Analysis of potential genomic confounding in genetic association studies and an online genomic confounding browser (GCB).
Genome-wide association studies have transformed genetic studies of disease susceptibility, identifying many variants that may tag functional polymorphism nearby. Variants are often ascribed to a physically close gene exhibiting plausible functionality for a causal pathway. However, more physically remote genes may be at a lesser linkage or linkage disequilibrium (LD) distance from the tested SNP and could therefore contain the functional variant tagged. This analysis aims to identify instances where research may be misled by misassociation of a variant with a gene and develop tools to analyse genomic confounding. A catalogue of reported associations was systematically analysed for unreported genes which may represent the true functionality ascribed to a reported variant, calculating physical and genetic distances for all genes within 1 cM of the tagging polymorphism. Results revealed 55 SNPs where recombination was lower between the identified SNP and a physically more remote gene than initially reported, and 374 where an alternative gene was genetically and physically closer than the reported gene. Analyses show potential for genomic confounding through false inferences of variant association to a gene. An online visualization tool (http://gcb.genes.org.uk/) was developed to plot genes by physical and genetic distance relative to a variant, along with LD data.


A functional and morphological study of cells adjacent to ectopic bone implants in rats.
Subcutaneous implantation of bone chips into normal and osteopetrotic (ia) rats results in the formation of multinucleate giant cells (MNGCs) adjacent to the bone surface. In this study the resorptive and morphological characteristics of the cells surrounding these implants were assessed to determine if the bone-resorbing defects seen in ia animals would be mimicked, thus giving validity to the use of this system as a model for the study of osteoclastic lineage and function. Direct measurement of in vivo bone resorption was achieved through the use of (45)Ca-labelled bone-chip pairs that were primarily osteoid-exposed and freeze-thawed (FT), primarily mineral-exposed and bleached (B), or primarily mineral-exposed and collagenase-treated (CT). Comparison of the (45)Ca content of implanted chips to that of controls indicated the total (45)Ca release during a two-week implantation period. There was no significant difference in the amount of label released between normal and ia animals. Both normal and ia rats showed 23% greater total (45)Ca release from mineral- versus osteoid-exposed matrix. Cellular events occurring on the bony substrate were evaluated by light and electron microscopy. At 3 days, bone chips were surrounded primarily by mononuclear cells. By 14 days, MNGCs were present at the bone surface in both ia and normal animals. In mineral-exposed implants, 40-50% of the bone surface was covered by MNGCs as compared to 20% of the osteoid-exposed surface. These MNGCs possessed occasional clear zones, but did not exhibit ruffled borders; therefore, they could not be classified as osteoclasts. Thus, the defects seen in ia mutants were not reproduced in this implant system. The (45)Ca release that occurred was probably due to the action of mononuclear phagocytes and macrophage polykaryons rather than to true osteoclastic bone resorption.


Click-chemistry-derived triazole ligands of arginine-glycine-aspartate (RGD) integrins with a broad capacity to inhibit adhesion of melanoma cells and both in vitro and in vivo angiogenesis.
A click chemistry approach was applied for the discovery of triazole-based arginine-glycine-aspartate (RGD) mimetics by Cu(I)-catalyzed 1,3-dipolar alkyne-azide coupling reaction, which showed binding affinity properties toward α(v)β(3)/α(v)β(5) integrins. Biological assays showed compound 18 capable of binding α(v)β(3) integrin with nanomolar affinity according to a two-sites model, and molecular modeling studies revealed a peculiar π-stacking interaction between the triazole ring and Tyr178 side chain. Accordingly, compound 18 inhibited the adhesion of integrin-expressing human melanoma cells to RGD-containing proteins of the extracellular matrix, such as vitronectin, fibronectin, and osteopontin, and also angiogenesis in in vitro and in vivo experimental models. The relevant biological effects exerted by compound 18 suggest its potential application as an antiangiogenic agent in the diagnosis and therapy of tumors where α(v)β(3) integrin expression is up-regulated.


Spermicide effects questioned by FDA panel.
Spermicides clearly work better than no contraceptive at all, but there's no way to say how effective they are or whether women should opt for a gel, foam, suppository, or film, a scientific panel concluded. The advisors to the US Food and Drug Administration (FDA) heard disturbing evidence that women who rely on spermicides as their sole contraceptive have anywhere from a 9% to a 57% chance of becoming pregnant within a year. Spermicides appear to work best when used just 30 minutes before intercourse, said Dr. James Trussell of Princeton University. Nobody knows for sure how well spermicides work, and which work best, because the only studies ever performed were flawed. The manufacturers never proved how well their products work because they hit the market in 1950, before the FDA required such proof. The US National Institutes of Health (NIH) plans to begin a study comparing one brand of each type of commercially available spermicide to see how effective they are. The results will not be available for at least 4 years, so the FDA is debating what to tell consumers in the meantime and if that study will be enough. The FDA may soon change spermicide labels at least to rank the products as less effective than other contraceptives, Dr. Lisa Rarick, FDA reproductive health chief, said. Spermicides also should bear a warning that they may cause vaginal irritation, a condition one study suggests might increase a woman's chances of catching HIV, the scientific panel told the FDA. The same panel agreed, however, that spermicides can help protect women against two other sexually transmitted diseases, gonorrhea and chlamydia.


Kinetic study of the addition of trihalides to unsaturated compounds in ionic liquids. Evidence of a remarkable solvent effect in the reaction of ICl2-.
The kinetic constants and activation parameters for the reactions of Br(3)(-) and ICl(2)(-) with some alkenes and alkynes have been determined in the ionic liquids [bmim][PF(6)], [emim][Tf(2)N], [bmim][Tf(2)N], [hmim][TF(2)N], [bm(2)im][Tf(2)N], and [bpy][TF(2)N] (where emim = 1-ethyl-3-methylimidazolium, bmim = 1-butyl-3-methylimidazolium, hmim = 1-hexyl-3-methylimidazolium, bm(2)im = 1-butyl-2,3-dimethylimidazolium, bpy = butylpyridinium, PF(6) = hexafluorophosphate, and Tf(2)N = bis(trifluoromethylsulfonyl)imide) and in 1,2-dichloroethane. The rates of both reactions increase on going from 1,2-dichloroethane to ILs. Evidence suggests that, while the hydrogen bonding ability of the imidazolium cation is probably the main factor able to increase the rate of the addition of ICl(2)(-) to double and triple bonds, this property has no effect on the electrophilic addition of Br(3)(-) to alkenes and alkynes. Furthermore, in the case of the ICl(2)(-) reaction, the hydrogen bonding ability of ILs can be exploited to suppress the unwanted nucleophilic substitution reaction on the products by the Cl(-) anion.


Metamagnetic quantum criticality in metals.
We present a renormalization group treatment of metamagnetic quantum criticality in metals. We show that for clean systems the universality class is that of the overdamped, conserving (dynamical exponent z = 3) Ising type. We obtain detailed results for the field and temperature dependence of physical quantities including the differential susceptibility, resistivity, and specific heat. Our results are shown to be in quantitative agreement with data on Sr3Ru2O7 except very near to the critical point itself.


Oral manifestations of granulomatosis with polyangiitis - Clinical and radiological assessment.
Granulomatosis with polyangiitis (GPA) is a type of primary systemic vasculitis associated with the presence of anti-neutrophil cytoplasmic antibodies (ANCA). GPA mainly affects medium and small vessels and may manifest in different organs, most commonly upper respiratory tract. Oral lesions occur in 6-13% of GPA patients and might be the first symptom observed by the patient. This study presents the evaluation of orofacial manifestations of patients with GPA. Prospective study was conducted between June 2014 and November 2017 in Department of Allergy and Immunology at University Hospital in Krakow. Patients diagnosed with GPA, after medical assessment, were examined including precise dental inspection, bacterial and fungal flora of oral cavity evaluation and Cone Beam Computed Tomography (CBCT) imaging. Nine patients were enrolled in the study. Characteristic for GPA strawberry gingivitis was observed in one patient. Bone destruction and inflammatory lesions in paranasal sinuses was confirmed by CBCT in 55.6% of patients. Fungal infection was revealed in 66.7% of patients. These findings oblige dentists to consult patient with laryngologist or internal medicine physician to establish further diagnostic approach, because early diagnosis of GPA is crucial for implementing appropriate treatment and preventing chronic organ damage.


A low energy electron microscopy study of the initial growth, structure, and thermal stability of 4,4'-biphenyldicarboxylic acid domains on Cu(001).
The growth of 4,4(')-biphenyldicarboxylic acid (BDA) on Cu(001) has been studied using low energy electron microscopy and selective area low energy electron diffraction. The emergence of large islands and hydrogen bonding to perpendicularly oriented, adjacent molecules is confirmed. The two benzene rings of adsorbed BDA are twisted along the molecular axis. Unconventional growth of the domains, followed by a second nucleation stage, is observed at room temperature. This unanticipated feature is attributed to the accumulation of stress in the islands. Ostwald ripening in the films and the decay of BDA domains at 448 K exhibits features that are consistent with diffusion limited behavior.


Yolk thiobarbituric acid reactive substances and n-3 fatty acids in response to whole and ground flaxseed.
n-3 fatty acid (FA)-enriched shell eggs have been proposed as an economical source for increasing consumption of n-3 FA. As dietary n-3 FA sources vary with respect to fatty acid profile and lipid stability, the comparison of resulting egg products is warranted. This study was designed to determine yolk thiobarbituric acid reactive substances (TBARS) and n-3 FA when hens were fed whole or ground flaxseed. Gold flaxseed (whole or ground) fed at levels of 5 or 15% were compared to a 1.5% menhaden oil or a typical control layer ration. A 5-wk feeding trial was used with 21 hens per treatment. Eggs were collected and FA composition determined during Weeks 4 and 5 of the feeding trial. The TBARS content of yolk was also determined. All flaxseed treatments increased total n-3 FA (C18:3 + C20:5 + C22:6) marine n-3 FA (C20:5 + C22:6) did not increase proportionately with increasing dietary flaxseed. Seed form did not influence n-3 FA deposition at the 5% level of flaxseed; however, ground 15% flaxseed resulted in greater total n-3 FA deposition than whole 15% flaxseed. Yolk TBARS were not different due to diet, indicating that seed form did not influence the oxidative quality of these n-3 FA-rich lipids.


Inhibition of metmyoglobin/H2O2-dependent low density lipoprotein lipid peroxidation by naturally occurring phenolic acids.
The ferrylmyoglobin <==> metmyoglobin redox transitions promoted by hydrogen peroxide and dietary phenolic acids and their potential role in the oxidation of LDL were studied. The use of parinaric acid incorporated in LDL as a probe for radicals (detected by fluorescence quenching of the probe) revealed an oxidative stress inside LDL shortly ( < 1 min) after addition of hydrogen peroxide to metmyoglobin in the aqueous phase outside the particle, reflecting an efficient access of the oxidant to LDL lipids. However, the propagation step of peroxidation only occurs after a lag phase, as detected by the kinetics of oxygen consumption. Triton X-100 decreases but does not suppress the lag phase of oxidation. Addition of metmyoglobin (without peroxide) to LDL was not followed by significant oxidation during the time of the experiment, unless Triton X-100 was present in the medium. When dietary phenolic acids were present in the medium before peroxide addition, an inhibition of parinaric acid fluorescence quenching and oxygen consumption was recorded as a function of concentration and substitution pattern on the phenol ring of the phenolic acids. This was associated with a conversion of ferrylmyoglobin to metmyoglobin. The results indicate that the naturally occurring phenolic acids prevent ferrylmyoglobin-dependent LDL oxidation in a way strongly dependent on the substitution pattern on the phenol ring. Among the phenolic compounds studied, the o-dihydroxy derivatives of cinnamic and benzoic acids (caffeic, chlorogenic, and protocatechuic acids), in a molar ratio of 1 to metmyoglobin, efficiently blocked LDL oxidation initiated by ferrylmyoglobin. Replacement of one OH group from catecholic structure with an H (p-coumaric acid) or methoxy group (ferulic acid) decreased the antioxidant activity. Also, the catechol structure fused in heterocyclic rings with adjacent carbonyl groups (ellagic acid) resulted in decreased antioxidant activity. These observations correlate with the efficiency of phenolic acids to reduce ferrylmyoglobin to metmyoglobin. Therefore, the protection of LDL against oxidation is assigned to the reduction of the oxoferryl moiety of the hemoprotein to the ferric form. Additionally, it is suggested that an access constraint of oxidants plays a minor role in the ferrylmyoglobin-induced oxidation against LDL.


Randomized clinical trial of ganciclovir vs acyclovir for prevention of cytomegalovirus antigenemia after allogeneic transplantation.
Cytomegalovirus (CMV) disease remains a major cause of morbidity following allogeneic stem cell transplantation (SCT). In a prospective randomized trial, we tested prophylactic therapy with ganciclovir or acyclovir for patients at high risk of disease. Ninety-one CMV seropositive recipients of related (n = 53) and unrelated (n = 38) donor transplants were enrolled. All patients received intravenous (i.v.) ganciclovir 5 mg/kg every 12 h days -7 to -2, followed by acyclovir 10 mg/kg i.v. every 8 h from day -1 until neutrophil engraftment. Patients were then randomly assigned to either ganciclovir (n = 45) or acyclovir (n = 46) until day 100 post transplant. Any degree of antigenemia was treated with ganciclovir 5 mg/kg i.v. twice a day for 2 weeks, followed by 5 mg/kg i.v. each weekday for 6 weeks. At day 100, the cumulative incidence of antigenemia was 31% (95% CI 17-45%) for ganciclovir and 41% (95% CI 26-56%) (P = 0.22) for acyclovir prophylaxis, respectively. The assigned prophylaxis cohort did not predict for CMV antigenemia. The cumulative incidence of CMV disease at 12 months was 13% (95% CI 3-23%) and 17% (95% CI 6-28%) (P = 0.59) for the ganciclovir- and acyclovir-treated groups, respectively. An absolute neutrophil count (ANC) <or=1500 x 10(6)/l at randomization (P < 0.01) and grade II-IV acute graft-versus-host-disease (P = 0.01), but not the assigned prophylaxis cohort (P = 0.62), were independent risk factors for CMV disease. The incidence of fungal infections and renal insufficiency was similar across treatment groups; however, bacterial infections and secondary neutropenia occurred more frequently in the ganciclovir group. With our study powered to detect a 60% reduction in antigenemia with ganciclovir prophylaxis, we did not find a statistically significant difference between ganciclovir and acyclovir when used as part of an overall strategy for prevention of CMV antigenemia and disease in SCT, although fewer side-effects occurred with acyclovir treatment.


Ethical issues in registry research: in-hospital resuscitation as a case study.
RESEARCH BASED ON REGISTRY STUDIES involves significant ethical issues. Using detailed information about one registry concerning in-hospital resuscitation, we present issues concerning informed consent, access to identifiable medical information, and benefit for participants. In addition, multiple methodological difficulties have indirect implications for the ethical conduct of registry research, including consensus about variable definitions, validity and reliability for clinical decisions, sample sizes, and sources of data. Both direct and indirect ethical issues are examined from the viewpoint of accepted regulations and codes regarding ethical conduct of research; specific examples of more or less ethical solutions to the problems are presented from published research.


CO2 Complexes with Five-Membered Heterocycles: Structure, Topology, and Spectroscopic Characterization.
In a first step toward the rational design of macrocyclic structures optimized for CO2 capture, we systematically explored the potential of 30 five-membered aromatic heterocycles to establish coordinating complexes with this pollutant. The interactions between the two moieties were studied in several orientations, and the obtained complexes were analyzed in terms of electron density and vibrational fingerprint. The former is an aid to provide an in-depth knowledge of the interaction, whereas the latter should help to select structural motifs that have not only good complexation properties but also diagnostic spectroscopic signals.


Subarachnoid hemorrhage: evaluation with MR imaging.
Thirty-seven magnetic resonance (MR) examinations were performed at 0.5 T in 33 patients with subarachnoid hemorrhage (SAH) caused by a ruptured aneurysm. Images were obtained 2 hours to 75 days after the ictus. Twenty-four proton-density-weighted (long repetition time [TR], short echo time [TE]) images were obtained in the acute stage (< 72 hours after the ictus) of SAH; SAH was hyperintense to brain parenchyma and cerebrospinal fluid in all cases. The detectability of acute SAH on T1- (short TR, short TE) and T2- (long TR, long TE) weighted images was 36% and 50%, respectively. In the subacute and chronic stages (> 3 days after the ictus), the detectability of SAH on T1-, T2-, and proton-density-weighted images was 73%, 31%, and 83%, respectively. Although computed tomography is still the modality of choice for evaluating acute SAH, the authors emphasize that even acute SAH can be reliably demonstrated with MR imaging with the appropriate parameters.


Development of potent non-carbohydrate imidazole-based small molecule selectin inhibitors with antiinflammatory activity.
A novel series of non-carbohydrate imidazole-based selectin inhibitors has been discovered via high-throughput screening using a P-selectin ELISA-based assay system. The initial lead 1 had an IC(50) of 17 microM in the P-selectin ELISA; this potency was significantly improved via an extensive SAR exploration. One of the current lead compounds (29) has an IC(50) of 300 nM in a P-selectin ELISA; it also has good activity in P- and E-selectin cell adhesion assays and shows efficacy in vivo. These compounds represent a novel series of sLe(X) mimetics with antiinflammatory activity. Their unique profile supports our interest in their further evaluation as drug candidates for the treatment of inflammation. Herein we describe the syntheses, optimization, and SAR of this series of novel potent selectin antagonists.


Oedema in the lower limb of patients with chronic critical limb ischaemia (CLI).
approximately 70% of patients with chronic critical limb ischaemia (CLI) show clinical signs of oedema in the distal leg and foot. The primary aim of the present investigation was to quantify this oedema. In addition we investigated whether oedema formation could be due to deep venous thrombosis (DVT). fifteen patients with unilateral CLI and oedema were studied, four males and 11 females, with a mean age of 77+/-10.3 years. Water displacement volumetry (WDV) was used to measure limb volume. Colour duplex ultrasound (CDU) and venous occlusion plethysmography (VOP) were applied to exclude functionally significant DVT. Blood chemistry was analysed to screen for some causative factors of generalised oedema formation. the mean volume of the limbs with CLI was 9% greater than the contralateral limbs (1279+/-325 ml vs. 1179+/-298 ml). None of the patients had functionally significant DVT. The mean plasma albumin concentration was reduced at 28.5+/-6.6 g/l. a significantly reduced plasma albumin concentration cannot be regarded as a causative factor, since the oedema is unilateral. The aetiology of oedema formation is probably multifactorial, and further investigations are under progress to elucidate relevant pathogenetic factors.


Detection and identification of Listeria monocytogenes in cooked sausage products and in milk by in vitro amplification of haemolysin gene fragments.
Recent outbreaks of listeriosis have emphasized the urgent need for rapid and reliable detection methods for Listeria spp., especially in food. Haemolysin production is a major factor in the pathogenesis of listeriosis and the polymerase chain reaction (PCR) was used to amplify two specific DNA fragments of the alpha- and the beta-haemolysin genes. The amplification system specifically recognized L. monocytogenes strains. The detection limit determined with pure cultures was 10 bacteria when estimated with alpha-haemolysin primers. In the analysis of 50 samples of cooked sausage products, bacterial colonies suspected to be Listeria spp. were isolated by conventional methods from six samples. PCR analysis identified three of six as L. monocytogenes. Subsequent serotyping showed perfect agreement with the PCR results. Since enrichment is the most time consuming step in conventional methods a PCR procedure which allows the direct detection of L. monocytogenes in milk was developed. Pasteurized milk was artificially contaminated with various levels of L. monocytogenes. The detection limit was determined to be 10 bacteria/10 ml milk and direct detection and identification of L. monocytogenes took less than two working days. These results show that this haemolysin gene amplification system is very rapid and reliable and therefore avoids cumbersome and lengthy cultivation steps.


Synergy of pandemics-social isolation is associated with worsened Parkinson severity and quality of life.
Social isolation and its deleterious effects on health increases with age in the general population. People with Parkinson's Disease (PWP) are no exception. Social isolation is a risk factor for worsened health outcomes and increased mortality. Symptoms such as depression and sleep dysfunction are adversely affected by loneliness. There is a paucity of research on social isolation in Parkinson's disease (PD), which is all the more critical now in the setting of social distancing due to COVID-19. The goal of this study was to survey individuals with PD to evaluate whether social isolation is associated with PD symptom severity and quality of life. Only individuals reporting a diagnosis of idiopathic PD were included in this analysis. The primary outcome measures were the Patient-Reported Outcomes in PD (PRO-PD) and questions from PROMIS Global related to social health. PRO-PD scores increased as social performance and social satisfaction scores diminished. Individuals who reported being lonely experienced a 55% greater symptom severity than those who were not lonely (P < 0.01). Individuals who documented having a lot of friends had 21% fewer symptoms than those with few or no friends (P < 0.01). Social isolation was associated with greater patient-reported PD severity and lower quality of life, although it is unclear whether this is the cause and/or a consequence of the disease. In essence, the Parkinson pandemic and the pandemic of social isolation have been further compounded by the recent COVID-19 pandemic. The results emphasize the need to keep PWP socially connected and prevent loneliness in this time of social distancing. Proactive use of virtual modalities for support groups and social prescribing should be explored.


Role of probiotics in health and disease: a review.
For some decades now, bacteria known as probiotics have been added to various foods because of their beneficial effects for human health. The mechanism of action of probiotics is related to their ability to compete with pathogenic microorganisms for adhesion sites, to antagonize these pathogens or to modulate the host's immune response. The potential application of probiotics includes prevention and treatment of various health conditions and diseases such as gastrointestinal infections, inflammatory bowel disease, lactose intolerance, allergies, urogenital infections, cystic fibrosis, various cancers, reduction of antibiotic side effects, in oral health such as prevention of dental caries, periodontal diseases and oral malodour and many other effects which are under investigation. The results of many of these clinical investigations suggests that probiotics may be useful in preventing and treating various health conditions and diseases. However, many of these clinical studies require validation so as to apply these results to clinical realm. The role of clinical trials is instrumental in such investigations and in near future the results of such trials will decide the usefulness of probiotics in health and disease. This article strives to summarize the currently available data on the potential benefits of probiotics in health and disease.


Origin of glutamate dehydrogenase from the mucosa of the sheep rumen.
Glutamate dehydrogenase [GDH] from the mucosa of sheep rumen was partly purified and characterized. In chromatography on DEAE cellulose, GDH activity separated into two fractions. Fraction I, isolated at a low NaCl gradient concentration, was not affected by Zn2+ or by the purine nucleotides GTP and AMP. The chromatographic behaviour of fraction II was the same as the parallel fraction isolated from the liver by the same technique. The activity of fraction II and the liver fraction was strongly inhibited by Zn2+ in 10(-6) to 3.10(-5) mol/1 concentration and by GTP in 10(-5) to 2.10(-5) mol/1 concentration. It was activated by AMP in 5.10(-6) to 6.10(-5) mol/1 concentration and by leucine in 3.10(-3) to 10(-2) mol/1 concentration. The coenzyme specificity of fraction I was greater for NADPH than for NADH. In the case of fraction II, like the fraction isolated from sleep liver, it was greater for NADH than for NADPH. It is concluded from the different effect of Zn2+ and of purine nucleotides on the enzyme activity of the fractions isolated from rumen mucosa that the mucosa of the sheep rumen contains two enzymes with GDH activity, one of which [probably adsorbed] is of bacterial origin and the other is a constitutive tissue enzyme of the rumen wall.


Characterization of the Drosophila melanogaster ribosomal proteome.
We have combined high-resolution two-dimensional (2-D) gel electrophoresis with mass spectrometry to identifying proteins represented in a 2-D gel database of Drosophila melanogaster ribosomes. First, we purified ribosomes from third instar Drosophila larvae and constructed a high-resolution 2-D gel database containing 58 Coomassie blue stained polypeptides. Next, we carried out preparative 2-D PAGE to isolate some of the polypeptides and characterize them by MALDI-TOF. Using this strategy we identified 52 ribosomal spots in the database, and in each case confirmed their identity by MALDI-TOF/TOF. The database can be used to analyze Minute mutants of Drosophila.


Structural basis of Vps33A recruitment to the human HOPS complex by Vps16.
The multisubunit homotypic fusion and vacuole protein sorting (HOPS) membrane-tethering complex is required for late endosome-lysosome and autophagosome-lysosome fusion in mammals. We have determined the crystal structure of the human HOPS subunit Vps33A, confirming its identity as a Sec1/Munc18 family member. We show that HOPS subunit Vps16 recruits Vps33A to the human HOPS complex and that residues 642-736 are necessary and sufficient for this interaction, and we present the crystal structure of Vps33A in complex with Vps16(642-736). Mutations at the binding interface disrupt the Vps33A-Vps16 interaction both in vitro and in cells, preventing recruitment of Vps33A to the HOPS complex. The Vps33A-Vps16 complex provides a structural framework for studying the association between Sec1/Munc18 proteins and tethering complexes.


Involvement of the adherens junction-actin filament system in acantholytic dyskeratosis of Hailey-Hailey disease. A histological, ultrastructural, and histochemical study of lesional and non-lesional skin.
Hailey-Hailey disease is a blistering genodermatosis that shows acantholytic dyskeratosis throughout the epidermis. The aim of our study was to investigate the involvement of adherens structures and cytofilaments in this particular type of acantholysis. Both lesional and non-lesional skin from 18 patients was studied histologically and ultrastructurally. Additionally, the samples were stained for desmosomes, adherens junctions, keratin filaments, actin filaments, and actin-associated proteins, and finally investigated with an electron and a confocal laser scanning microscope (CLSM), respectively. Acantholytic dyskeratosis was not only confined to lesions, but was also focally detectable in clinically unaffected skin. Despite disruption and internalization of the desmosomes, keratinocytes remained linked together by well-preserved adherens junctions. Staining for actin filaments with fluorochrome-labeled phalloidin showed a remarkable formation of actin stress fibers in these keratinocytes. Thus, incomplete acantholysis, as demonstrable in both lesional and non-lesional skin of Hailey-Hailey patients, may be due to a cohesive function of the adherens junction-actin system succeeding the dissolution of desmosomes. Most remarkably, none of the adnexal epithelia expressed the intrinsic defect of cell adhesion. This finding offers an explanation for the successful treatment of Hailey-Hailey disease by dermabrasion, which after complete removal of the involved epidermis results in reepithelialization from skin appendages.


High tibial valgus osteotomy in unicompartmental medial osteoarthritis of the knee: a retrospective follow-up study over 13-21 years.
High tibial osteotomy is a well-established method for the treatment of medial unicompartmental osteoarthritis of the knee. We analysed retrospectively the long-term outcome of open and closing wedge valgisation high tibial osteotomies. Out of 71 patients, 54 (76%) were available for the study. Survival rates and the influence of the osteotomy type were investigated. Secondary outcome measures were the course of radiological leg axis and osteoarthritis as well as score outcomes. During a median follow-up of 16.5 years (IQR 14.5-17.9; range 13-21), 13 patients (24%) underwent conversion to total knee arthroplasty; the other 41 patients (76%, survivor group) were studied by score follow-up as well as clinical and radiological examinations. Osteotomy survival was of 98% after 5 years, 92% after 10 years and 71% after 15 years. Comparison between open and closing wedge high tibial osteotomy showed no significant difference in survival and score outcome. The median Visual Analogue Score (VAS) was 0 (IQR 0-1; range 0-4), the Satisfaction Index was 80% (IQR 63-89; range 30-100), the median Knee Injury and Osteoarthritis Outcome Score was 71 (IQR 49-82; range 9-100) and the median Western Ontario and McMaster Universities Osteoarthritis index was 84 (IQR 66-96; range 9-100). Radiological evaluation showed only a slight progression of the degree of osteoarthritis following the Kellgren and Lawrence classification. In each case, the axis passed through the healthy compartment or at least through the centre of the knee. Open and closing wedge high tibial osteotomies are a successful choice of treatment for unicompartmental degenerative diseases with associated varus in active patients. Survival of both techniques is comparable in our series and is associated with low pain scores, high satisfaction and high activity levels of the survivors.


Brucella-Induced Ruptured Infrarenal Dissecting Abdominal Aortic Aneurysm.
Mycotic aneurysms, often saccular, accounting for approximately 2.5% of all abdominal aortic aneurysms, possess increased risk of rupture, uncontrolled sepsis, and protracted hospital stay and are associated with high morbidity and mortality. The authors report the case of a 49-year-old female with no known comorbidities who presented with free rupture of an infrarenal dissecting mycotic aneurysm and underwent emergent open repair successfully. The etiological agent, Brucella melitensis, a Gram-negative zoonotic coccobacillus, is rarely reported to cause mycotic aneurysm.


Health-Related Quality of Life after Hip and Knee Arthroplasty Operations.
The aim of this study was to investigate the health-related quality of life before and after a hip and a knee arthroplasty operation using a 15D instrument and to compare these scores to the Finnish control population 15D scores. The pre- and post-operative data of 15D were prospective collected from the patients undergoing total hip arthroplasty or total knee arthroplasty at the Kuopio University Hospital. Post-operative data were collected at 6 and 12 months after the operation. The mean change of the 15D score after hip arthroplasty was +0.062 and after knee arthroplasty, it was +0.033 at the 12-month follow-up (p < 0.001). Total hip arthroplasty patients of all ages reached the control population 15D scores at the 12-month follow-up. Of the total knee arthroplasty patients, only patients aged >75 years and males aged 55-64 years did reach control population 15D scores at the 12-month follow-up. Patients experienced a statistically significant improvement in mobility, vision, sleeping, usual activities, discomfort and symptoms, distress, and vitality (p < 0.05). Successful hip and knee arthroplasty operations improve patients' health-related quality of life. According to this study, hip arthroplasty improves the health-related quality of life more than knee arthroplasty.


Exemptions and waivers from cost sharing: ineffective safety nets in decentralized districts in Uganda.
The introduction of user-payment for health services is frequently followed by concern about the impact on equity of access for poor people. Decentralizing governments often try to remedy the created inequities by putting in place safety nets in the form of exemptions and waivers in the user-fee systems. However, where user payments merely operate as local government strategies for health financing, without national policy they are likely to be self-defeating, as local governments are frequently more interested in raising revenue to meet recurrent costs of devolved services than in promoting equity. Thus guidelines put in place by the central government to operationalize safety nets are seen by local governments as being contradictory to this goal, and are thus ignored or altered to suit the district revenue aims. This study was carried out to investigate the context and the constraints in implementing exemption schemes. Data were collected in two selected administrative districts of Uganda (Mbarara and Mukono). Qualitative approaches to data collection were adopted, namely focus group discussions and key informant interviews with policy-makers, health administrators, service providers and community members. These methods were combined with document review. We found little evidence of safety-net guidelines initiated by decentralized/local governments, since district local governments had little motivation to extend exemptions, waivers or credits. The conclusion is that safety nets such as waivers and exemptions will only be effective if they are backed by a national health financing policy, they reconcile the often competing demands of local government revenue needs, and are strictly enforced and supervised by both the local and central governments. The implications of the findings for remedying the tension between the needs for cost recovery and for attainment of equity goals through exemption policies for the poor and indigent are discussed.


Screening for alcohol abuse among urban Native Americans in a primary care setting.
The authors examined the rate of and factors associated with alcohol abuse among 754 urban American Indian and Alaska Native primary care patients. Data were collected through a self-administered survey and by abstracting medical records. A total of 423 respondents (56 percent) screened positive for lifetime alcohol abuse, and 202 (27 percent) reported current alcohol abuse. A diagnosis of alcohol abuse was found in the medical records of 68 of the patients who screened positive (16 percent). Logistic regression analysis showed that being male, having been a victim of violence, and feeling depressed were associated with alcohol abuse. Further research is warranted to help clinicians detect and treat alcohol abuse in this special population.


HIV prevalence in the Israeli tuberculosis cohort, 1999-2011.
Tuberculosis (TB) patients who are co-infected with HIV are at greater risk of mortality. Nevertheless, not all countries achieved sustainable and TB and HIV collaboration to describe the burden of both diseases at a national scale. This study aims to describe HIV prevalence among TB-patients in Israel and identify variable associated with TB/HIV co-infection. This retrospective study was conducted by cross-matching the National HIV and TB Registries to describe TB/HIV epidemiology during the last 13-years and define variables predicting TB/HIV co-infection. Between 1999 and 2011, 5,502 TB-patients were reported: 779 (14.2%) were Israeli-born and 4,723 (85.8%) non-Israeli born. Of all TB patients, 254 (4.6%) were HIV-infected. The trend of HIV/TB co-infection among non-Israeli born has generally decreased since 2003 (trend analysis p < 0.001).TB/HIV co-infected patients were mostly males, their TB diagnosis had been performed relatively in shorter time following their arrival in Israel, more likely to be in the 35-44 and 25-34 age groups, non-Israeli born (mostly Africa born), more likely to be culture positive, have multi-drug resistant strains, had worse treatment outcomes and more likely to die treatment than HIV-negative tuberculosis patient. In a multivariate analysis, short time after arrival in Israel, older age, being born in Ethiopia, having positive sputum, positive culture and multi-drug resistant TB predicted TB/HIV co-infection.TB/HIV co-infected patients with extra-pulmonary TB had a higher proportion of infection in lymphatic, miliary and abdominal sites than those with extra-pulmonary TB who were HIV-negative. Most TB/HIV co-infected patients were migrants originating in high-burden countries. Despite the moderate 4.6% TB/HIV co-infection rate in Israel, these patients had worse treatment outcomes and higher mortality rates. This study illustrates the importance of integrating TB with HIV in surveillance and treatment components, which should be employed in other countries, as it has a positive impact on disease control.


Deterministic growth of AgTCNQ and CuTCNQ nanowires on large-area reduced graphene oxide films for flexible optoelectronics.
We describe a synchronous reduction and assembly procedure to directly produce large-area reduced graphene oxide (rGO) films sandwiched by a high density of metal nanoparticles (silver and copper). Further, by using the sandwiched metal NPs as sources, networks consisting of AgTCNQ and CuTCNQ nanowires were deterministically grown from the rGO films, forming structurally and functionally integrated rGO/metal-TCNQ hybrid films with outstanding flexibility, bending endurance, and electrical stability. Interestingly, due to the p-type nature of the rGO film and the n-type nature of the metal-TCNQ NWs, the hybrid films are essentially thin-film p-n junctions which are useful in ubiquitous electronics and optoelectronics. Measurements of the optoelectronic properties demonstrate that the rGO/metal-TCNQ hybrid films exhibit substantial photoconductivity and highly reproducible photoswitching behaviours. The present approach may open the door to the versatile and deterministic integration of functional nanostructures into flexible conducting substrates and provide an important step towards producing low-cost and high-performance soft electronic and optoelectronic devices.


Vaginal squamous cells in follicular aspirates following transvaginal puncture.
Squamous cells are common findings on cytological examination of fluid obtained following transvaginal aspiration of simple ovarian cysts. This prospective study confirms the hypothesis that these cells are commonly introduced as contaminants during puncture of the vaginal wall. The occasional finding of bacteria also suggests a possible mechanism for post-operative pelvic infection.


Chip-based CE for rapid separation of 8-aminopyrene-1,3,6-trisulfonic acid (APTS) derivatized glycans.
Fluorescently labeled carbohydrates released from glycoproteins were separated using a commercially available microfluidic chip electrophoresis system. While the instrumentation was primarily designed for DNA analysis it was found that the application base can be easily expanded using the development software provided by the manufacturer. The carbohydrates were released by enzymatic digestion (PNGase F) from glycoproteins present in human plasma after boronic acid - lectin affinity enrichment. After fluorescent labeling with 8-aminopyrene-1,3,6-trisulfonic acid the carbohydrates were separated based on capillary gel electrophoresis mechanism and detected by a fluorescence detector using a blue (470 nm) LED. The separation was completed in 40 s in a microfluidic channel of 14 mm length. Glucose ladder carbohydrate oligomers differing by one glucose unit were baseline separated up to a 20-mer with the main limitation being the detection sensitivity. As expected, the observed resolution in these experiments did not approach that of standard CE with 20 times longer separation distance; however, the chip-based analysis excelled in the speed of the separation. Similar electrophoretic profiles of glycans released from plasma glycoproteins were obtained using a standard CE equipment with 35 cm separation length and microfluidic chips with a separation distance of only 14 mm.


Knowledge, attitudes, and behavior related to pregnancy in a rural teenage population.
Despite recent declining national trends, teenage pregnancy continues to be a community health problem warranting attention, especially in rural areas of the southern United States. Communities are unique and require assessment of risk populations to develop appropriate initiatives. This pilot study examined consent strategies and survey procedures and tools. Additionally, descriptive statistics related to pregnancy and pregnancy prevention knowledge, attitudes, and behaviors were examined. The study sample consisted of 52 boys and girls who perceived themselves to be in the middle or upper levels of their 9th-grade class and who were predominantly college bound and fairly active in extracurricular activities. Often groups with similar demographics are not perceived as at risk for pregnancy and early sexual intercourse, but the findings suggest otherwise. Many girls were sexually active, several had already experienced pregnancy, and some were at risk for intentional pregnancy. Knowledge regarding pregnancy prevention was modest. School-based and peer-focused programs are promising intervention strategies in response to identified attitudes and reported information sources related to pregnancy and pregnancy prevention.


Transcriptome-module phenotype association study implicates extracellular vesicles biogenesis in Plasmodium falciparum artemisinin resistance.
Plasmodium falciparum malaria is still an important disease in sub-Saharan Africa (sSA). Great strides have been made in its control spear-headed by artemisinin (ART)-based combination therapies (ACTs). However, concerns about the imminent spread of ART-resistant (ARTr) malaria parasites to sSA threaten gains already made. Attempts to mitigate this risk have highlighted the need to discover novel P. falciparum drug targets. Therefore, studies to deepen our understanding of the biology of P. falciparum are needed. The role of extracellular vesicles (EVs) in the biology of malaria parasites is not fully understood. Recently, the ART resistance-associated transcriptional profile has been reported to involve several biological processes connected to vesicular trafficking, proteotoxic stress, erythrocyte remodelling, and mitochondrial metabolism. We explored a role for EVs in developing the P. falciparum ARTr phenotype using bulk RNA sequencing of unsynchronized parasite cultures under untreated, 0.1% dimethyl sulfoxide and 700nM dihydroartemisinin treated conditions for six hours. As pathway and gene ontology analysis is limited in its curated knowledge repertoire on EVs biogenesis in P. falciparum, we used a modular (gene set) analysis approach to explore whether an EVs biogenesis module is associated with the ARTr phenotype in P. falciparum. We first generated well-defined EVs modules of interest and used statistical tools to determine differences in their expression among the parasite and treatment conditions. Then we used gene set enrichment analysis to determine the strength of the association between each EVs module of interest and the ARTr phenotype. This transcriptome-module phenotype association study (TMPAS) represents a well-powered approach to making meaningful discoveries out of bulk gene expression data. We identified four EVs module of interest and report that one module representing gene sets with correlated expression to PF3D7_1441800 - involved with EVs biogenesis in P. falciparum - is associated with the ARTr phenotype (R539T_DHA_treated versus R539T_untreated: normalized enrichment score (NES) = 1.1830174, FDR q-value < 0.25; C580R_DHA_treated versus C580R_untreated: NES = 1.2457103, FDR q-value < 0.25). PF3D7_1441800 has been reported to reduce EVs production when knocked out in P. falciparum. Altogether, our findings suggest a role for EVs in developing ART resistance and warrant further studies interrogating this association.


Phosphorylcholine esterase is critical for Dolichos biflorus and Helix pomatia agglutinin binding to pneumococcal teichoic acid.
Streptococcus pneumoniae (the pneumococcus) has wall teichoic acid (WTA) and lipoteichoic acid (LTA) expressing the Forssman antigen (FA). Two lectins, Dolichos biflorus agglutinin (DBA) and Helix pomatia agglutinin (HPA), are known to bind FA. To determine the molecular structure targeted by these two lectins, different pneumococcal strains were studied for DBA/HPA binding with flow cytometry and fluorescence microscopy. Genetic experiments were used to further examine the lectins' molecular target. Twelve strains were positive for DBA binding, whereas three were negative. Super-resolution microscopy showed that DBA stained only the subcapsular area of pneumococci. The three DBA nonbinders showed no phosphorylcholine esterase (Pce) activity in vitro, whereas 10 DBA binders displayed Pce activity (the remaining two strains were DBA binders with no Pce activity in vitro). The pcegene sequence for 10 representative strains revealed two functional pce alleles, the previously recognized "allele A" and a newly discovered "allele B" (with 12 additional nucleotides). Isolates with allele B showed no Pce activity in vitro but did bind to DBA, indicating allele B Pce is functional in vivo. Genetic transfer experiments confirmed that either allele is sufficient (and necessary) for DBA binding. The three DBA nonbinders had various mutations that affected Pce function. Observations with HPA were identical to those with DBA. We show that DBA and HPA bind only to the WTA/LTA of pneumococcal isolates with a functional Pce enzyme. A newly discovered Pce variant (allele B) is functional in vivo but nonfunctional when assayed in vitro.


Natural environmental cues and circadian rhythms of behaviour--a perspective.
Natural environmental cycles are often extremely difficult to reproduce under laboratory conditions. Laboratory light-dark cycles differ from natural light-dark cycles in terms of intensity and spectral distribution, whilst simulated temperature cycles may differ from natural temperature cycles in waveform. The expression of a free-running rhythm depends upon the 'level' of constant conditions provided. Environmental cues affect the period, phasing, amplitude and activity-rest ratios of circadian rhythms and, if inappropriate, may result in aberrant behaviour patterns which are unlike those observed in nature.


Interaction of Benzylaminopurine with Electron Transport in Plant Mitochondria during Malate Oxidation.
The effect of 6-benzylaminopurine (BA) was assayed on malate oxidation in mitochondria isolated from fresh and aged potato (Solanum tuberosum L.) slices. Depending on the experimental pH, two pathways for malate oxidation were selected. A pH of 7.7 favored the activity of malate dehydrogenase, which is connected with a rotenone-sensitive NADH dehydrogenase, whereas at pH 6.5 malic enzyme, linked to a rotenone-resistant NADH dehydrogenase, was more active.Experimental results indicate the existence of two sites of inhibition for BA. The first site is common with the site of inhibition of rotenone. The second site is on the classical cyanide-resistant alternative pathway, but is different from the site of salicylhydroxamic acid (SHAM) inhibition, as in succinate oxidation.Moreover, a distinct cyanide-resistant pathway, sensitive to SHAM but resistant to BA, is found to coexist with the well-known alternative pathway which is sensitive to SHAM and BA. This outlet of electrons can accommodate 10% of the total electron flow in mitochondria from fresh slices, and up to 30% in mitochondria from aged slices.


Gramine Accumulation in Leaves of Barley Grown under High-Temperature Stress.
The indole alkaloid gramine is toxic to animals and may play a defensive role in plants. Under certain conditions, shoots of barley cultivars such as ;Arimar' and CI 12020 accumulate gramine (N,N-dimethyl-3-aminomethylindole) and lesser amounts of its precursors 3-aminomethylindole (AMI) and N-methyl-3-aminomethylindole (MAMI); other cultivars such as ;Proctor' do not. When grown at optimal temperatures (21 degrees C/16 degrees C, day/night), Arimar contained a high level of gramine in the first leaf (approximately 6 milligrams per gram dry weight), but progressively less accumulated in successive leaves so that the gramine level in the shoot as a whole fell sharply with age. In Arimar and CI 12020 plants transferred at the two- to three-leaf stage from 21 degrees C/16 degrees C to supra-optimal temperatures (>/=30 degrees C/25 degrees C), there was massive gramine accumulation in leaves which developed at high temperature, so that gramine level in the whole shoot remained high (about 3-8 milligrams per gram dry weight).Proctor lacked both constitutive gramine accumulation in the first leaf and heat-induced gramine accumulation in later leaves. The following evidence indicates that this results from a lesion in the pathway of synthesis (tryptophan -->--> AMI --> MAMI --> gramine) between tryptophan and AMI. (a) Proctor and Arimar leaves readily absorbed [(14)C]gramine, but neither cultivar degraded it extensively. (b) Arimar leaf tissue incorporated [(14)C]formate label into the N-methyl groups of gramine and MAMI, and converted [methylene-(14)C]tryptophan to AMI, MAMI, and gramine; Proctor leaf tissue did not, even when a trapping pool of unlabeled gramine was supplied. (c) Proctor converted [(14)C]MAMI to gramine as actively as Arimar. (d) Proctor incorporated [(14)C]formate label into gramine and MAMI when supplied with AMI; the ratio [(14)C]gramine/[(14)C]MAMI fell with leaf age, suggesting that the two N-methylations involve different enzymes. Inasmuch as Proctor leaf tissue did not methylate added tryptamine or tyramine, the N-methyltransferase(s) of gramine synthesis may be substrate specific.In sterile culture at optimal temperatures, 10 millimolar gramine did not affect autotrophic growth of Arimar or Proctor plantlets or heterotrophic growth of callus. At supra-optimal temperature, plantlet growth was reduced by gramine although callus growth was not. We speculate that gramine-accumulating cultivars may suffer autotoxic effects at high leaf temperatures.


An investigation of the possible scenarios for the optimal locating of quality sensors in the water distribution networks with uncertain contamination.
One of the ways to reduce the risk of contaminated water consumption is to optimally locate the quality sensors. These sensors warn users in the case of contamination detection. Analyzing the actual conditions of the contamination which enters the network is faced with many uncertainties. These uncertainties include the dose of contamination, time and location of its entry which have received less attention. Also, the uncertainty in the nodes' water demand causes changes in the distribution and contamination diffusion within the network. The main impetus of the present study is to determine the optimal quality sensor locations in the water distribution network in order to reduce the damage caused by contaminated water consumption prior to the contamination event detection. For this purpose, a parameter is defined as the maximum possible damage for calculating which the vulnerability and importance of the nodes have been considered in addition to the uncertainties in the location and time of the contamination entry. The importance of each node differs from that of other ones. Ranking the importance of the nodes is influenced by both land use and covered population ratio. In this study, six scenarios are defined for the contamination event in the water distribution network. These scenarios consider the effects of varying pollutant dose and the contamination input from nodes which are prone to its entry. Also, the NSGA-II has been utilized in order to minimize the damage with minimum number of sensors. The proposed model is evaluated on a real network in Iran. The results indicate that adding only one or two contamination warning sensors to the proposed locations can lead to the decreasing damage caused by the contaminated water consumption from 54 to 82%. According to the proposed method, the best answer for scenarios 1-6 was obtained for 7, 6, 6, 2, 2 and 2 sensors, respectively. The results showed that the slope of the pollution rate diagram does not change much from 6 sensors upwards in the first three scenarios, and from 4 sensors upwards in the second three scenarios. In scenarios 1-3, with 7, 6 and 6 sensors, respectively, in different nodes, the best placement is for 203-224 equivalent attack population, and in scenarios 4-6, with sensors in nodes 4 and 43, the best placement is for 225-279 equivalent attack population.


[Effect of rolling needle therapy on quality of life in the patient of non-organic chronic insomnia: a randomized controlled trial].
To probe therapeutic effect of rolling needle therapy on insomnia and standardize the therapeutic program. Multi-central randomized blind controlled trials were used, and 180 cases were randomly divided into a rolling needle group and a medication group, 90 cases in each group. The rolling needle group were treated with rolling needle therapy at the Urinary Bladder Channel line 1 and 2 at the back and the Governor Vessel, and the medication group with clonopin 4-6 mg, for 4 weeks. Effective rates for sleep disturbance improvement and Spitzer index of life quality were evaluated after 4 weeks' treatment and 3 months' follow-up. The effective rate was 82.2% in the rolling needle group and 60.0% in the medication group with a significant difference between the two groups (P < 0.05), and with a significant difference between the two groups in Spitzer index of life quality (P < 0.05). After 3 months' follow-up, the effective rate was 40.0% in the rolling needle group and 30.3% in the medication group with no significant difference between the two groups (P > 0.05), and there was a significant difference between the two groups in Spitzer index of life quality. Rolling needle therapy can treat chronic insomnia and increase life quality of the patient.


Plant gametogenesis: conservation and contrasts in development.
Although the process of gamete formation in plants has many unique features, much has been learnt from the comparative analysis between plants and other eukaryotic systems. Plants have a number of factors that have made them desirable for the analysis of gamete development; these include late germline specification, the non-lethality of mutations affecting gamete development and the large size of their chromosomes. The availability of the fully annotated Arabidopsis genome and comparative analysis using yeast, animal and E. coli has led to the identification and functional characterisation of many genes with roles in gamete development, principally those associated with meiosis, recombination and DNA repair. The advantages that plants give with the use of mutant screens to identify genes associated with gamete formation have also provided access to genes that are difficult to characterise by alternative routes. This has yielded novel information regarding the processes of gamete formation in higher plants. The times may now be changing with the advantages that plants provide serving to advance knowledge of gamete formation in other eukaryotic systems.


Accelerating proliferation of neural stem/progenitor cells in collagen sponges immobilized with engineered basic fibroblast growth factor for nervous system tissue engineering.
Neural stem/progenitor cells (NS/PCs) play a therapeutic role in nervous system diseases and contribute to functional recovery. However, their efficacy is limited as the majority of cells die post-transplantation. In this study, collagen sponges were utilized as carriers for NS/PCs. Basic fibroblast growth factor (bFGF), a mitogen for NS/PCs, was incorporated into the collagen sponges to stimulate NS/PC proliferation. However, the effect of native bFGF is limited because it diffuses into the culture medium and is lost following medium exchange. To overcome this problem, a collagen-binding polypeptide domain, which has high affinity to collagen, was fused with bFGF to sustain the exposure of NS/PCs within the collagen sponges to bFGF. The results indicated that the number of NS/PCs was significantly higher in collagen sponges incorporating engineered bFGF than in those with native bFGF or the PBS control after 7 days in culture. Here, we designed a natural biological neural scaffold consisting of collagen sponges, engineered bFGF, and NS/PCs. In addition to the effect of proliferated NS/PCs, the engineered bFGF retained in the natural biological neural scaffolds could have a direct effect on nervous system reconstruction. The two aspects of the natural biological neural scaffolds may produce synergistic effects, and so they represent a promising candidate for nervous system repair.


Meiotic cell cycle in Xenopus oocytes is independent of cdk2 kinase.
In vertebrates, M phase-promoting factor (MPF), a universal G2/M regulator in eukaryotic cells, drives meiotic maturation of oocytes, while cytostatic factor (CSF) arrests mature oocytes at metaphase II until fertilization. Cdk2 kinase, a G1/S regulator in higher eukaryotic cells, is activated during meiotic maturation of Xenopus oocytes and, like Mos (an essential component of CSF), is proposed to be involved in metaphase II arrest in mature oocytes. In addition, cdk2 kinase has been shown recently to be essential for MPF activation in Xenopus embryonic mitosis. Here we report injection of Xenopus oocytes with the cdk2 kinase inhibitor p21Cip in order to (re)evaluate the role of cdk2 kinase in oocyte meiosis. Immature oocytes injected with p21Cip can enter both meiosis I and meiosis II normally, as evidenced by the typical fluctuations in MPF activity. Moreover, mature oocytes injected with p21Cip are retained normally in metaphase II for a prolonged period, whereas those injected with neutralizing anti-Mos antibody are released readily from metaphase II arrest. These results argue strongly against a role for cdk2 kinase in MPF activation and its proposed role in metaphase II arrest, in Xenopus oocyte meiosis. We discuss the possibility that cdk2 kinase stored in oocytes may function, as a maternal protein, solely for early embryonic cell cycles.


Natural Product Based Antibody Drug Conjugates: Clinical Status as of November 9, 2020.
As of early November 2020, there are 10 approved antibody drug conjugates (ADCs) plus two others that are not usually listed. In addition, there are 70 ADCs at stages from phase I to phase III and 23 that are at the preclinical stage. The warheads of all of these drugs and drug candidates have their origins in natural product structures. The sources and modifications are discussed in general and then specifically commented on in each case with either the generic name if known and/or the ADC's ID names. Interestingly, almost all warheads listed are from microbial sources though initially a number were thought to have been from plants. The latest NCT numbers from Clintrials.gov of all phase I to phase III candidates are also given. Three unusual ADCs are also discussed, two of which (an antitumor agent and one directed against autoimmune diseases) are not usually listed as ADCs, with the third being an anti-infective.


Family patterns of decision-making in pediatric clinical trials.
The decision-making process related to a child's participation in clinical trials often involves multiple family members. The aim of this study was to compare family patterns of decision-making within and across family units in pediatric clinical trials. Participants for this secondary analysis included 14 families from a larger study of informed consent. Four distinct patterns of decision-making were identified: Exclusionary, informative, collaborative, and delegated. These patterns varied with regard to three dimensions of parents' decision-making goals, child level of involvement, and the parental role. These patterns of decision-making affect how parents and children communicate with health professionals and influence the effectiveness of health care providers interactions with the family related to the decision-making process.


Coccidioidomycosis causing massive cervical lymphadenopathy.
Coccidioidomycosis involving the head and neck is uncommon. We present a case of a patient with disseminated coccidioidomycosis and massive submandibular cystic lymphadenopathy unresponsive to systemic antifungal treatment. It is our impression that significant clinical improvement resulted from concomitant needle aspiration of the cystic nodes to decrease fungal load and modification of systemic medication. Although reduction of fungal burden may provide an improved response to antifungal therapy, disseminated coccidioidomycosis is a systemic disease that requires appropriate systemic therapy.


Thermal blooming of pulsed focused gaussian laser beams.
Thermal blooming of a pulsed, focused Gaussian laser beam in a gas is studied for a specific model of gaseous heating. Analytic results in the limit of times both long and short compared with the sound transit time across any part of the beam are presented as irradiance profiles at any range as a function of time. Experiments that can be done with present laser technology to test the theoretical predictions are suggested.


PROLink-Single Step Circularization and Purification Procedure for the Generation of an Improved Variant of Human Growth Hormone.
Human growth hormone (hGH) plays an important role during human development and is also an approved therapeutic for the treatment of several diseases. However, one major drawback of hGH is its short circulating half-life requiring frequent administration, which is inconvenient and painful for the patients. Recent publications indicate that circularization greatly increases the stability of proteins due to their protection from exoproteolytic attack and a higher thermal stability of the circular form. Using sortase A, a transpeptidase isolated from Staphylococcus aureus, we developed a single step solid-phase circularization and purification procedure resulting in a circular version of hGH with improved properties. We could show that circular hGH binds to the recombinant hGH receptor with binding kinetics similar to those of linear hGH and that circularization does not alter the biological activity of hGH in vitro. Besides, circular hGH showed almost complete resistance toward exoproteolytic attack and slightly increased thermal stability which could possibly translate into an extended plasma half-life. The single step solid-phase circularization and purification procedure is in principle a generic process, which could also be applied for other proteins that meet the requirements for circularization.


Diffusion-weighted MRI of denervated muscle: a clinical and experimental study.
The aim of this study was to investigate skeletal muscle denervation using diffusion-weighted magnetic resonance imaging (DWMRI). Sciatic nerve axotomy was performed in a group of nine New Zealand White rabbits, and electromyographic (EMG), pathological, and DWMRI studies were conducted on ipsilateral hamstring muscles 1 and 8 days after axotomy. In addition, DWMRI studies were carried out on leg muscles of ten patients with acute and subacute lumbosacral radiculopathy. High intensity signals on short tau inversion recovery (STIR) magnetic resonance imaging and an increased apparent diffusion coefficient (ADC) were observed in denervated muscles of the animals 1 and 8 days after axotomy as well as in denervated muscles of the patients with radiculopathy. In the clinical study, ADC was 1.26 +/- 0.18 x 10(-9) m(2)/s in normal muscle and increased to 1.56 +/- 0.23 x 10(-9) m(2)/s in denervated muscles (p = 0.0016). In animals, EMG and muscle pathological studies were normal 1 day after axotomy, and the muscles demonstrated spontaneous activity on EMG and neurogenic atrophy on histological studies 7 days later. This DWMRI study demonstrates that enlargement of extracellular fluid space in muscle denervation is an early phenomenon occurring several days before the appearance of EMG and histological abnormalities.


Clinical performance of the Reichert AT550: a new non-contact tonometer.
The aim of the present study was to assess the level of accuracy for measurements of intra-ocular pressure (IOP) obtained with a new non-contact tonometer (NCT) the Reichert AT550. Measurements were compared against those obtained with the Reichert Xpert Plus, Goldmann applanation tonometer and Perkins tonometer. Thirty-five university students were assessed with the four tonometers in a randomised order, with non-contact tonometry performed first. Each of the four measurement devices had its own trained clinical observer. Plots of differences of IOP as a function of the mean for each pair of instruments were obtained. No statistically significant differences were found when comparing the AT550 NCT with contact applanation tonometry (AT) (p> 0.05), displaying the closest level of agreement (as represented by the lowest mean difference and the narrowest confidence interval) with the Goldmann tonometer (limits of agreement, 0.12+/-2.17). In conclusion, readings of IOP with the AT550 NCT are clinically comparable with those obtained with Goldmann tonometry in a population with IOP within the normal range.


Tuberculosis in healthcare workers, Scotland.
In an attempt to explore healthcare worker acquisition of tuberculosis infection, we conducted population-based surveillance of all cases recorded as healthcare workers reported to Enhanced Surveillance of Mycobacterial Infection from 2000 to 2015. Over the study period, the mean incidence rate of tuberculosis among all healthcare workers was 15.4 per 100,000 healthcare workers. However, the incidence rate of tuberculosis amongst those healthcare workers born outside the UK was 164.8 per 100,000 compared with 5.0 per 100,000 UK-born healthcare workers. Fifty-seven per cent of all non-UK-born healthcare workers were diagnosed within five years of their arrival in the UK and would have been new entrants to the NHS. An effective new entrant occupational health screening programme for latent tuberculosis infection may have prevented some of these active cases of infection.


Circulating CD4+CD25high regulatory T cells and expression of PD-1 and BTLA on CD4+ T cells in patients with chronic hepatitis B virus infection.
The roles of regulatory T cells (Tregs) and PD-1 in hepatitis B have not been clearly described. Also, the role of B and T lymphocyte attenuator (BTLA), which serves as a negative regulator of T-cell activation, is still unknown in hepatitis B. In this study, we analyzed the frequency of circulating CD4(+)CD25(high) Tregs in patients with chronic hepatitis B (CHB), and subsequently investigated expression of PD-1 and BTLA on CD4(+) T cells, as well as their relationships with the clinical index of CHB patients. A total of 39 CHB patients and 19 healthy persons as controls were enrolled in the study. We found that the frequency of CD4(+)CD25(high) Tregs and PD-1 expression on CD4(+) T cells was significantly increased in CHB patients compared with normal controls. However, BTLA expression on CD4(+) T cells showed no significant difference between the two groups. The frequency of Tregs was significantly higher in patients with HBV DNA titers >or=10(8) than in those with HBV DNA titers <10(8). Circulating CD4(+)CD25(high) Treg frequency and PD-1 expression on CD4(+) T cells correlated positively with serum HBV DNA load in CHB patients. Our findings suggest that the increased frequency of CD4(+)CD25(high) Tregs and PD-1 expression on CD4(+) T lymphocytes may inhibit the cellular immune response against HBV and affect viral clearance, leading to the persistence of chronic HBV infection.


[Diagnosis and principles of complex therapy of sarcoidosis].
The diagnosis and therapy of 87 patients with sarcoidosis admitted to the Central Military Clinical Tubercular Hospital were analyzed. The diagnosis was based on the data of clinicoroentgenologic investigation and histologic examination of the specimen taken from the affected tissue. The differential diagnosis with tuberculosis is obligatory. The complex therapy includes corticosteroids, antioxydants, phytotherapy, physiotherapeutic methods and in some cases plasmapheresis.


H-NMR characterization of the human myocardium myoglobin and erythrocyte hemoglobin signals.
The 1H-NMR signal of deoxy Mb provides a unique opportunity to measure tissue oxygenation in vivo. To utilize the technique for human application, however, requires a specific spectral characterization of both human Mb and erythrocyte Hb. We report that the proximal histidyl-NH signal of human deoxy Mb resonates at 80.3 ppm at 25 degrees C and maintains a 3.9 ppm separation with the corresponding Hb A signal throughout the physiological temperature range. In the particular case of the human thenar muscle, the deoxy Mb signal is clearly detectable without any interference from Hb.


Gramene, a tool for grass genomics.
Gramene (http://www.gramene.org) is a comparative genome mapping database for grasses and a community resource for rice (Oryza sativa). It combines a semi-automatically generated database of cereal genomic and expressed sequence tag sequences, genetic maps, map relations, and publications, with a curated database of rice mutants (genes and alleles), molecular markers, and proteins. Gramene curators read and extract detailed information from published sources, summarize that information in a structured format, and establish links to related objects both inside and outside the database, providing seamless connections between independent sources of information. Genetic, physical, and sequence-based maps of rice serve as the fundamental organizing units and provide a common denominator for moving across species and genera within the grass family. Comparative maps of rice, maize (Zea mays), sorghum (Sorghum bicolor), barley (Hordeum vulgare), wheat (Triticum aestivum), and oat (Avena sativa) are anchored by a set of curated correspondences. In addition to sequence-based mappings found in comparative maps and rice genome displays, Gramene makes extensive use of controlled vocabularies to describe specific biological attributes in ways that permit users to query those domains and make comparisons across taxonomic groups. Proteins are annotated for functional significance using gene ontology terms that have been adopted by numerous model species databases. Genetic variants including phenotypes are annotated using plant ontology terms common to all plants and trait ontology terms that are specific to rice. In this paper, we present a brief overview of the search tools available to the plant research community in Gramene.


Pharmacodynamics of Finafloxacin, Ciprofloxacin, and Levofloxacin in Serum and Urine against TEM- and SHV-Type Extended-Spectrum-β-Lactamase-Producing Enterobacteriaceae Isolates from Patients with Urinary Tract Infections.
The pharmacodynamics of finafloxacin, ciprofloxacin, and levofloxacin against extended-spectrum-β-lactamase (ESBL)-producing Enterobacteriaceae isolates were compared. Since quinolones lose activity in acidic media, and particularly in urine, their activities were tested in parallel under conventional conditions and in acidic artificial urine. For this purpose, TEM- and SHV-type ESBL-producing Escherichia coli and Klebsiella pneumoniae strains and their wild-type counterparts were exposed in a modified Grasso model to simulated concentrations of drugs in serum and urine following oral doses of either finafloxacin at 800 mg once a day (q.d.), immediate-release ciprofloxacin at 500 mg twice a day (b.i.d.), extended-release ciprofloxacin at 1,000 mg q.d., or levofloxacin at 500 or 750 mg q.d. The concentrations of the drugs in urine were fitted by compartmental modeling. Bacteria were cultivated in Mueller-Hinton broth (MHB) at pH 7.2 or 5.8 or in artificial urine at pH 5.8. Bacteria were counted every 2 h until 10 h and at 24 h; the areas under the bacterial-count-versus-time curves were calculated. It was found that finafloxacin eliminated all strains within 2 h under all the conditions studied. At all doses studied, ciprofloxacin and levofloxacin were highly active against wild-type strains in MHB at pH 7.2 but lost activity in MHB, and particularly in urine, at pH 5.8. Viable counts of ESBL producers were reduced for 6 to 8 h by 3 log10 titers, but the bacteria regrew thereafter. Ciprofloxacin and levofloxacin were almost inactive against the SHV producer grown in artificial urine. We conclude that pharmacodynamic models using artificial urine may mirror the physiology of urinary tract infections more closely than those using conventional media. In contrast to ciprofloxacin and levofloxacin, finafloxacin gained activity in this model at an acidic pH, maintained activity in artificial urine, and was active against TEM and SHV producers.


Measurement of phosphocreatine in cutaneous tissue by high pressure liquid chromatography.
A high pressure liquid chromatography technique for measuring phosphocreatine and adenine nucleotides in human cutaneous tissue is described. The presence of phosphocreatine in human skin was confirmed by this analytic method. Molar concentration of phosphocreatine and adenine nucleotides were determined in normal skin and in benign and malignant skin lesions. The preliminary results suggest that absolute amounts of phosphocreatine and adenine nucleotides and phosphocreatine/adenosine triphosphate ratios in malignant skin neoplasms and benign cutaneous lesions are different from those measured in normal nonischemic human skin.


Amiodarone and mortality among elderly patients with acute myocardial infarction with atrial fibrillation.
Amiodarone has been shown to be safe in patients with acute myocardial infarction (AMI) who are at risk for sudden cardiac death. However, there is limited data concerning the safety of amiodarone in patients who experience AMI complicated by atrial fibrillation. To determine the safety of amiodarone therapy, we conducted a retrospective analysis of elderly patients hospitalized with AMI who experienced atrial fibrillation and had survived to hospital discharge (n = 17,597). Amiodarone prescribed at discharge was evaluated for its association with short-term and long-term mortality in crude and adjusted analyses employing propensity score methods. Of the 17,597 patients, 550 patients (3.1%) were prescribed amiodarone, 2317 patients (13.2%) were prescribed other antiarrhythmic agents (excluded from analysis), and 14,730 (83.7%) were prescribed no antiarrhythmic medication at discharge. Thirty-day mortality rates were similar for patients prescribed amiodarone and those not prescribed amiodarone (6.8% amiodarone vs 5.4% no amiodarone, P =.21), but mortality at 1 year was higher among patients prescribed amiodarone (35.6% vs 31.6%, P =.001). However, amiodarone was not associated with mortality at 30 days (odds ratio 0.80, 95% CI 0.53-1.20) or at long-term follow-up (mean duration 612 days, hazard ratio 1.04, 95% CI 0.92-1.18) after multivariable modeling. Amiodarone was not independently associated with short-term or long-term mortality in elderly patients discharged after a hospitalization for AMI complicated by atrial fibrillation. Although our data suggest that amiodarone may be safe to use in this population, randomized controlled trial data are needed to confirm this finding.


Systematic review and meta-analysis of radiotherapy in various head and neck cancers: comparing photons, carbon-ions and protons.
To synthesize and compare available evidence considering the effectiveness of carbon-ion, proton and photon radiotherapy for head and neck cancer. A systematic review and meta-analyses were performed to retrieve evidence on tumor control, survival and late treatment toxicity for carbon-ion, proton and the best available photon radiotherapy. In total 86 observational studies (74 photon, 5 carbon-ion and 7 proton) and eight comparative in-silico studies were included. For mucosal malignant melanomas, 5-year survival was significantly higher after carbon-ion therapy compared to conventional photon therapy (44% versus 25%; P-value 0.007). Also, 5-year local control after proton therapy was significantly higher for paranasal and sinonasal cancer compared to intensity modulated photon therapy (88% versus 66%; P-value 0.035). No other statistically significant differences were observed. Although poorly reported, toxicity tended to be less frequent in carbon-ion and proton studies compared to photons. In-silico studies showed a lower dose to the organs at risk, independently of the tumor site. For carbon-ion therapy, the increased survival in mucosal malignant melanomas might suggest an advantage in treating relatively radio-resistant tumors. Except for paranasal and sinonasal cancer, survival and tumor control for proton therapy were generally similar to the best available photon radiotherapy. In agreement with included in-silico studies, limited available clinical data indicates that toxicity tends to be lower for proton compared to photon radiotherapy. Since the overall quantity and quality of data regarding carbon-ion and proton therapy is poor, we recommend the construction of an international particle therapy register to facilitate definitive comparisons.


MMPI profiles of impaired nurses.
The MMPI profiles of 42 nurses presenting themselves for treatment of alcoholism or opiate addiction in a residential setting were examined. When comparisons were made, no significant differences in this sample emerged based on sex, level of professional training, or drug of choice. Results suggested that a significant elevation emerged on the Pd scale, and the D scale approached clinical significance. The clinical implications of these data as they impact the treatment of impaired nurses are discussed.


Episogram: Visual Summarization of Egocentric Social Interactions.
The key challenges of visualizing social interaction data include the difficulties of understanding the general structure of social interactions and representing the data in the context of various user activities to reveal different behavior patterns. The design of the proposed interactive visualization tool Episogram is based on an anatomy of social interaction process in which the actors and objects involved can be formally represented as a time-varying tripartite network. The authors show the effectiveness of the proposed technique using real-world datasets and user studies.


Effects of aging on audio-visual speech integration.
This study investigated the impact of aging on audio-visual speech integration. A syllable identification task was presented in auditory-only, visual-only, and audio-visual congruent and incongruent conditions. Visual cues were either degraded or unmodified. Stimuli were embedded in stationary noise alternating with modulated noise. Fifteen young adults and 15 older adults participated in this study. Results showed that older adults had preserved lipreading abilities when the visual input was clear but not when it was degraded. The impact of aging on audio-visual integration also depended on the quality of the visual cues. In the visual clear condition, the audio-visual gain was similar in both groups and analyses in the framework of the fuzzy-logical model of perception confirmed that older adults did not differ from younger adults in their audio-visual integration abilities. In the visual reduction condition, the audio-visual gain was reduced in the older group, but only when the noise was stationary, suggesting that older participants could compensate for the loss of lipreading abilities by using the auditory information available in the valleys of the noise. The fuzzy-logical model of perception confirmed the significant impact of aging on audio-visual integration by showing an increased weight of audition in the older group.


Sharp boundary formation and invasion between spatially adjacent periodical cicada broods.
Periodical cicadas, Magicicada spp., are a useful model system for understanding the population processes that influence range boundaries. Unlike most insects, these species typically exist at very high densities (occasionally >1000/ m2) and have unusually long life-spans (13 or 17 years). They spend most of their lives underground feeding on plant roots. After the underground period, adults emerge from the ground to mate and oviposit over a period of just a few days. Collections of populations that are developmentally synchronized across large areas are known as "broods". There are usually sharp boundaries between spatially adjacent broods and regions of brood overlap are generally small. The exact mechanism behind this developmental synchronization and the sharp boundary between broods remain unknown: previous studies have focused on the impacts of predator-driven Allee-effects, competition among nymphs, and their impacts on the persistence of off-synchronized emergence events. Here, we present a nonlinear Leslie-type matrix model to additionally consider cicada movement between spatially separated broods, and examine its role in maintaining brood boundaries and within-brood developmental synchrony that is seen in nature. We successfully identify ranges of competition and dispersal that lead to stable coexistence of broods that differ between spatial patches.


Population Analyses Reveal Preenrichment Method and Selective Enrichment Media Affect Salmonella Serovars Detected on Broiler Carcasses.
Poultry is a major Salmonella reservoir, but conventional culture-based methods typically identify the most abundant serovars while those less abundant remain undetected. Choice of enrichment procedure also introduces bias, and for broiler carcasses, a 1-min rinse before preenrichment is insufficient to release all Salmonella present. The inability to assess serovar diversity means that serovars more often associated with human illness may be masked by more abundant Salmonella. CRISPR-SeroSeq (serotyping by sequencing clustered regularly interspaced short palindromic repeats), an amplicon-based, next-generation sequencing tool, allows detection of multiple serovars and maps the relative serovar frequencies in a sample. To address the preceding limitations, CRISPR-SeroSeq was used on broiler carcasses collected prechilled at a commercial plant. Standard carcass rinse aliquot preenrichments and whole carcass preenrichments that were enriched in Rappaport-Vassiliadis (RV) and tetrathionate (TT) broths were compared. On average, five serovars were observed per carcass, including nine on one carcass. CRISPR-SeroSeq detected serovars comprising as little as 0.005% of the population. CRISPR-SeroSeq data matched (28 of 32) standard culture analysis for abundant serovars. Salmonella serovars Kentucky, Typhimurium, and Schwarzengrund were found on each carcass. Overall, serovar diversity was higher in whole carcass preenrichments that were enriched in RV (P < 0.05). Serovar Schwarzengrund was present at higher frequencies in whole carcass preenrichments compared with rinse aliquot preenrichments (t test, P < 0.05), suggesting it adheres more strongly to the carcass. Salmonella serovar Enteritidis was enriched eightfold more in TT than in RV, and serovars Schwarzengrund and Reading were preferentially enriched in RV. Comparison of preenriched and enriched samples suggests that selective enrichment in RV or TT was inhibitory to some serovars. This article addresses limitations of Salmonella surveillance protocols and provides information related to Salmonella population dynamics.


Bone Scintigraphy: A Review of Technical Aspects and Applications in Orthopedic Surgery.
Due to its high sensitivity, low cost, accessibility, and ease of use, bone scintigraphy is used in orthopedic surgery for the diagnosis and management of varied pathology. It is commonly used for insufficiency fractures, metastatic neoplasia, staging and surveillance of sarcoma, and nonaccidental trauma. It augments diagnoses, including stress or occult fractures, musculoskeletal neoplasia or infection, and chronic regional pain syndrome, in patients presenting with normal results on radiographs. Bone scan images are resistant to metal-based implant artifact, allowing effective evaluation of failed total joint prostheses. Bone scintigraphy remains an underused tool in the evaluation and management of orthopedic patients. [Orthopedics. 2019; 42(1):e14-e24.].


How to WHO: lessons from aviation in checklists and debriefs.
The World Health Organization (WHO) surgical safety checklist (SSC) has had an overall positive impact; however, it has not completely prevented adverse events and compliance with the checklist varies. The aviation industry is considered to have better engagement with their safety checklists, reporting not only safety improvements, but also a cultural shift in their checklist philosophy over recent years. We explored the personal attitudes of pilots working in the aviation industry to identify principles of an effective checklist philosophy that could be transposed to the healthcare setting to empower more effective, consistent and ultimately successful implementation of the WHO SSC. A questionnaire was developed by the authors. Three airline pilots were interviewed via telephone, and asked questions regarding the logistics of and attitudes to checklists in the aviation industry. Several key factors for successful checklist implementation were identified. These include regular training and education on human factors and the checklist's purpose, and institution of an atmosphere that is receptive, engaged and welcoming. Much can be learned from the aviation industry, where not only has the incidence of adverse events decreased, but the attitudes of people working in the industry have also transformed. The WHO SSC is an invaluable tool used in healthcare settings worldwide. However, it is not a standalone commodity. To be effective, it necessitates steadfast engagement from the team members involved in its implementation. Human and checklist must work in partnership, using each other's strengths and fallibilities, to optimise outcomes and prevent risks to patient safety.


Interleukin-17 promotes proliferation, migration, and invasion of trophoblasts via regulating PPAR-γ/RXR-α/Wnt signaling.
To investigate the effect of Interleukin 17 (IL-17) on the invasive capacity of trophoblast cells and the underlying mechanism, we collected placental tissues samples from pregnant women with preeclampsia (PE) and healthy pregnant women. The expression levels of IL-17 mRNA and protein in tissue samples were determined using qRT-PCR and Western blot, respectively. Cell viability and cell proliferation was determined using CCK-8 assay, and colony formation assay, respectively. Cell migration and invasion capacity were determined using transwell cell migration assay. Our results showed that the mRNA expression of IL-17 was significantly increased in PE patients and may be used as a sensitive biomarker for PE (P < 0.01). IL-17 overexpression promoted cell viability, migration, and invasion of human extravillous trophoblast cell line, HTR8/SVneo; however, IL-17 knockdown inhibited these effects. Additionally, IL-17 activated PPAR-γ/RXR-α signaling pathway, which promoted proliferation, migration, and invasion of trophoblast cells. Moreover, PPAR-γ/RXR-α heterodimers activated Wnt signaling. In conclusion, our study provides evidence that IL-17 is overexpressed in PE and promotes proliferation, migration and invasion of trophoblast cells via activating PPAR-γ/RXR-α/Wnt signaling.


Genotyping of two Neisseria gonorrhoeae fluroquinolone-resistant strains in the Brazilian Amazon Region.
We report two ciprofloxacin and ofloxacin-resistant Neisseria gonorrhoeae strains that were isolated from the urethral discharge of male patients at the sexually transmitted diseases outpatient clinic of the Alfredo da Matta Foundation (Manaus, state of Amazonas, Brazil). The gonococci displayed minimal inhibitory concentrations (> 32.00 µg/mL) and three mutations in the quinolone resistance-determining region (S91F and D95G in GyrA and S87R in ParC). Both isolates were genotyped using N. gonorrhoeae multi-antigen sequence typing and the analysis showed that the ST225 which represented an emerging widespread multi-resistant clone that has also been associated with reduced susceptibility to ceftriaxone. We recommend continued surveillance of this pathogen to assess the efficacy of anti-gonococcal antibiotics in Brazil.


Identification and characterization of two CD40-inducible enhancers in the mouse TRAF1 gene locus.
We have shown that CD40 engagement induces TRAF1 gene expression in B lymphocytes. Here we report that CD40-dependent TRAF1 gene transcription in murine B cells is controlled by two enhancer regions. One region is located approximately 2 kb upstream of the transcription start site and the other lies in the intron between exons 5 and 6. The upstream enhancer contains a single NF-kappaB site in addition to sites that bind constitutive transcription factors. Mutation of this NF-kappaB site completely abrogates CD40-driven TRAFl transcription. The intronic enhancer contains two sites that strongly bind the CD40-inducible factors NF-kappaB and AP-1. Simultaneous mutation of the AP-1 site and of the NF-kappaB site abolishes transcription driven by this enhancer. When cloned together into reporter constructs, the two TRAF1 enhancers do not synergize, suggesting that each enhancer may separately participate in the induction of TRAF1 transcription in B cells following CD40 activation.


Prevalence and Associations of Hypertension in Sri Lankan Adults: Estimates from the SLHAS 2018-19 Survey Using JNC7 and ACC/AHA 2017 Guidelines.
Sri Lanka lacks robust estimates of hypertension (HTN) prevalence owing to few national studies, hindering optimization of control strategies. Evidence on how the revised 2017 American College of Cardiology/American Heart Association (ACC/AHA) HTN definition affects prevalence in low- and middle-income countries (LMICs) is also limited. To make robust estimates of HTN prevalence in the Sri Lankan adult population, and to assess impact of the ACC/AHA 2017 definitions. Data were sourced from the 2018-2019 first wave of the Sri Lanka Health and Ageing Study (SLHAS), a nationally representative longitudinal study of the noninstitutionalized adult population. After excluding those with missing data and aged <18 years, 6,342 participants (95.1%) were included in the analysis. HTN was defined using either the traditional threshold of systolic BP (SBP) ≥140 mmHg or a diastolic BP (DBP) ≥90 mmHg, or the ACC/AHA 2017 threshold of SBP ≥130 mmHg or DBP ≥80 mmHg, or if taking antihypertensive medication. Estimated prevalence of HTN in all Sri Lankan adults was 28.2% using the traditional definition, and it doubled to 51.3% when applying the ACC/AHA 2017 definition. Of those classified as hypertensive according to the older and ACC/AHA 2017 definitions, 53.4% and 31.2%, respectively, were previously diagnosed. Of the 23.2% of adults reclassified as hypertensive by the ACC/AHA 2017 definition, 16.6% had a history of CVD or diabetes. Increased prevalence was associated with urban residence, socioeconomic status, obesity, and Muslim ethnicity. Prevalence increased with age, but the increase was steeper in women from their 30s. Nearly one in three adult Sri Lankans are hypertensive, requiring antihypertensive treatment. Applying the ACC/AHA 2017 definitions almost doubles numbers, but many of those reclassified would require treatment under recent WHO guidelines. Study findings also suggest that design effects in HTN surveys may be higher than usually assumed.


Total blood mercury and serum measles antibodies in US children, NHANES 2003-2004.
Environmental toxins, pathogens and host susceptibility cofactors may interact to contribute to disease. In vitro mercury exposure inhibited antiviral cytokines in human cells; however, little is known about the relationship between mercury and viruses in children. Children are susceptible to mercury toxicity; lower vitamin B-12 and folate levels and higher homocysteine levels may represent susceptibility cofactors. This study aimed to evaluate associations between total blood mercury (Hg) and measles antibodies in children, and the influence of these susceptibility cofactors. Cross-sectional data on serum measles antibodies, Hg, homocysteine, methylmalonic acid (MMA, indicator of B-12 deficiency), and folate were obtained from the 2003-2004 NHANES for children aged 6-11 years with measles seropositivity (n=692). We used linear regression to evaluate relationships between measles antibodies and Hg, stratified by sex, MMA ≥, folate <, and homocysteine≥sample medians, adjusted for demographic, nutritional and environmental cofactors. Hg (range: 0.10-19.10μg/L) was inversely associated with measles antibodies (range: 1.00-28.24 units) in non-stratified analysis (n=692), yet positively associated among the subset of boys with higher MMA and lower folate (n=98). Among this subset with higher homocysteine levels (n=61), correlations were positive across all Hg quartiles relative to Q1 (Hg≤0.20μg/L): Q2:β=6.60 (3.02, 10.19); Q3:β=8.49 (6.17, 10.81); Q4 (Hg>0.80μg/L):β=4.90 (2.12, 7.67) (p(trend)=0.077). Stratification by susceptibility cofactors revealed opposing directionality for correlations between Hg and measles antibodies, with positive effect estimates at lowest exposures only among boys with higher MMA, lower folate and higher homocysteine levels.


Analysis of gamma-ray spectra with spectral unmixing, Part II: Recalibration for the quantitative analysis of HPGe measurements.
In the context of radioactivity measurements, the quantitative analysis of a gamma-ray spectrum depends on the analysis algorithm. To that end, we recently introduced a Poisson statistics-based spectral unmixing approach. However, it also relies on a proper instrument recalibration as well as on an uncertainty estimation, for which no solution has been proposed so far. The goal of this article is twofold: i) we first present a novel method to correct for the instrument calibration of an HPGe detection system, which is tailored to spectral unmixing algorithms, and ii) we apply this new approach to the quantitative analysis of real data as well as on the evaluation of the uncertainty. Along with the characteristic limits determination investigated, this paper introduces the first full metrological analysis sequence of aerosol filter measurements based on spectral unmixing, which allows to quantify both the radionuclides' activities and their associated uncertainties.


SCFA Receptors in Pancreatic β Cells: Novel Diabetes Targets?
Nutrient sensing receptors are key metabolic mediators of responses to dietary and endogenously derived nutrients. These receptors are largely G-protein-coupled receptors (GPCRs) and many are gaining significant interest as drug targets with a potential therapeutic role in metabolic diseases. A distinct subclass of nutrient sensing GPCRs, two short chain fatty acid (SCFA) receptors (FFA2 and FFA3) are uniquely responsive to gut microbiota derived nutrients (such as acetate, propionate, and butyrate). Pharmacological, molecular, and genetic studies have investigated their role in organismal glucose metabolism and recently in pancreatic β cell biology. Here, we summarize the present knowledge on the role of these receptors as metabolic sensors in β cell function and physiology, revealing new therapeutic opportunities for type 2 diabetes.