Toll-like receptor signaling and regulation of intestinal immunity.
The intestine is a complex organ that must maintain tolerance to innocuous food antigens and commensal microbiota while being also able to mount inflammatory responses against invading pathogenic microorganisms. The ability to restrain tolerogenic responses while permitting inflammatory responses requires communication between commensal bacteria, intestinal epithelial cells and immune cells. Disruption or improper signaling between any of these factors may lead to uncontrolled inflammation and the development of inflammatory diseases. Toll-like receptors (TLR) recognize conserved molecular motifs of microorganisms and, not surprisingly, are important for maintaining tolerance to commensal microbiota, as well as inducing inflammation against pathogens. Perturbations in individual TLR signaling can lead to a number of different outcomes and illustrate a system of regulation within the intestine in which each TLR plays a largely non-redundant role in mucosal immunity. This review will discuss recent findings on the roles of individual TLRs and intestinal homeostasis.


Quantifying Bufo boreas connectivity in Yellowstone National Park with landscape genetics.
A major objective of ecology is to understand how ecological processes limit population connectivity and species' distributions. By spatially quantifying ecological components driving functional connectivity, we can understand why some locally suitable habitats are unoccupied, resulting in observed discontinuities in distribution. However, estimating connectivity may be difficult due to population stochasticity and violations of assumptions of parametric statistics. To address these issues, we present a novel application of Random Forests to landscape genetic data. We address the effects of three key ecological components on Bufo boreas connectivity in Yellowstone National Park: ecological process, scale, and hierarchical organization. Habitat permeability, topographic morphology, and temperature-moisture regime are all significant ecological processes associated with B. boreas connectivity. Connectivity was influenced by growing-season precipitation, 1988 Yellowstone fires, cover, temperature, impervious surfaces (roads and development), and topographic complexity (56% variation explained). We found that habitat permeability generally operates on fine scales, while topographic morphology and temperature-moisture regime operate across multiple scales, thus demonstrating the importance of cross-scale analysis for ecological interpretation. In a hierarchical analysis, we were able to explain more variation within genetic clusters as identified using Structure (a Bayesian algorithm) (74%; dispersal cover, growing-season precipitation, impervious surfaces) as opposed to between genetic clusters (45%; ridgelines, hot, dry slopes, length of hot season, and annual precipitation). Finally, the analytical methods we developed are powerful and can be applied to any species or system with appropriate landscape and genetic data.


Expressions of beta-defensins in human keratinocyte cell lines.
Defensins, a major family of antimicrobial peptides, are small cationic, cysteine-rich peptides with a wide range of antimicrobial activity. In human, beta-defensin-1 was isolated from urine and cervical mucous suggesting that this peptide plays an antimicrobial role in the genitourinary tract. Beta-defensin-2 was identified in psoriatic scale produced by keratinocytes suggesting that this peptide contributes to defend the expansive surface of the integuments. Current research was done to investigate the expression and modulation of beta-defensin mRNA in human keratinocyte cell lines. HaCaT and A431 cell lines were used to all culture experiments. Cultured human keratinocytes were stimulated with ultraviolet (UV) B irradiation or tumor necrosis factor-alpha (TNF-alpha) or lipopolysaccharide (LPS) to determine whether defensin mRNA production occurred. Reverse transcription-polymerase chain reaction (RT-PCR) was performed to amplify defesin cDNA from stimulated keratinocytes, and southern blots were used to verify the specificity of RT-PCR amplication products. Expression of human beta-defensins was upregulated with UVB irradiation, TNF-alpha and LPS in HaCaT cells and in comparison to the control, significantly higher at 6 h post stimulation with UVB 100 mJ/cm2 and peak at 12 to 18 h post stimulation with UVB 30 mJ/cm2, TNF-alpha and LPS. A431 cells did not show expression of human beta-defensins in unstimulated state, even after irradiation with UVB or TNF-alpha or LPS. This report demonstrates the presence of defensin in human keratinocytes and capacity of human keratinocytes to produce defensin mRNA in response to UVB irradiation, TNF-alpha and LPS. Release of defensins by keratinocytes in response to cytokines elaborated in inflammation may contribute to the host defense responses.


Inferior Vena Cava Torsion and Stenosis Complicated by Compressive Pericaval Regional Ascites following Orthotopic Liver Transplantation.
Inferior vena cava (IVC) stenosis and torsion are well-described rare complications following orthotopic liver transplantation (OLT). We present a case of inferior vena cava intermittent torsion and stenosis complicated by compressive regional ascites. To the best of our knowledge, this is the second case of post-OLT regional ascites related compressive IVC stenosis reported and the first reported case of torsion complicated by regional ascites compression.


Ultrashort pulse characterization by spectral shearing interferometry with spatially chirped ancillae.
We report a new version of spectral phase interferometry for direct electric field reconstruction (SPIDER), in which two spatially chirped ancilla fields are used to generate a spatially encoded SPIDER interferogram. We dub this new technique Spatially Encoded Arrangement for Chirped ARrangement for SPIDER (SEA-CAR-SPIDER). The single shot interferogram contains multiple shears, the spectral amplitude of the test pulse, and the reference phase, which is accurate for broadband pulses. The technique enables consistency checking through the simultaneous acquisition of multiple shears and offers a simple and precise calibration method. All calibration parameters--the shears, and the upconversionfrequency--can be accurately obtained from a single calibration trace.


TNFα secretion of monocytes exposed to pulsed radiofrequency treatment: a possible working mechanism of PRF chronic pain management.
Pulsed radiofrequency treatment (PRF) is a promising new technique increasingly used in treatment of chronic pain. The molecular working mechanism of PRF is not exactly known and is currently being investigated. This study investigates a possible role of PRF-induced modulation of TNFα secretion by differentiated monocytes in chronic pain management. The results show no significant PRF-induced change in TNFα secretion of lipopolysaccharides (LPS)-stimulated monocytes. However, PRF does significantly increase TNFα secretion of differentiated monocytes that have not been stimulated with LPS. This may indicate a possible role of PRF treatment in increasing TNFα production of nonstimulated monocytes. More research is needed to determine whether this is truly a part of the working mechanism of PRF in chronic pain management and which other factors are involved.


Development of an efficient host-vector system of Ruminiclostridium josui.
Although Ruminiclostridium josui (formerly Clostridium josui), a strictly anaerobic mesophilic cellulolytic bacterium, is a promising candidate for biomass utilization via consolidated bioprocessing, its host-vector system has not yet been established. The existence of a restriction and modification system is a significant barrier to the transformation of R. josui. Here, we partially purified restriction endonuclease RjoI from R. josui cell extract using column chromatography. Further characterization showed that RjoI is an isoschizomer of DpnI, recognizing the sequence 5'-Gmet ATC-3', where the A nucleotide is Dam-methylated. RjoI cleaved the recognition sequence between the A and T nucleotides, producing blunt ends. We then successfully introduced plasmids prepared from Escherichia coli C2925 (dam- /dcm- ) into R. josui by electroporation. The highest transformation efficiency of 6.6 × 103 transformants/μg of DNA was obtained using a square-wave pulse (750 V, 1 ms). When the R. josui cel48A gene, devoid of the dockerin-encoding region, cloned into newly developed plasmid pKKM801 was introduced into R. josui, a truncated form of RjCel48A, RjCel48AΔdoc, was detected in the culture supernatant but not in the intracellular fraction. This is the first report on the establishment of fundamental technology for molecular breeding of R. josui.


Overexpression of miR-203 increases the sensitivity of NSCLC A549/H460 cell lines to cisplatin by targeting Dickkopf-1.
The number of new lung cancer cases diagnosed yearly is high, and the mortality rate has not substantially declined. Non-small cell lung cancer (NSCLC) is the most common type of lung cancer, and adenocarcinoma accounts for the largest proportion of NSCLC. Currently, platinum-based combined chemotherapy, particularly cisplatin (DDP) is still the main form of treatment for advanced NSCLC. However, cisplatin resistance often occurs in patients who receive chemotherapy. Previous studies offer various explanations for how miRNAs affect cisplatin resistance, but the underlying mechanism remains largely unknown. The present study was designed to focus on miR-203 and Dickkopf-1 (DKK1), investigating the potential mechanisms involved in cisplatin resistance in tissues of lung adenocarcinoma and A549/H460 cell lines. In DDP-sensitive NSCLC samples, miR-203 was expressed at a higher level when compared with this level in DDP-insensitive samples, while DKK1 mRNA was expressed at a relatively low level as indicated by qRT-PCR. Dual luciferase reporter assay revealed that DKK1 is a target gene of miR-203 in A549 and H460 cells. Upregulation of miR-203 reduced the IC50 value of cisplatin in the A549 and H460 cells by inhibiting cell growth and promoting cell apoptosis. Similar effects of tumor inhibition and cisplatin sensitization were verified in vivo. Further research showed that both overexpression of miR-203 and knockdown of DKK1 increased the sensitization to DDP with a lower IC50 value. Upon DKK1 knockdown, overexpression of miR-203 had no added effects on the sensitivity of the cells. In addition, miR-203 was unable to sensitize cells with DKK1 that lacked the 3' untranslated region (3'UTR). We conclude that miR-203 targets the 3'UTR of DKK1, and increases cisplatin sensitivity in A549/H460 cell lines.


New guanidine-containing nanocomposites impeding the growth of Staphylococcus epidermidis 33 and the biofilm formation.
New water-soluble nanocomposites (AgNCs) based on Ag and copolymers of 2,2-diallyl-1,1,3,3-tetraethylguanidiniumchloride with N-vinylpyrrolidone [poly(AGC-VP)] and vinylacetate [poly(AGC-VA)] have been developed. Antibacterial action of new silver nanocomposites on S. epidermidis 33 (planctonic cells and biofilms) is reported in this study. AgNCs strongly inhibited biofilms formation of S. epidermidis 33. The viability of S. epidermidis 33 cells in biofilms was considerably reduced by new AgNCs. It has been shown that S. epidermidis 33 inactivation in biofilms occurs at AgNC concentrations > 5 times higher as compared to those inhibiting completely the planktonic cells. © 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 104A: 630-638, 2016.


Protein clearances and selectivity determinations in childhood nephrosis: a reappraisal.
To critically evaluate the clinical utility of determining specific proteins in patients with extensive proteinuria, we used immunonephelometric methods to measure albumin, transferrin, IgG, and alpha 2-macroglobulin in serum and in 24-h urine specimens from 37 children with idiopathic nephrotic syndrome. Renal biopsy demonstrated minimal change disease (I) in 15, focal glomerulosclerosis (II) in 15, and membranoproliferative glomerulonephritis (III) in seven patients. A three-group nonparametric rank test and three-group discriminant function analysis of the protein excretion and clearances of the four proteins we measured revealed significant differences in the excretion of IgG and the clearance of alpha 2-macroglobulin among the three groups of patients (p less than 0.05). Only patients with III had low serum complement C3 concentrations. Patients with I or II were best discriminated by differences in the excretion of transferrin and IgG, the clearance of alpha 2-macroglobulin, and the selectivity index (the clearance ratio of IgG/transferrin). These data indicate that measurement of specific urinary proteins and selectivity determinations may be helpful in predicting the type of histopathology and the prognosis of nephrotic children who have normal complement concentrations.


Silencing of FANCD2 enhances the radiosensitivity of metastatic cervical lymph node-derived head and neck squamous cell carcinoma HSC-4 cells.
Fanconi anemia complementation group D2 (FANCD2) is involved in the key steps of the Fanconi anemia (FA) pathway, which plays a role in the repair of DNA crosslink damage. However, the role of FANCD2 during radiotherapy for head and neck squamous cell carcinoma (HNSCC) is unclear. In this study, the HNSCC cell line HSC-4 was used. Western blotting was used to evaluate the expression of the FANCD2 in HSC-4 cells. We investigated the impact of FANCD2 on the radiosensitivity of HSC-4 cells in vitro and in vivo. TUNEL, western blotting and immunohistochemistry were used to analyze the apoptosis and proteins involved in apoptosis-related pathways after radiotherapy to investigate the relevant mechanism. The present study showed that shRNA interference could effectively and stably silence FANCD2 expression in HSC-4 cells. In vitro, the silencing of FANCD2 inhibited cell proliferation, decreased the survival rate, increased apoptosis and induced S phase arrest in HSC-4 cells after radiotherapy. In vivo, the silencing of FANCD2 could prolong the tumor-forming time and slow tumor growth. In addition, the tumor volume was significantly reduced, the weight was deceased, and the tumor inhibition rate was increased after radiotherapy. TUNEL showed that the silencing of FANCD2 significantly increased apoptosis in HSC-4 cells induced by radiotherapy. Both in vitro and in vivo esperiments revealed that the expression of the Bax and p-p38 proteins in HSC-4 cells, in which FANCD2 had been silenced, was increased after radiotherapy, whereas the expression of the p38 and Bcl2 proteins was decreased. Our results suggested that the silencing of FANCD2 enhanced the radiosensitivity of HSC-4 cells, and its mechanism involves the activation of the p38 MAPK signaling pathway and the regulation of the expression of Bax and Bcl2 proteins. This study provides a novel candidate target for HNSCC therapy.


Dominant hereditary inclusion-body myopathy gene (IBM3) maps to chromosome region 17p13.1.
We recently described an autosomal dominant inclusion-body myopathy characterized by congenital joint contractures, external ophthalmoplegia, and predominantly proximal muscle weakness. A whole-genome scan, performed with 161 polymorphic markers and with DNA from 40 members of one family, indicated strong linkage for markers on chromosome 17p. After analyses with additional markers in the region and with DNA from eight additional family members, a maximum LOD score (Zmax) was detected for marker D17S1303 (Zmax=7.38; recombination fraction (theta)=0). Haplotype analyses showed that the locus (Genome Database locus name: IBM3) is flanked distally by marker D17S945 and proximally by marker D17S969. The positions of cytogenetically localized flanking markers suggest that the location of the IBM3 gene is in chromosome region 17p13.1. Radiation hybrid mapping showed that IBM3 is located in a 2-Mb chromosomal region and that the myosin heavy-chain (MHC) gene cluster, consisting of at least six genes, co-localizes to the same region. This localization raises the possibility that one of the MHC genes clustered in this region may be involved in this disorder.


Antithrombin III activity and other coagulation changes in proliferative diabetic retinopathy.
Antithrombin III, factor VIII, fibrinogen and fibrinolytic activity were measured in two groups of long-standing insulin-dependent diabetic subjects (24 with proliferative retinopathy, 24 without detectable retinopathy) and 24 non-diabetic controls. Mean antithrombin III (+/- 1 SD) was 115.9 (+/- 15.1), 109.8 (+/- 18.1) and 101.4% (+/- 12.5), respectively, in the retinopathy, non-retinopathy and control groups. Statistical significance was obtained when comparing the retinopathy and control groups (p < 0.001) and when comparing all 48 diabetics collectively with controls (p < 0.01). Mean factor VIII coagulant activity was 137.5 (+/- 37.0), 126.2 (+/- 58.2) and 97.0% (+/- 38.7), respectively, in the three groups. Again the differences between all diabetics and controls (p < 0.01) were statistically significant. Similar increases were observed for other modalities of factor VIII activity. Fibrinolytic activity was significantly increased in both diabetic groups but fibrinogen levels, although increased, were not statistically different from levels in controls. It is suggested that the observed changes are more likely to be secondary to the development of retinopathy and that the increase in antithrombin III activity is due to an increase in alpha 2-macroglobulin.


Wais-III matrix reasoning: instruction effects on task perception and performance.
Instructions for the Matrix Reasoning subtest of the WAIS-III do not communicate to examinees that the subtest is untimed. The present study examined what percentage of participants (34 women, 26 men, M age = 20.1; 55 Caucasian, 3 African American, 2 Hispanic) made the assumption that Matrix Reasoning was timed, and its effect on examinees' scores. 55% of participants receiving standard instructions retrospectively reported assuming the subtest was timed, and those who did not assume Matrix Reasoning was timed scored significantly higher than participants who did. Participants receiving additional instructions that clarified the untimed nature of Matrix Reasoning scored significantly higher than those receiving standard instructions who believed the subtest was timed.


Approved Antiviral Drugs over the Past 50 Years.
Since the first antiviral drug, idoxuridine, was approved in 1963, 90 antiviral drugs categorized into 13 functional groups have been formally approved for the treatment of the following 9 human infectious diseases: (i) HIV infections (protease inhibitors, integrase inhibitors, entry inhibitors, nucleoside reverse transcriptase inhibitors, nonnucleoside reverse transcriptase inhibitors, and acyclic nucleoside phosphonate analogues), (ii) hepatitis B virus (HBV) infections (lamivudine, interferons, nucleoside analogues, and acyclic nucleoside phosphonate analogues), (iii) hepatitis C virus (HCV) infections (ribavirin, interferons, NS3/4A protease inhibitors, NS5A inhibitors, and NS5B polymerase inhibitors), (iv) herpesvirus infections (5-substituted 2'-deoxyuridine analogues, entry inhibitors, nucleoside analogues, pyrophosphate analogues, and acyclic guanosine analogues), (v) influenza virus infections (ribavirin, matrix 2 protein inhibitors, RNA polymerase inhibitors, and neuraminidase inhibitors), (vi) human cytomegalovirus infections (acyclic guanosine analogues, acyclic nucleoside phosphonate analogues, pyrophosphate analogues, and oligonucleotides), (vii) varicella-zoster virus infections (acyclic guanosine analogues, nucleoside analogues, 5-substituted 2'-deoxyuridine analogues, and antibodies), (viii) respiratory syncytial virus infections (ribavirin and antibodies), and (ix) external anogenital warts caused by human papillomavirus infections (imiquimod, sinecatechins, and podofilox). Here, we present for the first time a comprehensive overview of antiviral drugs approved over the past 50 years, shedding light on the development of effective antiviral treatments against current and emerging infectious diseases worldwide.


Characterization of Mycobacterium tuberculosis mycothiol S-conjugate amidase.
Mycothiol is comprised of N-acetylcysteine (AcCys) amide linked to 1D-myo-inosityl 2-amino-2-deoxy-alpha-D-glucopyranoside (GlcN-Ins) and is the predominant thiol found in most actinomycetes. Mycothiol S-conjugate amidase (Mca) cleaves the amide bond of mycothiol S-conjugates of a variety of alkylating agents and xenobiotics, producing GlcN-Ins and a mercapturic acid that can be excreted from the cell. Mca of Mycobacterium tuberculosis (Rv1082) was cloned and expressed as a soluble protein in Escherichia coli. The protein contained 1.4 +/- 0.1 equiv of zinc after purification, indicating that Mca is a metalloprotein with zinc as the native metal. Kinetic studies of Mca activity with 14 substrates demonstrated that Mca is highly specific for the mycothiol moiety of mycothiol S-conjugates and relatively nonspecific for the structure of the sulfur-linked conjugate. The deacetylase activity of Mca with GlcNAc-Ins is small but significant and failed to saturate at up to 2 mM GlcNAc-Ins, indicating that Mca may contribute modestly to the production of GlcN-Ins when GlcNAc-Ins levels are high. The versatility of Mca can be seen in its ability to react with a broad range of mycothiol S-conjugates, including two different classes of antibiotics. The mycothiol S-conjugate of rifamycin S was produced under physiologically relevant conditions and was shown to be a substrate for Mca in both oxidized and reduced forms. Significant activity was also seen with the mycothiol S-conjugate of the antibiotic cerulenin as a substrate for Mca.


Two- and three-phase mixing in a concentric draft tube gas-lift fermentor.
Two- and three-phase mixing studies were carried out in a 44-L concentric draft tube gas-lift fermentor. It was proposed to use the fermentor for the production of solvents using immobilized bacteria. Bubble size, gas holdup, liquid velocities, circulation, and mixing times were determined for various superficial gas velocities in distilled water, starch, carboxymethyl cellulose, and ethanol solutions. The observed trends for two phase mixing were similar to other studies but the results were found to be more sensitive to liquid properties. This was possibly due to the large value of downcomer to riser area used in this study. Mixing in three phases highlighted the difficulty in predicting the effect of adding solids to the gas-liquid system. Results showed that the gas-lift fermentor was ideally suited to dealing with three phases but more work is necessary before accurate models can be developed to account for the effect of solids.


High frequency of CD4+ CXCR5+ TFH cells in patients with immune-active chronic hepatitis B.
T follicular helper (TFH) cells are a special subpopulation of T helper cells and can regulate humoral immune responses. This study examined whether the frequency of CD4(+)CXCR5(+) TFH cells could be associated with active immunity in chronic hepatitis B (CHB) patients. The frequencies of peripheral blood CD4(+)CXCR5(+) TFH cells, inducible T cell costimulator (ICOS), and/or programmed death 1 (PD-1) positive CD4(+)CXCR5(+) TFH cells in immune-active (IA), immune-tolerant (IT) CHB, and healthy controls (HC) were characterized by flow cytometry analysis. The effect of adevofir dipivoxil treatment on the frequency of CD4(+)CXCR5(+) TFH cells, the concentrations of serum IL-2, IFN-γ, TNF-α, IL-4, IL-6, IL-10, IL-21, ALT, AST, HBsAg, HBsAb, HBeAg, HBeAb and HBV loads in IA patients were determined. The potential association of the frequency of CD4(+)CXCR5(+) TFH cells with clinical measures was analyzed. In addition, the frequency of splenic and liver CD4(+)CXCR5(+) TFH cells in HBV-transgenic mice was examined. We found that the frequency of CD4(+)CXCR5(+) TFH cells in IA patients was significantly higher than that of IT patients and HC, and the percentages of CD4(+)CXCR5(+) TFH in IA patients were positively correlated with AST. Furthermore, the percentages of ICOS(+), PD-1(+), and ICOS(+)PD-1(+) in CD4(+)CXCR5(+) TFH cells in CHB patients were significantly higher than that of HC. Treatment with adefovir dipivoxil reduced the frequency of CD4(+)CXCR5(+) TFH, PD-1(+)CD4(+)CXCR5(+) TFH cells and the concentrations of HBsAg and HBeAg, but increased the concentrations of HBsAb, HBeAb, IL-2 and IFN-γ in IA patients. Moreover, the frequency of splenic and liver CD4(+)CXCR5(+) TFH cells in HBV-transgenic mice was higher than that of wild-type controls. These data indicate that CD4(+)CXCR5(+) TFH cells may participate in the HBV-related immune responses and that high frequency of CD4(+)CXCR5(+) TFH cells may be a biomarker for the evaluation of active immune stage of CHB patients.


Neonatal Fc Receptor Regulation of Lung Immunoglobulin and CD103+ Dendritic Cells Confers Transient Susceptibility to Tuberculosis.
The neonatal Fc receptor (FcRn) extends the systemic half-life of IgG antibodies by chaperoning bound Fc away from lysosomal degradation inside stromal and hematopoietic cells. FcRn also transports IgG across mucosal barriers into the lumen, and yet little is known about how FcRn modulates immunity in the lung during homeostasis or infection. We infected wild-type (WT) and FcRn-deficient (fcgrt(-/-)) mice with Pseudomonas aeruginosa or Mycobacterium tuberculosis to investigate whether recycling and transport of IgG via FcRn influences innate and adaptive immunity in the lung in response to bacterial infection. We found that FcRn expression maintains homeostatic IgG levels in lung and leads to preferential secretion of low-affinity IgG ligands into the lumen. Fcgrt(-/-) animals exhibited no evidence of developmental impairment of innate immunity in the lung and were able to efficiently recruit neutrophils in a model of acute bacterial pneumonia. Although local humoral immunity in lung increased independently of the presence of FcRn during tuberculosis, there was nonetheless a strong impact of FcRn deficiency on local adaptive immunity. We show that the quantity and quality of IgG in airways, as well as the abundance of dendritic cells in the lung, are maintained by FcRn. FcRn ablation transiently enhanced local T cell immunity and neutrophil recruitment during tuberculosis, leading to a lower bacterial burden in lung. This novel understanding of tissue-specific modulation of mucosal IgG isotypes in the lung by FcRn sheds light on the role of mucosal IgG in immune responses in the lung during homeostasis and bacterial disease.


Maternal mortality with cesarean delivery: a literature review.
We sought to determine the present-day risk of maternal death with cesarean delivery. We reviewed the recent literature (years in analysis: 1975-2001) identified in a literature search and included data from the Royal College of Obstetricians and Gynaecologists. There were no publications with an ideal trial design and adequate power to establish the relationship between maternal mortality and method of delivery. Three studies, including the one randomized control trial included in analysis, and the Royal College of Obstetricians and Gynaecologists data suggest no significant difference in maternal mortality with cesarean delivery as compared with vaginal delivery. The strongest publications suggest there may not be an increased risk of maternal death with cesarean delivery as compared with vaginal delivery; however, there are inadequate data to accurately demonstrate the present-day risk of maternal death with cesarean delivery.


A Combinatorial Approach of High-Throughput Genomics and Mass Proteomics for Understanding the Regulation and Expression of Secondary Metabolite Production in Actinobacteria.
Secondary metabolites produced by Actinobacteria are an important source of antibiotics, drugs, and antimicrobial peptides. However, the large genome size of actinobacteria with high gene coding density makes it difficult to understand the complex regulation of biosynthesis of such critically and economically important products. In the last few decades, apart from genomics sequences, high-throughput proteomics has proven beneficial to understand the key players regulating the expression pattern of secondary metabolite and antibiotic production in different experimental set-ups. In the past, we have been analyzing the genomics data and mass spectrometry-based proteomics to predict the regulation dynamics and crucial regulatory hubs in Actinobacteria. The multidirectional regulation and expression of the biosynthetic gene cluster responsible for the production of important metabolite take their cue from the other primary metabolism pathways with which they show intricate interactions in the interactome. The regulation occurs by not only the action and expression of the biosynthetic gene cluster but also the role of transcription factors and primary metabolic pathways. Using the key players of these interactomes, we can regulate the synthesis/production of these valuable peptides/metabolites. Simultaneously, the multi-omics approach has now opened new gateways in investigation, screening, and identification of naturally occurring antimicrobial peptides from actinobacteria which are beneficial for humans and also provide economic and industrial benefits to humankind.


Arterial hypertension and noise: a cross-sectional study.
The occurrence of permanent arterial hypertension was studied among the workers of a textile plant (490 subjects) exposed to high frequency noise, at levels often exceeding 100 dBA. The prevalence of the disease among these workers was compared to that observed in a reference series (450 subjects). A prevalence rate ratio equal to 1.34 (1.14-1.57) was estimated, taking into account several determinants of disease acting as confounding factors in the study. The results, which are in good agreement with those obtained in similar studies, are discussed.


Why patients with inflammatory bowel disease use or do not use complementary and alternative medicine: a Canadian national survey.
The use of complementary and alternative medicine (CAM) is common in patients with inflammatory bowel disease (IBD). To determine the factors associated with use of CAM, the reasons commonly cited for use or nonuse of CAM, and the correlations between the factors associated with use of CAM and reasons for CAM use. The study included 2828 members of the Crohn's and Colitis Foundation of Canada. Subjects were mailed a questionnaire that included items on demographic characteristics, disease and treatment history, health attitudes and behaviours, and reasons for use or nonuse of CAM. Logistical regression was used to determine significant associations with current CAM use. In patients with Crohn's disease and ulcerative colitis, CAM use was associated with more severe disease activity, use of CAM for other purposes, use of exercise and prayer for IBD, and a desire for an active role in treatment decisions. CAM use was also associated with younger age in those with Crohn's disease, and less confidence in their IBD physician in those with ulcerative colitis. The most common reasons for CAM use were a desire for greater control, having heard or read that CAM might help, and the emphasis CAM places on treating the whole person. The most common reasons for not using CAM were that conventional treatments were successful, that not enough was known about CAM and a belief that CAM would not help. Disease activity and health attitudes and behaviours, but not demographic characteristics, are associated with CAM use by those with IBD.


Ending the Competition to Better Understand Emotional Processes in Borderline Personality Pathology: Commentary on Gunderson et al.
In reviewing four prominent theories, Gunderson et al. (2018) incidentally highlight for the reader the need for new collaborative research efforts that draw together scholars representing these various theories o borderline personality pathology. Many avenues for such research exist that have the potential to improve o r overall understanding of the development, maintenance and possible treatment of borderline personality pathology in ways that research grounded in just one theory does not. Herein, the similarities and differences among these theories in their assumptions with regard to emotional baselines and sequences of emotion (i.e., primary and secondary emotional reactions) are discussed as one important direction for cross-theory research collaborations.


Clinical impact of tamsulosin on generic and symptom-specific quality of life for benign prostatic hyperplasia patients: using international prostate symptom score and Rand Medical Outcomes Study 36-item Health Survey.
To examine the efficiency of alpha1-blocker treatment on disease-specific and generic quality of life (QOL) in men with clinically diagnosed benign prostatic hyperplasia (BPH), the improvement of QOL scores with International prostate symptom score (I-PSS) and Rand Medical Outcomes Study 36-item Health Survey (SF-36) was prospectively analyzed. A total of 68 newly diagnosed patients with symptomatic BPH that satisfied all inclusion and none of the exclusion criteria were prospectively recruited. All patients received 0.2 mg/day of tamsulosin for 12 weeks. All patients underwent pretreatment documentation of lower urinary tract symptoms (LUTS) and assessment of symptom-specific QOL. Symptoms and general health-related QOL (HRQOL) were assessed using the I-PSS and SF-36, respectively. Also, other objective variables, such as prostate volume, maximal urinary flow and postvoid residual urine volume, were evaluated. After 12 weeks, decrease in I-PSS was 27% compared with baseline (from 16.4 +/- 7.18 to 11.9 +/- 7.56). All questionnaires in the I-PSS showed improvement after tamsulosin treatment and the I-PSS QOL score was improved from 4.51 +/- 1.14 to 3.17 +/- 1.38 (P < 0.0001) at 12 weeks after tamsulosin administration. In intragroup comparisons of HRQOL scores with age-gender adjusted SF-36 Japanese national norms, three SF-36 subscales (bodily pain, BP; social function, SF; and mental health, MH) were worse in the BPH group aged over 70 years, while younger BPH groups aged <70 had better mean SF-36 physical function (PF) scores compared with age-gender adjusted Japanese national norms. In the BPH group with a prostatic volume > or =20 mL, three mean SF-36 scales (BP, SF and MH) were significantly improved after tamsulosin treatment. It is noteworthy that these SF-36 subscales were identical to those observed to worsen in the older BPH group compared to Japanese national norms. Treatment with tamsulosin for symptomatic BPH patients is associated with significant improvement in the generic HRQOL, in addition to disease-specific QOL and symptoms, at 3 months after drug administration. In particularly, for generic HRQOL with SF-36, tamsulosin treatment can efficiently improve three mean SF-36 subscales (BP, SF and MH) that are decreased in older BPH patients.


Proteomic approaches to decipher cancer cell secretome.
In this review, we give an overview of the actual proteomic approaches used in the study of cancer cells secretome. In particular, we describe the proteomic strategies to decipher cancer cell secretome initially focusing on the different aspects of sample preparation. We examine the issues related to the presence of low abundant proteins, the analysis of secreted proteins in the conditioned media with or without the removal of fetal bovine serum and strategies developed to reduce intracellular protein contamination. As regards the identification and quantification of secreted proteins, we described the different proteomic approaches used, i.e. gel-based, MS-based (label-based and label-free), and the antibody and array-based methods, together with some of the most recent applications in the field of cancer research. Moreover, we describe the bioinformatics tools developed for the in silico validation and characterization of cancer cells secretome. We also discuss the most important available tools for protein annotation and for prediction of classical and non-classical secreted proteins. In summary in this review advances, concerns and challenges in the field of cancer secretome analysis are discussed.


[Wound microbial sampling methods in surgical practice, imprint techniques].
The wound is a damage of tissue. The process of healing is influenced by many systemic and local factors. The most crucial and the most discussed local factor of wound healing is infection. Surgical site infection in the wound is caused by micro-organisms. This information is known for many years, however the conditions leading to an infection occurrence have not been sufficiently described yet. Correct sampling technique, correct storage, transportation, evaluation, and valid interpretation of these data are very important in clinical practice. There are many methods for microbiological sampling, but the best one has not been yet identified and validated. We aim to discuss the problem with the focus on the imprint technique.


Effects of bone marrow-derived mesenchymal stem cells on the autophagic activity of alveolar macrophages in a rat model of silicosis.
The aim of the present study was to evaluate the effects of bone marrow-derived mesenchymal stem cells (BMSCs) on the expression of the autophagy-associated proteins, microtubule-associated protein light chain 3 (LC-3) and autophagy-related gene Beclin-1 (Beclin-1), in alveolar macrophages (AMs) in a rat model of silicosis. Furthermore, the study investigated the molecular mechanisms underlying the effects of BMSC treatment. A population of 60 adult female Sprague-Dawley (SD) rats were allocated at random into three groups, namely the control, model and BMSC treatment groups (n=20 per group). BMSCs were isolated from five male SD rats (age, 6-8 weeks) and cultured in vitro. The silicosis model was established using a single 1.0-ml infusion of silicon dioxide suspension administered via non-exposed tracheal intubation. Rats in the BMSC treatment group received a 1.0-ml transplantation of BMSCs (1×106/ml). The rats were sacrificed on days 1, 7, 14 and 28 after modeling, and AMs were extracted from the rats using bronchoalveolar lavage. Third-generation BMSCs were identified using flow cytometry with fluorescein isothiocyanate staining, and the morphological characteristics of the AMs were observed using hematoxylin and eosin staining. The expression levels of LC-3 and Beclin-1 were determined using immunocytochemistry sand western blot analysis. The expression levels of LC-3 and Beclin-1 were found to be increased at all the time points in the model group. LC-3 and Beclin-1 levels began to increase at day 1, peaked at day 14 and decreased after day 28; however, the levels remained elevated compared with the basal expression levels. The AMs of the BMSC treatment group exhibited significantly alleviated pathological symptoms compared with the model group AMs, as indicated by significantly decreased expression levels of LC-3 and Beclin-1 at each time point. Therefore, the results indicated that autophagy was promoted in the AMs of the silicosis model rats. Furthermore, treatment with BMSCs was demonstrated to reduce the expression levels of LC-3 and Beclin-1, subsequently inhibiting autophagic activity and mitigating the damage associated with silicosis.


Diverse wild mouse origins of xenotropic, mink cell focus-forming, and two types of ecotropic proviral genes.
We analyzed wild mouse DNAs for the number and type of proviral genes related to the env sequences of various murine leukemia viruses (MuLVs). Only Mus species closely related to laboratory mice carried these retroviral sequences, and the different subclasses of viral env genes tended to be restricted to specific taxonomic groups. Only Mus musculus molossinus carried proviral genes which cross-reacted with the inbred mouse ecotropic MuLV env gene. The ecotropic viral env sequence associated with the Fv-4 resistance gene was found in the Asian mice M. musculus molossinus and Mus musculus castaneus and in California mice from Lake Casitas (LC). Both M. musculus castaneus and LC mice carried many additional Fv-4 env-related proviruses, two of which are common to both mouse populations, which suggests that these mice share a recent common ancestry. Xenotropic and mink cell focus-forming (MCF) virus env sequences were more widely dispersed in wild mice than the ecotropic viral env genes, which suggests that nonecotropic MuLVs were integrated into the Mus germ line at an earlier date. Xenotropic MuLVs represented the major component of MuLV env-reactive genes in Asian and eastern European mice classified as M. musculus molossinus, M. musculus castaneus, and Mus musculus musculus, whereas Mus musculus domesticus from western Europe, the Mediterranean, and North America contained almost exclusively MCF virus env copies. M. musculus musculus mice from central Europe trapped near the M. musculus domesticus/M. musculus musculus hybrid zone carried multiple copies of both types of env genes. LC mice also carried both xenotropic and MCF viral env genes, which is consistent with the above conclusion that they represent natural hybrids of M. musculus domesticus and M. musculus castaneus.


Livestock-wildlife joint land use in dry lands of Kenya: a case study of the Lolldaiga Hills ranch.
This study aimed to examine livestock-wildlife interactions at the micro level and to quantify how resources are shared in joint land use by comparing the monitoring records collected on the Lolldaiga Hills ranch in Laikipia, Kenya from 1990s onwards. Livestock and wildlife distributions together with existing water points were geo-referenced; by air and road census total animal biomass densities were estimated. Through 38-h observation at a water point, livestock-wildlife interaction was recorded. During this period, water decline has been identified as an acute factor for farming and ranching. It was found that distributions of livestock and wildlife were related to water and pasture availability during the severe drought in 2009. Although there is seasonality in densities of both livestock and wildlife populations, results of air census indicated that the stable resident populations of wildlife have resided on the ranch. In this paper, we describe how livestock and wildlife interact at a water point and on pastures on the ranch in terms of biomass density. Such resources shared at different times need to be investigated further as a key factor to improve productivity of livestock-wildlife joint land use.


The impact of oleuropein on miRNAs regulating cell death signaling pathway in human cervical cancer cells.
Cervical cancer is known as the second most pervasive malignancy in women across the globe. The role played by microRNAs (miRNAs) in the initiation, progression, and metastasis of this cancer has received specific attention. The use of natural compounds leading cancer cells toward apoptosis is a feasible strategy for cancer therapy. Oleuropein, an olive-extracted phenolic substance, displays anticancer properties. Here, it was attempted to assess the role played by oleuropein in cell viability in cervical cancer and changes in the expression of some miRNAs associated with cervical cancer as well as some of their possible target genes selected using bioinformatics analysis. For this purpose, HeLa cell line was exposed to several oleuropein concentrations for 48 and 72 h. After that, 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide assay and flow cytometry were employed to assess cell viability and apoptosis, respectively. In addition, to conduct bioinformatics analysis, Cytoscape computer program was used based on STRING database. Furthermore, to examine the role played by oleuropein in the expression of miRNAs of interest as well as their potential target genes, real-time PCR was employed. The findings indicated that oleuropein reduced cell viability through inducing apoptosis. As a result of treatment with oleuropein, miR-34a, miR-125b, and miR-29a showed increased expression levels, whereas miR-181b, miR-221, and miR-16 showed decreased expression levels. Furthermore, oleuropein reduced the expression of the anti-apoptotic genes Bcl-2 and Mcl1, whereas it elevated the expression of the pro-apoptotic Bid, Fas, and TNFRSF10B genes and the p53 tumor suppressor. Our results indicate that the apoptosis induction is a mechanism of action of oleuropein in HeLa cells. Because of its effect on the reflation of the expression of genes and miRNAs effective in the pathogenesis of cervical cancer, oleuropein shows potential as an effective research tool for developing new natural drugs for treating cervical cancer.


Pyruvate Dehydrogenase Activity and Quantity Decreases After Coronary Artery Bypass Grafting: a Prospective Observational Study.
Pyruvate dehydrogenase (PDH) is a key gatekeeper enzyme in aerobic metabolism. The main purpose of this study was to determine if PDH activity is affected by major stress in the form of coronary artery bypass grafting (CABG), which has previously been used as a model of critical illness. We conducted a prospective, observational study of patients undergoing CABG at an urban, tertiary care hospital. We included adult patients undergoing CABG with or without concomitant valve surgery. Measurements of PDH activity and quantity and thiamine were obtained prior to surgery, at the completion of surgery, and 6 h after surgery. Fourteen patients were enrolled (aged 67 ± 10 years, 21% female). Study subjects had a mean 41.7% (SD, 27.7%) reduction in PDH activity after surgery and a mean 32.0% (SD, 31.4%) reduction 6h after surgery (P < 0.001). Eight patients were thiamine deficient (≤ 7 nmol/L) after surgery compared with none prior to surgery (P = 0.002). Thiamine level was significantly associated with PDH quantity at all time points (P = 0.01). Postsurgery lactate levels were inversely correlated with postsurgery thiamine levels (r = -0.58 and P = 0.04). The stress of major surgery causes decreased PDH activity and quantity and depletion of thiamine levels.


Fetal Doppler to predict cesarean delivery for non-reassuring fetal status in the severe small-for-gestational-age fetuses of late preterm and term.
To evaluate the significance of fetal Doppler parameters in predicting adverse neonatal outcomes and the risk of cesarean delivery due to non-reassuring fetal status, in severe small for gestational age (SGA) fetuses of late preterm and term gestation. Fetal brain and umbilical artery (UmA) Doppler parameters of cerebroplacental ratio (CPR) and UmA pulsatility index (PI) were evaluated in a cohort of 184 SGA fetuses between 34 and 41 weeks gestational age, who were less than the 5th percentile. The risks of neonatal morbidities and cesarean delivery due to non-reassuring fetal status were analyzed. Univariate analysis revealed that abnormal CPR was significantly associated with cesarean delivery due to non-reassuring fetal status (P=0.018), but not with neonatal morbidities. However, abnormal CPR did not increase the risk of cesarean delivery due to non-reassuring fetal status in multivariate logistic regression analysis. Abnormal CPR with abnormal PI of UmA was associated with low Apgar score at 1 minute (P=0.048), mechanical ventilation (P=0.013) and cesarean delivery due to non-reassuring fetal status (P<0.001), in univariate analysis. It increased risk of cesarean delivery for non-reassuring fetal status (adjusted odds ratio, 7.0; 95% confidence interval, 1.2-41.3; P=0.033), but did not increase risk of low Apgar score or mechanical ventilation in multivariate logistic regression analysis. Abnormal CPR with abnormal PI of UmA increases the risk of cesarean delivery for non-reassuring fetal status, in severe SGA fetuses of late preterm and term. Monitoring of CPR and PI of UmA can help guide management including maternal hospitalization and fetal monitoring.


Construction of modified ribosomes for incorporation of D-amino acids into proteins.
While numerous biologically active peptides contain D-amino acids, the elaboration of such species is not carried out by ribosomal synthesis. In fact, the bacterial ribosome discriminates strongly against the incorporation of D-amino acids from D-aminoacyl-tRNAs. To permit the incorporation of D-amino acids into proteins using in vitro protein-synthesizing systems, a strategy has been developed to prepare modified ribosomes containing alterations within the peptidyltransferase center and helix 89 of 23S rRNA. S-30 preparations derived from colonies shown to contain ribosomes with altered 23S rRNAs were found to exhibit enhanced tolerance for D-amino acids and to permit the elaboration of proteins containing D-amino acids at predetermined sites. Five specific amino acids in Escherichia coli dihydrofolate reductase and Photinus pyralis luciferase were replaced with D-phenylalanine and D-methionine, and the specific activities of the resulting enzymes were determined.


Novel Flexible Wearable Sensor Materials and Signal Processing for Vital Sign and Human Activity Monitoring.
Advances in flexible electronic materials and smart textile, along with broad availability of smart phones, cloud and wireless systems have empowered the wearable technologies for significant impact on future of digital and personalized healthcare as well as consumer electronics. However, challenges related to lack of accuracy, reliability, high power consumption, rigid or bulky form factor and difficulty in interpretation of data have limited their wide-scale application in these potential areas. As an important solution to these challenges, we present latest advances in novel flexible electronic materials and sensors that enable comfortable and conformable body interaction and potential for invisible integration within daily apparel. Advances in novel flexible materials and sensors are described for wearable monitoring of human vital signs including, body temperature, respiratory rate and heart rate, muscle movements and activity. We then present advances in signal processing focusing on motion and noise artifact removal, data mining and aspects of sensor fusion relevant to future clinical applications of wearable technology.


Some insights about 1,6-diphenyl-1,3,5-hexatriene-lipid supramolecular assemblies by steady-state fluorescence measurements.
1,6-Diphenyl-1,3,5-hexatriene (DPH) is the most widely proposed molecular probe for the post-column fluorescence derivatization of lipids after liquid chromatography separation. This kind of detection consists of a supramolecular combination of DPH and eluted lipids. The detection is optimally performed in a mainly aqueous environment (over 80% v/v) because the weak fluorescence of DPH in water is drastically enhanced upon formation of supramolecular assemblies with lipids. In the present study, and in order to obtain better spectroscopic insights into the nature of these supramolecular assemblies, two different lipids were tested, 1,2,3-tridodecanoylglycerol (LLL) as a model triglyceride (nonpolar lipid) and dimyristoylphosphatidylcholine (DMPC) as a model phosphatidylcholine (charged amphiphilic lipid). Stoichiometry and association constants were determined on the basis of the variation of fluorescence intensity in the presence of various concentrations of lipids. LLL(60)-DPH(2) and DMPC(200)-DPH(2) complexes were identified with association constants as high as K(2) = (5.8 +/- 0.5) x 10(13) M(-2) and (17.3 +/- 2.0) x 10(13) M(-2) for LLL and DMPC, respectively. The fluorescence intensity of DPH in the presence of LLL is greater than in the presence of DMPC. An attempt to characterize the insertion mode of DPH in the lipidic supramolecular assemblies is also made.


Characteristics of the melatonin signal that provide the photoperiodic code for timing seasonal reproduction in the ewe.
The following is a progress report of our studies to identify important features of the circadian pattern of melatonin secretion which provide the photoperiodic code for daylength in regulating seasonal breeding in the Suffolk ewe. The first series of experiments evaluated two conceptual models of how melatonin codes for daylength: the circadian timing of the melatonin elevation as opposed to the length of the time melatonin is elevated during each 24-hr period (phase vs duration). Strong support has been gathered for the duration hypothesis. No evidence was obtained to support a role for phase; nevertheless, this hypothesis could not be discounted definitively. A second series of studies evaluated the importance of the previous melatonin pattern to the interpretation of a given melatonin signal. Evidence is presented that a fixed melatonin pattern can maintain a given reproductive response only for a limited length of time and that this response can be prolonged by appropriate changes in the melatonin pattern. Thus, change is an important feature of the melatonin signal. Further, the nature of the melatonin change appears to be crucial, specifically whether the nocturnal elevation increases or decreases in duration. Thus, transfer to a common photoperiod can promote either reproductive induction or arrest, depending upon whether the transfer leads to a decrease or increase in daylength. This has important ramifications to the photoperiodic timekeeping process in those species of mammals which utilize daylength to time their seasonal reproductive cycle.


The Role of Nonglycolytic Glucose Metabolism in Myocardial Recovery Upon Mechanical Unloading and Circulatory Support in Chronic Heart Failure.
Significant improvements in myocardial structure and function have been reported in some patients with advanced heart failure (termed responders [R]) following left ventricular assist device (LVAD)-induced mechanical unloading. This therapeutic strategy may alter myocardial energy metabolism in a manner that reverses the deleterious metabolic adaptations of the failing heart. Specifically, our previous work demonstrated a post-LVAD dissociation of glycolysis and oxidative-phosphorylation characterized by induction of glycolysis without subsequent increase in pyruvate oxidation through the tricarboxylic acid cycle. The underlying mechanisms responsible for this dissociation are not well understood. We hypothesized that the accumulated glycolytic intermediates are channeled into cardioprotective and repair pathways, such as the pentose-phosphate pathway and 1-carbon metabolism, which may mediate myocardial recovery in R. We prospectively obtained paired left ventricular apical myocardial tissue from nonfailing donor hearts as well as R and nonresponders at LVAD implantation (pre-LVAD) and transplantation (post-LVAD). We conducted protein expression and metabolite profiling and evaluated mitochondrial structure using electron microscopy. Western blot analysis shows significant increase in rate-limiting enzymes of pentose-phosphate pathway and 1-carbon metabolism in post-LVAD R (post-R) as compared with post-LVAD nonresponders (post-NR). The metabolite levels of these enzyme substrates, such as sedoheptulose-6-phosphate (pentose phosphate pathway) and serine and glycine (1-carbon metabolism) were also decreased in Post-R. Furthermore, post-R had significantly higher reduced nicotinamide adenine dinucleotide phosphate levels, reduced reactive oxygen species levels, improved mitochondrial density, and enhanced glycosylation of the extracellular matrix protein, α-dystroglycan, all consistent with enhanced pentose-phosphate pathway and 1-carbon metabolism that correlated with the observed myocardial recovery. The recovering heart appears to direct glycolytic metabolites into pentose-phosphate pathway and 1-carbon metabolism, which could contribute to cardioprotection by generating reduced nicotinamide adenine dinucleotide phosphate to enhance biosynthesis and by reducing oxidative stress. These findings provide further insights into mechanisms responsible for the beneficial effect of glycolysis induction during the recovery of failing human hearts after mechanical unloading.


Evolution of coordinated alternating reciprocity in repeated dyadic games.
A genetic algorithm incorporating mutation and crossing-over was used to investigate the evolution of social behaviour in repeated Prisoner's Dilemma, Chicken (Hawk-Dove), Battle of the Sexes, and Leader games. The results show that the strategic structure of an interaction has a crucial determining effect on the type of social behaviour that evolves. In particular, simulations using repeated Prisoner's Dilemma and Chicken (Hawk-Dove) games lead to the emergence of genes coding for symmetric reciprocity and the evolution of mutual cooperation, whereas simulations using repeated Battle of the Sexes and Leader games lead to near-fixation of genes coding for asymmetric strategic choices and the evolution of coordinated alternating reciprocity. A mechanism is suggested whereby, in games with asymmetric equilibrium points, coordinated alternating reciprocity might evolve without insight or communication between players.


IL-1RA promotes oral squamous cell carcinoma malignancy through mitochondrial metabolism-mediated EGFR/JNK/SOX2 pathway.
Interleukin-1 receptor antagonist (IL-1RA), a member of the IL-1 family, has diverse roles in cancer development. However, the role of IL-1RA in oral squamous cell carcinoma (OSCC), in particular the underlying mechanisms, remains to be elucidated. Tumor tissues from OSCC patients were assessed for protein expression by immunohistochemistry. Patient survival was evaluated by Kaplan-Meier curve analysis. Impact of differential IL-1RA expression on cultured OSCC cell lines was assessed in vitro by clonogenic survival, tumorsphere formation, soft agar colony formation, and transwell cell migration and invasion assays. Oxygen consumption rate was measured by Seahorse analyzer or multi-mode plate reader. PCR array was applied to screen human cancer stem cell-related genes, proteome array for phosphorylation status of kinases, and Western blot for protein expression in cultured cells. In vivo tumor growth was investigated by orthotopic xenograft in mice, and protein expression in xenograft tumors assessed by immunohistochemistry. Clinical analysis revealed that elevated IL-1RA expression in OSCC tumor tissues was associated with increased tumor size and cancer stage, and reduced survival in the patient group receiving adjuvant radiotherapy compared to the patient group without adjuvant radiotherapy. In vitro data supported these observations, showing that overexpression of IL-1RA increased OSCC cell growth, migration/invasion abilities, and resistance to ionizing radiation, whereas knockdown of IL-1RA had largely the opposite effects. Additionally, we identified that EGFR/JNK activation and SOX2 expression were modulated by differential IL-1RA expression downstream of mitochondrial metabolism, with application of mitochondrial complex inhibitors suppressing these pathways. Furthermore, in vivo data revealed that treatment with cisplatin or metformin-a mitochondrial complex inhibitor and conventional therapy for type 2 diabetes-reduced IL-1RA-associated xenograft tumor growth as well as EGFR/JNK activation and SOX2 expression. This inhibitory effect was further augmented by combination treatment with cisplatin and metformin. The current study suggests that IL-1RA promoted OSCC malignancy through mitochondrial metabolism-mediated EGFR/JNK activation and SOX2 expression. Inhibition of this mitochondrial metabolic pathway may present a potential therapeutic strategy in OSCC.


Chlorogenic Acid Protects Against Indomethacin-Induced Inflammation and Mucosa Damage by Decreasing Bacteroides-Derived LPS.
Background: Chlorogenic acid (CGA), a natural bioactive polyphenol, exerts anti-inflammatory, antioxidant, and antibacterial effects that support the maintenance of intestinal health. However, the influence of CGA on gut microbiota and their metabolites, as well as its potential effects and mechanism of action in inflammatory bowel disease, remain to be elucidated. Methods: First, an oral gavage was used to administer CGA to indomethacin-treated mice. Then, fecal microbiota transplantation was performed to explore the role of intestinal microbiota in indomethacin-induced inflammation. Results: CGA treatment protected against body weight loss, damage to intestinal morphology and integrity, inflammation, and alteration of microbiota composition in indomethacin-treated mice. Interestingly, CGA failed to inhibit inflammation or protect intestine integrity in mice treated with antibiotics. Notably, mice who had been colonized with intestinal microbiota from CGA-treated or CGA-and-indomethacin-treated mice, through the fecal microbiota transplantation program, were protected from indomethacin-induced inflammation, growth of Bacteroides, and the accumulation of Bacteroides-derived LPS, in congruence with those who had been treated with CGA. Conclusion: The results suggest that CGA may protect intestine integrity and alleviate inflammatory responses, primarily by inhibiting the growth of Bacteroides and the accumulation of Bacteroides-derived LPS, in indomethacin-induced colitis. This newly identified mechanism broadens our knowledge of how CGA exerts protective effects on intestinal inflammation and provides strategies for the prevention of gastrointestinal mucosal damage in patients treated with indomethacin.


Novel approaches in pain management in cardiac surgery.
Open cardiac surgery may cause severe postoperative pain and the activation of a perioperative stress response. If not treated adequately, the patient may suffer increased morbidity, a longer hospital stay, and higher overall costs. This article reviews the literature regarding various modalities for management of postoperative pain after cardiac surgery. Paravertebral block of the spinal nerve roots provides similar analgesia to thoracic epidural without the risk of hypotension or epidural hematoma. Continuous α-2 agonist infusion reduces opioid requirements in the immediate postoperative period and may convey a morbidity and mortality benefit in cardiac surgery patients that persists for 12 months. Antiepileptics may significantly decrease opioid requirements and improve pain scores. Finally, complementary and alternative practices such as acupuncture, music, and behavioral exercises both pre and postoperatively may improve acute pain and lessen conversion to chronic pain. Although published data remain limited, recent evidence indicates that patients may benefit from the addition of a variety of novel pain-management strategies currently under investigation. Selection of a multimodal approach to perioperative pain management is advocated, including selective application of regional analgesia, non-narcotic medications, and complimentary alternative options to improve patient comfort and overall outcome.


Crown ether appended cyclam receptors for cationic guests.
A series of crown ether appended macrocyclic amines has been prepared comprising benzo-12-crown-4, benzo-15-crown-5, or benzo-18-crown-6 attached to a diamino-substituted cyclam. The Co(III) complexes of these three receptors have been prepared and characterized spectroscopically and structurally. Crystal structures of each receptor in complex with an alkali metal ion and structures of the benzo-12-crown-4 and benzo-15-crown-5-receptors without guest ions are reported. 2D NMR and molecular mechanics modeling have been used to examine conformational variations upon guest ion complexation. Addition of cations to these receptors results in an appreciable anodic shift in the Co(III/II) redox potential, even in aqueous solution, but little cation selectivity is observed. Evidence for complex formation has been corroborated by (23)Na and (7)Li NMR spectroscopy and electrospray mass spectrometry.


Hesitant steps from the artificial skin to organ regeneration.
This is a historical account of the steps, both serendipitous and rational, that led my group of students and colleagues at MIT and Harvard Medical School to discover induced organ regeneration. Our research led to methods for growing back in adult mammals three heavily injured organs, skin, peripheral nerves and the conjunctiva. We conclude that regeneration in adults is induced by a modification of normal wound healing.


Promoting the integration of genetic counseling education and research across the spectrum of learners at a large academic institution.
Genetic counselors are valuable members of research teams and are increasingly taking leadership roles in major research trials. While the field of genetic counseling has explored a doctoral degree several times, the terminal degree is still a master's level. Genetic counseling training programs provide research training, but many genetic counselors desire additional opportunities to develop expertise. To address the gaps in the availability of research training before and after obtaining genetic counseling degrees, we applied for and received a grant centered around three aims: (1) develop an undergraduate minor in genomics and human health with a research component, (2) promote a rigorous genetic counseling research program for training to practice genetic counselors, and (3) initiate a combined PhD program in Molecular, Cellular, Developmental Biology, and Genetics (MCDB&G) with a track in genetic counseling. We describe each of these components and early progress toward these aims. As each individual aim is bearing fruit, the next challenges are implementing more crossover between people in each aim and distilling the lessons learned to see if this program could be replicated at other sites.


Progress in pretreatment of methadone: an update since 2015.
Methadone, a µ-opioid receptor agonist, is widely used in pain-relieving and treating opioid dependence. If not strictly controlled, as an opioid substitute, it can lead to abuse and cause more severe withdrawal responses than heroin. Also, overdose or abuse of this drug in clinical use may provide severe side effects such as apnea, circulatory collapse, cardiac arrest, and even death. For these reasons, simple, rapid, and efficient methods have been developed for the pretreatment of methadone. This review presents a comprehensive conclusion of the pretreatment methods used for methadone in various sample matrices, focusing on the developments since 2015. Traditionally used pretreatment methods like solid-phase extraction and liquid-liquid extraction are discussed and newly developed methods like solid-phase microextraction and liquid-liquid microextraction along with new materials applied are focused.


The influence of different degrees of chronic lymphocytic thyroiditis on thyroid function after surgery for benign, non-toxic goitre.
Of 220 patients, surgically treated for benign, non-toxic goitre with unilateral procedures during a six-year period, 201 could be followed up, on average, 8 years postoperatively. Twenty-four patients were treated with thyroxine immediately postoperatively ("recurrence prophylaxis"); in the other patients thyroxine was only given in cases of hypothyroidism (significant increase of S-TSH). Occurrence of lymphoid infiltration in the removed lobe was subjectively quantified according to a five point scale. Five of the 15 patients with pronounced inflammation developed hypothyroidism whereas 5 of the 137 patients without inflammation had hypothyroidism (p less than 0.05). There was a significant difference in S-TSH postoperatively between patients with no or, only a slight degree of, chronic inflammation, and patients with pronounced inflammation. This study indicates that histologic grading of lymphocytic infiltration in the thyroid gland may be useful for predicting the risk of postoperative hypothyroidism.


Molecular screening of potential HNPCC patients using a multiplex microsatellite PCR system.
Microsatellite instability (MSI) is a molecular hallmark of the H ereditary N on- P olyposis C olorectal C ancer (HNPCC) syndrome occurring in about 80-90% of the tumours and also in sporadic tumours of different organs, albeit at lower frequency. Highly unstable colorectal tumours (MSI-H) have different histopathological features and tend to have a better prognosis compared to neoplasms without (MSS) or with low levels of microsatellite instability (MSI-L). Since MSI classification allows the identification of potential HNPCC patients and might represent a valuable diagnostic parameter an increasing demand for high-throughput microsatellite analysis will arise. Therefore, we have adapted five diagnostic microsatellites, m(odified) ACTC, mBAT26, mD5S107, mD5S406 and mD13S153, to allow coamplification. Using this multiplex polymerase chain reaction (PCR) system 29 colorectal tumour tissues with known MSI status could be unambiguously identified as MSI-H (13 cases) or MSI-L/MSS (16 cases). Highly unstable colorectal tumour detection frequency of individual markers reached 77% (mD5S406), 85% (mACTC), 85% (mD5S107), 92% (D13S153) and 100% (mBAT26) showing similar sensitivity but improved specificity as compared with a microsatellite reference panel. In a prospective analysis of 31 colorectal tumours, the multiplex PCR system identified five MSI-H cases. Multiplex MSI PCR is a time saving and cost-effective method not restricted to specific technical equipment and applicable to a variety of microsatellite-based genotyping approaches.


Knowledge, attitudes and sources of information regarding HIV/AIDS in Iranian adolescents.
The World Health Organization recently reported an 'alarming trend' of increasing Human Deficiency Virus (HIV) infection in the Islamic Republic of Iran. Adolescents form a particularly important target group for primary prevention. An anonymous questionnaire, derived from standard surveys such as the Safer Choices questionnaire and the 2001 Youth Risk Behavior Survey, was distributed to 1227 Iranian students attending 19 randomly selected high schools in Hashtgerd in 2002. Students reported that television (84%) and school teachers (66%) were the best sources of HIV/AIDS information, while parents (27%) and school books (15%) were least informative. Most students knew that heterosexual intercourse (90%) and shared intravenous needles (94%) can cause HIV infection; however, salient misconceptions were revealed. Only 53% were aware that condoms protect against infection through sexual intercourse. More effective school-based HIV/AIDS education is needed in Iran.


Noninvasive pressure support ventilation.
In appropriately selected patients, NPPV decreases intubation rate, improves survival, and is cost-effective. Pressure support ventilation is commonly used successfully for NPPV. An issue with the use of pressure support is the ability of the ventilator to detect the end of inhalation in the presence of leaks or in patients with severe airflow obstruction. A concern with portable pressure ventilators is the potential for rebreathing of carbon dioxide. Despite these issues, pressure support ventilation has been used successfully for noninvasive ventilation in thousands of patients with acute respiratory failure.


[Ru(phen)2(PHEHAT)]2+ and [Ru(phen)2(HATPHE)]2+: two ruthenium(II) complexes with the same ligands but different photophysics and spectroelectrochemistry.
The properties of two mononuclear Ru(II) complexes formed with the extended planar ligand PHEHAT depend drastically on the chelation site by the metallic ion. When the chelation takes place on the HAT site of the ligand (case of the novel complex [Ru(phen)2(HATPHE)]2+), the emission behavior is quite similar to that of [Ru(phen)2(HAT)]2+. In contrast, when the chelation is on the phen motif of the ligand (case of [Ru(phen)2(PHEHAT)]2+), the spectroscopic (absorption and emission) and electrochemical data for the complex do not obey the linear spectroelectrochemical correlation and the emission behavior is comparable to that of the extensively studied dppz complex ([Ru(phen/bpy)2(dppz)]2+). Thus, for [Ru(phen)2(PHEHAT)]2+, the emission lifetimes and intensities as a function of temperature exhibit a maximum for nitrile solvents. However, in contrast to the dppz case, at least three different states (two emitting and one dark) participate in the deactivation with different contributions depending on the temperature. These different contributions explain the observed maximum. Moreover, the fact that the solvent is liquid or frozen also influences the nature of the luminescent species.


Heavy metal and trace elements in riparian vegetation and macrophytes associated with lacustrine systems in Northern Patagonia Andean Range.
Vegetation associated with lacustrine systems in Northern Patagonia was studied for heavy metal and trace element contents, regarding their elemental contribution to these aquatic ecosystems. The research focused on native species and exotic vascular plant Salix spp. potential for absorbing heavy metals and trace elements. The native species studied were riparian Amomyrtus luma, Austrocedrus chilensis, Chusquea culeou, Desfontainia fulgens, Escallonia rubra, Gaultheria mucronata, Lomatia hirsuta, Luma apiculata, Maytenus boaria, Myrceugenia exsucca, Nothofagus antarctica, Nothofagus dombeyi, Schinus patagonicus, and Weinmannia trichosperma, and macrophytes Hydrocotyle chamaemorus, Isöetes chubutiana, Galium sp., Myriophyllum quitense, Nitella sp. (algae), Potamogeton linguatus, Ranunculus sp., and Schoenoplectus californicus. Fresh leaves were analyzed as well as leaves decomposing within the aquatic bodies, collected from lakes Futalaufquen and Rivadavia (Los Alerces National Park), and lakes Moreno and Nahuel Huapi (Nahuel Huapi National Park). The elements studied were heavy metals Ag, As, Cd, Hg, and U, major elements Ca, K, and Fe, and trace elements Ba, Br, Co, Cr, Cs, Hf, Na, Rb, Se, Sr, and Zn. Geochemical tracers La and Sm were also determined to evaluate contamination of the biological tissues by geological particulate (sediment, soil, dust) and to implement concentration corrections.


Hypercalcaemia in a patient with tuberculous lymphadenitis.
This case report presents the clinical details, investigations, diagnosis, treatment and outcomes of a male patient in his 50s who presented with weight loss and fatigue. On evaluation, he had axillary lymphadenopathy, along with hypercalcaemia and elevated serum creatinine levels. The patient was diagnosed with tuberculous lymphadenitis based on lymph node biopsy and positive tuberculosis (TB)-PCR results. Treatment involved hydration, salmon calcitonin and zoledronic acid, leading to symptomatic improvement. This case highlights the rarity of hypercalcaemia and renal dysfunction in TB and underscores the importance of considering this entity in the differential diagnosis.


Measurement of spinal sagittal curvatures using the laser triangulation method.
The purpose of the first part of the study was to establish the variability of repeated measurements in different measuring conditions. In the second part, we performed in a large number of patients, a measurement of thoracic kyphosis and lumbar lordosis and compared them to age, gender, and level of nourishment. In the first part, measurements were performed on a plastic model of the back of a patient with a rigid and a normal spine. In the second part, 250 patients participated in the study (126 men and 124 women). For measuring spinal curvatures we used an apparatus for laser triangulation constructed at the Faculty of Mechanical Engineering, University of Ljubljana. A comparison of 30 repeated measurements was shown as the average value +/- 2 SD which included 95% of the results. Thirty repeated readings of one 3D measurement: thoracic kyphosis 41.2 degrees +/- 0.6 degrees, lumbar lordosis 4.4 degrees +/- 1.2 degrees; 30 measurements on a plastic model: thoracic kyphosis 36.8 degrees +/- 1.2 degrees, lumbar lordosis 30.9 degrees +/- 2.0 degrees; 30 measurements on a patient with a rigid spine: thoracic kyphosis 41.5 degrees +/- 2.4 degrees, lumbar lordosis 4.0 degrees +/- 1.8 degrees; 30 measurements on a patient with a normal spine: thoracic kyphosis 48.8 degrees +/- 7.4 degrees, lumbar lordosis 21.1 degrees +/- 4.4 degrees. The average size of thoracic kyphosis in 250 patients was 46.8 degrees (SD 10.1 degrees) and lumbar lordosis 31.7 degrees (SD 12.5 degrees). The angle size was statistically significantly correlated to gender (increased thoracic kyphosis and lumbar lordosis in women) and body mass index (increased thoracic kyphosis and lumbar lordosis in more nourished patients). Age was not significantly correlated to the observed angles. During measurements of the spinal angles it was important to pay attention to relaxation and the patient's position as well as to perform more measurements providing the average value. The age and the level of nourishment influence the size of the sagittal spinal angles. In the observed sample the effect of age was not confirmed.


Facile morphology-controlled synthesis of nickel-coated graphite core-shell particles for excellent conducting performance of polymer-matrix composites and enhanced catalytic reduction of 4-nitrophenol.
We have developed a novel seed-mediated growth method to fabricate nickel-coated graphite composite particles (GP@Ni-CPs) with controllable shell morphology by simply adjusting the concentration of sodium hydroxide ([NaOH]). The fabrication of two kinds of typical GP@Ni-CPs includes adsorption of Ni2+ via electrostatic attraction, sufficient heterogeneous nucleation of Ni atoms by an in situ reduction, and shell-controlled growth by regulating the kinetics of electroless Ni plating in turn. High [NaOH] results in fast kinetics of electroless plating, which causes heterogeneous nuclei to grow isotropically. After fast and uniform growth of Ni nuclei, GP@Ni-CPs with dense shells can be achieved. The first typical GP@Ni-CPs exhibit denser shells, smaller diameters and higher conductivities than the available commercial ones, indicating their important applications in the conducting of polymer-matrix composites. On the other hand, low [NaOH] favors slow kinetics. Thus, the reduction rate of Ni2+ slows down to a relatively low level so that electroless plating is dominated thermodynamically instead of kinetically, leading to an anisotropic crystalline growth of nuclei and finally to the formation of GP@Ni-CPs with nanoneedle-like shells. The second typical samples can effectively catalyze the reduction of p-nitrophenol into p-aminophenol with NaBH4 in comparison with commercial GP@Ni-CPs and RANEY® Ni, owing to the strong charge accumulation effect of needle-like Ni shells. This work proposes a model system for fundamental investigations and has important applications in the fields of electronic interconnection and catalysis.


Biphasic Effect of Sildenafil on Energy Sensing is Mediated by Phosphodiesterases 2 and 3 in Adipocytes and Hepatocytes.
Sirt1 (Sirtuin 1), AMPK (AMP-activated protein kinase), and eNOS (endothelial nitric oxide synthase) modulate hepatic energy metabolism and inflammation and play a major role in the development of NASH. Cyclic nucleotide phosphodiesterases (PDEs) play an important role in signal transduction by modulating intracellular levels of cyclic nucleotides. We previously found the PDE5 inhibitor sildenafil to synergize with leucine and leucine-metformin combinations in preclinical studies of NASH and obesity. However, efficacy is diminished at higher sildenafil concentrations. Herein, we have successfully modeled the U-shaped sildenafil dose-response in vitro and utilized this model to assess potential mechanisms of this dose-response relationship. Adipocytes and liver cells were treated with leucine (0.5 mM) and different concentrations of sildenafil (1 nM to 100 µM). cAMP, cGMP, and P-AMPK protein expression were used to demonstrate the biphasic response for increasing concentrations of sildenafil. The reversal with higher sildenafil levels was blunted by PDE2 inhibition. These data indicate that sildenafil-mediated increases in cGMP inhibits PDE3 at lower concentrations, which increases cAMP. However, further increases in cGMP from higher sildenafil concentrations activate PDE2 and consequently decrease cAMP, which demonstrates crosstalk between cAMP and cGMP via PDE2, PDE3, and PDE5. These changes in cAMP concentration are further reflected in downstream effects, including AMPK activation.


Effect of superparamagnetic iron oxide nanoparticles on fluidity and phase transition of phosphatidylcholine liposomal membranes.
Superparamagnetic iron oxide nanoparticles (SPIONs) with multifunctional properties have shown great promise in theranostics. The aim of our work was to compare the effects of SPIONs on the fluidity and phase transition of the liposomal membranes prepared with zwitterionic phosphatidylcholine lipids. In order to study if the surface modification of SPIONs has any influence on these membrane properties, we have used four types of differently functionalized SPIONs, such as: plain SPIONs (primary size was shown to bê11 nm), silica-coated SPIONs, SPIONs coated with silica and functionalized with positively charged amino groups or negatively charged carboxyl groups (the primary size of all the surface-modified SPIONs was ~20 nm). Small unilamellar vesicles prepared with 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine lipids and multilamellar vesicles prepared with 1,2-dipalmitoyl-sn-glycero-3-phosphocholine lipids were encapsulated or incubated with the plain and surface-modified SPIONs to determine the fluidity and phase transition temperature of the bilayer lipids, respectively. Fluorescent anisotropy and differential scanning calorimetric measurements of the liposomes that were either encapsulated or incubated with the suspension of SPIONs did not show a significant difference in the lipid ordering and fluidity; though the encapsulated SPIONs showed a slightly increased effect on the fluidity of the model membranes in comparison with the incubated SPIONs. This indicates the low potential of the SPIONs to interact with the nontargeted cell membranes, which is a desirable factor for in vivo applications.


Asymmetry of cerebral gray and white matter and structural volumes in relation to sex hormones and chromosomes.
Whilst many studies show sex differences in cerebral asymmetry, their mechanisms are still unknown. This report describes the potential impact of sex hormones and sex chromosomes by comparing MR data from 39 male and 47 female controls and 33 men with an extra X-chromosome (47,XXY). Regional asymmetry in gray and white matter volumes (GMV and WMV) was calculated using voxel based moprhometry (SPM5), by contrasting the unflipped and flipped individual GMV and WMV images. In addition, structural volumes were calculated for the thalamus, caudate, putamen, amygdala, and hippocampus, using the FreeSurfer software. Effects of plasma testosterone and estrogen on the GMV and WMV, as well on the right/left ratios of the subcortical volumes were tested by multi-regression analysis. All three groups showed a leftward asymmetry in the motor cortex and the planum temporale, and a rightward asymmetry of the middle occipital cortex. Both asymmetries were more pronounced in 46,XY males than 46,XX females and 47,XXY males, and were positively correlated with testosterone levels. There was also a rightward asymmetry of the vermis and leftward GMV asymmetry in the cerebellar hemispheres in all groups. Notably, cerebellar asymmetries were larger in 46,XX females and 47,XXY males, but were not related to sex hormone levels. No asymmetry differences between 46,XX females and 47,XXY males, and no overall effects of brain size were detected. The asymmetry in the planum temporale area and the occipital cortex seem related to processes associated with testosterone, whereas the observed cerebellar asymmetries suggest a link with X-chromosome escapee genes. Sex differences in cerebral asymmetry are moderated by sex hormones and X-chromosome genes, in a regionally differentiated manner.


The effect of midazolam on implicit and explicit memory in category exemplar production and category cued recall.
Transfer-appropriate processing theory (Roediger, Weldon, & Challis, 1989) proposes that dissociations between performance on explicit and implicit memory tests arise because these tests often rely on different types of information processing (e.g., perceptual processing vs conceptual processing). This perspective predicts that implicit and explicit memory tasks that rely primarily on conceptual processing should show comparable results, not dissociations. Numerous studies have demonstrated such similarities. It is, however, possible that these results arise from explicit memory contamination of performance on implicit memory tasks. To address this issue, an experiment was conducted in which participants were administered the sedative midazolam prior to study. Midazolam is known to create a temporary, but dense, period of anterograde amnesia. The effects of blocking stimulus materials by semantic category at study and generation at study were investigated on category exemplar production and category-cued recall. The results of this study demonstrated a dissociation of the effects of midazolam on category exemplar production and category-cued recall. Specifically, midazolam reduced the effect of blocking stimulus materials in category-cued recall, but not in category exemplar production. The differential effect of midazolam on explicit and implicit memory is at odds with transfer-appropriate processing theory and suggests that theories of memory must distinguish the roles of different types of conceptual processing on implicit and explicit memory tests.


Structure and visible light luminescence of 3D flower-like Co3O4 hierarchical microstructures assembled by hexagonal porous nanoplates.
A two-step strategy has been developed to fabricate 3D flower-like Co3O4 hierarchical microstructures assembled by hexagonal porous nanoplates. The synthetic procedure was described as (1) 3D flower-like α-Co(OH)2 microstructures were prepared by a facile surfactant-free low-temperature hydrothermal process; (2) 3D flower-like Co3O4 hierarchical microstructures were fabricated by annealing the obtained 3D flower-like α-Co(OH)2 microstructures. X-ray diffraction and Raman spectrum analyses demonstrate that the hierarchical microstructures formed from 3D flower-like α-Co(OH)2 microstructures are composed of pure cubic phase Co3O4. Scanning electronic microscopy demonstrates that the as-prepared Co3O4 microstructures exhibit 3D flower-like hierarchical structures assembled by hexagonal porous nanoplates. Photoluminescence demonstrates that these novel 3D flower-like Co3O4 hierarchical microstructures display a broad strong emission in the visible range of 650 to 800 nm with a peak at around 710 nm (1.75 eV), which is very close to the indirect optical band gap of 1.60 eV for Co3O4 thin film. The result indicates that the photoluminescence emission likely originates from the indirect optical band gap emission. The broad photoluminescence emission may be resulted from a wide size distribution of porous nanoplates in 3D hierarchical microstructures. These 3D flower-like Co3O4 hierarchical microstructures with unique optical properties may find new potential applications in visible light emitting materials.


Severe pulmonary metastasis in obese and diabetic mice.
Although obesity is known as a risk factor for several human cancers, the association of obesity with cancer recurrence and metastasis remains to be characterized. Here, B16-BL6 melanoma and Lewis lung carcinoma cells were intravenously injected into diabetic (db/db) and obese (ob/ob) mice. The number of experimental lung colonies was markedly promoted in these mice when compared with C57BL/6 mice. In contrast, tumor growth at the implanted site was comparable when cells were inoculated orthotopically. The use of B16-BL6 cells stably transfected with the luciferase gene revealed that the increased metastasis reflected a difference mainly within 6 hr after the intravenous inoculation of tumor cells. Administration of recombinant leptin in ob/ob mice abolished the increase in metastasis early on as well as the decrease in the splenic NK cell number. In addition, depletion of NK cells by an anti-asialo-GM1 antibody abrogated the enhanced metastasis in db/db mice. These results demonstrate that metastasis is markedly promoted in diabetic and obese mice mainly because of decreased NK cell function during the early phase of metastasis.


Bilirubin binding activity of cytokeratin 18 isolated from the porcine liver.
A porcine liver 40 kDa protein designated SBP40 isolated by affinity chromatography with agarose-linked spermine was identified as a porcine cytokeratin 18 on the basis of partial amino acid sequences of peptides derived by lysylendopeptidase digestion and by its reactivity with two commercially available preparations of monoclonal antibody. Immunohistochemistry revealed that SBP40 is localized at the hepatocyte membranes, preferentially in the bile canalicular area in accordance with the previously reported localization of cytokertain 18 in the murine liver. Affinity chromatography with agarose-linked bilirubin, a solubilization experiment of bilirubin from bilirubin-Sephadex G-10 complex, and gel-filtration of a mixture with bilirubin demonstrated that SBP40 or porcine cytokeratin 18 has binding affinity for bilirubin. These results suggest that cytokeratin 18 may play a role as a membrane reservoir in the event of transport and secretion of bile pigments in the liver.


Work conditions and their repercussions on the quality of life of rural workers.
To assess the context of work and analyze their repercussions on the quality of life of rural workers in soybean agribusiness. Cross-sectional study, with an intentional sample of rural workers, conducted between the months of October and December 2019, with the application of the instruments World Health Organization Quality Life-bref and Scales of Assessment of Context and Human Cost of Work. The data were analyzed through bivariate and multivariate descriptive statistics. The participants amounted to 299 rural workers. The scores of Work Conditions and Socioprofessional Relations were satisfactory and Organization of Work was critical. The scores of Physical and Cognitive costs were considered critical and of the Affective Cost was satisfactory. The Socioprofessional Relations have negatively impacted the overall quality of life and the Physical and Psychological, the Social Relations and the Environmental Domains, whereas Cognitive Cost positively impacted the Physical and the Environmental Domain and the Affective Cost positively impacted the Social Relations Domain. Although the Work Context and Human Cost indicated adverse conditions, only the Socioprofessional Relations exerted negative influences on the quality of life of rural workers.


Cyclic modulation of H-reflex depression in ipsilateral and contralateral soleus muscles during rhythmic arm swing.
The objective of the present study was to investigate the effects of rhythmic arm swing on ipsilateral and contralateral soleus motoneuron pool excitability. Ten healthy human subjects participated in this study. Soleus H-reflexes were recorded from the ipsilateral and contralateral soleus muscles while the subject swung the right arm anteroposteriorly as if during gait. The soleus H-reflex was depressed throughout the whole arm swing cycle except in the ipsilateral leg during the onset of the backward arm swing, and in the contralateral leg during the last half of the backward arm swing and the onset of the forward arm swing. The depression was cyclically modulated in accordance with the time course of the arm swing periods, and the pattern of the modulation was reciprocal between the ipsilateral and contralateral legs. This cyclical and reciprocal modulation may be related to the regulation of soleus motoneuron pool excitability during gait.


A Multimode 28 GHz CMOS Fully Differential Beamforming IC for Phased Array Transceivers.
A 28 GHz fully differential eight-channel beamforming IC (BFIC) with multimode operations is implemented in 65 nm CMOS technology for use in phased array transceivers. The BFIC has an adjustable gain and phase control on each channel to achieve fine beam steering and beam pattern. The BFIC has eight differential beamforming channels each consisting of the two-stage bi-directional amplifier with a precise gain control circuit, a six-bit phase shifter, a three-bit digital step attenuator, and a tuning bit for amplitude and phase variation compensation. The Tx and Rx mode overall gains of the differential eight-channel BFIC are around 11 dB and 9 dB, respectively, at 27.0-29.5 GHz. The return losses of the Tx mode and Rx mode are >10 dB at 27.0-29.5 GHz. The maximum phase of 354° with a phase resolution of 5.6° and the maximum attenuation of 31 dB, including the gain control bits with an attenuation resolution of 1 dB, is achieved at 27.0-29.5 GHz. The root mean square (RMS) phase and amplitude errors are <3.2° and <0.6 dB at 27.0-29.5 GHz, respectively. The chip size is 3.0 × 3.5 mm2, including pads, and Tx mode current consumption is 580 mA at 2.5 V supply voltage.


[Lesions of the gastric mucosa caused by blunt abdominal injury].
Within a casuistic is reported on an 8-year-old boy who suffered from an intestinal haemorrhage due to a blunt abdominal trauma. Due to the endoscopic possibilities which are nowadays at our disposal appeared an alteration of the gastric mucous membrane in our opinion conditioned by contusion which from the macroscopic standpoint let us think of an ulceration. Theoretical fundaments as well as the anamnesis and the clinical course are correlated with our opinion to this case.


Morbidity, mortality and long term survival in patients with vascular resection in pancreatic cancer - single center experience.
Although venous resection in pancreatic cancer is widely used method, recently published data about its safety and survival benefit showed conflicting results. A retrospective case matched study was performed to compare the results of patients who underwent venous resection to those with no venous resection during radical surgery in pancreatic cancer.From January 2010 to December 2015, 297 pancreatic resections due to pancreatic tumor were performed in the National Cancer Institute (NCI). Fifty-three patients with venous resection were identified and enrolled into the study and matched with 66 patients without vascular resection during radical resection of pancreatic head/body cancer. Both groups matched for age, ASA score, need for preoperative biliary drainage and clinical staging of the tumor. Morbidity was determined according Clavien and Dindo classification [1] and was similar in both groups of patients (p = 0.48). Thirty day postoperative mortality was also equal in both groups 5.6 vs 4.5% (p > 0.99) and long term survival was with no significant difference. Median overall survival was 18.8 vs 20.7 months (p = 0.33) for patients with/without venous resection.Therefore we consider venous resection in pancreatic cancer safe procedure with equal morbidity and perioperative mortality as in patients with no need for vascular resection and with the same long term survival if R0 resection is achieved.


Inhibitors of BRD4 Protein from a Marine-Derived Fungus Alternaria sp. NH-F6.
Bromodomains (BRD) are readers of the epigenetic code that regulate gene transcription through their recognition of acetyl-lysine modified histone tails. Recently, bromodomain-containing proteins such as BRD4 have been demonstrated to be druggable through the discovery of potent inhibitors. These protein-protein interaction inhibitors have the potential to modulate multiple diseases by their profound anti-inflammatory and antiproliferative effects. In order to explore new BRD4 inhibitors as well as lead compounds for the development of new drugs, the secondary metabolites of Alternaria sp. NH-F6, a fungus isolated from deep-sea sediment samples, were analyzed systematically. Five new compounds including two new perylenequinones (1-2), one new alternaric acid (3), 2-(N-vinylacetamide)-4-hydroxymethyl-3-ene-butyrolactone (4), one new cerebroside (5), together with 19 known compounds (6-24) were isolated from the ethyl acetate extracts of this strain. Their structures were elucidated using nuclear magnetic resonance (NMR) and high resolution electrospray ionization mass spectrometry (HR-ESI-MS) analyses. Finally, all these compounds were evaluated for their inhibitory activity against BRD4 protein, and compound 2 exhibited a potent inhibition rate of 88.1% at a concentration of 10 µM. This research provides a new BRD4 inhibitor which may possess potential antitumoral, antiviral, or anti-inflammatory pharmaceutical values.


Ultra-broad bandwidth low-dispersion mirror with smooth dispersion and high laser damage resistance.
Low-dispersion mirrors (LDMs), which require a broad bandwidth, low dispersion, and high damage threshold, are essential optics in ultra-intense and ultra-short laser devices. Bragg mirrors and chirped LDMs do not satisfy these requirements simultaneously. We propose a novel LDM (NLDM) based on the hump-like structure and quarter wavelength optical thickness (QWOT) structure to achieve a broad bandwidth, smooth dispersion, and high robustness. The spectral and dispersion characteristics of the two structures compensate for each other, which makes up for the deficiency that the dispersion bandwidth of the sinusoidal modulation structure cannot be broadened. Based on this structure, the LDM can achieve a design bandwidth of 240 nm and support the transmission of sub-11-fs pulses. The accuracy of the NLDM is experimentally evaluated. The structure shows the potential for broad-spectrum laser damage performance due to the low electric field intensity. The NLDM improves the mirror performance and paves the way for a new generation of ultra-intense and ultra-short laser devices.