Methodological improvements for the analysis of domain movements in large biomolecular complexes.
Domain movements play a prominent role in the function of many biomolecules such as the ribosome and F0F1-ATP synthase. As more structures of large biomolecules in different functional states become available as experimental techniques for structure determination advance, there is a need to develop methods to understand the conformational changes that occur. DynDom and DynDom3D were developed to analyse two structures of a biomolecule for domain movements. They both used an original method for domain recognition based on clustering of "rotation vectors". Here we introduce significant improvements in both the methodology and implementation of a tool for the analysis of domain movements in large multimeric biomolecules. The main improvement is in the recognition of domains by using all six degrees of freedom required to describe the movement of a rigid body. This is achieved by way of Chasles' theorem in which a rigid-body movement can be described as a screw movement about a unique axis. Thus clustering now includes, in addition to rotation vector data, screw-axis location data and axial climb data. This improves both the sensitivity of domain recognition and performance. A further improvement is the recognition and annotation of interdomain bending regions, something not done for multimeric biomolecules in DynDom3D. This is significant as it is these regions that collectively control the domain movement. The new stand-alone, platform-independent implementation, DynDom6D, can analyse biomolecules comprising protein, DNA and RNA, and employs an alignment method to automatically achieve the required equivalence of atoms in the two structures.


Surface-enhanced fluorescence from silver fractallike nanostructures decorated with silver nanoparticles.
Fluorescence emission of fluorophore molecules in the close vicinity of a nanostructured metal surface can be enhanced through a local electromagnetic field with the help of surface plasmon resonance. The fluorescence enhancement effect is very sensitive to the topography and dielectric property of the metal substrate. In the current work, metal substrates with complex structures, which are made of silver fractallike structures and nanoparticles (NPs), are prepared through electrochemical reduction followed by physical deposition. The surface-enhanced fluorescence of Rhodamine 6G monolayer molecules deposited on the prepared complex substrates are investigated with the laser spectroscopic technique. The experimental results show that the fractallike structure decorated with silver NPs presents stronger fluorescence enhancement, compared with silver NPs or pure silver fractallike structures.


Investigating perceptions of the academic educational environment across six undergraduate health care courses in the United Kingdom.
To compare how health care professional students perceive their academic learning environment in one Higher Education Institution (HEI). The Dundee Ready Education Environment Measure (DREEM), a scale that measures students' perceptions of their academic classroom learning environments, and demographic items were completed by 673 undergraduate students enrolled in health professional courses at the University of Huddersfield, United Kingdom. Respondent scores, partitioned by demographic variables, were obtained on the total DREEM scale and on the individual sub-scales. Analysis of variance techniques were utilized to investigate associations between demographic variables on the total DREEM score and sub-scale scores. All students from all year levels reported total DREEM scores between 101 - 150, indicating a more positive than negative perception of their academic learning environments. Both profession and year of study were found to be significantly associated with total DREEM scores, with occupational therapists, physiotherapists and nurses scoring higher than operating department practice students; and first year students scoring higher than third year students. A significant interaction between these two factors was found to exist, indicating that the impact of the year of study does not apply equally over the different professional groups of students. Students in this study reported positive perceptions about their academic classroom learning environments and teaching delivered. Further interrogation of the DREEM sub-scale dimensions at a programme/cohort level may offer course teams' further information to inform continuous quality improvement.


Manipulation of visual mental images in children and adults.
Recently, there has been a debate on whether visual patterns can be transformed and reinterpreted in mental imagery. In the present study, task demands and age of subjects were manipulated to see whether children and adults were able to discover a novel visual pattern after transforming a mental image. Two tasks, called combination and subtraction, were devised. They consist of either compounding or taking away parts of images to discover a new construal. Results indicate that not only adults, but also children aged 6 and 10, are able to transform a mental image so as to yield another image with a different interpretation. Task demands had a greater effect on children than adults, consistent with the suggestion that the ease of manipulating mental images is a function of the efficiency of control processes.


Adaptation at specific loci. IV. Differential mating success among glycolytic allozyme genotypes of Colias butterflies.
Male mating success as a function of genotype is an important fitness component. It can be studied in wild populations, in species for which a given group of progeny has exactly one father, by determining genotypes of wildcaught mothers and of sufficient numbers of their progeny. Here, we study male mating success as a function of allozyme genotype at two glycolytic loci in Colias butterflies, in which sperm precedence is complete, so that the most recent male to mate fathers all of a female's subsequent progeny.--For the phosphoglucose isomerase, PGI, polymorphism, we predict mating advantage and disadvantage of male genotypes based on evaluation of their biochemical functional differences in the context of thermal-physiological-ecological constraints on the insects' flight activity. As predicted, we find major, significant advantage in mating success for kinetically favored genotypes, compared to the genotype distribution of males active with the sampled females in the wild. These effects are repeatable among samples and on different semispecies' genetic backgrounds.--Initial study of the phosphoglucomutase, PGM, polymorphism in the same samples reveals heterozygote advantage in male-mating success, compared to males active with the females sampled. This contrasts with a lack of correspondence between PGI and PGM genotypes in other fitness index or component differences.--Epistatic interactions in mating success between the two loci are absent.--There is no evidence for segregation distortion associated with the alleles of either primary locus studied, nor is there significant assortative mating.--These results extend our understanding of the specific variation studied and suggest that even loci closely related in function may have distinctive experience of evolutionary forces. Implications of the specificity of the effects seen are briefly discussed.


First Report of "Candidatus Liberibacter solanacearum" on Tomato in Honduras.
Tomato (Lycopersicum esculentum) crops grown in several departments of Honduras and heavily infested with the psyllid Bactericera cockerelli were observed in April of 2012 with plant symptoms suggestive of "Candidatus Liberibacter solanacearum" infection. B. cockerelli is a serious pest of potato, tomato, and other solanaceous plants and a vector of "Ca. L. solanacearum" (1,2,3,4). The symptoms included overall chlorosis, severe stunting, leaf cupping, excessive branching of axillary shoots, and leaf purpling and scorching (2,3). Disease incidence ranged from 5 to 50% symptomatic plants per field. Tomato (cv. Pony) plant samples were collected from two psyllid-infested commercial fields in the municipalities of Danli and Comayagua in the departments of El-Paraiso and Comayagua, respectively. Total DNA was extracted from leaf tissues of 50 and 20 symptomatic and asymptomatic plants, respectively, with the cetyltrimethylammonium bromide (CTAB) buffer extraction method (1,3). The DNA samples were tested for "Ca. L. solanacearum" by PCR with primer pairs specific for 16S rDNA (OA2 and OI2c) and the outer membrane protein gene (OMB 1482f and 2086r) of the bacterium (1,2). Ten (20%) of the 50 symptomatic tomato samples were positive for "Ca. L. solanacearum" using both primer pairs and the remaining samples were negative for the bacterium with both primer sets. None of the 20 asymptomatic plants tested positive for "Ca. L. solanacearum". Amplicons from DNA of two plant samples (one plant/municipality) with each primer pair were cloned and four clones of each of the four amplicons were sequenced. BLASTn analysis of the 16S rDNA consensus sequences from the clones (deposited in GenBank as Accession Nos. KC768321 and KC768322) were identical for both locations and showed 99 to 100% identity to several "Ca. L. solanacearum" sequences in GenBank (e.g., JN848753, JN84856, and HM246509). The OMB consensus sequences from the two tomato plants (deposited in GenBank as KC768329 and KC768330) were 100% identical to OMB sequences of Lso in GenBank (CP002371 and JN48754, respectively). To our knowledge, this is the first report of "Ca. Liberibacter solanacearum" associated with tomato crops in Honduras. This bacterium has caused millions of dollars in losses to the tomato industry in the United States, Mexico, and New Zealand (2,3,4). Serious damages to tomato crops due to "Ca. L. solanacearum" could expand throughout Central America, especially in those countries where B. cockerelli occurs. References: (1) J. M. Crosslin. Southwest. Entomol. 36:125, 2011. (2) L. W. Liefting et al. Plant Dis. 93:208, 2009. (3) J. E. Munyaneza et al. Plant Dis. 93:1076, 2009. (4) J. E. Munyaneza. Am. J. Pot. Res. 89:329, 2012.


Variants in the APOE gene are associated with improved outcome after anti-VEGF treatment for neovascular AMD.
Anti-vascular endothelial growth factor (anti-VEGF) drugs have dramatically improved the treatment of neovascular AMD. In pivotal studies, almost 90% of patients maintain vision, with approximately 30% showing significant improvement. Despite these successes, 10% to 15% of patients continue to lose vision, even with treatment. It has been reported that variants in some AMD-associated genes influence treatment outcome. This study showed an association of treatment outcome with variants in the apolipoprotein E (APOE) gene. One hundred ninety-two patients receiving anti-VEGF treatment for subfoveal choroidal neovascularization secondary to AMD were enrolled. Information on demographics, lesion characteristics, delay until treatment, visual acuity (VA), and number of treatments was collected, and variants of APOE were assessed in all patients at baseline. Best corrected logarithm of the minimum angle of resolution (logMAR) VA was recorded in all patients. The presence of the APOE ε4 allele was associated with improved treatment outcome at 3 (P = 0.02) and 12 (P = 0.06) months, compared with the presence of the ε2 allele, after adjustment for baseline acuity, treatment delay after first symptoms, age, and sex. Patients with an APOE ε4 allele had an odds ratio (OR) of 4.04 (95% confidence interval [CI], 1.11-14.70) for a 2-line gain in vision from baseline at 3 months (P = 0.03) and an OR of 2.54 (95% CI, 0.61-10.52; P = 0.20) at 12 months after treatment, based on multivariate analysis. In patients with neovascular AMD, the presence of the APOE ε4 allele conferred significantly better visual outcomes after anti-VEGF treatment than did the ε2 allele. These findings suggest a possible role for a personalized approach to treatment with anti-VEGF.


A Qualitative Study of African American/Black MSM's Experiences of Participating in a Substance Use and Sexual Risk Reduction Intervention.
The majority of new HIV infections in the United States are among men who have sex with men (MSM), and African American/Black MSM are especially affected. Employing a grounded theory approach, this study presents qualitative data from 21 African American/Black MSM who participated in a substance use and sexual risk reduction intervention trial (Project ROOM [men Reaching Out to Other Men]) in South Florida. African American/Black MSM from Project ROOM reduced their substance use and sexual risk behaviors at a faster rate than other men in the study. The present study examines how the experiences of participation in Project ROOM influenced the behavior change among African American/Black MSM. In-depth interviews indicate that study assessments enhanced African American/Black men's mindfulness and self-realization of behaviors leading to behavior modification and changes in social relationships. Furthermore, these findings suggest that interventions tailored to the social environment of HIV transmission and substance use behaviors are key to reducing risk behaviors among this population.


Disinfection of Blood Pressure Cuffs and Electrocardiographic Telemetry Leads With 0.5% Hydrogen Peroxide Wipes.
Disinfection of frequently touched surfaces in the hospital is critical for providing safe care. Because of their complex intricate surfaces, blood pressure cuffs and electrocardiographic telemetry leads may be harder than other equipment to disinfect. To determine the effectiveness of 0.5% hydrogen peroxide wipes in cleaning and disinfecting inpatient hospital surfaces, including hook and loop (Velcro) fasteners of blood pressure cuffs and electrocardiographic telemetry leads. A prospective study of an inpatient telemetry unit was designed to measure the persistence of a UV indicator and the presence of bacterial colonization on electrocardiographic telemetry leads and blood pressure cuffs. Call buttons and patient trays were used as control surfaces, because they are often touched but are easy to disinfect via standard practices. A total of 392 samples were collected between July 11 and August 3, 2016, and cultured for microorganisms. Among the cultures, 247 (63%) had at least 1 colony-forming unit. After adjustments in a multivariable analysis, odds of a contaminated surface (≥ 10 colony-forming units) after final cleaning with 0.5% hydrogen peroxide wipes were 3.70 times greater for patient trays than for blood pressure cuffs (P = .03) and 3.80 times greater for telemetry leads than for blood pressure cuffs (P = .04). The UV indicator persisted longer on blood pressure cuffs and telemetry leads than on patient trays or call buttons (P < .001). Wipes with 0.5% hydrogen peroxide adequately disinfect blood pressure cuffs but not telemetry leads.


Geographic Differences in the Dietary Quality of Food Purchases among Participants in the Nationally Representative Food Acquisition and Purchase Survey (FoodAPS).
Objective grocery transactions may reflect diet, but it is unclear whether the diet quality of grocery purchases mirrors geographic and racial/ethnic disparities in diet-related diseases. This cross-sectional analysis of 3961 households in the nationally representative Food Acquisition and Purchase Survey evaluated geographic and racial/ethnic disparities in grocery purchase quality. Respondents self-reported demographics and recorded purchases over 7 days; the Healthy Eating Index (HEI) 2015 assessed diet quality. Survey-weighted multivariable-adjusted regression determined whether there were geographic and racial/ethnic differences in HEI-15 scores. Respondents were, on average, 50.6 years, non-Hispanic white (NHW) (70.3%), female (70.2%), and had attended some college (57.8%). HEI-15 scores differed across geographic region (p < 0.05), with the highest scores in the West (57.0 ± 0.8) and lowest scores in the South (53.1 ± 0.8), and there was effect modification by race/ethnicity (p-interaction = 0.02). Regionally, there were diet disparities among NHW and non-Hispanic black (NHB) households; NHWs in the South had HEI-15 scores 3.2 points lower than NHWs in the West (p = 0.003). Southern NHB households had HEI-15 scores 8.1 points lower than Western NHB households (p = 0.013). Racial/ethnic disparities in total HEI-15 by region existed in the Midwest and South, where Hispanic households in the Midwest and South had significantly lower diet quality than NHW households. Heterogeneous disparities in the diet quality of grocery purchases by region and race/ethnicity necessitate tailored approaches to reduce diet-related disease.


Biodistribution and persistence of an MVA-vectored candidate HIV vaccine in SIV-infected rhesus macaques and SCID mice.
Recombinant modified vaccinia virus Ankara (MVA) is together with a few other attenuated viral vectors on the forefront of human immunodeficiency virus type 1 (HIV-1) vaccine development. As such, MVA-vectored vaccines are likely to be administered into immunocompromized individuals. Here, we demonstrated in a good laboratory practice study safety and biological clearance of candidate HIV-1 vaccine MVA.HIVA in simian immunodeficiency virus (SIV)-infected rhesus macaques and mice with a severe combined immunodeficiency (SCID) following an intradermal vaccine administration. In SIV-infected macaques, MVA.HIVA DNA was undetectable by nested PCR 6 weeks after dosing. In SCID mice, the MVA.HIVA vaccine was well tolerated and a positive PCR signal was only observed at the site of injection 49 days after dosing in four out of six mice, but even these sites were negative by day 81 post-injection. Therefore, the MVA.HIVA vaccine is considered safe for application in phase I clinical trials in HIV-1-infected human subjects. These results also contribute to the confidence of using MVA as a smallpox vaccine.


Cell Line-Specific Direct Irradiation and Bystander Responses are Influenced by Fetal Bovine Serum Serotonin Concentrations.
The radiation-induced bystander effect is mechanistically complex, involving many different signaling components. Serotonin, present in fetal bovine serum (FBS), has been implicated in the modulation of cellular responses to radiation. However, the role of this ubiquitous signaling molecule has yet to be elucidated with regard to cell line-specific radiation responses. In this study, cell survival was measured in HCT116 p53 wild-type (HCT116+/+) and HaCaT cell cultures treated with media containing serotonin-depleted FBS and compared to our standard FBS-supplemented media, using clonogenic assays. We utilized an enzyme-linked immunosorbent assay to quantify the difference (4.3 ± 1.3 ng/ml) in serotonin concentrations among the media. Serotonin-depleted media significantly reduced survival in both nonirradiated cell lines. Furthermore, we sought to determine the effects to cells in this media exposed to direct irradiation as well as bystander media from irradiated cells. Cell survival was significantly increased when HCT116+/+ cells were directly irradiated in serotonin-depleted media, while HaCaT cells showed no significant difference in survival between the media. Bystander investigations demonstrated that HCT116+/+ cells were only able to generate a bystander effect when cultured in standard media conditions containing greater serotonin levels. Conversely, HaCaT cells were unaffected by the different media in terms of producing a bystander response, generating bystander effects irrespective of the media. Previous research linking serotonin receptors to the bystander effect, together with our results, indicate that receptor heterogeneity among cell types may underlie serotonin sensitivity in direct irradiation and bystander responses through serotonin receptor-mediated cell signaling cascades.


Hypoxia and sterile inflammation in cystic fibrosis airways: mechanisms and potential therapies.
Cystic fibrosis is one of the most common autosomal recessive genetic diseases in Caucasian populations. Diagnosis via newborn screening and targeted nutritional and antibiotic therapy have improved outcomes, however respiratory failure remains the key cause of morbidity and mortality. Progressive respiratory disease in cystic fibrosis is characterised by chronic neutrophilic airway inflammation associated with structural airway damage leading to bronchiectasis and decreased lung function. Mucus obstruction is a characteristic early abnormality in the cystic fibrosis airway, associated with neutrophilic inflammation often in the absence of detectable infection. Recent studies have suggested a link between hypoxic cell death and sterile neutrophilic inflammation in cystic fibrosis and other diseases via the IL-1 signalling pathway. In this review, we consider recent evidence regarding the cellular responses to respiratory hypoxia as a potential driver of sterile neutrophilic inflammation in the lung, current knowledge on hypoxia as a pathogenic mechanism in cystic fibrosis and the potential for current and future therapies to alleviate hypoxia-driven sterile inflammation.


The role of B cells and humoral immunity in Mycobacterium tuberculosis infection.
Mycobacterium tuberculosis remains a major public health burden. It is generally thought that while B cell- and antibody-mediated immunity plays an important role in host defense against extracellular pathogens, the primary control of intracellular microbes derives from cellular immune mechanisms. Studies on the immune regulatory mechanisms during infection with M. tuberculosis, a facultative intracellular organism, has established the importance of cell-mediated immunity in host defense during tuberculous infection. Emerging evidence suggest a role for B cell and humoral immunity in the control of intracellular pathogens, including obligatory species, through interactions with the cell-mediated immune compartment. Recent studies have shown that B cells and antibodies can significantly impact on the development of immune responses to the tubercle bacillus. In this review, we present experimental evidence supporting the notion that the importance of humoral and cellular immunity in host defense may not be entirely determined by the niche of the pathogen. A comprehensive approach that examines both humoral and cellular immunity could lead to better understanding of the immune response to M. tuberculosis.


Estimating the effect of COVID-19 on trial design characteristics: a registered report.
There have been reports of poor-quality research during the COVID-19 pandemic. This registered report assessed design characteristics of registered clinical trials for COVID-19 compared to non-COVID-19 trials to empirically explore the design of clinical research during a pandemic and how it compares to research conducted in non-pandemic times. We did a retrospective cohort study with a 1 : 1 ratio of interventional COVID-19 registrations to non-COVID-19 registrations, with four trial design outcomes: use of control arm, randomization, blinding and prospective registration. Logistic regression was used to estimate the odds ratio of investigating COVID-19 versus not COVID-19 and estimate direct and total effects of investigating COVID-19 for each outcome. The primary analysis showed a positive direct and total effect of COVID-19 on the use of control arms and randomization. It showed a negative direct effect of COVID-19 on blinding but no evidence of a total effect. There was no evidence of an effect on prospective registration. Taken together with secondary and sensitivity analyses, our findings are inconclusive but point towards a higher prevalence of key design characteristics in COVID-19 trials versus controls. The findings do not support much existing COVID-19 research quality literature, which generally suggests that COVID-19 led to a reduction in quality. Limitations included some data quality issues, minor deviations from the pre-registered plan and the fact that trial registrations were analysed which may not accurately reflect study design and conduct. Following in-principle acceptance, the approved stage 1 version of this manuscript was pre-registered on the Open Science Framework at https://doi.org/10.17605/OSF.IO/5YAEB. This pre-registration was performed prior to data analysis.


LRP1 functions as an atheroprotective integrator of TGFbeta and PDFG signals in the vascular wall: implications for Marfan syndrome.
The multifunctional receptor LRP1 controls expression, activity and trafficking of the PDGF receptor-beta in vascular smooth muscle cells (VSMC). LRP1 is also a receptor for TGFbeta1 and is required for TGFbeta mediated inhibition of cell proliferation. We show that loss of LRP1 in VSMC (smLRP(-)) in vivo results in a Marfan-like syndrome with nuclear accumulation of phosphorylated Smad2/3, disruption of elastic layers, tortuous aorta, and increased expression of the TGFbeta target genes thrombospondin-1 (TSP1) and PDGFRbeta in the vascular wall. Treatment of smLRP1(-) animals with the PPARgamma agonist rosiglitazone abolished nuclear pSmad accumulation, reversed the Marfan-like phenotype, and markedly reduced smooth muscle proliferation, fibrosis and atherosclerosis independent of plasma cholesterol levels. Our findings are consistent with an activation of TGFbeta signals in the LRP1-deficient vascular wall. LRP1 may function as an integrator of proliferative and anti-proliferative signals that control physiological mechanisms common to the pathogenesis of Marfan syndrome and atherosclerosis, and this is essential for maintaining vascular wall integrity.


Diversity of novel archaeal viruses infecting methanogens discovered through coupling of stable isotope probing and metagenomics.
Diversity of viruses infecting non-extremophilic archaea has been grossly understudied. This is particularly the case for viruses infecting methanogenic archaea, key players in the global carbon biogeochemical cycle. Only a dozen of methanogenic archaeal viruses have been isolated so far. In the present study, we implemented an original coupling between stable isotope probing and complementary shotgun metagenomic analyses to identify viruses of methanogens involved in the bioconversion of formate, which was used as the sole carbon source in batch anaerobic digestion microcosms. Under our experimental conditions, the microcosms were dominated by methanogens belonging to the order Methanobacteriales (Methanobacterium and Methanobrevibacter genera). Metagenomic analyses yielded several previously uncharacterized viral genomes, including a complete genome of a head-tailed virus (class Caudoviricetes, proposed family Speroviridae, Methanobacterium host) and several near-complete genomes of spindle-shaped viruses. The two groups of viruses are predicted to infect methanogens of the Methanobacterium and Methanosarcina genera and represent two new virus families. The metagenomics results are in good agreement with the electron microscopy observations, which revealed the dominance of head-tailed virus-like particles and the presence of spindle-shaped particles. The present study significantly expands the knowledge on the viral diversity of viruses of methanogens.


Functional polyoxometalates from solvothermal reactions of VOSO4 with tripodal alkoxides - a study on the reactivity of different "tris" derivatives.
We report a study on the structure directing effects of functional groups and counterions. The aim was to find a facile and high yielding synthetic procedure to obtain polyoxometalate (POM) building blocks for post-functionalisation. Therefore, solvothermal reactions of VOSO4 with various tris(hydroxymethyl)methane derivatives in alkaline methanolic solutions were investigated. In doing so, new POM fragments were isolated and characterised. The binding modes of the functionalised tripodal alkoxides turned out to be surprisingly different.


Terasakiispira papahanaumokuakeensis gen. nov., sp. nov., a gammaproteobacterium from Pearl and Hermes Atoll, Northwestern Hawaiian Islands.
A Gram-negative, helical bacterium designated PH27AT was cultivated from an anchialine pool on Pearl and Hermes Atoll, Northwestern Hawaiian Islands. The obligately halophilic strain was motile by bipolar tufts of flagella and grew optimally at pH 7, and microaerobically or aerobically. Closest neighbours based on 16S rRNA gene nucleotide sequence identity are Marinospirillum celere v1c_Sn-redT (93.31 %) and M. alkaliphilum Z4T (92.10 %) in the family Oceanospirillaceae, class Gammaproteobacteria. PH27AT is distinguished phenotypically from members of the genus Marinospirillum by its hydrolysis of gelatin, the absence of growth in media containing ≤ 1 % (w/v) NaCl and the ranges of temperature (12–40 °C) and pH (5–8) for growth. The major compound ubiquinone Q-9 distinguishes the quinone system of strain PH27AT from those in members of the genus Marinospirillum and other members of the Oceanospirillaceae, in which the major quinone is Q-8. Major polar lipids in PH27AT were phosphatidylethanolamine and phosphatidylglycerol, with moderate amounts of diphosphatidylglycerol and phosphatidylserine. Spermidine and cadaverine dominated the polyamine pattern; large proportions of cadaverine have not been reported in members of the genus Marinospirillum. Genotypic and chemotaxonomic data show that PH27AT does not belong in the genus Marinospirillum or other genera of the family Oceanospirillaceae or the Halomonadaceae. We propose a new genus, Terasakiispira gen. nov., be created to accommodate Terasakiispira papahanaumokuakeensis gen. nov., sp. nov. as the type species, with PH27AT ( = ATCC BAA-995T = DSM 16455T = DSM 23961T) as the type strain.


Health behaviours and mental and physical health status in older adults with a history of homelessness: a cross-sectional population-based study in England.
This study compared (1) levels of engagement in lifestyle risk behaviours and (2) mental and physical health status in individuals who have previously been homeless to those of individuals who have not. Cross-sectional. Data were from participants (n=6931) of the English Longitudinal Study of Ageing. : Measures : Participants reported whether they had ever been homeless. We used regression models to analyse associations between homelessness and (1) cigarette smoking, daily alcohol consumption and physical inactivity, adjusting for sociodemographic covariates (age, sex, ethnicity, highest level of education, marital status and household non-pension wealth) and (2) self-rated health, limiting long-standing illness, depressive symptoms, life satisfaction, quality of life and loneliness, adjusting for sociodemographics and health behaviours. 104 participants (1.5%) reported having been homeless. Individuals who had been homeless were significantly more likely to be physically inactive (OR 1.62, 95% CI 1.44 to 2.52), report fair/bad/very bad self-rated health (OR 1.75, 95% CI 1.07 to 2.86), have a limiting long-standing illness (OR 2.66, 95% CI 1.65 to 4.30) and be depressed (OR 3.06, 95% CI 1.85 to 5.05) and scored lower on measures of life satisfaction (17.34 vs 19.96, p<0.001) and quality of life (39.02 vs 41.21, p=0.013). Rates of smoking (20.2% vs 15.4%, p=0.436), daily drinking (27.6% vs 22.8%, p=0.385) and loneliness (27.1% vs 21.0%, p=0.080) were also elevated. Those who were once homeless have poorer mental and physical health outcomes and are more likely to be physically inactive. Interventions to improve their health and quality of life are required.


Sortase A-mediated site-specific labeling of camelid single-domain antibody-fragments: a versatile strategy for multiple molecular imaging modalities.
A generic site-specific conjugation method that generates a homogeneous product is of utmost importance in tracer development for molecular imaging and therapy. We explored the protein-ligation capacity of the enzyme Sortase A to label camelid single-domain antibody-fragments, also known as nanobodies. The versatility of the approach was demonstrated by conjugating independently three different imaging probes: the chelating agents CHX-A"-DTPA and NOTA for single-photon emission computed tomography (SPECT) with indium-111 and positron emission tomography (PET) with gallium-68, respectively, and the fluorescent dye Cy5 for fluorescence reflectance imaging (FRI). After a straightforward purification process, homogeneous single-conjugated tracer populations were obtained in high yield (30-50%). The enzymatic conjugation did not affect the affinity of the tracers, nor the radiolabeling efficiency or spectral characteristics. In vivo, the tracers enabled the visualization of human epidermal growth factor receptor 2 (HER2) expressing BT474M1-tumors with high contrast and specificity as soon as 1 h post injection in all three imaging modalities. These data demonstrate Sortase A-mediated conjugation as a valuable strategy for the development of site-specifically labeled camelid single-domain antibody-fragments for use in multiple molecular imaging modalities. Copyright © 2016 John Wiley & Sons, Ltd.


Prognostic accuracy of the quick Sequential Organ Failure Assessment (qSOFA)-lactate criteria for mortality in adults with suspected bacterial infection in the emergency department of a hospital with limited resources.
Routine use of the Sequential Organ Failure Assessment (SOFA) score to prognosticate patients with sepsis is challenged by the requirement to perform numerous laboratory tests. The prognostic accuracy of the quick SOFA (qSOFA) without or with lactate criteria has not been prospectively investigated in low and middle income countries. We assessed the performance of simplified prognosis criteria using qSOFA-lactate criteria in the emergency department of a hospital with limited resources, in comparison with SOFA prognosis criteria and systemic inflammatory response syndrome (SIRS) screening criteria. This prospective cohort study was conducted between March and December 2017 in adult patients with suspected bacterial infection visiting the emergency department of the Indonesian National Referral Hospital. Variables from sepsis prognosis and screening criteria and venous lactate concentration at enrolment were recorded. Patients were followed up until hospital discharge or death. Prognostic accuracy was measured using area under the receiver operating characteristic curve (AUROC) of each criterion in the prediction of in-hospital mortality. Of 3026 patients screened, 1213 met the inclusion criteria. The AUROC of qSOFA-lactate criteria was 0.74 (95% CI 0.71 to 0.77). The AUROC of qSOFA-lactate was not statistically significantly different to the SOFA score (AUROC 0.75, 95% CI 0.72 to 0.78; p=0.462). The qSOFA-lactate was significantly higher than qSOFA (AUROC 0.70, 95% CI0.67 to 0.74; p=0.006) and SIRS criteria (0.57, 95% CI0.54 to 0.60; p<0.001). The prognostic accuracy of the qSOFA-lactate criteria is as good as the SOFA score in the emergency department of a hospital with limited resources. The performance of the qSOFA criteria is significantly lower than the qSOFA-lactate criteria and SOFA score.This abstract has been translated and adapted from the original English-language content. Translated content is provided on an "as is" basis. Translation accuracy or reliability is not guaranteed or implied. BMJ is not responsible for any errors and omissions arising from translation to the fullest extent permitted by law, BMJ shall not incur any liability, including without limitation, liability for damages, arising from the translated text.


Identification of a mutation in the ileal sodium-dependent bile acid transporter gene that abolishes transport activity.
The ileal Na+/bile acid cotransporter plays a critical role in the reabsorption of bile acids from the small intestine. In the course of cloning and characterizing the human ileal Na+/bile acid cotransporter cDNA, a dysfunctional isoform was identified in a patient diagnosed with Crohn's disease. Expression studies using hamster-human ileal Na+/bile acid cotransporter cDNA chimeras narrowed the location of the defect to the carboxyl-terminal 94 amino acids. Comparison of the sequence of the dysfunctional isoform to that of a wild-type human ileal Na+/bile acid cotransporter genomic clone revealed a single C to T transition resulting in a proline to serine substitution at amino acid position 290. The inheritance of this mutation in the proband's family was confirmed by single-stranded conformation polymorphism analysis and DNA sequencing. In transfected COS-1 cells, the single amino acid change abolished taurocholate transport activity but did not alter the transporter's synthesis or subcellular distribution. This dysfunctional mutation represents the first known molecular defect in a human sodium-dependent bile acid transporter.


A controlled Phase II trial assessing three doses of enfuvirtide (T-20) in combination with abacavir, amprenavir, ritonavir and efavirenz in non-nucleoside reverse transcriptase inhibitor-naive HIV-infected adults.
Enfuvirtide is a novel antiretroviral that blocks HIV-1 cell fusion and viral entry. This Phase II, controlled, open-label, randomized, multicentre dose-ranging trial explored the safety, antiviral activity and pharmacokinetics of enfuvirtide, administered by subcutaneous (s.c.) injection, in 71 HIV-1-infected, protease inhibitor-experienced, non-nucleoside reverse transcriptase inhibitor (NNRTI)-naive adults for 48 weeks. Study participants were randomized to receive enfuvirtide at a deliverable dose of 45, 67.5 or 90 mg twice daily; the 45 mg twice daily dose required 2 injections/day, while the higher doses required 4 injections/day. A background oral antiretroviral (ARV) regimen of abacavir (300 mg twice daily), amprenavir (1200 mg twice daily), ritonavir (200 mg twice daily) and efavirenz (600 mg once daily) was provided with enfuvirtide. A control group received the background ARV regimen alone. All potential participants underwent an HIV genotype at screen to ensure a homogenous population and to exclude patients with evidence of genotypic resistance to NNRTIs. Overall, the tolerability of the combination of abacavir, amprenavir, ritonavir, efavirenz and enfuvirtide was generally comparable to control through 48 weeks. No enfuvirtide dose-dependent adverse events (AEs) were observed across treatment groups. Injection site reactions (ISRs) occurred at least once in 68.5% of the enfuvirtide-treated population, and most ISRs were mild to moderate in severity, with no apparent dose relationship. Excluding ISRs, the most common treatment-emergent AEs were nausea, diarrhoea, dizziness and fatigue; with no clinically significant differences in the incidence of AEs observed between the control and enfuvirtide groups. Each treatment group benefited from ARV therapy, with a trend of increasing antiviral and immunological activity associated with increasing enfuvirtide dose. At 48 weeks, the median HIV-1 RNA change from baseline for the ITT population was -2.24 log10 copies/ml for the combined enfuvirtide groups compared with -1.87 log10 copies/ml for the control group. In addition, 54.9% of patients in the enfuvirtide group achieved HIV-1 RNA < or = 400 copies/ml versus 36.8% of patients in the control group. These results indicate that enfuvirtide has a favourable safety profile and is a promising new antiviral agent for HIV-infected patients who have been on previously failing ARV regimens.


Prevalence of malnutrition in a tertiary care hospital in India.
Malnutrition adversely affects clinical outcome of hospitalized patients. This observational prospective study was done to assess the prevalence of malnutrition and its grade among patients admitted in a mixed intensive care unit (ICU) of a tertiary care hospital in order to help devise a comprehensive nutrition program for the malnourished. A total of 500 sequential patients admitted to the ICU were screened on admission over a year period for malnutrition using the Subjective Global Nutritional Assessment (SGNA) score. Distribution of the degree of malnutrition according to co-morbidities was also documented. Of the total, 198 (39.6%) patients were malnourished, including one patient qualifying as severely malnourished; 68% patients were male, however, there was no statistically significant difference between nutrition status between sexes. Hypertension, diabetes, and cancer were the three most commonly encountered co-morbidities among the malnourished. A total of 86% of all cancer patients admitted were malnourished against only 12% of trauma patients. This study showed that almost two-fifth of the patients admitted were malnourished in this tertiary care hospital and that there is an urgent need to develop a comprehensive nutritional care program in many such Indian ICUs.


Oral captopril versus placebo among 13,634 patients with suspected acute myocardial infarction: interim report from the Chinese Cardiac Study (CCS-1)
13,634 patients entering 650 Chinese hospitals up to 36 h after the onset of suspected acute myocardial infarction (MI) were randomised between one month of oral captopril (6.25 mg initial dose, 12.5 mg 2 h later, and then 12.5 mg three times daily) or matching placebo. Captopril was associated with a non-significant reduction in 4-week mortality (617 [9.05%] captopril-allocated vs 654 [9.59%] placebo-allocated deaths; 2p = 0.3). There was a significant excess of hypotension, mostly early after the start of treatment, but no evidence of any adverse effect on early mortality (even among patients who were hypotensive at entry). Taken together with the other trials of converting enzyme inhibitors started early in acute MI, these results indicate that such therapy is generally safe and typically prevents about 5 deaths per 1000 patients treated for the first month.


Medical comorbidity in women and men with schizophrenia: a population-based controlled study.
Persons with persistent mental illness are at risk for failure to receive medical services. In order to deliver appropriate preventive and primary care for this population, it is important to determine which chronic medical conditions are most common. We examined chronic medical comorbidity in persons with schizophrenia using validated methodologies. Retrospective analysis of longitudinal administrative claims data from Wellmark Blue Cross/Blue Shield of Iowa. Subjects with schizophrenia or schizoaffective disorder (N=1,074), and controls (N=726,262) who filed at least 1 claim for medical services, 1996 to 2001. Case subjects had schizophrenia as the most clinically predominant psychotic disorder, based on psychiatric hospitalization, psychiatrist diagnoses, and outpatient care. Controls had no claims for any psychiatric comorbidity. Using a modified version of the Elixhauser Comorbidity Index, inpatient and outpatient claims were used to determine the prevalence of 46 common medical conditions. Odds ratios (ORs) were adjusted for age, gender, residence, and nonmental health care utilization using logistic regression. Subjects with schizophrenia were significantly more likely to have 1 or more chronic conditions compared with controls. Adjusted OR (95% confidence interval [CI]) were 2.62 (2.09 to 3.28) for hypothyroidism, 1.88 (1.51 to 2.32) for chronic obstructive pulmonary disease, 2.11 (1.36 to 3.28) for diabetes with complications, 7.54 (3.55 to 15.99) for hepatitis C, 4.21 (3.25 to 5.44) for fluid/electrolyte disorders, and 2.77 (2.23 to 3.44) for nicotine abuse/dependence. Schizophrenia is associated with substantial chronic medical burden. Familiarity with conditions affecting persons with schizophrenia may assist programs aimed at providing medical care for the mentally ill.


2-chlorodeoxyadenosine (2-CDA) therapy in previously untreated patients with follicular stage III-IV non-Hodgkin's lymphoma.
This phase II multi-institutional trial was designed to assess response and toxicity of 2-chlorodeoxyadenosine (2-CDA) in patients with previously untreated follicular lymphoma. The clinical significance of detecting cells carrying the t(14;18) translocation (bcl-2/JH rearrangement) in peripheral blood and bone marrow by polymerase chain reaction (PCR) before, during and after treatment was also examined. Between May 1993 and October 1995, 37 patients were accrued: male/female: 15/22, median age 51 years (range: 20-78), stage III/IV: 9/28. Patients received a total 2-CDA dose of 0.7 mg/kg as continuous s.c. or i.v. infusions over 7 days, every 28 days for a maximum of 5 cycles. A total of 165 cycles were administered. In 25 patients, blood and bone marrow before, during and after treatment were available for PCR analysis of the bcl-2/JH rearrangements. All 37 patients were evaluable for response and toxicity. The overall response rate was 84% (95% confidence interval, 68%-94%) with 14% CR (n = 5) and 70% PR (n = 26) and a median time to treatment failure of 15.7 months. bcl-2/JH rearrangement in peripheral blood and/or bone marrow was found in 10/25 of patients (40%) before treatment and 5 of these became repeatedly negative after 2-CDA therapy. There was no apparent association between bcl-2/ JH result and response. In 11 patients, 2-CDA was stopped because of progressive disease (n = 4), myelotoxicity (grade 2-3, n = 4), and other causes (n = 3, pulmonary embolism, metabolic disorder, and patient's decision). Four patients (11%) suffered from infections (grade 2-3). In 6 patients, persistent thrombocytopenia of 7.5 months (range: 3-21) occurred after completion of the 5 cycles. 2-CDA is active in untreated follicular lymphomas, but time to treatment failure suggests no advantage compared with standard treatment and toxicity on haematopoietic stem cells appears to be more pronounced. Molecular remission is induced in a considerable proportion of patients with disappearance of the bcl-2/JH rearrangement, and its possible significance as a predictive factor for quality of response and relapse warrants further study.


Subtrochanteric valgus osteotomy for chronically dislocated, painful spastic hips.
Many treatments have been proposed for a painful chronically dislocated hip in a child with spasticity who is unable to walk. The valgus subtrochanteric femoral osteotomy was originally described as a treatment for a missed congenital hip dislocation in a young adult who was able to walk. The purpose of this study was to investigate the outcomes of a modified Hass subtrochanteric valgus osteotomy for the treatment of symptomatic chronic hip dislocations in patients with spasticity due to neuromuscular disorders. Patients were identified by surgical codes. Medical records were reviewed for surgical indications, complications, and results. Preoperative and postoperative radiographs were assessed. Caregivers were contacted and asked to complete a brief survey detailing the pain that the patient experienced with daily activities before and after surgery as well as other information. Thirty-one Hass osteotomies in twenty-four consecutive patients were performed by one surgeon between 1995 and 2005. Although fifteen patients (63%) had at least one complication (including urinary tract infection, pneumonia, hardware failure, and heterotopic bone formation), the majority of the patients were doing well at an average of forty-four months postoperatively. The duration for which the patient could sit, ease of transfers, and ease of diaper changes had all improved according to the caregivers. Fourteen of fifteen caregivers who replied to the question stated that they were satisfied with the operative results and would have the procedure done again if they had the decision to make. The modified Hass osteotomy provides children who have symptomatic hip dislocations due to neuromuscular disorders with reproducible pain relief and improves ease of positioning by their caregivers. The complication rate, although high, was comparable with that of similar surgical procedures in this patient population. Concurrent femoral head resection at the time of the proximal femoral osteotomy was not necessary in this group of patients.


Forty years on: Uta Frith's contribution to research on autism and dyslexia, 1966-2006.
Uta Frith has made a major contribution to our understanding of developmental disorders, especially autism and dyslexia. She has studied the cognitive and neurobiological bases of both disorders and demonstrated distinctive impairments in social cognition and central coherence in autism, and in phonological processing in dyslexia. In this enterprise she has encouraged psychologists to work in a theoretical framework that distinguishes between observed behaviour and the underlying cognitive and neurobiological processes that mediate that behaviour.


Setting of import tolerances for metalaxyl-M in oil palms fruits and peppercorn.
In accordance with Article 6 of Regulation (EC) No 396/2005, the applicant Syngenta Crop Protection AG submitted a request to the competent national authority in Belgium to set an import tolerance for the active substance metalaxyl-M in oil palms fruits and peppercorn (black, green and white). The data submitted in support of the request were found to be sufficient to derive MRL proposals for oil palms fruits and peppercorn. Adequate analytical methods for enforcement are available to control the residues of metalaxyl-M in the commodities under consideration at the validated limit of quantification (LOQ) of 0.01 mg/kg. Based on the risk assessment results, EFSA concluded that the short-term and long-term intake of residues resulting from the use of metalaxyl-M according to the authorised agricultural practices is unlikely to present a risk to consumer health.


Insulin-like growth factor- I and factors affecting it in thalassemia major.
Despite improvement of blood transfusion regimens and iron chelation therapy growth and maturational delay, cardiomyopathy, endocrinopathies and osteoporosis still occur in good number of thalassemic patients. Decreased IGF-1 secretion occurs in the majority of the thalassemic patients particularly those with growth and pubertal delay. Many factors contribute to this decreased synthesis of IGF-I including disturbed growth hormone (GH) - insulin-like growth factor - I (IGF-I) axis. The possible factors contributing to low IGF-I synthesis in thalassemia and the possible interaction between low IGF-I secretion and the occurrence of these complications is discussed in this mini-review. Improvement of IGF-I secretion in thalassemic patients should be intended to improve linear growth and bone mineral accretion in thalassemic patients. This can be attained through adequate correction of anemia and proper chelation, nutritional supplementation (increasing caloric intake), correction of vitamin D and zinc deficiencies, induction of puberty and correction of hypogonadism at the proper time and treating GH deficiency. This review paper provides a summary of the current state of knowledge regarding IGF-I and factors affecting it in patients with thalassaemia major (TM). Search on PubMed and reference lists of articles with the term 'IGF-I, GH, growth, thalassemia, thyroxine, anemia, vitamin D, and zinc' was carried out. A hundred and forty-eight articles were found and used in the write up and the data analyzed was included in this report.


Control of central self-tolerance induction by autoreactive CD4+ thymocytes.
Medullary thymic epithelial cells (mTECs) are crucial for the selection of a T-cell-receptor (TCR) repertoire purged of self-reactive specificities, because these cells activate a promiscuous gene-expression program that leads to the synthesis of a wide array of peripheral tissue-restricted self-antigens. This review summarizes recent progress in our understanding of the cellular interactions, ligands, receptors and signal-transduction pathways that control mature-mTEC development. The particular focus is on new findings supporting the model that mature-mTEC development in the postnatal thymus depends on nuclear factor-kappaB (NF-kappaB) signaling induced by CD40-CD40 ligand, and receptor-activator-of-NF-kappaB (RANK)-RANK ligand interactions, and that these signals are delivered in the context of antigen-specific interactions between CD4(+) thymocytes carrying autoreactive TCRs and mTECs displaying cognate autoantigen-MHC-class-II complexes.


Secondary structure analysis of the putative membrane-associated domains of the inward rectifier K+ channel ROMK1.
The inward rectifier K+ channels contain two putative membrane-spanning domains per subunit (M1, M2) and a 'pore' (P) region, which is similar to the H5 domain of voltage-gated K+ channels. Here we have used Fourier transform infrared (FTIR) and CD spectroscopy to analyse the secondary structures of synthetic peptides corresponding to the M1, M2 and P regions of ROMK1 in aqueous solution, in organic solvents and in phospholipid membranes. A previous CD study was unable to provide any structural data on a similar P peptide [Ben-Efraim and Shai (1997) Biophys. J. 72, 85-96]. However, our FTIR and CD spectroscopic analyses indicate that this peptide adopts an alpha-helical structure when reconstituted into dimyristoyl phosphatidylcholine vesicles and lysophosphatidyl choline (LPC) micelles as well as in trifluoroethanol (TFE) solvent. This result is in good agreement with a previous study on a peptide corresponding to the pore domain of a voltage-gated K+ channel [Haris, Ramesh, Sansom, Kerr, Srai and Chapman (1994) Protein Eng. 7, 255-262]. FTIR spectra of the M1 peptide in LPC micelles displayed a strong absorbance characteristic of an intermolecular beta-sheet structure, suggesting aggregation of the M1 peptide. Sucrose gradient centrifugation was used to separate aggregated peptide from peptide incorporated into micelles in an unaggregated manner; subsequent analysis by FTIR suggested that the M1 peptide adopted an alpha-helical structure when incorporated into phospholipid membranes. FTIR and CD spectra of the M2 peptide in phospholipids and high concentrations of TFE suggest that this peptide adopts an alpha-helical structure. The structural data obtained in these experiments have been used to propose a model for the structure of the membrane-associated core (M1-P-M2) of the inward rectifier K+ channel protein.


Relationship between calcium, lactose, vitamin D, and dairy products and ovarian cancer.
Few prospective studies of the relationship between intake of dairy foods, calcium, vitamin D, and lactose and ovarian cancer have been conducted, and results have been largely inconsistent. Two recent studies found significant increased risk with frequent dairy consumption and perhaps with high intakes of calcium or lactose. The authors investigated the association between these foods and nutrients and ovarian cancer risk among 31,925 subjects in the Breast Cancer Detection Demonstration Project follow-up cohort. Multivariable (MV) relative risks (RRs) adjusted for age, parity, and other factors were estimated using Cox proportional hazards models. Over an average follow-up of 8.3 yr, 146 incident ovarian cancer cases were confirmed. Higher intakes of total dairy food (comparing four or more servings per day vs. less than one serving per day) were associated with a statistically significant decreased risk of ovarian cancer, although the trend was not significant (MV RR = 0.42; 95% confidence interval (CI) = 0.20-0.89; P for trend = 0.07). Comparing extreme quartiles, we observed a statistically nonsignificant inverse association between high dietary calcium intake and ovarian cancer (RR = 0.67; 95% CI = 0.43, 1.04; P for trend = 0.08). No statistically significant relations were found for consumption of specific dairy foods, lactose, or vitamin D and ovarian cancer risk. The possibility of a decreased risk of ovarian cancer for dietary calcium merits further evaluation.


Pathology survey on a captive-bred colony of the Mexican Goodeid, nearly extinct in the wild, Zoogoneticus tequila (Webb & Miller 1998).
The Mexican Goodeid, Zoogoneticus tequila, is considered nearly extinct in the wild and it is maintained in captivity by the nonprofit international "Goodeid Working Group." The unique Italian colony has produced about 180 fish so far. The observable diseases were registered and some fish were submitted, immediately after spontaneous death, to necroscopic and histopathologic exams. Encountered diseases included the following: 7 cases of scoliosis (2 males and 5 females); 2 fish with a similar congenital deviation of ocular axis; 1 adult male with left corneal opacity, presumably of traumatic origin; 1 female fish with a large subocular fluid-filled sac, histologically referable to a lymphatic cyst, similarly to the eye sacs of a Goldfish variety (Carassius auratus) called bubble eye; and 1 female fish with recurrent abdominal distension consequent to distal bowel dilation and thinning, associated with complete mucosal atrophy, and comparable to intestinal pseudo-obstruction syndromes described in humans and various animal species. The absence of infectious or parasitic diseases, as well as the low incidence of diseases potentially related to environmental alterations or nutritional disorders such as spinal deformities, suggests the adequacy of breeding management techniques of Z. tequila for its conservation and reintroduction in to the original habitat in the near future.


Reproductive hormone profile and pubertal development in 14-year-old boys prenatally exposed to polychlorinated biphenyls.
Because polychlorinated biphenyls (PCBs) are thought to cause endocrine disruption, we examined 438 adolescent boys from a birth cohort in the Faroe Islands, where PCB exposures are elevated. We measured PCBs and p,p'-dichlorodiphenyldichloroethylene (DDE) in cord blood and in serum from clinical examination at age 14. Higher prenatal PCB exposure was associated with lower serum concentrations of both luteinizing hormone (LH) and testosterone. In addition, sex hormone binding globulin (SHBG) was positively associated with both prenatal and concurrent PCB exposures. The PCB-SHBG association was robust to covariate adjustment. In a structural equation model, a doubling in prenatal PCB exposure was associated with a decrease in LH of 6% (p=0.03). Prenatal exposure to PCB and DDE showed weak, non-significant inverse associations with testicular size and Tanner stage. DDE was highly correlated with PCB and showed slightly weaker associations with the hormone profile. These findings suggest that delayed puberty with low serum-LH concentrations associated with developmental exposure to non dioxin-like PCBs may be due to a central hypothalamo-pituitary mechanism.


EPIC-DB: a proteomics database for studying Apicomplexan organisms.
High throughput proteomics experiments are useful for analyzing the protein expression of an organism, identifying the correct gene structure of a genome, or locating possible post-translational modifications within proteins. High throughput methods necessitate publicly accessible and easily queried databases for efficiently and logically storing, displaying, and analyzing the large volume of data. EPICDB is a publicly accessible, queryable, relational database that organizes and displays experimental, high throughput proteomics data for Toxoplasma gondii and Cryptosporidium parvum. Along with detailed information on mass spectrometry experiments, the database also provides antibody experimental results and analysis of functional annotations, comparative genomics, and aligned expressed sequence tag (EST) and genomic open reading frame (ORF) sequences. The database contains all available alternative gene datasets for each organism, which comprises a complete theoretical proteome for the respective organism, and all data is referenced to these sequences. The database is structured around clusters of protein sequences, which allows for the evaluation of redundancy, protein prediction discrepancies, and possible splice variants. The database can be expanded to include genomes of other organisms for which proteome-wide experimental data are available. EPICDB is a comprehensive database of genome-wide T. gondii and C. parvum proteomics data and incorporates many features that allow for the analysis of the entire proteomes and/or annotation of specific protein sequences. EPICDB is complementary to other -genomics- databases of these organisms by offering complete mass spectrometry analysis on a comprehensive set of all available protein sequences.


Temperature, by Controlling Growth Rate, Regulates CRISPR-Cas Activity in Pseudomonas aeruginosa.
Clustered regularly interspaced short palindromic repeat (CRISPR)-associated (CRISPR-Cas) systems are adaptive defense systems that protect bacteria and archaea from invading genetic elements. In Pseudomonas aeruginosa, quorum sensing (QS) induces the CRISPR-Cas defense system at high cell density when the risk of bacteriophage infection is high. Here, we show that another cue, temperature, modulates P. aeruginosa CRISPR-Cas. Increased CRISPR adaptation occurs at environmental (i.e., low) temperatures compared to that at body (i.e., high) temperature. This increase is a consequence of the accumulation of CRISPR-Cas complexes, coupled with reduced P. aeruginosa growth rate at the lower temperature, the latter of which provides additional time prior to cell division for CRISPR-Cas to patrol the cell and successfully eliminate and/or acquire immunity to foreign DNA. Analyses of a QS mutant and synthetic QS compounds show that the QS and temperature cues act synergistically. The diversity and level of phage encountered by P. aeruginosa in the environment exceed that in the human body, presumably warranting increased reliance on CRISPR-Cas at environmental temperatures.IMPORTANCEP. aeruginosa is a soil dwelling bacterium and a plant pathogen, and it also causes life-threatening infections in humans. Thus, P. aeruginosa thrives in diverse environments and over a broad range of temperatures. Some P. aeruginosa strains rely on the CRISPR-Cas adaptive immune system as a phage defense mechanism. Our discovery that low temperatures increase CRISPR adaptation suggests that the rarely occurring but crucial naive adaptation events may take place predominantly under conditions of slow growth, e.g., during the bacterium's soil dwelling existence and during slow growth in biofilms.


Experimental reconstitution of chronic ER stress in the liver reveals feedback suppression of BiP mRNA expression.
Endoplasmic reticulum (ER) stress is implicated in many chronic diseases, but very little is known about how the unfolded protein response (UPR) responds to persistent ER stress in vivo. Here, we experimentally reconstituted chronic ER stress in the mouse liver, using repeated injection of a low dose of the ER stressor tunicamycin. Paradoxically, this treatment led to feedback-mediated suppression of a select group of mRNAs, including those encoding the ER chaperones BiP and GRP94. This suppression was due to both silencing of the ATF6α pathway of UPR-dependent transcription and enhancement of mRNA degradation, possibly via regulated IRE1-dependent decay (RIDD). The suppression of mRNA encoding BiP was phenocopied by ectopic overexpression of BiP protein, and was also observed in obese mice. Our findings suggest that persistent cycles of UPR activation and deactivation create an altered, quasi-stable setpoint for UPR-dependent transcriptional regulation-an outcome that could be relevant to conditions such as metabolic syndrome.


Definitive chemoradiation for oesophageal cancer--a standard of care in patients with non-metastatic oesophageal cancer.
A retrospective analysis was carried out of 291 cases of oesophageal cancer treated with definitive chemoradiotherapy (dCRT) at a single UK cancer centre between 1995 and 2009. Our protocol consisted of two cycles of neoadjuvant platinum-based chemotherapy followed by two further cycles given concurrently with 50Gy of external beam radiotherapy delivered in 25 fractions over 5 weeks. Demographic, patient and outcome data were recorded prospectively through an electronic health record and retrospectively analysed, using appropriate statistical tools. Data on 266 patients were available for analysis. The median age was 66.6 years, 53% were adenocarcinomas. dCRT was used instead of surgery because of age/co-morbidity in 44% and disease extent in 39%. Ninety-three per cent of patients completed treatment according to protocol. Grade 3 and 4 toxicities were seen in 42 and 7%, respectively. Median survival was 20.6 months; 2, 3 and 5 year survival rates were 43.6, 32.9 and 19.5%, respectively. Advanced disease was associated with a worse outcome. Shorter disease length was associated with a better median survival, but some patients with disease >10cm had long-term disease control. The effect of other patient- and disease-related factors was also analysed. We present data showing that dCRT is well tolerated and should be considered as an alternative to surgery for all patients with locally advanced oesophageal cancer, not only those with co-morbidity. Furthermore, the benefits of dCRT are not confined to carcinomas with squamous histology.


Socioeconomic position, not race, is linked to death after cardiac surgery.
Health disparities have been associated with the prevalence of cardiovascular disease. In cardiac surgery, association has been found between race, sex, and poorer prognosis after surgery. However, there is a complex interplay between race, sex, and socioeconomic position (SEP). In our investigation we sought to identify which of these was the driver of risk-adjusted survival. From January 1, 1995, and December 30, 2005, 23 330 patients (15 156 white men, 6932 white women, 678 black men, and 564 black women) underwent isolated coronary artery bypass grafting, valve, or combined coronary artery bypass grafting and valve procedures. Median follow-up was 5.8 years (25th and 75th percentiles: 3 and 8.6 years). Effect of race, sex, and SEP on all-cause mortality was examined with 2-phase Cox model and generalized propensity score technique. As expected, blacks and women had lower SEP as compared with whites and men for all 6 SEP indicators. Patients with lower SEP had more atherosclerotic disease burden, more comorbidity, and were more symptomatic. Lower SEP was associated with a risk-adjusted dose-dependent reduction in survival after surgery (men, P<0.0001; women, P=0.0079), but black race, once adjusted for SEP, was not. Our large investigation demonstrates that disparities in SEP are present and significantly affect health outcomes. Although race per se was not the driver for reduced survival, patients of low SEP were predominantly represented by blacks and women. Socioeconomically disadvantaged patients had significantly higher risk-adjusted mortality after surgery. Further investigation and targeted intervention should focus specifically on patients of low SEP, their health behaviors, and secondary prevention efforts.


Changes in radiotherapy fractionation-breast cancer.
Conventional fractionation for half a century has been justified on the basis that 2.0 Gy fractions spare dose-limiting late-responding normal tissues to a greater degree than cancerous tissues. Early indications that breast cancer responds more strongly to fraction size than many other common cancers were followed several decades of investigation, but there is now reliable Level I evidence that this is the case. Four randomised trials testing fraction sizes in the range 2.7-3.3 Gy have reported 10-year follow up in almost 8000 patients, and they provide robust estimates of α/β in the range of 3 Gy. The implication is that there are no advantages in terms of safety or effectiveness of persisting with 2.0 Gy fractions in patients with breast cancer. 15- or 16-fraction schedules are replacing the conventional 25-fraction regimen as a standard of care for adjuvant therapy in an increasing number of countries. A number of concerns relating to the appropriateness of hypofractionation in patient subgroups, including those treated post-mastectomy, advanced local-regional disease and/or to lymphatic pathways are addressed. Meanwhile, hypofractionation can be exploited to modulate dose intensity across the breast according to relapse risk by varying fraction size across the treatment volume. The lower limits of hypofractionation are currently being explored, one approach testing a 5-fraction schedule of local-regional radiotherapy delivered in 1 week.


Evaluation of quality of life in PKU before and after introducing tetrahydrobiopterin (BH4); a prospective multi-center cohort study.
Phenylketonuria (PKU) is a rare inborn error of metabolism caused by phenylalanine hydroxylase enzyme (PAH) deficiency. Treatment constitutes a strict Phe restricted diet with unpalatable amino acid supplements. Residual PAH activity enhancement with its cofactor tetrahydrobiopterin (BH4) is a novel treatment which increases dietary tolerance in some patients and permits dietary relaxation. Relaxation of diet may improve health related quality of life (HRQoL). This prospective cohort study aims to evaluate HRQoL of patients with PKU and effects of BH4 treatment on HRQoL. Patients aged 4years and older, diagnosed through newborn screening and early and continuously treated, were recruited from eight metabolic centers. Patients and mothers completed validated generic and chronic health-conditions HRQoL questionnaires (PedsQL, TAAQOL, and DISABKIDS) twice: before and after testing BH4 responsivity. Baseline results were compared to the general population. Data collected after BH4 testing was used to find differences in HRQoL between BH4 unresponsive patients and BH4 responsive patients after one year of treatment with BH4. Also a within patient comparison was performed to find differences in HRQoL before and after treatment with BH4. 69/81 (85%) patients completed the questionnaires before BH4 responsivity testing, and 45/69 (65%) participated again after testing. Overall PKU patients demonstrated normal HRQoL. However, some significant differences were found when compared to the general population. A significantly higher (thus better) score on the PedsQL was reported by children 8-12 years on physical functioning and by children 13-17 years on total and psychosocial functioning. Furthermore, adult patients reported significantly lower (thus worse) scores in the TAAQOL cognitive domain. 10 patients proved to be responsive to BH4 treatment; however improvement in their HRQoL after relaxation of diet could not be demonstrated.


Three-dimensional modeling of nitrate-N transport in vadose zone: Roles of soil heterogeneity and groundwater flux.
Contamination of groundwater from nitrogen fertilizers in agricultural lands is an important environmental and water quality management issue. It is well recognized that in agriculturally intensive areas, fertilizers and pesticides may leach through the vadose zone and eventually reach groundwater. While numerical models are commonly used to simulate fate and transport of agricultural contaminants, few models have considered a controlled field work to investigate the influence of soil heterogeneity and groundwater flow on nitrate-N distribution in both root zone and deep vadose zone. In this work, a numerical model was developed to simulate nitrate-N transport and transformation beneath a center pivot-irrigated corn field on Nebraska Management System Evaluation area over a three-year period. The model was based on a realistic three-dimensional sediment lithology, as well as carefully controlled irrigation and fertilizer application plans. In parallel, a homogeneous soil domain, containing the major sediment type of the site (i.e. sandy loam), was developed to conduct the same water flow and nitrate-N leaching simulations. Simulated nitrate-N concentrations were compared with the monitored nitrate-N concentrations in 10 multi-level sampling wells over a three-year period. Although soil heterogeneity was mainly observed from top soil to 3 m below the surface, heterogeneity controlled the spatial distribution of nitrate-N concentration. Soil heterogeneity, however, has minimal impact on the total mass of nitrate-N in the domain. In the deeper saturated zone, short-term variations of nitrate-N concentration correlated with the groundwater level fluctuations.


Integrating threshold concepts with reflective practice: Discussing a theory-based approach for curriculum refinement in dental education.
The theory of threshold concepts (TCs) is a burgeoning area of educational research and curriculum reform. This study describes and reviews the TCs literature, describes the differences between TCs and threshold learning outcomes (TLOs) and seeks to promote discussion on using an integrated reflective practice approach to facilitate the translation of TCs into fulfilment of TLOs. A worked example is included in this study to explore the challenges inherent in this task but also the learning gains to be made for both staff and students in this endeavour. INTERSECTION OF REFLECTIVE CLINICAL PRACTICE AND THRESHOLD CONCEPTS: Reflection is intrinsic to all aspects of learning and is a vital characteristic of developing competence in the health professions. Further, the need for explicit teaching and assessment of reflection in health professions education is recognised. We suggest that the identification of TCs can help to scaffold reflective processes in dentistry and argue that TCs may achieve a bridging function between theory and practice when used in conjunction with reflective writing and dialogue. This study provides a discussion of the complexities and challenges involved as well as an explicit example of the process involved for 1 TC in 1 particular discipline-removable prosthodontics. Reflective practice is recognised by the authors as a pedagogical tool to support the acquisition of threshold concepts. An example is provided of integrating reflective practice tasks with identified TCs in curricula which shows the potential of reflective dialogue and discourse as vehicles to make TCs explicit as well as facilitate the attainment of threshold learning outcomes or graduate attributes. This is an ongoing process, and although the fully revised curriculum is not ready to be translated to other contexts, we suggest it is beneficial to start working towards the integration of reflective practice tasks and identified TLOs as part of curriculum refinement-rather than adding to a crowded curriculum. We suggest such an integrated approach can harness the challenges inherent in discipline-specific contexts to bridge the theory-practice nexus for students and permit staff greater flexibility in the teaching and assessment of such traditionally difficult areas. We propose a way of thinking about curriculum change by integrating threshold concepts with the explicit use and assessment of reflective practice tasks as a means to achieve threshold learning outcomes and professional attributes identified by accrediting bodies. We provide an overview of the literature in this area and an example of our approach in linking reflective practice with newly identified threshold concepts in our School. We welcome further discussion, debate and collaboration in this area.


Discovery of deoxyceramides and diacylglycerols as CD1b scaffold lipids among diverse groove-blocking lipids of the human CD1 system.
Unlike the dominant role of one class II invariant chain peptide (CLIP) in blocking MHC class II, comparative lipidomics analysis shows that human cluster of differentiation (CD) proteins CD1a, CD1b, CD1c, and CD1d bind lipids corresponding to hundreds of diverse accurate mass retention time values. Although most ions were observed in association with several CD1 proteins, ligands binding selectively to one CD1 isoform allowed the study of how differing antigen-binding grooves influence lipid capture. Although the CD1b groove is distinguished by its unusually large volume (2,200 Å(3)) and the T' tunnel, the average mass of compounds eluted from CD1b was similar to that of lipids from CD1 proteins with smaller grooves. Elution of small ligands from the large CD1b groove might be explained if two small lipids bind simultaneously in the groove. Crystal structures indicate that all CD1 proteins can capture one antigen with its hydrophilic head group exposed for T-cell recognition, but CD1b structures show scaffold lipids seated below the antigen. We found that ligands selectively associated with CD1b lacked the hydrophilic head group that is generally needed for antigen recognition but interferes with scaffold function. Furthermore, we identified the scaffolds as deoxyceramides and diacylglycerols and directly demonstrate a function in augmenting presentation of a small glycolipid antigen to T cells. Thus, unlike MHC class II, CD1 proteins capture highly diverse ligands in the secretory pathway. CD1b has a mechanism for presenting either two small or one large lipid, allowing presentation of antigens with an unusually broad range of chain lengths.


Anorectal Disorders: An Update.
Anorectal disorders encompass structural, neuromuscular, and functional disorders. They are common, often distressing, and in some cases debilitating, and significantly add to the health care burden. They present with multiple, overlapping symptoms that can often obscure the underlying pathology and can pose significant diagnostic and management dilemmas. A meticulous history and comprehensive digital rectal examination can provide clarity on the diagnosis, appropriate testing, and management of these conditions. Today, with the development of sophisticated diagnostic tools such as high-resolution and high-definition (3-D) anorectal manometry, 3-D anal ultrasonography, magnetic resonance defecography and imaging, and neurophysiological tests such as translumbosacral anorectal magnetic stimulation, it is possible to more accurately define and characterize the underlying structural and functional abnormalities. In this review, we present a succinct update on the latest knowledge with regards to the pathophysiology, diagnosis and management of anal fissure, hemorrhoids, rectal prolapse, intussusception, rectocele, solitary rectal ulcer syndrome, levator ani syndrome, dyssynergic defecation and fecal incontinence.


The beta subunit of human granulocyte-macrophage colony-stimulating factor receptor forms a homodimer and is activated via association with the alpha subunit.
Human granulocyte-macrophage colony-stimulating factor (hGM-CSF) receptor (hGMR) consists of alpha and beta subunits, and the precise stoichiometry of these subunits has remained to be determined. In this work, oligomerization of the beta subunit was studied using a chemical cross-linker. In Ba/F3, a mouse interleukin-3-dependent cell line expressing both subunits of hGMR (Ba/F3-alpha,beta), a protein with a molecular mass corresponding to that of a homodimer of the beta subunit (beta homodimer) was detected only when cells were treated with the cross-linker. Dimerization of the beta subunit was confirmed by coimmunoprecipitation of a tagged beta subunit with the wild type beta subunit COS7 cells. The beta homodimer had already formed in the absence of hGM-CSF, whereas stimulation with the ligand brought both alpha and beta subunits into a complex, the result being tyrosine phosphorylation of the beta homodimer. Tyrosine phosphorylation of the subunit was impaired by deletion of the cytoplasmic domain of the alpha subunit without interfering with the association of both subunits. These results indicate that the beta homodimer, which alone is insufficient for signaling, forms the functional hGMR with the alpha subunit in response to hGM-CSF.


Digital quantification and selection of high-lipid-producing microalgae through a lateral dielectrophoresis-based microfluidic platform.
Microalgae are promising alternatives to petroleum as renewable biofuel sources, however not sufficiently economically competitive yet. Here, a label-free lateral dielectrophoresis-based microfluidic sorting platform that can digitally quantify and separate microalgae into six outlets based on the degree of their intracellular lipid content is presented. In this microfluidic system, the degree of cellular lateral displacement is inversely proportional to the intracellular lipid level, which was successfully demonstrated using Chlamydomonas reinhardtii cells. Using this functionality, a quick digital quantification of sub-populations that contain different intracellular lipid level in a given population was achieved. In addition, the degree of lateral displacement of microalgae could be readily controlled by simply changing the applied DEP voltage, where the level of gating in the intracellular lipid-based sorting decision could be easily adjusted. This allowed for selecting only a very small percentage of a given population that showed the highest degree of intracellular lipid content. In addition, this approach was utilized through an iterative selection process on natural and chemically mutated microalgal populations, successfully resulting in enrichment of high-lipid-accumulating microalgae. In summary, the developed platform can be exploited to quickly quantify microalgae lipid distribution in a given population in real-time and label-free, as well as to enrich a cell population with high-lipid-producing cells, or to select high-lipid-accumulating microalgal variants from a microalgal library.


Genetic separation of southern and northern soybean breeding programs in North America and their associated allelic variation at four maturity loci.
North American soybean breeders have successfully developed a large number of elite cultivars with diverse maturity groups (MG) from a small number of ancestral landraces. To understand molecular and genetic basis underlying the large variation in their maturity and flowering times, we integrated pedigree and maturity data of 166 cultivars representing North American soybean breeding. Network analysis and visualization of their pedigree relationships revealed a clear separation of southern and northern soybean breeding programs, suggesting that little genetic exchange occurred between northern (MG 0-IV) and southern cultivars (MG V-VIII). We also analyzed the transcript sequence and expression levels of four major maturity genes (E1 to E4) and revealed their allelic variants in 75 major ancestral landraces and milestone cultivars. We observed that e1-as was the predominant e mutant allele in northern genotypes, followed by e2 and e3. There was no allelic variation at E4. Transcript accumulation of the e2 mutant allele was significantly reduced, which might be caused by its premature stop codon triggering the nonsense-mediated mRNA decay pathway. The large DNA deletion generating the e3 mutant allele also created a gene fusion transcript. The e alleles found in milestone cultivars were traced through pedigrees to their ancestral landraces and geographic origins. Our analysis revealed an approximate correlation between dysfunctional alleles and maturity groups for most of the 75 cultivars. However, single e mutant alleles and their combinations were not sufficient to fully explain their maturity diversity, suggesting that additional genes/alleles are likely involved in regulating maturity time.


A newly developed tetraplex real-time RT-PCR for simultaneous screening of influenza virus types A, B, C and D.
Human- or avian-to-swine transmissions have founded several autonomously circulating influenza A virus (IAV) lineages in swine populations that cause economically important respiratory disease. Little is known on other human influenza virus types, like B (IBV) and C (ICV) in European swine, and of the recently detected novel animal influenza virus type D (IDV). Development of a cost-effective diagnostic tool for large-scale surveillance programmes targeting all four influenza virus types. An influenza ABCD tetraplex real-time RT-PCR (RT-qPCR) was developed in the frame of this study. A selection of reference virus strains and more than 4000 porcine samples from a passive IAV surveillance programme in European swine with acute respiratory disease were examined. Two IBV, a single IDV but no ICV infections were identified by tetraplex RT-qPCR. IBV and IDV results were confirmed by conventional RT-PCR and partial sequence analysis. The tetraplex RT-qPCR proved fit for purpose as a sensitive, specific and high-throughput tool to study influenza virus transmission at the human-animal interface. Complementing close-meshed active virological and serological surveillance is required to better understand the true incidence and prevalence of influenza virus type B, C and D infections in swine.


The value of noncoronary atherosclerosis for identifying coronary artery disease: results of the Leipzig LIFE Heart Study.
Despite the widespread use of noninvasive testing prior to invasive coronary diagnostic the diagnostic yield of elective coronary angiography has been reported low in subjects with suspected obstructive CAD. To determine the predictive value of noncoronary atherosclerosis (NCA) in subjects with suspected stable coronary artery disease (CAD) intended to invasive coronary angiography. Ultrasound-based assessment of carotid artery plaque (CAP), carotid intima-media thickness (CIMT) and ankle-brachial index (ABI) was performed in 2216 subjects with suspected CAD prior to coronary angiography. Logistic regression and c-statistics were used to analyze the diagnostic value of NCA for the presence of obstructive CAD and the intention to revascularization. Percentage of positive results of elective coronary angiography was low but comparable to other studies (41% obstructive CAD). We identified 1323 subjects (60%) with NCA, most of them were characterized by CAP (93%). CAP independently predicted obstructive CAD in addition to traditional risk factors and clinical factors while CIMT and ABI failed to improve the prediction. The presence of NCA and typical angina were the strongest predictors for obstructive CAD (OR 4.0 and 2.4, respectively). A large subgroup of patients (n = 703, 32%) with atypical clinical presentation and lack of NCA revealed a low indication for revascularization <15% indicating a large proportion of subjects with non-obstructive CAD in this subgroup. The evaluation of noncoronary atherosclerosis has the potential to impact clinical decision making and to direct subsequent diagnostic procedures in subjects with suspected coronary artery disease. NCT00497887.


CXCR2 expression and postoperative complications affect long-term survival in patients with esophageal cancer.
Esophagectomy is one of the most invasive surgical treatments for digestive tract cancer, and the blood levels of inflammatory cytokines such as interleukin-1, interleukin-6, and interleukin-8 are increased for several hours after surgery or in patients experiencing postoperative complications. CXCR2, an interleukin-8 receptor, is reportedly expressed in several carcinomas, and interleukin-8 signaling promotes cancer cell proliferation. The impact of postoperative complications following esophagectomy on long-term survival is controversial. In this study, we demonstrate the significance of CXCR2 expression and validate the effects of CXCR2 expression and postoperative complications on long-term prognosis of esophageal squamous cell carcinoma using resected specimens. Eighty-two specimens were sectioned from archived, paraffin-embedded tumor tissues obtained from patients with esophageal squamous cell carcinoma who underwent esophagectomy and extended lymphadenectomy for complete resection of cancer in our institute between 1997 and 2002. Immunohistochemistry was performed using a polyclonal antibody to CXCR2, and the correlation of stainability with clinicopathological factors and long-term survival was examined. CXCR2 was expressed in 33 of 82 (40.2 %) specimens. In the CXCR2-positive group, the recurrence-free survival and overall survival rates of patients who developed postoperative complications were both significantly lower than those for patients who did not develop any complications. In contrast, in the CXCR2-negative group, there was no significant difference in long-term prognosis between patients with and without complications. CXCR2 positivity combined with postoperative complications was an independent risk factor for subsequent tumor recurrence, showing the highest hazard ratio. Our results suggest that the patients with CXCR2-positive esophageal cancer who develop postoperative complications have a poor prognosis and should be carefully followed. This study was approved by Keio University School of Medicine Ethics Committee with a trial registration number of 2011-241.


Glycine stimulates protein synthesis and inhibits oxidative stress in pig small intestinal epithelial cells.
Glycine has recently been classified as a nutritionally essential amino acid for maximal growth in young pigs. Currently, little is known about the metabolism or function of glycine in the neonatal intestine. This work was conducted to test the hypothesis that glycine has a protective effect against oxidative stress in intestinal epithelial cells. Jejunal enterocytes isolated from newborn pigs were cultured in the presence of 0.0-2 mmol/L glycine for measurements of glycine metabolism, cell proliferation, protein turnover, apoptosis, and antioxidative response. Compared with 0.0-0.5 mmol/L glycine, 1.0 mmol/L glycine enhanced (P < 0.05) cell growth (by 8-24% on day 2 and by 34-224% on day 4, respectively) and protein synthesis (by 36-419%) while reducing (P < 0.05) protein degradation (by 7-28%). This effect of glycine was associated with activation of the mammalian target of rapamycin signaling pathway in enterocytes. By using a model of oxidative stress induced by 30 μmol/L 4-hydroxynonenal (4-HNE), which was assessed by flow cytometry analysis, 1.0 mmol/L glycine inhibited (P < 0.05) activation of caspase 3 by 25% and attenuated (P < 0.05) 4-HNE-induced apoptosis by 38% in intestinal porcine epithelial cell line 1 cells through promotion of reduced glutathione synthesis and expression of glycine transporter 1 while reducing the activation of extracellular signal-regulated kinases, c-Jun amino-terminal kinases, and p38 protein in the mitogen-activated protein kinase signaling pathway. These novel findings provide a biochemical mechanism for the use of dietary glycine to improve intestinal health in neonates under conditions of oxidative stress and glycine deficiency.


Holographic molecularly imprinted polymers for label-free chemical sensing.
Holographic molecularly imprinted polymer films for the use in chemical sensors are obtained in one step through photopolymerization with interfering laser beams. This results in hierarchical structuring at four length scales: micrometer-scale patterning of millimeter- to centimeter- size polymer objects with holographic optical properties, exhibiting nanometer-scale porosity and specific molecular recognition properties at the molecular scale through self-assembly. Specific binding of the target analyte testosterone is measured by diffraction analysis.


Primary care provider perspectives on virtual and in-person depression management during the COVID-19 pandemic.
During the COVID-19 pandemic, primary care providers (PCPs), nurses, and integrated mental health specialists continued to collaboratively manage depression among patients using both in-person and virtual (i.e., hybrid) modalities. Few studies have characterized how hybrid services are currently delivered within interdisciplinary primary care teams. This study aimed to understand frontline PCPs' perspectives on providing hybrid virtual and in-person depression care during the pandemic. From September to November 2020, 12 semistructured individual interviews focused on depression management were conducted with PCPs in two Veterans Health Administration (VA) clinics in Los Angeles, which resumed in-person services while balancing rising COVID-19 cases. Interviews were audio-recorded, transcribed, and coded for depression management patterns. Themes were derived using a team-based constant comparative analytic approach. The pandemic and subsequent expanded use of virtual care necessitated clinic adaptations to depression assessments and procedures. PCPs perceived increased depression and anxiety among patients with existing psychiatric conditions, attributed to social distancing and isolation restrictions. They expressed acceptance of virtual care modalities for patients' depression management. PCPs did not perceive a delay in mental health care delivery in the shift to virtual care but noted the possibility of patients being lost to follow-up. During the pandemic, there has been heightened PCP concern for patients' emotional well-being and adaptations of clinic processes to meet needs for depression care. While PCPs were optimistic about new virtual care options for depression management, virtual care transfers remained poorly defined and the extent to which patient care experiences and health outcomes have been disrupted remains unknown. (PsycInfo Database Record (c) 2023 APA, all rights reserved).


SIRT2 deficiency modulates macrophage polarization and susceptibility to experimental colitis.
SIRT2 belongs to a highly conserved family of NAD+-dependent deacylases, consisting of seven members (SIRT1-SIRT7), which vary in subcellular localizations and have substrates ranging from histones to transcription factors and enzymes. Recently SIRT2 was revealed to play an important role in inflammation, directly binding, deacetylating, and inhibiting the p65 subunit of NF-κB. A Sirt2 deficient mouse line (Sirt2-/-) was generated by deleting exons 5-7, encoding part of the SIRT2 deacetylase domain, by homologous recombination. Age- and sex-matched Sirt2-/- and Sirt2+/+ littermate mice were subjected to dextran sulfate sodium (DSS)-induced colitis and analyzed for colitis susceptibility. Sirt2-/- mice displayed more severe clinical and histological manifestations after DSS colitis compared to wild type littermates. Notably, under basal condition, Sirt2 deficiency does not affect the basal phenotype and intestinal morphology Sirt2 deficiency, however, affects macrophage polarization, creating a pro-inflammatory milieu in the immune cells compartment. These data confirm a protective role for SIRT2 against the development of inflammatory processes, pointing out a potential role for this sirtuin as a suppressor of colitis. In fact, SIRT2 deletion promotes inflammatory responses by increasing NF-κB acetylation and by reducing the M2-associated anti-inflammatory pathway. Finally, we speculate that the activation of SIRT2 may be a potential approach for the treatment of inflammatory bowel disease.


Familial testicular cancer and urogenital developmental anomalies.
In a case-control study of testicular cancer, 6 of 269 cases (2.2%) reported a first-degree relative with testicular cancer, compared to 1 of 259 controls (0.4%). Fathers and brothers of testicular cancer cases had a six-fold elevated risk of developing a testicular malignancy compared to men in the general population. Cryptorchidism was reported in a first-degree relative in 1 (17%) of the familial cases versus 7 of 259 (2.7%) controls and 14 of 263 (5.3%) cases with a negative family history for testicular cancer. One half of the 6 familial cases reported a first-degree relative with a groin hernia (all surgically repaired before age 12), compared to 12.7% of 259 controls and 10.3% of 263 nonfamilial cases. Three familial clusters identified through the case-control study were selected for clinical evaluation. One of the 6 surviving males with testicular cancer in these 3 families had undergone orchiopexy and inguinal herniorrhaphy at age 6 years, and one had a hydrocele associated with his testicular tumor. Of the 12 living fathers and brothers of these 6 men, 3 reported childhood inguinal hernias, two with coexisting hydroceles. One additional hernia and two additional hydroceles were detected during urologic evaluation of these healthy relatives. The high prevalence of cryptorchidism, inguinal hernias, and hydroceles among men in these families suggests that an underlying alteration in urogenital embryogenesis may be associated with the familial predisposition to testicular neoplasia.


Rheumatoid arthritis susceptibility and interleukin 10: a study of two ethnically diverse populations.
IL-10 is an immunoregulatory cytokine which may modulate disease expression in rheumatoid arthritis (RA). The IL-10 gene is highly polymorphic with a number of single nucleotide polymorphisms in the promoter region and two microsatellite loci, IL10.R and IL10.G, 4 kb and 1.1 kb 5' of the transcription initiation site. It has been reported that allele 2 of the IL10.R microsatellite (IL10.R2) is associated with increased IL-10 secretion and IL10.R3 with reduced secretion. Subsequently, over-representation of IL10.R2 and under-representation of IL10.R3 in three independent RA groups has been reported. The aim of the current study is to determine whether there is an association between the IL10.R2 allele and RA in two ethnically distinct populations. IL10.R genotypes were determined by semi-automated DNA sequencing technology in 186 UK Caucasians and 138 South Africans of Zulu or Sotho origin, fulfilling the 1987 American College of Rheumatology (ACR) criteria for RA. The Caucasian patients had relatively severe disease and comprised 75 patients with RA vasculitis, 22 with Felty's syndrome and 89 who had undergone a joint replacement (hip or knee) within 15 years of the onset of disease. Allele frequencies were compared with 296 Caucasians and/or 73 South Africans. The frequency of the IL10.R2 allele was significantly greater in the South Africans (RA and controls) than in the Caucasians (0.78 vs 0.66, P=1 x 10(-6)), while the frequency of IL10.R3 was less common (0.16 vs 0.3, P=1 x 10(-8)). No differences were observed in either IL10.R2 or IL10.R3 frequencies between patients and controls in either population. We were unable to confirm any association between IL10.R alleles and RA in this study. However, significant differences were demonstrated in the frequency of IL10.R2 and IL10.R3 between the two ethnic groups. The relatively high frequency of IL10.R2 in the South African population (0.78) would have reduced the power to detect an association with RA.