and military settings, and there is increased need to reduce acute lung injury and delayed clinical complications from exposures. This minireview highlights our current understanding of acute toxicity and pathophysiology of a select number of chemicals of concern. It discusses potential early-stage therapeutic development, as well as challenges in developing countermeasures applicable for administration in mass casualty situations.


Sensitivity of periparturient breakdown of immunity to parasites to dietary protein source.
Effects of increased MP supply on the degree of periparturient relaxation of immunity (PPRI) in sheep may be dependent on quality of supplied MP. Here we tested the hypothesis that additional MP supply from rations based on xylose-treated soybean meal would be more effective than from rations based on faba beans in reducing the degree of PPRI, as indicated by nematode egg excretion. Twenty-four multiple-bearing ewes were trickle infected with Teladorsagia circumcincta larvae from d -56 to d 31 relative to start of lactation (d 0). From d -26 onwards, ewes were fed at either 0.8 (LP) or at 1.2 times their respective calculated MP requirements using either xylose-treated soybean (HPS) or faba beans (HPB). Litter size was adjusted to 2 lambs at parturition. Feeding treatments did not affect nematode egg excretion, ewe BW or BCS during late pregnancy (P > 0.10), but HPS and HPB ewes had reduced plasma pepsinogen concentrations (P = 0.003). During lactation, HPS and HPB feeding increased ewe BW gain (P < 0.001) and BCS (P = 0.017), and reduced plasma pepsinogen concentrations (P = 0.008) to the same extent, compared with LP feeding. However, only HPS feeding increased litter weight gain (P = 0.017) and reduced nematode egg excretion (P = 0.015), which were both similar between HPB and LP (P > 0.10). The results support the view that extra MP supply from xylose-treated soybean based rations is more effective in reducing parasitism than MP from faba bean-based rations, suggesting that protein source and/or quality are important factors to consider for the nutritional control of parasitism.


The business case as a strategic tool for change.
The authors discuss the clinically focused business case, when it is and is not needed, and the knowledge and skills the nurse executive must master to use the business case effectively as a strategic tool. Necessary skills include translating nursing practice proposals into cost-effective change initiatives and marketing those changes to colleagues.


Technical advances in monitoring human motility patterns.
Abnormal motor patterns are implicated in many motility disorders. However, for many regions of the gut, our knowledge of normal and abnormal motility behaviors and mechanisms remains incomplete. There have been many recent advances in the development of techniques to increase our knowledge of gastrointestinal motility, some readily available while others remain confined to research centers. This review highlights a range of these recent developments and examines their potential to help diagnose and guide treatment for motility disorders.


How is female mate choice affected by male competition?
The plethora of studies devoted to the topics of male competition and female mate choice belie the fact that their interaction remains poorly understood. Indeed, on the question of whether competition should help or hinder the choice process, opinions scattered throughout the sexual selection literature seem unnecessarily polarised. We argue, in the light of recent theoretical and empirical advances, that the effect of competition on mate choice depends on whether it results in the choosy sex attaining high breeding value for total fitness, considering both direct and indirect fitness benefits. Specifically, trade-offs may occur between different fitness benefits if some are correlated with male competitive ability whilst others are not. Moreover, the costs and benefits of mating with competitive males may vary in time and/or space. These considerations highlight the importance of injecting a life-history perspective into sexual selection studies. Within this context, we turn to the sexual selection literature to try to offer insights into the circumstances when competition might be expected to have positive or negative implications for pre-copulatory female choice. In this regard, we elaborate on three stages where competition might impact upon the choice process: (i) during mate detection, (ii) mate evaluation, and (iii) in dictating actual mating outcomes. We conclude by offering researchers several potentially rewarding avenues for future research.


Relative quantification of human β-defensins by a proteomics approach based on selected reaction monitoring.
A targeted proteomics method based on selected reaction monitoring (SRM) is a relevant approach for the analysis of multiple analytes in biological samples. Defensins are phylogenetically conserved small antimicrobial peptides contributing to innate host defense and exhibiting low immunogenicity, resistance to proteolysis and a broad range of antimicrobial activities. The goal of the present study was to develop and optimize SRM-based targeted proteomics methods for the detection of human β-defensins 1-4 in various biological fluids. An SRM-based targeted proteomics method was developed and validated for the detection of human β-defensins 1-4. The supernatants of resting and IL-1β-stimulated Caco2, HT-29 and SW-1116 colonic epithelial cells (CEC), cell lysates of CECs and tear samples of human healthy individuals were analyzed and the feasibility of the developed method was validated by ELISA and dot-blot analysis complemented by RT-qPCR. Our results demonstrate that the developed SRM method offers an alternative approach for the cost-effective and rapid analysis of human β-defensins in samples with biological relevance. A semi-quantitative targeted mass spectrometry method was developed and validated for the relative quantification of β-defensins 1-4 in cell culture supernatants and body fluid analyses.


Fisetin Ameliorates Alcohol-Induced Liver Injury through Regulating SIRT1 and SphK1 Pathway.
Alcoholic liver disease (ALD) often leads to hepatitis, hepatic cirrhosis, and even hepatocellular carcinoma. Fisetin has been shown to confer protection against liver injury. Herein, we investigated whether fisetin could prevent ethanol-induced hepatotoxicity. Mice were fed on 5% (v/v) Lieber-DeCarli ethanol diet. Human primary hepatic stellate cells (HSCs) co-cultured with ethanol were used to verify the therapeutic effect of fisetin. The results of alanine/aspartate aminotransferase (ALT/AST), Triglyceride (TG), total cholesterol (TC) in serum, Oil O Red and Masson staining revealed that fisetin (80[Formula: see text]mg/kg) ameliorated ethanol-induced mice liver injury and fibrosis. Besides, immunofluorescence results of [Formula: see text]-SMA revealed that fisetin suppressed HSCs activation. The suppression was dose-dependent. Furthermore, fisetin promoted SIRT1-mediated autophagy and inhibited Sphk1-mediated endoplasmic reticulum stress (ER stress) both in vitro and in vivo. Molecular docking results indicated potential interaction of fisetin with SIRT1 and SphK1. The inhibitory effect of fisetin on HSCs activation was reversed on co-culturing with EX-527, a specific inhibitor against STIR1 overexpression. Thus, fisetin has the potential to ameliorate alcohol-induced liver injury through suppression of HSCs activation, SIRT1-mediated autophagy and Sphk1-mediated ER stress.


[Visceral adhesion after intraperitoneal ventral hernia treatment: monocentric study comparative of protected versus unprotected meshes].
Intraperitoneal (IP) ventral hernia repair is advantageous because of reduced dissection, shorter operative time and less postoperative pain. However, the IP positioning of the mesh is suspected to increase the risk of visceral adhesion and induce complications. To overcome these drawbacks, an innovative mesh: Parietex Composite (RC) was developed with one side protected by a hydrophilic resorbable film. The purpose of this study was to compare using ultrasonography the rate of visceral adhesions after IP placement of a conventional mesh (RP) versus RC mesh. Twenty-six patients who received a Parietex Composite were prospectively compared to a retrospective series of 26 consecutive asymptomatic patients who received a non protected polyester mesh (RP). In order to objectively assess visceral adhesion toward the abdominal wall, an ultrasound specific examination was used after previous validation by comparison of preoperative ultrasonographic data with peroperative gross appearance in both groups. With a mean follow up of 33 months, both groups were equivalent in term of inclusion criteria excepted for age which was older the RP group. US validation data were: sensitivity 72%, accuracy 69%, negative predictive value 67%. Using this procedure, 81% of the patients exhibit visceral adhesion to the mesh in the RP group, versus 27% in the PC group (P =0.0002, chi2). US examination represents a suitable tool to evaluate postoperative adhesions to the abdominal wall. A significant reduction of visceral adhesion in the RC group was shown.


Relative efficacy of T cell stimuli as inducers of productive HIV-1 replication in latently infected CD4 lymphocytes from patients on suppressive cART.
Quantification of cell-associated replication-competent HIV, in blood samples from patients with undetectable plasma viremia, requires specialized culture conditions that include exogenous pan T cell stimulation. Different research groups have used several stimuli for this purpose; however, the relative efficacies of these T cell stimuli to induce productive HIV replication from latently infected cells ex vivo have not been systematically evaluated. To this end, we compared four commonly used T cell stimuli: 1) irradiated allogeneic cells plus phytohaemagglutinin (PHA); 2) PHA alone; 3) phorbol myristate acetate plus Ionomycin; and 4) immobilized αCD3 plus αCD28 antibodies. End-point dilutions of patient CD4 T cells were performed, using virion RNA production to quantify HIV induction. Our results demonstrated that these activation approaches were not equivalent and that antibody cross-linking of CD3 and CD28 membrane receptors was the most effective means to activate HIV replication from a resting cell state, closely followed by stimulation with irradiated allogeneic cells plus PHA.


Quantitative phase maps denoising of long holographic sequences by using SPADEDH algorithm.
We propose a denoising method for digital holography mod 2π wrapped phase map by using an adaptation of the SPArsity DEnoising of Digital Holograms (SPADEDH) algorithm. SPADEDH is a l(1) minimization algorithm able to suppress the noise components on digital holograms without any prior knowledge or estimation about the statistics of noise. We test our algorithm with either general numerical simulated wrapped phase, quantifying the performance with different efficiency parameters and comparing it with two popular denoising strategies, i.e., median and Gaussian filters, and specific experimental tests, by focusing our attention on long-sequence wrapped quantitative phase maps (QPMs) of in vitro cells, which aim to have uncorrupted QPMs. In addition, we prove that the proposed algorithm can be used as a helper for the typical local phase unwrapping algorithms.


An analysis of horseradish peroxidase enzyme for effluent treatment.
The present study explains computational methods to design thermostable horseradish peroxidase enzyme using the crystal structure available from Protein Data Bank (PDB ID: 6ATJ). Multiple mutations were introduced to the original enzyme and developed a model by using Modeler9.14. After designing the model functional effect was confirmed in terms of protein ligand binding by molecular docking using Autodock 4.2. The implementation of modeling steps is demonstrated in the context of performing mutations for particular amino acid residue on the ligand pocket of the horseradish peroxidase, to derive the desired ligand binding properties. The docking investigation of modelled HRP with Quercetindihydroxide using Autodock 4.2 software that six amino acid residues, P139, H42, A31, L174, A38, and G169 are involved in hydrogen bonding. More importantly, it provides insight into understanding and properly interpreting the data produced by these methods. The 3D model was docked with Quercetindihydroxide (a known horseradish modulator) to understand molecular interactions at the active site region.


Sarcoid Pericarditis and Large Vessel Vasculitis Detected on FDG PET/CT.
A 65-year-old woman with history of sarcoidosis, hypertension, and peripheral vascular disease presented with lower extremity claudication and low left ventricular ejection fraction of 45% on echocardiogram. Further FDG PET/CT revealed hypermetabolic bilateral chest lymphadenopathy and pulmonary nodules in a butterfly-shaped distribution pattern, which was typically seen in patients with sarcoidosis. In addition, abnormal increased radiotracer uptake was present in pericardium, and along the walls of inferior vena cava, aorta, and pulmonary artery, which correlated with delayed enhancement on further cardiac MRI. These findings were suggestive of sarcoid-related pericarditis and great vessel vasculitis.


MicroRNAs in the etiology of colorectal cancer: pathways and clinical implications.
MicroRNAs (miRNAs) are small single-stranded RNAs that repress mRNA translation and trigger mRNA degradation. Of the ∼1900 miRNA-encoding genes present in the human genome, ∼250 miRNAs are reported to have changes in abundance or altered functions in colorectal cancer. Thousands of studies have documented aberrant miRNA levels in colorectal cancer, with some miRNAs reported to actively regulate tumorigenesis. A recurrent phenomenon with miRNAs is their frequent participation in feedback loops, which probably serve to reinforce or magnify biological outcomes to manifest a particular cellular phenotype. Here, we review the roles of oncogenic miRNAs (oncomiRs), tumor suppressive miRNAs (anti-oncomiRs) and miRNA regulators in colorectal cancer. Given their stability in patient-derived samples and ease of detection with standard and novel techniques, we also discuss the potential use of miRNAs as biomarkers in the diagnosis of colorectal cancer and as prognostic indicators of this disease. MiRNAs also represent attractive candidates for targeted therapies because their function can be manipulated through the use of synthetic antagonists and miRNA mimics.


Ligand Selectivity Mechanism and Conformational Changes in Guanine Riboswitch by Molecular Dynamics Simulations and Free Energy Calculations.
Riboswitches regulate gene expression through direct and specific interactions with small metabolite molecules. Binding of a ligand to its RNA target is high selectivity and affinity and induces conformational changes of the RNA's secondary and tertiary structure. The structural difference of two purine riboswitches aptamers is caused by only one single mutation, where cytosine 74 in the guanine riboswitch is corresponding to a uracil 74 in adenine riboswitch. Here we employed molecular dynamics (MD) simulation, molecular mechanics Poisson-Boltzmann surface area (MM-PBSA) and thermodynamic integration computational methodologies to evaluate the energetic and conformational changes of ligands binding to purine riboswitches. The snapshots used in MM-PBSA calculation were extracted from ten 50 ns MD simulation trajectories for each complex. These free energy results are in consistent with the experimental data and rationalize the selectivity of the riboswitches for different ligands. In particular, it is found that the loss in binding free energy upon mutation is mainly electrostatic in guanine (GUA) and riboswitch complex. Furthermore, new hydrogen bonds are found in mutated complexes. To reveal the conformational properties of guanine riboswitch, we performed a total of 6 μs MD simulations in both the presence and the absence of the ligand GUA. The MD simulations suggest that the conformation of guanine riboswitch depends on the distance of two groups in the binding pocket of ligand. The conformation is in a close conformation when U51-A52 is close to C74-U75.


Evaluation of an elective ultrasound course for medical students.
To evaluate the educational benefits of teaching ultrasound in an elective radiological course for medical students. We conducted a single-center retrospective analysis of a one-week elective ultrasound course in which 39 medical students (25 female; median age 25.8 ± 2.8 years, range 21-35, 4th and 5th years of undergraduate medical education) participated as part of their radiological training. The students completed a pre- and post-course questionnaire using a five-point Likert scale for self-assessment of technical knowledge, confidence in the procedure regarding different organs, motivation for performing ultrasound, and the importance of ultrasound in medical education. The students also assessed the value of ultrasound teaching in the context of their general medical degree. Pre-test and post-test scores showed significantly improved radiology knowledge (p ≤ 0.001) concerning all abdominal organs (liver, spleen, gallbladder, kidney, urinary bladder, aorta and retroperitoneum). A significant improvement was registered in performing ultrasound of peripheral arteries and venous structures (each p = 0.001), and also of the thyroid gland (p = 0.006). General surveys at the end of the elective were overwhelmingly positive, with constructive criticism and complimentary comments. Systematic training in ultrasound markedly improved the students' skills. This was especially pronounced for solid organ structures, while students requested more education regarding the retroperitoneum and vascular structures. Teaching ultrasound in an elective hands-on training course improves anatomical understanding and practical skills associated with increased motivation. The results affirm the necessity and clinical relevance of the course during the radiological training of medical students.


Foetal anasarca in Awassi sheep.
Two anasarcous foetuses of Awassi sheep are described. The foetuses were removed from the dams by caesarean section because of dystocia due to failure of cervical dilation. Uterine incision was made in situ because uteri were so distended they could not be brought out from the site of incision. Large quantities of uterine fluids and abnormal thick placentas were found. One foetus weighed about 7 kg and the other 13 kg. The foetal heads were deformed: the upper jaw was prognathic and the left ear of the small foetus was cystic. Necropsy revealed subcutaneous musculature was soft and flabby and abdominal and thoracic cavities contained serosanguinous fluid. Histopathological examination revealed that only the larger foetus had focal aggregates of basophilic nucleated red blood cells and scattered megakaryocytes in the liver. We conclude that anasarca can occur in Awassi sheep, with and without associated extramedullary haematopoiesis.


A safer framework to evaluate characterization technologies of exhaled biologic materials using electrospun nanofibers.
Exhaled biologic material is the source for the spread of many respiratory tract infections. To avoid the high-level of biosafety required to manage dangerous pathogens, we developed a safer framework using the endogenous surrogate targets RNase P and Streptococcus mitis as a means to sample exhaled biologics. Our exhalation collection scheme uses nanoscale fibrous poly(vinyl alcohol) substrates as facemask inserts. After a period of breathing or speaking, the inserts are removed and dissolved. RNase P RNA and S. mitis DNA are extracted for quantification by multiplexed RT-qPCR. Both surrogate biomarkers were detected in all samples obtained during breathing for at least five minutes or speaking for one minute. Phrases repeated 30 times had the most copies with 375 ± 247 of S. mitis and 54 ± 33 of RNase P. When the phrases were repeated just 5 times, the S. mitis copies collected were still detectable but at a significantly lower level of 11 ± 5 for S. mitis and 12 ± 9 for RNase P. These results demonstrate a collection and quantification framework that can be readily adapted to further characterize the exhalation of nanoscale biologic materials from healthy individuals, explore new collection designs safely, and serve as a method to incorporate sample controls for future pathogen exhalation studies.


Does single-stranded DNA pass through the inner channel of the protein hexamer in the complex with the Escherichia coli DnaB Helicase? Fluorescence energy transfer studies.
The structure of the complex of the Escherichia coli primary replicative helicase DnaB protein with single-stranded (ss) DNA and replication fork substrates has been examined using the fluorescence energy transfer method. In these experiments, we used the DnaB protein variant, R14C, which has arginine 14 replaced by cysteine in the small 12-kDa domain of the protein using site-directed mutagenesis. The cysteine residues have been modified with a fluorescent marker which serves as a donor or an acceptor to another fluorescence label placed in different locations on the DNA substrates. Using the multiple fluorescence donor-acceptor approach, we provide evidence that, in the complex with the enzyme, ssDNA passes through the inner channel of the DnaB hexamer. This is the first evidence of the existence of such a structure of a hexameric helicase-ssDNA complex in solution. In the stationary complex with the 5' arm of the replication fork, without ATP hydrolysis, the distance between the 5' end of the arm and the 12-kDa domains of the hexamer (R = 47 A) is the same as in the complex with the isolated ssDNA oligomer (R = 47 A) having the same length as the arm of the fork. These data indicate that both ssDNA and the 5' arm of the fork bind in the same manner to the DNA binding site. Moreover, in the complex with the helicase, the length of the ssDNA is similar to the length of the ssDNA strand in the double-stranded DNA conformation. In the stationary complex, the helicase does not invade the duplex part of the fork beyond the first 2-3 base pairs. This result corroborates the quantitative thermodynamic data which showed that the duplex part of the fork does not contribute to the free energy of binding of the enzyme to the fork. Implications of these results for the mechanism of a hexameric helicase binding to DNA are discussed.


Nitrogen-doped graphene and its application in electrochemical biosensing.
Chemical doping with foreign atoms is an effective method to intrinsically modify the properties of host materials. Among them, nitrogen doping plays a critical role in regulating the electronic properties of carbon materials. Recently, graphene, as a true two-dimensional carbon material, has shown fascinating applications in bioelectronics and biosensors. In this paper, we report a facile strategy to prepare N-doped graphene by using nitrogen plasma treatment of graphene synthesized via a chemical method. Meanwhile, a possible schematic diagram has been proposed to detail the structure of N-doped graphene. By controlling the exposure time, the N percentage in host graphene can be regulated, ranging from 0.11 to 1.35%. Moreover, the as-prepared N-doped graphene has displayed high electrocatalytic activity for reduction of hydrogen peroxide and fast direct electron transfer kinetics for glucose oxidase. The N-doped graphene has further been used for glucose biosensing with concentrations as low as 0.01 mM in the presence of interferences.


Recognition DNA sequence of a novel putative transcription factor, BCL6.
The BCL6 gene involved in the 3q27 translocation associated with B-cell lymphomas encodes a novel Cys2-His2 zinc finger protein. We generated a fusion protein of glutathione S-transferase and zinc finger domain of BCL6 to determine recognition sequences of BCL6 with polymerase chain reaction using random oligonucleotides of 26 bases as a ligand. A consensus of 14 nucleotides consisting of (T/A)NCTTTCNAGG(A/G)AT was identified in the recognition sequences. In a gel mobility shift assay, the probe containing the 14-nucleotide recognition sequence formed a complex with the fusion protein and nuclear proteins from Burkitt's cell lines overexpressing the BCL6 transcripts. The consensus sequence was protected from the digestion by nuclease in a DNase I footprinting assay. In conclusion, BCL6 may be involved in tumorigenesis by binding to the consensus sequences of the other genes.


Copy number variations in normal karyotype acute myeloid leukaemia and their association with treatment response.
Copy number variation (CNV) has been reported to be associated with chemotherapy response, which affects disease prognosis. Here, we determined the frequency of genome-wide cytogenetic CNV aberrations in Korean patients with normal karyotype (NK) acute myeloid leukaemia (AML) and tested whether these genomic variations contribute to differences in Ara-C and anthracycline-based chemotherapy responses. Bone marrow aspirates and blood from 30 previously untreated de novo NK-AML patients were provided at the time of diagnosis for copy number analysis. Possible associations between cytogenetic aberrations and clinical parameters were analysed. CNVs were identified in 23 (76.7%) of the 30 cases tested. Multivariate analyses controlled for other clinical co-variates showed that patients having copy number loss had a decreased probability of complete remission (OR, 0.015 [95% CI, 0-0.737], p = 0.035). Patients who had a copy number gain of more than four regions tended to have shorter event-free survival (EFS) (p = 0.083) with multivariate analysis showing that CNV increase is an independent predictive factor for shorter EFS (HR, 22.104 [95% CI, 1.644-297.157], p = 0.020). In addition, we identified candidate genes that may be involved in Ara-C and anthracycline drug response in Korean patients with NK-AML. These results suggest that CNVs may affect the success of Ara-C and anthracycline-based chemotherapy in Korean patients with NK-AML.


Particulate matter containing environmentally persistent free radicals induces AhR-dependent cytokine and reactive oxygen species production in human bronchial epithelial cells.
Particulate matter (PM) is emitted during the combustion of fuels and wastes. PM exposure exacerbates pulmonary diseases, and the mechanism may involve oxidative stress. At lower combustion temperatures such as occurs in the cool zone of a flame, aromatic compounds chemisorb to the surface of metal-oxide-containing PM, resulting in the formation of surface-stabilized environmentally persistent free radicals (EPFR). Prior studies showed that PM-containing EPFR redox cycle to produce reactive oxygen species (ROS), and after inhalation, EPFR induce pulmonary inflammation and oxidative stress. Our objective was to elucidate mechanisms linking EPFR-induced oxidant injury with increased cytokine production by pulmonary epithelial cells. We thus treated human bronchial epithelial cells with EPFR at sub-toxic doses and measured ROS and cytokine production. To assess aryl hydrocarbon receptor (AhR) activity, cells were transfected with a luciferase reporter for xenobiotic response element activation. To test whether cytokine production was dependent upon AhR activation or oxidative stress, some cells were co-treated with an antioxidant or an AhR antagonist. EPFR increased IL-6 release in an ROS and AhR- and oxidant-dependent manner. Moreover, EPFR induced an AhR activation that was dependent upon oxidant production, since antioxidant co-treatment blocked AhR activation. On the other hand, EPFR treatment increased a cellular ROS production that was at least partially attenuated by AhR knockdown using siRNA. While AhR activation was correlated with an increased expression of oxidant-producing enzymes like cytochrome P450 CYP1A1, it is possible that AhR activation is both a cause and effect of EPFR-induced ROS. Finally, lipid oxidation products also induced AhR activation. ROS-dependent AhR activation may be a mechanism for altered epithelial cell responses after EPFR exposure, potentially via formation of bioactive lipid or protein oxidation products.


Glucocorticoids synergize with tumor necrosis factor alpha in the induction of HIV expression from a chronically infected promonocytic cell line.
In this study we have investigated the effects of glucocorticoids (GCs) on the expression of human immunodeficiency virus (HIV) in a chronically infected promonocytic cell line, U1. Although no increase in virus production was observed in U1 cells stimulated with physiological concentrations of GC alone, costimulation with dexamethasone plus tumor necrosis factor alpha (TNF-alpha) synergistically enhanced TNF-alpha-dependent HIV expression. Molecular analysis demonstrated that GCs plus TNF-alpha resulted in an accumulation of steady state HIV RNA secondary to either an increase in transcription or an increase in message stability. These findings may be of physiological relevance because GCs are used in the treatment of certain disorders associated with HIV infection and TNF-alpha levels have been reported to be elevated in the plasma and cerebrospinal fluid of certain HIV-infected individuals.


Vulvar varices: an uncommon entity in surgical pathology.
Varicose veins in the vulvar and perivulvar area are seen in 4% of women. Most of them are secondary to pregnancy and usually regress spontaneously. Vulvar varicose veins are rare in nonpregnant women. When present, they can be seen alone, associated with leg varices or associated with venous malformations of the labia, clitoral area, or vagina with or without arteriovenous malformations on the limbs or trunk (Klippel-Trenaunay-Weber syndrome and Parkes-Weber syndrome). In some cases, vulvar varices are seen as part of the so-called "pelvic congestion syndrome." Clinically, vulvar varices may present as small isolated protrusions, mainly in the labia majora, or as large masses, involving the vulva and even the perivulvar area. The treatment of choice of vulvar varices seen during pregnancy is conservative and symptomatic. Surgical pathologists need to be aware of the existence of vulvar varicose veins and its possible presence in biopsy specimens. Vulvar varicose veins can be misdiagnosed clinically as cysts or masses mainly in the Bartholin gland area. Correct diagnosis of the lesion is important to determine appropriate therapy and to recognize the possibility of associated anatomical or pathological problems.


Voltage-dependent currents in isolated vestibular afferent calyx terminals.
Na(+) currents were studied by whole cell patch clamp of chalice-shaped afferent terminals attached to type I hair cells isolated from the gerbil semicircular canal and utricle. Outward K(+) currents were blocked with intracellular Cs(+) or with extracellularly applied 20 microM linopirdine and 2.5 mM 4-aminopyridine (4-AP). With K(+) currents blocked, inward currents activated and inactivated rapidly, had a maximum mean peak amplitude of 0.92 +/- 0.60 (SD) nA (n = 24), and activated positive to -60 mV from holding potentials of -70 mV and more negative. The transient inward currents were blocked almost completely by 100 nM TTX, confirming their identity as Na(+) currents. Half-inactivation of Na(+) currents occurred at -82.6 +/- 0.9 mV, with a slope factor of 9.2 +/- 0.8 (n = 7) at room temperature. In current clamp, large overshooting action potential-like events were observed only after prior hyperpolarizing current injections. However, spontaneous currents consistent with quantal release from the hair cell were observed at holding potentials close to the zero-current potential. This is the first report of ionic conductances in calyx terminals postsynaptic to type I hair cells in the mammalian vestibular system.


Impact of Medication Adherence on Healthcare Resource Utilization, Work Loss, and Associated Costs in a Privately Insured Employed Population Treated With Adalimumab in the United States.
The aim of this study was to evaluate the impact of adherence to adalimumab on all-cause work loss, healthcare resource utilization (HRU), and direct medical and indirect costs over 2 years using real-world data. This was a retrospective cohort study using a large, United States administrative claims database. Adult patients treated with adalimumab were grouped into adherent and non-adherent cohorts and followed for up to 2 years. Outcomes were compared between cohorts. Over 2 years, adherent patients had $10,214 lower per patient medical and indirect costs compared to non-adherent patients, resulting from lower HRU, fewer days of absenteeism, and lower rates of work loss events. Patient and societal benefits of adherence to adalimumab are significant over 2 years. These findings highlight the importance of policies aimed at improving adherence to self-administrated medications.


Phase diagram of the Weeks-Chandler-Andersen potential from very low to high temperatures and pressures.
A combination of two molecular simulation algorithms has been used to determine the solid-liquid coexistence of the Weeks-Chandler-Andersen (WCA) fluid from low temperatures up to very high temperatures. Values are reported for the coexistence pressure, temperature, energy, enthalpy change, and densities of both the liquid and solid phases. At very high temperatures, the coexistence pressure approaches the same 12th-power soft-sphere asymptote as the 12-6 Lennard-Jones potential. However, in contrast to the Lennard-Jones potential, which shows a discontinuity of pressure at low temperatures, the coexistence pressure of the WCA potential approaches the zero-temperature limit. Empirical relationships are determined to accurately reproduce the coexistence pressure and both solid and liquid phase densities from near zero temperature to very high temperatures. The simulation data are used to improve the accuracy of a WCA equation of state. The validity of common melting and freezing rules is tested.


ELECTRON MICROSCOPE STUDIES ON THE ORIGIN OF ANNULATE LAMELLAE IN OOCYTES OF NECTURUS.
Developing oocytes, ranging from approximately 0.1 to 1.0 mm in diameter, in Necturus were studied with the electron microscope. The outer layer of the nuclear envelope is actively engaged in the formation of vesicular elements along most of its surface, especially in smaller oocytes. Groups of vesicles appear to be released into the ooplasm at about the same time, resulting in long chains of individual vesicles immediately adjacent to the nuclear membrane. This process is repeated so that chains of vesicles grouped in rather ordered ranks extend progressively into the surrounding cytoplasm. Eventually, the cytoplasm becomes more concentrated with chains of vesicles and the distance between the individual rows becomes less. Very soon after a chain of vesicles has been budded off from the nuclear membrane, fine intervesicular connections appear between certain of the vesicles comprising the rows. Several of the vesicles in a row may then fuse, forming short, flattened cisternae. Fusion of vesicles continues, individual rows of vesicles become more closely packed and, finally, regions appear in the cytoplasm which have the appearance of annulate lamellae. Further growth of the lamellae appears to occur by the progressive fusion of vesicles at the ends of those lamellae already present, as well as by the addition of other fusing rows of vesicles.


Colostomy closure promotes cell proliferation and dimethylhydrazine-induced carcinogenesis in rat distal colon.
The influence of adaptive cell proliferation on colonic carcinogenesis was studied in male Fischer rats with a defunctioning transverse colostomy that was closed 4 wk later. Control observations were made in other rats after colonic transection, repeated at 4 wk, after laparotomy alone, or after permanent colostomy. Tumors were induced by 1,2-dimethylhydrazine (total dose, 300 mg/kg) over 11 wk, starting 2 days after the second operation. After creation of the colostomy, amounts of protein, RNA, and DNA in the distal colon halved in 4 wk (p less than 0.001), but returned to normal 7 days after restoration of colonic continuity. This reactive hyperplasia promoted the development of distal colonic carcinomas, as compared with rats having repeated transection of the bowel (incidence 32% vs. 6%; p less than 0.03). Although the amounts of protein and nucleic acid in the proximal colon were unchanged by transverse colostomy, values increased by 18%-59% 4 wk after colostomy closure (p = 0.05-0.002); nonetheless, the yield of tumors in this segment was unaltered. Suture-line cancers were commoner after repeat transection than after colostomy closure (76% vs. 39%; p less than 0.01). These data confirm the promotional effect of increased cell proliferation on intestinal carcinogenesis.


Enhancing recovery and sensitivity studies in an unconventional tight gas condensate reservoir.
The recovery factor from tight gas reservoirs is typically less than 15%, even with multistage hydraulic fracturing stimulation. Such low recovery is exacerbated in tight gas condensate reservoirs, where the depletion of gas leaves the valuable condensate behind. In this paper, three enhanced gas recovery (EGR) methods including produced gas injection, CO2 injection and water injection are investigated to increase the well productivity for a tight gas condensate reservoir in the Montney Formation, Canada. The production performance of the three EGR methods is compared and their economic feasibility is evaluated. Sensitivity analysis of the key factors such as primary production duration, bottom-hole pressures, and fracture conductivity is conducted and their effects on the well production performance are analyzed. Results show that, compared with the simple depletion method, both the cumulative gas and condensate production increase with fluids injected. Produced gas injection leads to both a higher gas and condensate production compared with those of the CO2 injection, while waterflooding suffers from injection difficulty and the corresponding low sweep efficiency. Meanwhile, the injection cost is lower for the produced gas injection due to the on-site available gas source and minimal transport costs, gaining more economic benefits than the other EGR methods.


IL-17RB enhances thyroid cancer cell invasion and metastasis via ERK1/2 pathway-mediated MMP-9 expression.
IL-17RB, a member of the IL-17 receptor family that can be activated by IL-17B, has been proved to be involved in inflammatory diseases and cancers. However, the function of IL-17RB in thyroid cancer is still unknown. In this study, IL-17RB expression in thyroid cancer cell lines and tissues was examined by real-time PCR and western blot. The effects of IL-17RB on cell invasion and migration were determined by in vitro invasion and migration assays, while the effects of IL-17RB on cell metastasis were analyzed by in vivo experiments. The results showed that IL-17RB expression was upregulated in both thyroid cancer cells and tissues. IL-17B dose-dependently promoted the invasion, growth and migration of thyroid cancer cells, whereas knockdown of IL-17RB attenuated the effects of IL-17B in vitro. Moreover, IL-17RB was involved in the metastasis and growth of thyroid cancer cells in vivo. In addition, IL-17RB induced ERK1/2 activation and increased MMP-9 expression in vitro and in vivo. Inhibition of ERK1/2 pathway blocked the IL-17RB-mediated thyroid cancer cell invasion and MMP-9 expression. Together, our findings demonstrate that IL-17RB can enhance thyroid cancer cell invasion and metastasis via ERK1/2 pathway-mediated MMP-9 expression, suggesting that IL-17RB may act as a potential therapeutic target for thyroid cancer therapy.


Cloning and expression of the Bacillus sphaericus 2362 mosquitocidal genes in a non-toxic unicellular cyanobacterium, Synechococcus PCC6301.
Genes encoding the mosquitocidal binary toxin of Bacillus sphaericus 2362 were introduced into Synechococcus PCC6301, a cyanobacterium that can tolerate a number of potential variations in the mosquito breeding environment, and can serve as a food source for mosquito larvae. The toxin genes, preceded by a Synechococcus rbcL promoter, were located on a mobilizable Escherichia coli Synechococcus shuttle vector, which was introduced into Synechococcus PCC6301 at frequencies of 10(-5)-10(-7) exconjugants/recipient, depending on the selective conditions used. Recombinant Synechococcus exhibited significant toxicity against 2-day-old and 6-day-old Culex quinquefasciatus larvae, the concentration required to kill 50% of larvae (LC50) being 2.1 x 10(5) and 1.3 x 10(5) cells/ml respectively. Mosquitocidal activity decreased tenfold after 20 generations of non-selective growth.


Adaptations of guest and host in expanded self-assembled capsules.
Reversible encapsulation complexes create spaces where two or more molecules can be temporarily isolated. When the mobility of encapsulated molecules is restricted, different arrangements in space are possible, and new forms of isomerism ("social isomerism") are created: the orientation of one encapsulated molecule influences that of the other in the confined space. Expansion of a capsule's length is possible through addition of small-molecule spacer elements. The expanded capsules have dimensions that permit the observation of social isomerism of two identical guests, and they adopt arrangements that properly fill the host's space. The host also can adapt to longer guests by incorporating additional spacers, much as protein modules are added to a viral capsid in response to larger genomes. Arachidonic and related fatty acid derivatives act in this way to induce the assembly of further extended capsules having sufficient length to accommodate them.


The 3'-untranslated region of the Ste20-like kinase SLK regulates SLK expression.
Ste20-like kinase, SLK, a germinal center kinase found in kidney epithelial cells, signals to promote apoptosis. Expression of SLK mRNA and protein and kinase activity are increased during kidney development and recovery from ischemic acute renal failure. The 3'-untranslated region (3'-UTR) of SLK mRNA contains multiple adenine and uridine-rich elements, suggesting that 3'-UTR may regulate mRNA stability. This was confirmed in COS cell transient transfection studies, which showed that expression of the SLK open-reading frame plus 3'-UTR mRNA was reduced by 35% relative to the open-reading frame alone. To further characterize the SLK-3'-UTR, this nucleotide sequence was subcloned downstream of enhanced green fluorescent protein (EGFP) cDNA. In COS, 293T, and glomerular epithelial cells, expression of EGFP mRNA and protein was markedly reduced in the presence of the SLK-3'-UTR. After transfection and subsequent addition of actinomycin D, EGFP mRNA remained stable in cells for at least 6 h, whereas EGFP-SLK-3'-UTR mRNA decayed with a half-life of approximately 4 h. A region containing five AUUUA motifs within the SLK-3'-UTR destabilized EGFP mRNA. Deletion of this region from the SLK-3'-UTR, in part, restored mRNA stability. By UV cross-linking and SDS-PAGE, the SLK-3'-UTR bound to protein(s) of approximately 30 kDa in extracts of COS cells, glomerular epithelial cells, and kidney. Cotransfection of HuR (a RNA binding protein of approximately 30 kDa) increased the steady-state mRNA level of EGFP-SLK-3'-UTR but not EGFP. Thus the SLK-3'-UTR may interact with kidney RNA-binding proteins to regulate expression of SLK mRNA during kidney development and after ischemic injury.


Evidence-based practice and setting basic standards for stroke rehabilitation in Canada.
It is a concern that the consistency of care in stroke rehabilitation appears to vary significantly among practice settings within Canada. Based upon an evidence-based review of stroke rehabilitation, a group of physiatrists from across Canada agreed to some basic standards for stroke rehabilitation. The following standards were deemed to be critical to providing effective stroke rehabilitation care: early assessment and quick access to specialized interdisciplinary rehabilitation, appropriate intensity of therapies, availability of outpatient therapy, appropriate community supports, and careful attention to secondary prevention of stroke. Providing stroke rehabilitation care consistently within these standards remains a challenge, particularly given the potential initial costs, despite significant improvements in patient outcomes and quality of life. Nevertheless, these standards should be the expectation of stroke patients, essentially forming a stroke rehabilitation "bill of rights."


Analysis of quantitative relationship between viability determination in leprosy by MFP, ATP bioluminescence and gene amplification assay.
Two hundred twenty-one untreated, borderline lepromatous/lepromatous (BL/LL) leprosy patients have been investigated for viability by the mouse foot pad method (MFP), adenosine triphosphate (ATP) and polymerase chain reaction (PCR). The biopsies were collected at the beginning of and 12/24 months after treatment. The patient group was treated with a) immunotherapy (BCG/Mw) + MDT; b) MDT + pyrazinamide; c) control MDT; d) MDT + minocycline 100 mg once a month supervised + ofloxacin 400 mg once a month supervised. Biopsies were divided in three parts for use in the mouse foot pad, molecular and ATP investigations. In untreated and treated patients (at 12 and 24 months), there was a general agreement among all three techniques, and PCR and ATP showed higher positivity as compared to MFP. Further, there was good correlation among the viable biomass estimated by bacillary ATP levels, PCR assay and growth in mouse foot pads. The positivity was observed by MFP as well as PCR assay (18-kDa and 36-kDa) from all of the specimens when the ATP content was more than 3.6 pg/million. When the ATP content was below 3.5 pg/million, the positive takes in MFP decreased but the PCR positivity correlated with ATP bioluminescence up to 0.04 pg/million. When the ATP content was even lower, the uptake in the MFP was possibly a matter of chance, while PCR positivity was observed in 96% of the cases. For specimens with undetectable ATP, positivity was seen in 1% of the cases, showing the inability of ATP bioluminescence method to detect low background due to host ATP. PCR signals in some cases could be due to the higher sensitivity of the method or persistence of DNA after bacterial death in some cases. On the whole, the PCR methods even though targeting DNA have shown good correlations with biomass which confirm their usefulness in monitoring therapeutic responses in leprosy.


Comparative Analysis of HRTFs Measurement Using In-Ear Microphones.
The head-related transfer functions (HRTFs) describe the acoustic path transfer functions between sound sources in the free-field and the listener's ear canal. They enable the evaluation of the sound perception of a human being and the creation of immersive virtual acoustic environments that can be reproduced over headphones or loudspeakers. HRTFs are strongly individual and they can be measured by in-ear microphones worn by real subjects. However, standardized HRTFs can also be measured using artificial head simulators which standardize the body dimensions. In this paper, a comparative analysis of HRTF measurement using in-ear microphones is presented. The results obtained with in-ear microphones are compared with the HRTFs measured with a standard head and torso simulator, investigating different positions of the microphones and of the sound source and employing two different types of microphones. Finally, the HRTFs of five real subjects are measured and compared with the ones measured by the microphones in the ear of a standard mannequin.


Non-Angled Intercostal Percutaneous Access Under Full Expiration: Safety Is Not an Issue Anymore.
Percutaneous nephrolithotomy (PCNL) is a well-established procedure for the management of urinary calculi and can be performed intercostally or subcostally. Favoring one approach vs the other is still debatable, and literature has been inconclusive regarding the efficacy and safety of both approaches. Hence, this study aims to assess the safety and efficacy of direct non-angled intercostal technique performed under full expiration and to compare it to the subcostal approach. PCNL was conducted among 361 patients during 2010-2015 at Saint George Hospital University Medical Center in Beirut, Lebanon. PCNL was done by one operator and by following a standard technique. After reviewing the medical records, 304 patients were included. Data analysis was conducted using Stata/IC 10.0. Bivariate analysis was conducted using Pearson's Chi-square, and logistic regression model was run. Alpha level was set at 0.05. Of the total patients, 54.6% and 45.4% underwent intercostal (Group I) and subcostal (Group II) access, respectively. Mean drop in hemoglobin in Group II was 1.9 g/dL vs 1.48 g/dL in Group I (p-value = 0.0040). The mean difference in operation time between group I (88.61 minutes) and group II (102.58 minutes) was statistically significant (p-value = 0.0064). Patients were stone free in 88.05% of the intercostal cases and 78.52% of the subcostal cases. Group II patients were twice more likely to have residual stones compared to Group I (p-value = 0.029). No statistical significance was observed in postoperative complications among both groups. In addition, no cases of pneumothorax were reported. Compared to subcostal access, intercostal approach under full expiration is a safe technique that provides optimal approach to the intrarenal collecting system and allows less angulation, less bleeding, and yields higher stone clearance with minimal complications. When performed by a well-trained urologist, intercostal access should be advocated in PCNL to obtain a direct non-angled access to the tip of the desired posterior calix.


Delineating a Ca2+ binding pocket within the venus flytrap module of the human calcium-sensing receptor.
The Ca(2+)-sensing receptor (CaSR) belongs to the class III G-protein-coupled receptors (GPCRs), which include receptors for pheromones, amino acids, sweeteners, and the neurotransmitters glutamate and gamma-aminobutyric acid (GABA). These receptors are characterized by a long extracellular amino-terminal domain called a Venus flytrap module (VFTM) containing the ligand binding pocket. To elucidate the molecular determinants implicated in Ca(2+) recognition by the CaSR VFTM, we developed a homology model of the human CaSR VFTM from the x-ray structure of the metabotropic glutamate receptor type 1 (mGluR1), and a phylogenetic analysis of 14 class III GPCR VFTMs. We identified critical amino acids delineating a Ca(2+) binding pocket predicted to be adjacent to, but distinct from, a cavity reminiscent of the binding site described for amino acids in mGluRs, GABA-B receptor, and GPRC6a. Most interestingly, these Ca(2+)-contacting residues are well conserved within class III GPCR VFTMs. Our model was validated by mutational and functional analysis, including the characterization of activating and inactivating mutations affecting a single amino acid, Glu-297, located within the proposed Ca(2+) binding pocket of the CaSR and associated with autosomal dominant hypocalcemia and familial hypocalciuric hypercalcemia, respectively, genetic diseases characterized by perturbations in Ca(2+) homeostasis. Altogether, these data define a Ca(2+) binding pocket within the CaSR VFTM that may be conserved in several other class III GPCRs, thereby providing a molecular basis for extracellular Ca(2+) sensing by these receptors.


Paired related homeobox 1 is associated with the invasive properties of glioblastoma cells.
Glioblastoma is a highly proliferative and invasive tumor. Despite extensive efforts to develop treatments for glioblastoma, the currently available therapies have only limited effects. To develop novel strategies for glioblastoma treatment, it is crucial to elucidate the molecular mechanisms that promote the invasive properties of glioblastoma. In the present study, we showed that the paired related homeobox 1 (PRRX1) is associated with glioblastoma cell invasion. The depletion of PRRX1 suppressed the invasion and neurosphere formation of glioblastoma cells. Conversely, the exogenous expression of PRRX1 promoted invasion. The Notch signaling pathway, which is an evolutionarily conserved pathway that is essential for developmental processes, plays an important role in the tumorigenesis of glioblastoma. The expression of PRRX1 induced the activation of Notch signaling, and the inhibition of Notch signaling suppressed PRRX1-mediated cell invasion. Our results indicate that activation of Notch signaling by PRRX1 is associated with the promotion of glioblastoma cell invasion.


The value of pancuronium in prolonged intraocular operations.
We wished to determine the effectiveness of Pancuronium, a non-depolarizing muscle relaxant, in prolonged intraocular operations. We compared the variations in intraocular pressure when using four different methods of induction of general anesthesia. With Pancuronium the intraocular pressure fell initially and then rose to just below pre-induction levels and remained there. Our study suggests that pancuronium stabilizes intraocular pressure at a level which is highly desirable for intraocular surgery.


Prevention of postmenopausal bone loss using tibolone or conventional peroral or transdermal hormone replacement therapy with 17beta-estradiol and dydrogesterone.
Postmenopausal bone loss can be prevented by continuous or intermittent estradiol (E2) administration. Concomitant progestogen therapy is mandatory in nonhysterectomized women to curtail the risk of endometrial hyperplasia or cancer. However, the recurrence of vaginal bleeding induced by sequential progestogen therapy in addition to continuous estrogen administration is one of the reasons for noncompliance to hormone replacement therapy (HRT). Tibolone, a synthetic steroid with simultaneous weak estrogenic, androgenic, and progestational activity, which does not stimulate endometrial proliferation, has recently been proposed for the treatment of climacteric symptoms. To compare the efficacy of conventional oral and transdermal HRT with that of tibolone in the prevention of postmenopausal bone loss, 140 postmenopausal women (age, 52 +/- 0.6 years; median duration of menopause, 3 years) were enrolled in an open 2-year study. Volunteers had been offered a choice between HRT and no therapy (control group, CO). Patients selecting HRT were randomly allocated to one of the following three treatment groups: TIB, tibolone, 2.5 mg/day continuously, orally; PO, peroral E2, 2 mg/day continuously, plus sequential oral dydrogesterone (DYD), 10 mg/day, for 14 days of a 28-day cycle; TTS, transdermal E2 by patch releasing 50 microg/day, plus DYD as above. Bone densitometry of the lumbar spine, upper femur, and whole body was performed using dual-energy X-ray absorptiometry at baseline, and then 6, 12, 18, and 24 months after initiation of therapy. One hundred and fifteen women (82%) completed the 2 years of the study. The dropout rate was similar in each group. Over 2 years, bone preservation was observed in all three treatment groups as compared with controls, without significant differences among treatment regimens. In conclusion, tibolone can be regarded as an alternative to conventional HRT to prevent postmenopausal bone loss.


Phenolics of Arbutus unedo L. (Ericaceae) fruits: identification of anthocyanins and gallic acid derivatives.
Arbutus unedo L., the strawberry tree (Ericaceae family), is an evergreen shrub or small tree, typical of the Mediterranean fringe and climate. The aim of the present study was to evaluate the profile of the phenolic constituents of A. unedo fruits. Seven compounds were purified by Sephadex LH-20 column chromatography of the MeOH extract followed by HPLC and were characterized as arbutin, beta-D-glucogalline, gallic acid 4-O-beta-D-glucopyranoside, 3-O-galloylquinic acid, 5-O-galloylquinic acid, 3-O-galloylshikimic acid, and 5-O-galloylshikimic acid, by means of NMR and ESI-MS analyses. Moreover, LC-PDA-MS analysis of the red pigment of A. unedo fruits revealed the presence of three anthocyanins recognized as cyanidin 3-O-beta-D-galactopyranoside, delphinidin 3-O-beta-D-glucopyranoside, and cyanidin 3-O-beta-D-arabinopyranoside. These pigments were also quantified.


Measuring the rate of gross chromosomal rearrangements in Saccharomyces cerevisiae: A practical approach to study genomic rearrangements observed in cancer.
Gross chromosomal rearrangements (GCRs), including translocations, deletions, amplifications and aneuploidy are frequently observed in various types of human cancers. Despite their clear importance in carcinogenesis, the molecular mechanisms by which GCRs are generated and held in check are poorly understood. By using a GCR assay, which can measure the rate of accumulation of spontaneous GCRs in Saccharomyces cerevisiae, we have found that many proteins involved in DNA replication, DNA repair, DNA recombination, checkpoints, chromosome remodeling, and telomere maintenance, play crucial roles in GCR metabolism. We describe here the theoretical background and practical procedures of this GCR assay. We will explain the breakpoint structure and DNA damage that lead to GCR formation. We will also summarize the pathways that suppress and enhance GCR formation. Finally, we will briefly describe similar assays developed by others and discuss their potential in studying GCR metabolism.


Senescence in human AC16 cardiac cells is associated with thymidine kinase induction and histone loss.
AC16 cells are a transformed human cardiac cell line commonly used to study cardiomyocyte biology. We show that reduced proliferation and senescence markers can be robustly induced in AC16 cells cultured in low serum condition and treated with (i) low-dose doxorubicin, (ii) UV 254 nm, or (iii) H 2 O 2 exposure for up to 48 hours. Increased p21 (CDKN1A) and H2A.X variant histone (H2AX) levels serve as reliable molecular markers upon all three treatment conditions, but the up-regulation of another common senescence marker, p16 (CDKN2A) was not observed. A proteomics screen further shows that the loss of histones and the increased expression of thymidine kinases (TK1) are prominent features of AC16 cells under doxorubicin induced senescence.


A comparative study of responses to acute hypoglycaemia induced by human and porcine insulins in patients with Type 1 diabetes.
The effects of human and porcine insulins on the symptomatic, physiological, and counterregulatory hormonal responses to acute hypoglycaemia were compared in 40 patients with Type 1 diabetes, 20 of whom were newly diagnosed while 20 had been treated for between 5 and 20 years. In a double-blind, cross-over trial all patients were treated with human or porcine insulin, in random order, for two consecutive 3-month periods. At the end of each treatment period they were subjected to an acute episode of experimental hypoglycaemia induced by a continuous intravenous infusion (2.0 mU kg(-1)min(-1)) of the same insulin species. Haemodynamic, sweating, and tremor responses were measured during both studies, symptom scores were recorded and the arterialized plasma glucose thresholds for autonomic activation and the onset of subjective symptoms were identified. In all patients the glycaemic thresholds for the initiation of the autonomic physiological responses to hypoglycaemia and the onset of the symptomatic response were concurrent and did not differ with insulin species (plasma glucose 1.94 vs 1.96 mmol I(-1), human vs porcine studies). The onset, temporal pattern, nature, and magnitude of the physiological responses (sweating, heart rate, blood pressure, and tremor) during acute experimental hypoglycaemia were also identical with each insulin species. The magnitude and temporal pattern of the response of counterregulatory hormones (adrenaline, noradrenaline, glucagon, ACTH, and GH) to hypoglycaemia as induced by human and porcine insulins were indistinguishable, as were the total and individual scores of autonomic and neuroglycopenic symptoms. In conclusion, in patients who had newly diagnosed and intermediate duration (5-20 years) of diabetes, the symptomatic, physiological, and counterregulatory hormonal responses to acute insulin-induced hypoglycaemia did not differ between human and porcine insulins, and the plasma glucose thresholds at which the symptomatic and autonomic responses were initiated were identical with both insulin species. This study does not support the hypothesis that treatment with human insulin modifies the symptomatic, physiological, and counterregulatory hormonal responses to acute hypoglycaemia.