Species specificity in avian sperm:perivitelline interaction.
The interaction of chicken spermatozoa with the inner perivitelline layer from different avian species in vitro during a 5 min co-incubation was measured as the number of points of hydrolysis produced per unit area of inner perivitelline layer. The average degree of interaction, as a proportion of that between chicken spermatozoa and their homologous inner perivitelline layer, was: equal to or greater than 100% within Galliformes (chicken, turkey, quail, pheasant, peafowl and guineafowl); 44% within Anseriformes (goose, duck); and less than 30% in Passeriformes (Zebra Finch) and Columbiformes (collared-dove). The homologue of the putative chicken sperm-binding proteins, chicken ZP1 and ZP3, were identified by Western blotting with anti-chicken ZP1/ZP3 antibody in the perivitelline layers of all species. The functional cross-reactivity between chicken spermatozoa and heterologous inner perivitelline layer appeared to be linked to known phylogenetic distance between the species, although it was not related to the relative affinity of the different ZP3 homologues for anti-chicken ZP3. This work demonstrates that sperm interaction with the egg investment does not represent such a stringent species-specific barrier in birds as it does in mammals and marine invertebrates. This may be a factor in the frequency of hybrid production in birds.


Medication prescribing practices for older prisoners in the Texas prison system.
We sought to assess appropriateness of medication prescribing for older Texas prisoners. In this 12-month cross-sectional study of 13 117 prisoners (aged > or = 55 years), we assessed medication use with Zhan criteria and compared our results to prior studies of community prescribing. We assessed use of indicated medications with 6 Assessing Care of Vulnerable Elders indicators. Inappropriate medications were prescribed to a third of older prisoners; half of inappropriate use was attributable to over-the-counter antihistamines. When these antihistamines were excluded, inappropriate use dropped to 14% (> or = 55 years) and 17% (> or = 65 years), equivalent to rates in a Department of Veterans Affairs study (17%) and lower than rates in a health maintenance organization study (26%). Median rate of indicated medication use for the 6 indicators was 80% (range = 12%-95%); gastrointestinal prophylaxis for patients on nonsteroidal anti-inflammatories at high risk for gastrointestinal bleed constituted the lowest rate. Medication prescribing for older prisoners in Texas was similar to that for older community adults. However, overuse of antihistamines and underuse of gastrointestinal prophylaxis suggests a need for education of prison health care providers in appropriate prescribing practices for older adults.


Transcriptomic regulations in oligodendroglial and microglial cells related to brain damage following fetal growth restriction.
Fetal growth restriction (FGR) is a major complication of human pregnancy, frequently resulting from placental vascular diseases and prenatal malnutrition, and is associated with adverse neurocognitive outcomes throughout life. However, the mechanisms linking poor fetal growth and neurocognitive impairment are unclear. Here, we aimed to correlate changes in gene expression induced by FGR in rats and abnormal cerebral white matter maturation, brain microstructure, and cortical connectivity in vivo. We investigated a model of FGR induced by low-protein-diet malnutrition between embryonic day 0 and birth using an interdisciplinary approach combining advanced brain imaging, in vivo connectivity, microarray analysis of sorted oligodendroglial and microglial cells and histology. We show that myelination and brain function are both significantly altered in our model of FGR. These alterations, detected first in the white matter on magnetic resonance imaging significantly reduced cortical connectivity as assessed by ultrafast ultrasound imaging. Fetal growth retardation was found associated with white matter dysmaturation as shown by the immunohistochemical profiles and microarrays analyses. Strikingly, transcriptomic and gene network analyses reveal not only a myelination deficit in growth-restricted pups, but also the extensive deregulation of genes controlling neuroinflammation and the cell cycle in both oligodendrocytes and microglia. Our findings shed new light on the cellular and gene regulatory mechanisms mediating brain structural and functional defects in malnutrition-induced FGR, and suggest, for the first time, a neuroinflammatory basis for the poor neurocognitive outcome observed in growth-restricted human infants. GLIA 2016;64:2306-2320.


Mass spectrometry-based assay for the rapid detection of thiol-containing natural products.
Natural products are privileged scaffolds due to their high propensity to possess bioactivity. To expedite discovery of thiol-containing compounds, we devised a selective solid-supported reagent for their immobilization, followed by cleavage of a photocleavable linker to yield stable natural product conjugates for direct detection by mass spectrometry. Importantly, the natural products can also be tracelessly released to yield the native structures for chemical and biological evaluation.


Anticonvulsant mechanisms of piperine, a piperidine alkaloid.
Piperine, a natural compound isolated from the fruits of Piper, is known to modulate several neurotransmitter systems such as serotonin, norepinephrine, and GABA, all of which have been linked to the development of convulsions. Fruits of Piper species have been suggested as means for managing seizure disorders. The present study was designed to elucidate the anticonvulsant effect of piperine and its mechanisms of action using in-silico, in-vivo and in-vitro techniques.PASS software was used to determine its possible activity and mechanisms. Furthermore the latency for development of convulsions and mortality rate was recorded in different experimental mouse models of epilepsy (pentylenetetrazole, maximal electroshock, NMDA, picrotoxin, bicuculline, BAYK-8644, strychnine-induced convulsions) after administration of various doses of piperine (5, 10 and 20 mg/kg, i.p.). Finally, the effect of piperine on Na(+) and Ca(2+) channels were evaluated using the whole cell patch clamp techniqueOur results revealed that piperine decreased mortality in the MES-induced seizure model. Moreover, piperine (10 mg/kg) delayed the onset of tonic clonic convulsions in the pentylenetetrazole test and reduced associated mortality. Furthermore, an anticonvulsant dose of piperine also delayed the onset of tonic clonic seizures in strychnine, picrotoxin and BAY K-8644. Complete protection against mortality was observed in BAYK-8644 induced convulsions. Finally, whole cell patch clamp analysis suggested an inhibitory effect of piperine on Na(+) channels. Together, our data suggest Na(+) channel antagonist activity as a contributor to the complex anticonvulsant mechanisms of piperine.


Merosin-negative congenital muscular dystrophy: diffusion-weighted imaging findings of brain.
Merosin-negative congenital muscular dystrophy is a rare genetic disease of childhood involving the central and peripheral nervous system. There were high signal intensities throughout the centrum semiovale, periventricular, and sub-cortical white matters on T2-weighted images in a 4-year-old girl with merosin-negative congenital muscular dystrophy. An apparent diffusion coefficient map revealed increased signal intensity and apparent diffusion coefficient values in the periventricular and deep white matters. It may be attributable to increased water content in the white matter because of an abnormal blood-brain barrier rather than to decreased or abnormal myelination.


The interaction between impulsivity and neighborhood context on offending: the effects of impulsivity are stronger in poorer neighborhoods.
This research blends 2 traditions of theorizing on the causes of crime, one focused on the role of individual differences and the other focused on structural and contextual variables. Two related studies examined the relations among impulsivity, neighborhood context, and juvenile offending. The first, cross-sectional study uses a large sample of 13-year-old inner-city boys, whereas the second, longitudinal study offers a conceptual replication using 17-year-old inner-city boys who are a subset of the original sample. Across both studies, results indicate that the effects of impulsivity on juvenile offending are stronger in poorer neighborhoods. Furthermore, nonimpulsive boys in poor neighborhoods were at no greater risk for delinquency than nonimpulsive boys in better-off neighborhoods.


Genome-wide functional element detection using pairwise statistical alignment outperforms multiple genome footprinting techniques.
Comparative genomic sequence analysis is a powerful approach for identifying putative functional elements in silico. The availability of full-genome sequences from many vertebrate species has resulted in the development of popular tools, for example, the phastCons software package that search large numbers of genomes to identify conserved elements. While phastCons can analyze many genomes simultaneously, it ignores potentially informative insertion and deletion events and relies on a fixed, precomputed multiple sequence alignment. We have developed a new method, GRAPeFoot, which simultaneously aligns two full genomes and annotates a set of conserved regions exhibiting reduced rates of insertion, deletion and substitution mutations. We tested GRAPeFoot using the human and mouse genomes and compared its performance to a set of phastCons predictions hosted on the UCSC genome browser. Our results demonstrate that despite the use of only two genomes, GRAPeFoot identified constrained elements at rates comparable with phastCons, which analyzed data from 28 vertebrate genomes. This study demonstrates how integrated modelling of substitutions, indels and purifying selection allows a pairwise analysis to exhibit a sensitivity similar to a heuristic analysis of many genomes. The GRAPeFoot software and set of genome-wide functional element predictions are freely available to download online at http://www.stats.ox.ac.uk/ approximately satija/GRAPeFoot/.


Alcohol consumption over time and risk of death: a systematic review and meta-analysis.
The results from the few cohort studies that have measured usual alcohol consumption over time have not been summarized. We therefore conducted a systematic review and meta-analysis to quantify mortality risk. Pertinent studies were identified by searching the Medline, Web of Science, Cumulative Index to Nursing and Allied Health Literature (CINAHL) Plus, and Scopus databases through August 2012 using broad search criteria. Studies reporting relative mortality risks for quantitatively defined categories of alcohol consumption over time were eligible. Nine cohort studies published during 1991-2010 (comprising 62,950 participants and 10,490 deaths) met the inclusion criteria. For men, there was weak evidence of lower mortality risk with low levels of alcohol intake over time but higher mortality risk for those with intakes over 40 g/day compared with abstainers using a random-effects model (P for nonlinearity = 0.02). The pooled relative risks were 0.90 (95% confidence interval: 0.81, 0.99) for 1-29 g/day, 1.19 (95% confidence interval: 0.89, 1.58) for 30-59 g/day, and 1.52 (95% confidence interval: 0.78, 2.98) for 60 or more g/day compared with abstention. There was moderate between-study heterogeneity but no evidence of publication bias. Studies including women were extremely scarce. Our findings include a curvilinear association between drinking over time and mortality risk for men overall and widespread disparity in methods used to capture exposure and report results.


Segregation of ferritin in glomerular protein absorption droplets.
Ferritin was used as a tracer to study the mechanism by which proteins are segregated into droplets by the visceral epithelium of glomerular capillaries. In glomeruli from both normal and aminonucleoside-nephrotic rats ferritin molecules introduced into the general circulation penetrated the endothelial openings and were seen at various levels in the basement membrane. Striking differences between nephrotic and controls were seen only in the amount of ferritin incorporated into the epithelium. In normal animals, a few ferritin molecules were seen in small invaginations of the cell membrane limiting the foot processes, within minute vesicles in the epithelium, or within occasional large vacuoles and dense bodies. In nephrotics, epithelial pinocytosis was marked, and numerous ferritin molecules were seen within membrane invaginations and in small cytoplasmic vesicles at all time points. After longer intervals, the concentration of ferritin increased in vacuoles and particularly within the dense bodies or within structures with a morphology intermediate between that of vacuoles and dense bodies. In nephrotic animals cleft-like cavities or sinuses were frequently encountered along the epithelial cell surface facing the urinary spaces. Some of these sinuses contained material resembling that filling the dense bodies except that it appeared less compact. The findings suggest that ferritin molecules-and presumably other proteins which penetrate the basement membrane-are picked up by the epithelium in pinocytotic vesicles and transported via the small vesicles to larger vacuoles which are subsequently transformed into dense bodies by progressive condensation. The content of the dense bodies may then undergo partial digestion and be extruded into the urinary spaces where it disperses. The activity of the glomerular epithelium in the incorporation and segregation of protein is similar in normal and nephrotic animals, except that the rate is considerably higher in nephrosis where the permeability of the glomerular basement membrane is greatly increased.


The patch transform.
The patch transform represents an image as a bag of overlapping patches sampled on a regular grid. This representation allows users to manipulate images in the patch domain, which then seeds the inverse patch transform to synthesize modified images. Possible modifications include the spatial locations of patches, the size of the output image, or the pool of patches from which an image is reconstructed. When no modifications are made, the inverse patch transform reduces to solving a jigsaw puzzle. The inverse patch transform is posed as a patch assignment problem on a Markov random field (MRF), where each patch should be used only once and neighboring patches should fit to form a plausible image. We find an approximate solution to the MRF using loopy belief propagation, introducing an approximation that encourages the solution to use each patch only once. The image reconstruction algorithm scales well with the total number of patches through label pruning. In addition, structural misalignment artifacts are suppressed through a patch jittering scheme that spatially jitters the assigned patches. We demonstrate the patch transform and its effectiveness on natural images.


Both cyclin A and cyclin E have S-phase promoting (SPF) activity in Xenopus egg extracts.
Extracts of activated Xenopus eggs in which protein synthesis has been inhibited support a single round of chromosomal DNA replication. Affinity-depletion of cyclin dependent kinases (Cdks) from these extracts blocks the initiation of DNA replication. We define 'S-phase promoting factor' (SPF) as the Cdk activity required for DNA replication in these Cdk-depleted extracts. Recombinant cyclins A and E, but not cyclin B, showed significant SPF activity. High concentrations of cyclin A promoted entry into mitosis, which inhibited DNA replication. In contrast, high concentrations of cyclin E1 promoted neither nuclear envelope disassembly nor full chromosome condensation. In the early embryo cyclin E1 complexes exclusively with Cdk2 and cyclin A is complexed predominantly with Cdc2; only later in development does cyclin A associate with Cdk2. We show that baculovirus-produced complexes of cyclin A-Cd2, cyclin A-Cdk2 and cyclin E-Cdk2 could each provide SPF activity. These results suggest that although in the early Xenopus embryo cyclin E1-Cdk2 is sufficient to support entry into S-phase, cyclin A-Cdc2 provides a significant additional quantity of SPF as its levels rise during S phase.


Bark- and wood-borer colonization of logs and lumber after heat treatment to ISPM 15 specifications: the role of residual bark.
Wood packaging material (WPM) is a major pathway for international movement of bark- and wood-infesting insects. ISPM 15, the first international standard for treating WPM, was adopted in 2002 and first implemented in the United States in 2006. ISPM 15 allows bark to remain on WPM after treatment, raising concerns that insects could infest after treatment, especially if bark were present. We conducted field studies to evaluate insect infestation of green logs and lumber with varying amounts of bark after heat treatment. In a log study, Cerambycidae and Scolytinae (ambrosia beetles and bark beetles) readily infested and developed in logs with bark after heat treatment. In a lumber study, Cerambycidae and bark beetles laid eggs in all sizes of bark patches tested (approximately 25, 100, 250, and 1,000 cm2) after heat treatment but did not infest control or heat-treated lumber without bark. Cerambycidae completed development only in boards with bark patches of 1,000 cm2, whereas bark beetles completed development on patches of 100, 250, and 1,000 cm2. Survival of bark beetles was greater in square patches (10 by 10 cm) versus rectangular patches (2.5 by 40 cm) of the same surface area (100 cm2). In surveys at six U.S. ports in 2006, 9.4% of 5,945 ISPM 15-marked WPM items contained bark, and 1.2% of 564 ISPM 15-marked WPM items with bark contained live insects of quarantine significance under the bark. It was not possible to determine whether the presence of live insects represented treatment failure or infestation after treatment.


Emergence, Retention and Selection: A Trilogy of Origination for Functional De Novo Proteins from Ancestral LncRNAs in Primates.
While some human-specific protein-coding genes have been proposed to originate from ancestral lncRNAs, the transition process remains poorly understood. Here we identified 64 hominoid-specific de novo genes and report a mechanism for the origination of functional de novo proteins from ancestral lncRNAs with precise splicing structures and specific tissue expression profiles. Whole-genome sequencing of dozens of rhesus macaque animals revealed that these lncRNAs are generally not more selectively constrained than other lncRNA loci. The existence of these newly-originated de novo proteins is also not beyond anticipation under neutral expectation, as they generally have longer theoretical lifespan than their current age, due to their GC-rich sequence property enabling stable ORFs with lower chance of non-sense mutations. Interestingly, although the emergence and retention of these de novo genes are likely driven by neutral forces, population genetics study in 67 human individuals and 82 macaque animals revealed signatures of purifying selection on these genes specifically in human population, indicating a proportion of these newly-originated proteins are already functional in human. We thus propose a mechanism for creation of functional de novo proteins from ancestral lncRNAs during the primate evolution, which may contribute to human-specific genetic novelties by taking advantage of existed genomic contexts.


Kinetics of 1H-13C multiple-contact cross-polarization as a powerful tool to determine the structure and dynamics of complex materials: application to graphene oxide.
Hartmann-Hahn cross-polarization (HHCP) is the most widely used solid-state NMR technique to enhance the magnetization of dilute spins from abundant spins. Furthermore, as the kinetics of CP depends on dipolar interactions, it contains valuable information on molecular structure and dynamics. In this work, analytical solutions are derived for the kinetics of HHCP and multiple-contact CP (MC-CP) using both classical and non-classical spin-coupling models including the effects of molecular dynamics and several 1H, 13C relaxation and 1H-13C CP experiments are performed in graphene oxide (GO). HHCP is found to be inefficient in our GO sample due to very fast 1H T1ρ relaxation. By contrast, the MC-CP technique which alleviates most of the magnetization loss by 1H T1ρ relaxation leads to a much larger polarization transfer efficiency reducing the measuring time by an order of magnitude. A detailed analysis of the HHCP and MC-CP kinetics indicates the existence of at least two different kinds of hydroxyl (C-OH) functional groups in GO, the major fraction (∼90%) of these groups being in the unusual "slow CP regime" in which the rate of 1H T1ρ relaxation is fast compared to the rate of cross-polarization. This 13C signal component is attributed to mobile C-OH groups interacting preferentially with fast-relaxing water molecules while the remaining carbons (∼10%) in the usual "fast CP regime" are assigned to C-OH groups involved in hydrogen bonding with neighboring hydroxyl and/or epoxy groups.


Trends in domiciliary dental care including the need for oral appliances and dental technicians in Japan.
The need for domiciliary dental care (DDC) for people requiring long-term nursing care is increasing as the super-aged society of Japan grows still older. Dysphagia diagnosis and rehabilitation are becoming more important in DDC; thus, the need for prostheses used for dysphasia rehabilitation is presumed to be increasing. To identify DDC trends in Japan, as well as the need for prostheses and dental technicians for DDC, we sent a self-administered questionnaire to dentists providing DDC and analyzed responses from 138 dentists (valid response rate, 39.8%). The results showed that 37.7% of respondents reported treating ≥50 patients per month. The most frequently performed procedures were removable prosthetic treatment and oral care, followed by dysphagia rehabilitation. Use of palatal augmentation prostheses was experienced by 54.3% of respondents, and most indicated that the prostheses were effective for improvement of oropharyngeal function. The rates of cooperation with primary care doctors and nursing care professionals were 76.8% and 85.5%, respectively. Only 6.5% of respondents reported accompanying dental technicians to DDC. The present analysis of trends in DDC indicates that oral care and dysphagia rehabilitation have become more frequent and that cooperation with healthcare professionals other than dental technicians has increased in recent DDC.


Mortality and morbidity in diabetic and nondiabetic patients during a 2-year period after coronary artery bypass grafting.
To describe mortality and morbidity during a 2-year period after coronary artery bypass grafting (CABG) among diabetic and nondiabetic patients. All the patients in western Sweden in whom CABG was undertaken between June 1988 and June 1991 and in whom concomitant procedures were not performed were registered prospectively. The study was a prospective follow-up. Diabetic patients (n = 268) differed from nondiabetic patients (n = 1,859) in that more women were included, and the patients more frequently had a previous history of myocardial infarction (MI), hypertension, congestive heart failure, intermittent claudication, and obesity. Diabetic patients more frequently required reoperation and had a higher incidence of peri- and postoperative neurological complications. Mortality during the 30 days after CABG was 6.7% in diabetic patients versus 3.0% in nondiabetic patients (P < 0.01). Mortality between day 30 and 2 years was 7.8 and 3.6%, respectively (P < 0.01). During 2 years of follow-up, a history of diabetes appeared to be a significant independent predictor of death. Whereas the development of MI after discharge from the hospital did not significantly differ between the two groups; 6.3% of diabetic patients developed stroke versus 2.5% in nondiabetic patients (P < 0.001). Diabetic patients have a mortality rate during the 2-year period after CABG that is about twice that of nondiabetic patients during both the early and late phase after the operation.


Predicting time-resolved electrophysiological brain networks from structural eigenmodes.
How temporal modulations in functional interactions are shaped by the underlying anatomical connections remains an open question. Here, we analyse the role of structural eigenmodes, in the formation and dissolution of temporally evolving functional brain networks using resting-state magnetoencephalography and diffusion magnetic resonance imaging data at the individual subject level. Our results show that even at short timescales, phase and amplitude connectivity can partly be expressed by structural eigenmodes, but hardly by direct structural connections. Albeit a stronger relationship was found between structural eigenmodes and time-resolved amplitude connectivity. Time-resolved connectivity for both phase and amplitude was mostly characterised by a stationary process, superimposed with very brief periods that showed deviations from this stationary process. For these brief periods, dynamic network states were extracted that showed different expressions of eigenmodes. Furthermore, the eigenmode expression was related to overall cognitive performance and co-occurred with fluctuations in community structure of functional networks. These results implicate that ongoing time-resolved resting-state networks, even at short timescales, can to some extent be understood in terms of activation and deactivation of structural eigenmodes and that these eigenmodes play a role in the dynamic integration and segregation of information across the cortex, subserving cognitive functions.


Robust Linear Models for Cis-eQTL Analysis.
Expression Quantitative Trait Loci (eQTL) analysis enables characterisation of functional genetic variation influencing expression levels of individual genes. In outbread populations, including humans, eQTLs are commonly analysed using the conventional linear model, adjusting for relevant covariates, assuming an allelic dosage model and a Gaussian error term. However, gene expression data generally have noise that induces heavy-tailed errors relative to the Gaussian distribution and often include atypical observations, or outliers. Such departures from modelling assumptions can lead to an increased rate of type II errors (false negatives), and to some extent also type I errors (false positives). Careful model checking can reduce the risk of type-I errors but often not type II errors, since it is generally too time-consuming to carefully check all models with a non-significant effect in large-scale and genome-wide studies. Here we propose the application of a robust linear model for eQTL analysis to reduce adverse effects of deviations from the assumption of Gaussian residuals. We present results from a simulation study as well as results from the analysis of real eQTL data sets. Our findings suggest that in many situations robust models have the potential to provide more reliable eQTL results compared to conventional linear models, particularly in respect to reducing type II errors due to non-Gaussian noise. Post-genomic data, such as that generated in genome-wide eQTL studies, are often noisy and frequently contain atypical observations. Robust statistical models have the potential to provide more reliable results and increased statistical power under non-Gaussian conditions. The results presented here suggest that robust models should be considered routinely alongside other commonly used methodologies for eQTL analysis.


Are Working Memory Training Effects Paradigm-Specific?
A randomized controlled trial compared complex span and n-back training regimes to investigate the generality of training benefits across materials and paradigms. The memory items and training intensities were equated across programs, providing the first like-with-like comparison of transfer in these two widely used training paradigms. The stimuli in transfer tests of verbal and visuo-spatial n-back and complex span differed from the trained tasks, but were matched across the untrained paradigms. Participants were randomly assigned to one of three training groups: complex span training, n-back training, or no training. Pre- to- post training changes were observed for untrained n-back tasks following n-back training. Following complex span training there was equivocal evidence for improvements on a verbal complex span task, but no evidence for changes on an untrained visuo-spatial complex span activity. Relative to a no intervention group, the evidence supported no change on an untrained verbal complex span task following either n-back or complex span training. Equivocal evidence was found for improvements on visuo-spatial complex span and verbal and visuo-spatial n-back tasks following both training regimes. Evidence for selective transfer (comparing the two active training groups) was only found for an untrained visuo-spatial n-back task following n-back training. There was no evidence for cross-paradigm transfer. Thus transfer is constrained by working memory paradigm and the nature of individual processes executed within complex span tasks. However, within-paradigm transfer can occur when the change is limited to stimulus category, at least for n-back.


DNA specificities modulate the binding of human transcription factor A to mitochondrial DNA control region.
Human mitochondrial DNA (h-mtDNA) codes for 13 subunits of the oxidative phosphorylation pathway, the essential route that produces ATP. H-mtDNA transcription and replication depends on the transcription factor TFAM, which also maintains and compacts this genome. It is well-established that TFAM activates the mtDNA promoters LSP and HSP1 at the mtDNA control region where DNA regulatory elements cluster. Previous studies identified still uncharacterized, additional binding sites at the control region downstream from and slightly similar to LSP, namely sequences X and Y (Site-X and Site-Y) (Fisher et al., Cell 50, pp 247-258, 1987). Here, we explore TFAM binding at these two sites and compare them to LSP by multiple experimental and in silico methods. Our results show that TFAM binding is strongly modulated by the sequence-dependent properties of Site-X, Site-Y and LSP. The high binding versatility of Site-Y or the considerable stiffness of Site-X tune TFAM interactions. In addition, we show that increase in TFAM/DNA complex concentration induces multimerization, which at a very high concentration triggers disruption of preformed complexes. Therefore, our results suggest that mtDNA sequences induce non-uniform TFAM binding and, consequently, direct an uneven distribution of TFAM aggregation sites during the essential process of mtDNA compaction.


The effect of stance width on the electromyographical activity of eight superficial thigh muscles during back squat with different bar loads.
Many strength trainers believe that varying the stance width during the back squat can target specific muscles of the thigh. The aim of the present work was to test this theory measuring the activation of 8 thigh muscles while performing back squats at 3 stance widths and with 3 different bar loads. Six experienced lifters performed 3 sets of 10 repetitions of squats, each one with a different stance width, using 3 resistances: no load, 30% of 1-repetition maximum (1RM), and 70% 1RM. Sets were separated by 6 minutes of rest. Electromyographic (EMG) surface electrodes were placed on the vastus medialis, vastus lateralis, rectus femoris, semitendinosus, biceps femoris, gluteus maximus, gluteus medium, and adductor maior. Analysis of variance and Scheffè post hoc tests indicated a significant difference in EMG activity only for the gluteus maximus; in particular, there was a higher electrical activity of this muscle when back squats were performed at the maximum stance widths at 0 and 70% 1RM. There were no significant differences concerning the EMG activity of the other analyzed muscles. These findings suggest that a large width is necessary for a greater activation of the gluteus maximus during back squats.


BDNF evokes release of endogenous cannabinoids at layer 2/3 inhibitory synapses in the neocortex.
The neurotrophin brain-derived neurotrophic factor (BDNF) is a potent regulator of inhibitory synaptic transmission, although the locus of this effect and the underlying mechanisms are controversial. We explored a potential interaction between BDNF and endogenous cannabinoid (endocannabinoid) signaling because activation of type 1 cannabinoid (CB1) receptors potently regulates γ-aminobutyric acid (GABA) release and both trkB tyrosine kinase receptors and CB1 receptors are highly expressed at synapses in neocortical layer 2/3. Here, we found that the effects of BDNF at inhibitory cortical synapses are mediated by the release of endocannabinoids acting retrogradely at presynaptic CB1 receptors. Specifically, acute application of BDNF rapidly reduced the amplitude of inhibitory postsynaptic currents (IPSCs) via postsynaptic trkB receptor activation because intracellular delivery of the tyrosine kinase inhibitor K252a completely blocked the BDNF effect. Although triggered by postsynaptic trkB activation, BDNF exposure decreased presynaptic release probability, as evidenced by increases in the paired-pulse ratio and coefficient of variation of evoked responses. In addition, BDNF decreased the frequency but not the amplitude of action potential-independent miniature IPSCs and BDNF did not alter the postsynaptic response to locally applied GABA. These results suggest that BDNF induces the release of a retrograde messenger from the postsynaptic cell that regulates presynaptic neurotransmitter release. Consistent with a role for endocannabinoids as the retrograde signal, the effect of BDNF on IPSCs was blocked by CB1 receptor antagonists and was occluded by a cannabinoid receptor agonist. Furthermore, inhibiting endocannabinoid synthesis or transport also disrupted the BDNF effect, implicating postsynaptic endocannabinoid release triggered by BDNF.


Courses of helping alliance in the treatment of people with severe mental illness in Europe: a latent class analytic approach.
The helping alliance (HA) between patient and therapist has been studied in detail in psychotherapy research, but less is known about the HA in long-term community mental health care. The aim of this study was to identify typical courses of the HA and their predictors in a sample of people with severe mental illness across Europe over a measurement period of one year. Self-ratings of the HA by 588 people with severe mental illness who participated in a multicentre European study (CEDAR; ISRCTN75841675) were examined using latent class analysis. Four main patterns of alliance were identified: (1) high and stable (HS, 45.6 %), (2) high and increasing (HI, 36.9 %), (3) high and decreasing (HD, 11.3 %) and (4) low and increasing (LI, 6.1 %). Predictors of class membership were duration of illness, ethnicity, and education, receipt of state benefits, recovery, and quality of life. Results support findings from psychotherapy research about a predominantly stable course of the helping alliance in patients with severe mental illness over time. Implications for research and practice indicate to turn the attention to subgroups with noticeable courses.


Selective Targeting of Human and Animal Pathogens of the Helicobacter Genus by Flavodoxin Inhibitors: Efficacy, Synergy, Resistance and Mechanistic Studies.
Antimicrobial resistant (AMR) bacteria constitute a global health concern. Helicobacter pylori is a Gram-negative bacterium that infects about half of the human population and is a major cause of peptic ulcer disease and gastric cancer. Increasing resistance to triple and quadruple H. pylori eradication therapies poses great challenges and urges the development of novel, ideally narrow spectrum, antimicrobials targeting H. pylori. Here, we describe the antimicrobial spectrum of a family of nitrobenzoxadiazol-based antimicrobials initially discovered as inhibitors of flavodoxin: an essential H. pylori protein. Two groups of inhibitors are described. One group is formed by narrow-spectrum compounds, highly specific for H. pylori, but ineffective against enterohepatic Helicobacter species and other Gram-negative or Gram-positive bacteria. The second group includes extended-spectrum antimicrobials additionally targeting Gram-positive bacteria, the Gram-negative Campylobacter jejuni, and most Helicobacter species, but not affecting other Gram-negative pathogens. To identify the binding site of the inhibitors in the flavodoxin structure, several H. pylori-flavodoxin variants have been engineered and tested using isothermal titration calorimetry. An initial study of the inhibitors capacity to generate resistances and of their synergism with antimicrobials commonly used in H. pylori eradication therapies is described. The narrow-spectrum inhibitors, which are expected to affect the microbiota less dramatically than current antimicrobial drugs, offer an opportunity to develop new and specific H. pylori eradication combinations to deal with AMR in H. pylori. On the other hand, the extended-spectrum inhibitors constitute a new family of promising antimicrobials, with a potential use against AMR Gram-positive bacterial pathogens.


Great potential of a panel of multiple hMTH1, SPD, ITGA11 and COL11A1 markers for diagnosis of patients with non-small cell lung cancer.
Research on molecular mechanisms underlying the carcinogenesis of non-small cell lung cancer (NSCLC) may provide gene targets in critical pathways valuable for improving the efficacy of therapy and survival of patients with NSCLC. However, the molecular markers highly sensitive for the prognosis and treatment evaluation of NSCLC are not yet available. To explore candidates, we conducted an oligonucleotide microarray study with three pairs of NSCLC and normal lung tissue, and determined 8 differentially expressed genes including the Human MutT homologue (hMTH1), Surfactant protein D (SPD), Human hyaluronan binding protein 2 (HABP2), Crystalline-mu (CRYM), Ceruloplasmin (CP), Integrin alpha-11 subunit (ITGA11), Collagen type XI alpha I (COL11A1), and Lung-specific X protein (Lun X). Four lung cancer-related markers MUC-1, hTERT, hnRNP B1, and CK-19 were also incorporated for further analysis. The expression profiles of the twelve genes in seventy pairs of NSCLC tumor and normal lung tissue were then detected quantitatively by using membrane array and quantitative real-time PCR (qRT-PCR). The data of the membrane array and qRT-PCR were compared for consistency and the potential of these mRNA markers in clinical application. The results showed that membrane array and qRT-PCR obtained consistent data for the tested genes in both sensitivity and specificity (correlation coefficient 0.921, p<0.0001). For patients' clinicopathological characteristics, the overexpression of hMTH1, SPD, HABP 2, ITGA11, COL11A1, and CK-19 was significantly correlated with the pathological stage (p<0.05). In addition, the overexpression of hMTH1, SPD, ITGA11, and COL11A1 was correlated with lymph node metastasis and poor prognosis. This is the first report relating SPD to a prognosis marker for NSCLC. Moreover, the combined detection of these four mRNA markers by membrane array had a sensitivity of 89% and a specificity of 84% for NSCLC, significantly higher than these markers had achieved separately. In conclusion, we identified mRNA markers for NSCLC prognosis and therapy evaluation from differentially expressed genes determined by using micro-array. Further studies are needed to collect the data of the mRNA markers used in clinical practice.


Liraglutide increases FGF-21 activity and insulin sensitivity in high fat diet and adiponectin knockdown induced insulin resistance.
Liraglutide is a glucagon-like peptide-1 analogue that stimulates insulin secretion and improves β-cell function. However, it is not clear whether liraglutide achieves its glucose lowering effect only by its known effects or whether other as yet unknown mechanisms are involved. The aim of this study was to examine the effects of liraglutide on Fibroblast growth factor-21 (FGF-21) activity in High-fat diet (HFD) fed ApoE(-/-) mice with adiponectin (Acrp30) knockdown. HFD-fed ApoE(-/-) mice were treated with adenovirus vectors expressing shAcrp30 to produce insulin resistance. Hyperinsulinemic-euglycemic clamp studies were performed to evaluate insulin sensitivity of the mouse model. QRT-PCR and Western blot were used to measure the mRNA and protein expression of the target genes. The combination of HFD, ApoE deficiency, and hypoadiponectinemia resulted in an additive effect on insulin resistance. FGF-21 mRNA expressions in both liver and adipose tissues were significantly increased while FGF-21 receptor 1 (FGFR-1) and β-Klotho mRNA levels in adipose tissue, as well as FGFR-1-3 and β-Klotho mRNA levels in liver were significantly decreased in this model. Liraglutide treatment markedly improved insulin resistance and increased FGF-21 expression in liver and FGFR-3 in adipose tissue, restored β-Klotho mRNA expression in adipose tissue as well as FGFR-1-3, β-Klotho levels and phosphorylation of FGFR1 up to the levels observed in control mice in liver. Liraglutide treatment also further increased FGF-21 proteins in liver and plasma. In addition, as shown by hyperinsulinemic-euglycemic clamp, liraglutide treatment also markedly improved glucose metabolism and insulin sensitivity in these animals. These findings demonstrate an additive effect of HFD, ApoE deficiency, and adiponectin knockdown on insulin resistance and unveil that the regulation of glucose metabolism and insulin sensitivity by liraglutide may be partly mediated via increased FGF-21 and its receptors action.


Increasing educational inequalities in self-rated health in Brazil, 1998-2013.
The objectives of this study are to analyze the associations between educational levels and poor self-rated health (SRH) among adults in Brazil and to assess trends in the prevalence of poor self-rated health across educational groups between 1998 and 2013. Individual-level data came from the 1998, 2003 and 2008 Brazilian National Household Survey and the 2013 National Health Survey. We estimate prevalence rates of poor SRH by education. Using multivariable regressions, we assess the associations between educational levels and poor self-rated health. We use these regressions to predict the estimated ratios between the prevalence rates of those in low vs. high education in order to assess if relative changes in poor SRH have narrowed over time. Finally, we tested for statistically significant time trends in adult chronic disease inequalities by education. Results indicate a clear educational gradient in poor SRH. Prevalence ratios show that Brazilian adults with no education have levels of poor SRH that are 7 to 9 times higher than those with some college or more. The difference between those with lowest and highest education increased from 1998 to 2013. Compared to those with no education, there were increases in the prevalence of poor SRH among those with primary and secondary incomplete as well as among those with secondary complete in 2008 and 2013. In conclusion, there is a positive association between poor SRH and low education. Brazil has many social and geographic inequalities in health. Even though educational levels are increasing, there is no improvement in the general subjective health of Brazilians. Health inequalities by race and region highlight the need to improve the health of socially disadvantaged groups in Brazil. Addressing chronic conditions and mental health is needed to improve self-perceptions of health in Brazil as well.


Photoperiodic Reversibility of Diapause Induction in an Insect.
The diapause of the pink bollworm is under photoperiodic control. Diapause is prevented when the dark phases of the daily photoperiod are 8 to 10 hours in duration. If the dark period is extended to 12 hours, diapause is induced. Intercalation of 8- or 10-hour nights may reverse the diapause induction caused by exposure to photocycles having 12-hour dark phases. The 10-hour night was much more effective in reversing induction than the 8-hour night. The intensity of diapause, in part, appeared to be dependent on the previous photoperiodic experiences of the test animals.


Feasibility of SERS-Active Porous Ag Substrates for the Effective Detection of Pyrene in Water.
Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous pollutants that are typically released into the environment during the incomplete combustion of fossil fuels. Due to their relevant carcinogenicity, mutagenicity, and teratogenicity, it is urgent to develop sensitive and cost-effective strategies for monitoring them, especially in aqueous environments. Surface-enhanced Raman spectroscopy (SERS) can potentially be used as a reliable approach for this purpose, as it constitutes a valid alternative to traditional techniques, such as liquid and gas chromatography. Nevertheless, the development of an SERS-based platform for detection PAHs has so far been hindered by the poor adsorption of PAHs onto silver- and gold-based SERS-active substrates. To overcome this limitation, several research efforts have been directed towards the development of functionalized SERS substrates for the improvement of PAH adsorption. However, these strategies suffer from the interference that functionalizing molecules can produce in SERS detection. Herein, we demonstrate the feasibility of label-free detection of pyrene by using a highly porous 3D-SERS substrate produced by an inductively coupled plasma (ICP). Thanks to the coral-like nanopattern exhibited by our substrate, clear signals ascribable to pyrene molecules can be observed with a limit of detection of 23 nM. The observed performance can be attributed to the nanoporous character of our substrate, which combines a high density of hotspots and a certain capability of trapping molecules and favoring their adhesion to the Ag nanopattern. The obtained results demonstrate the potential of our substrates as a large-area, label-free SERS-based platform for chemical sensing and environmental control applications.


Measuring Inhibition and Cognitive Flexibility in Friedreich Ataxia.
Friedreich ataxia (FRDA) is an autosomal recessive neurodegenerative disorder with subtle impact on cognition. Inhibitory processes and cognitive flexibility were examined in FRDA by assessing the ability to suppress a predictable verbal response. We administered the Hayling Sentence Completion Test (HSCT), the Trail Making Test, and the Stroop Test to 43 individuals with FRDA and 42 gender- and age-matched control participants. There were no significant group differences in performance on the Stroop or Trail Making Test whereas significant impairment in cognitive flexibility including the ability to predict and inhibit a pre-potent response as measured in the HSCT was evident in individuals with FRDA. These deficits did not correlate with clinical characteristics of FRDA (age of disease onset, disease duration, number of guanine-adenine-adenine repeats on the shorter or larger FXN allele, or Friedreich Ataxia Rating Scale score), suggesting that such impairment may not be related to the disease process in a straightforward way. The observed specific impairment of inhibition and predictive capacity in individuals with FRDA on the HSCT task, in the absence of impairment in associated executive functions, supports cerebellar dysfunction in conjunction with disturbance to cortico-thalamo-cerebellar connectivity, perhaps via inability to access frontal areas necessary for successful task completion.


Anterior thigh composition measured using ultrasound imaging to quantify relative thickness of muscle and non-contractile tissue: a potential biomarker for musculoskeletal health.
This study aimed to use ultrasound imaging to provide objective data on the effects of ageing and gender on relative thickness of quadriceps muscle and non-contractile tissue thickness (subcutaneous fat, SF, combined with perimuscular fascia). In 136 healthy males and females (aged 18-90 years n = 63 aged 18-35 years; n = 73 aged 65-90) images of the anterior thigh (dominant) were taken in relaxed supine using B-mode ultrasound imaging. Thickness of muscle, SF and perimuscular fascia were measured, and percentage thickness of total anterior thigh thickness calculated. Independent t-tests compared groups. Correlation between tissue thickness and BMI was examined using Pearson's coefficient. Muscle thickness was: 39 ± 8 mm in young males, 29 ± 6 mm in females, 25 ± 4 mm in older males and 20 ± 5 mm in females. Percentage muscle to thigh thickness was greater in young participants (p = 0.001). Percentage SF and fascia was 17 ± 6% in young and 26 ± 8% in older males, 32 ± 7% in young and 44 ± 7% in older females. BMI was similar for age and correlated moderately with non-contractile tissue (r = 0.54; p < 0.001) and poorly with muscle (r = -0.01; p = 0.93). In conclusion, this novel application of ultrasound imaging as a simple and rapid means of assessing thigh composition (relative thickness of muscle and non-contractile tissue) may help inform health status, e.g. in older people at risk of frailty and loss of mobility, and aid monitoring effects of weight loss or gain, deconditioning and exercise.


Deaf students and their classroom communication: an evaluation of higher order categorical interactions among school and background characteristics.
This article investigated to what extent age, use of a cochlear implant, parental hearing status, and use of sign in the home determine language of instruction for profoundly deaf children. Categorical data from 8,325 profoundly deaf students from the 2008 Annual Survey of Deaf and Hard-of-Hearing Children and Youth were analyzed using chi-square automated interaction detector, a stepwise analytic procedure that allows the assessment of higher order interactions among categorical variables. Results indicated that all characteristics were significantly related to classroom communication modality. Although younger and older students demonstrated a different distribution of communication modality, for both younger and older students, cochlear implantation had the greatest effect on differentiating students into communication modalities, yielding greater gains in the speech-only category for implanted students. For all subgroups defined by age and implantation status, the use of sign at home further segregated the sample into communication modality subgroups, reducing the likelihood of speech only and increasing the placement of students into signing classroom settings. Implications for future research in the field of deaf education are discussed.


Soft drinks and in vitro dental erosion.
The purpose of this investigation was to determine to what extent the in vitro exposure of healthy teeth to various commonly consumed carbonated soft drinks may precipitate dental erosion. Forty-two healthy, extracted, previously unerupted human molars were weighed prior to, during, and after suspension in various sugared and diet or zero-calorie carbonated beverages for 20 days; the specimens were stored at room temperature while being stirred at 275 rpm. The percentage decrease in tooth weight from before to after exposure represented the weight loss due to enamel erosion; values in the experimental groups varied from 3.22% to 44.52% after 20 days' exposure. Data were subjected to analysis of variance and post hoc Scheffe testing at a level of α = 0.05. Nonsugared drinks (diet and zero-calorie) as a whole were more erosive than sugared beverages. A significant positive correlation was found between the amount of titratable acid and percentage of tooth erosion, while a significant negative correlation was revealed between the beverage pH and percentage of tooth erosion. No significant correlations were found between calcium or phosphate ion concentrations and the amount of erosion. It appears that enamel erosion is dependent on not only the beverage flow rate, pH, and amount of titratable acid, but also whether the soft drink is of the diet or zero-calorie variety, which reflects the type of artificial sweetener present.


Hair-loss epizootic in moose (Alces alces) associated with massive deer ked (Lipoptena cervi) infestation.
Deer keds (Lipoptena cervi) are blood-sucking flies in the family Hippoboscidae; moose (Alces alces) are their main host in Scandinavia. There are no detailed reports of the negative impacts of deer keds on moose. In 2006 and 2007, hunters in southeastern Norway and midwestern Sweden found several moose cadavers with severe alopecia; numerous moose had extensive hair loss. Between February 2006 and June 2007, materials from 23 moose were submitted for laboratory examination and large numbers of deer keds were found in the coat of most animals. The body condition of the moose varied but was poor in animals with severe alopecia. The findings of enormous numbers of deer keds in the coat of the majority of the affected animals and a consistent histologic image (acute to chronic, multifocal to coalescing, eosinophilic to lymphocytic dermatitis), concurrent with the absence of any other lesions, trace element deficiencies, or dermal infections which are known to cause alopecia, suggest that the hair-loss epizootic was linked to massive infestations with deer keds. The emergence of this hair-loss syndrome implies that the dynamics between parasite and host have been disrupted by a currently unknown environmental or ecological factor. A high moose density, combined with extraordinarily mild weather June 2006-June 2007 and a particularly long period with the absence of night-frost in autumn of 2006, may have been ideal for deer ked development, survival, and optimal host acquisition.


VOCs, pesticides, nitrate, and their mixtures in groundwater used for drinking water in the United States.
Samples of untreated groundwater from 1255 domestic drinking-water wells and 242 public supply wells were analyzed as part of the National Water-Quality Assessment Program of the U.S. Geological Survey between 1992 and 1999. Wells were sampled to define the regional quality of the groundwater resource and, thus, were distributed geographically across large aquifers, primarily in rural areas. For each sample, as many as 60 volatile organic compounds (VOCs), 83 pesticides, and nitrate were analyzed. On the basis of previous studies, nitrate concentrations as nitrogen > or = 3 mg/L were considered to have an anthropogenic origin. VOCs were detected more frequently (44%) than pesticides (38%) or anthropogenic nitrate (28%). Seventy percent of the samples contained at least one VOC, pesticide, or anthropogenic nitrate; 47% contained at least two compounds; and 33% contained at least three compounds. The combined concentrations of VOCs and pesticides ranged from about 0.001 to 100 microg/L, with a median of 0.02 microg/L. Water from about 12% of the wells contained one or more compounds that exceeded U.S. Environmental Protection Agency drinking-water standards or human health criteria, primarily because of nitrate concentrations exceeding the maximum contaminant level in domestic wells. A mixture is defined as a unique combination of two or more particular compounds, regardless of the presence of other compounds that may occur in the same sample. There were 100 mixtures (significantly associated with agricultural land use) that had a detection frequency between 2% and 19%. There were 302 mixtures (significantly associated with urban land use) that had a detection frequency between 1% and <2%. Only 14 compounds (seven VOCs, six pesticides, and nitrate) contributed over 95% of the detections in these 402 mixtures; however, most samples with these mixtures also contain a variety of other compounds.


Surface antigens of Plasmodium falciparum-infected erythrocytes as immune targets and malaria vaccine candidates.
Understanding the targets and mechanisms of human immunity to malaria caused by Plasmodium falciparum is crucial for advancing effective vaccines and developing tools for measuring immunity and exposure in populations. Acquired immunity to malaria predominantly targets the blood stage of infection when merozoites of Plasmodium spp. infect erythrocytes and replicate within them. During the intra-erythrocytic development of P. falciparum, numerous parasite-derived antigens are expressed on the surface of infected erythrocytes (IEs). These antigens enable P. falciparum-IEs to adhere in the vasculature and accumulate in multiple organs, which is a key process in the pathogenesis of disease. IE surface antigens, often referred to as variant surface antigens, are important targets of acquired protective immunity and include PfEMP1, RIFIN, STEVOR and SURFIN. These antigens are highly polymorphic and encoded by multigene families, which generate substantial antigenic diversity to mediate immune evasion. The most important immune target appears to be PfEMP1, which is a major ligand for vascular adhesion and sequestration of IEs. Studies are beginning to identify specific variants of PfEMP1 linked to disease pathogenesis that may be suitable for vaccine development, but overcoming antigenic diversity in PfEMP1 remains a major challenge. Much less is known about other surface antigens, or antigens on the surface of gametocyte-IEs, the effector mechanisms that mediate immunity, and how immunity is acquired and maintained over time; these are important topics for future research.


Antibody-mediated rejection despite inhibition of terminal complement.
Terminal complement blockade has been shown to decrease the incidence of early acute antibody-mediated rejection (eAMR) in the first month after positive cross-match kidney transplant recipients, yet some patients still develop eAMR. The current study investigated possible mechanisms of eAMR despite eculizumab treatment. Of the 26 patients treated with eculizumab, two developed clinical eAMR and another patient developed histologic signs of eAMR without graft dysfunction ('subclinical eAMR'). Twenty-three did not have histologic injury on early surveillance biopsies. All 26 patients had therapeutic levels of eculizumab and showed complete blockade of complement in hemolytic assays. High levels of donor-specific alloantibody (DSA) including total IgG, IgG3, and C1q+ DSA were present in patients with and without eAMR, and none correlated well with eAMR. In contrast, IgM DSA was present in only four patients after transplantation: the two patients with clinical eAMR, one patient with subclinical AMR, and one patient without eAMR (P = 0.006 correlation with eAMR). Both clinical eAMR episodes were easily treated with plasma exchange which removed IgM more completely and rapidly than IgG, resulting in normalization of function and histology. These data suggest a possible role of antidonor IgM DSA in the pathogenesis of eAMR in patients treated with terminal complement blockade (ClinicalTrials.gov Identifier: NCT00670774).


Improved detection of methicillin-resistant Staphylococcus aureus using phenyl mannitol broth containing aztreonam and ceftizoxime.
We tested a phenyl mannitol broth containing ceftizoxime and aztreonam (PHMB(+)) for detection of methicillin-resistant Staphylococcus aureus (MRSA) with reference MRSA strains and, subsequently, with clinical samples (n = 1,098). All reference MRSA strains induced color change in PHMB(+) after 24 to 72 h of incubation. In a clinical setting, 40 MRSA strains were detected with PHMB(+), compared with only 23 detected with a routine method. Thus, this selective broth significantly (P < 0.001) improved the rate of MRSA detection.


The Evolutionary Fates of a Large Segmental Duplication in Mouse.
Gene duplication and loss are major sources of genetic polymorphism in populations, and are important forces shaping the evolution of genome content and organization. We have reconstructed the origin and history of a 127-kbp segmental duplication, R2d, in the house mouse (Mus musculus). R2d contains a single protein-coding gene, Cwc22 De novo assembly of both the ancestral (R2d1) and the derived (R2d2) copies reveals that they have been subject to nonallelic gene conversion events spanning tens of kilobases. R2d2 is also a hotspot for structural variation: its diploid copy number ranges from zero in the mouse reference genome to >80 in wild mice sampled from around the globe. Hemizygosity for high copy-number alleles of R2d2 is associated in cis with meiotic drive; suppression of meiotic crossovers; and copy-number instability, with a mutation rate in excess of 1 per 100 transmissions in some laboratory populations. Our results provide a striking example of allelic diversity generated by duplication and demonstrate the value of de novo assembly in a phylogenetic context for understanding the mutational processes affecting duplicate genes.


Additive-Manufactured Gyroid Scaffolds of Magnesium Oxide, Phosphate Glass Fiber and Polylactic Acid Composite for Bone Tissue Engineering.
Composites of biodegradable phosphate glass fiber and polylactic acid (PGF/PLA) show potential for bone tissue engineering scaffolds, due to their ability to release Ca, P, and Mg during degradation, thus promoting the bone repair. Nevertheless, glass degradation tends to acidify the surrounding aqueous environment, which may adversely affect the viability and bone-forming activities of osteoblasts. In this work, MgO was investigated as a neutralizing agent. Porous network-phase gyroid scaffolds were additive-manufactured using four different materials: PLA, MgO/PLA, PGF/PLA, and (MgO + PGF)/PLA. The addition of PGF enhanced compressive properties of scaffolds, and the resultant scaffolds were comparably strong and stiff with human trabecular bone. While the degradation of PGF/PLA composite induced considerable acidity in degradation media and intensified the degradation of PGF in return, the degradation media of (MgO + PGF)/PLA maintained a neutral pH close to a physiological environment. The experiment results indicated the possible mechanism of MgO as the neutralizing agent: the local acidity was buffered as the MgO reacted with the acidic degradation products thereby inhibiting the degradation of PGF from being intensified in an acidic environment. The (MgO + PGF)/PLA composite scaffold appears to be a candidate for bone tissue engineering.


Patient-specific simulation as a basis for clinical decision-making.
Patient-specific medical simulation holds the promise of determining tailored medical treatment based on the characteristics of an individual patient (for example, using a genotypic assay of a sequence of DNA). Decision-support systems based on patient-specific simulation can potentially revolutionize the way that clinicians plan courses of treatment for various conditions, ranging from viral infections to arterial abnormalities. Basing medical decisions on the results of simulations that use models derived from data specific to the patient in question means that the effectiveness of a range of potential treatments can be assessed before they are actually administered, preventing the patient from experiencing unnecessary or ineffective treatments. We illustrate the potential for patient-specific simulation by first discussing the scale of the evolving international grid infrastructure that is now available to underpin such applications. We then consider two case studies, one concerned with the treatment of patients with HIV/AIDS and the other addressing neuropathologies associated with the intracranial vasculature. Such patient-specific medical simulations require access to both appropriate patient data and the computational resources on which to perform potentially very large simulations. Computational infrastructure providers need to furnish access to a wide range of different types of resource, typically made available through heterogeneous computational grids, and to institute policies that facilitate the performance of patient-specific simulations on those resources. To support these kinds of simulations, where life and death decisions are being made, computational resource providers must give urgent priority to such jobs, for example by allowing them to pre-empt the queue on a machine and run straight away. We describe systems that enable such priority computing.


[Acute pancreatitis. Analysis of 97 patients].
Acute pancreatitis (AP) is a disease with several causes and diverse clinical course. In our environment there are not many papers on this entity and they approach punctual features inside the broad spectrum picture. The aim of this study is to determinate clinical features, etiology, handling and evolution of patients diagnosed with AP admitted in our Hospital during a three years period. The clinical records of patients with AP admitted at the British Hospital of Buenos Aires between April 2004 and April 2007 were retrospectively analyzed. Demographic data, etiology, treatment, severity of the illness, morbidity and mortality were evaluated. Ninety seven patients were diagnosed with AP during the analyzed period. Forty nine were male (50.5%). The average age was 58.5 +/- 1.6 years (range 21-93). Ninety two patients presented only one episode of AP and five patients suffered two or more (recurrent pancreatitis). According to the etiology, 48 had a biliary origin, 23 were idiopathic, 11 post biliary tract procedures, 2 recognized an alcoholic origin and 13 other causes. The median hospital stay was 7 days. Twenty five percent required admission in Intensive Care Unit. Most of the patients (n: 66; 72%) presented mild episodes according to the Ranson Score. The median of beginning oral feeding was 3 days. Regarding to complications, 13% have multiple organ failure, 8% pancreatic necrosis (4 infected), only one patient had a pseudocist and another patient a peripancreatic abscess. Mortality rate was 7.2% (7 cases, all with Ranson = 3). Although this is a short series, the biliar origin is still the most prevalent etiology in our population. The low number of alcoholic origin caught our attention. Mortality was similar to that described in other series, but the number of local complications was lower. Our data are consistent with other papers regarding management and prognosis, and update the Argentine casuistic.


Wnt antagonist gene DKK2 is epigenetically silenced and inhibits renal cancer progression through apoptotic and cell cycle pathways.
Wnt/beta-catenin signaling is involved in renal cancer. DKK2, a Wnt antagonist, is silenced in some cancers, although its function has not been investigated. We hypothesized that DKK2 may be epigenetically silenced and inhibits progression of renal cell carcinoma (RCC). RCC cell lines and a normal kidney cell line were used for methylation and chromatin immunoprecipitation assays. To assess various functions of DKK2, we established stable DKK2-transfected cells and examined them with regard to cell viability, colony formation, apoptosis, cell cycle, and invasive capability. A total of 52 patients with confirmed conventional RCC were enrolled in this study. RCC cell lines had decreased levels of DKK2, which were significantly increased after treatment with 5-Aza-2'-deoxycytidine alone or 5-Aza-2'-deoxycytidine and trichostatin A. In chromatin immunoprecipitation assay, the levels of acetyl H3, acetyl H4, and dimethylated H3K4 were decreased, whereas the level of dimethylated H3K9 was increased in RCC cell lines compared with HK2 cells. Increased methylation in RCC tissues was associated with higher grades, pathologic stages, and pathologic tumor in RCC. Functional analysis showed that the numbers of viable A498 cells were significantly decreased in DKK2-transfected cells compared with mock cells. The number of apoptotic cells and S/G(2)-M phase cells was significantly increased and decreased after DKK2 transfection, respectively. Corresponding to these results, Bcl2 and cyclin D1 expression were also decreased in DKK2-overexpressing cells. DKK2 is epigenetically silenced by methylation in higher grades and stages of RCC. These results suggest that DKK2 inhibits renal cancer progression through apoptotic and cell cycle pathways.


Expression of heat shock protein 27 in human atherosclerotic plaques and increased plasma level of heat shock protein 27 in patients with acute coronary syndrome.
We intended to identify proteins that are differentially expressed in human atherosclerotic plaques. Comparative 2-dimensional electrophoretic analysis on carotid atherosclerotic endarterectomy specimens (n = 10) revealed that heat shock protein 27 (Hsp27) expression was significantly increased in the nearby normal-appearing area compared with the plaque core area from the same vessel specimen, which was further confirmed by Western blot analysis. The Hsp27 expression in the adjacent normal-appearing vessel areas was much higher than that in nonatherosclerotic reference arteries. The phosphorylation of Hsp27 showed a gradation in the degree of phosphorylation: greatest in the reference arteries, intermediate in the adjacent normal-appearing area, and lowest in plaque core area. Immunohistochemical analysis showed that the phosphorylation of Hsp27 of smooth muscle cells in the carotid endarterectomy specimens was decreased compared with that in the reference artery specimen. The mean plasma level of Hsp27 was significantly higher in patients with acute coronary syndrome (ACS) (n = 27; 106.1 +/- 74.1 ng/mL) than in the normal reference subjects (n = 29; 45.8 +/- 29.5 ng/mL; P < 0.005). The plasma levels of Hsp27 were significantly correlated with those of heat shock protein 70 (Hsp70) (r = 0.422, P < 0.0005), with adjustment for ACS/reference status. In the atherosclerotic lesion, Hsp27 expression is increased in the normal-appearing vessel adjacent to atherosclerotic plaque, whereas levels in the plaque itself are significantly decreased. Both plaque and adjacent artery show decreased Hsp27 phosphorylation compared with reference vessel. In ACS, plasma Hsp27 and Hsp70 are increased, and levels of Hsp27 correlate with Hsp70, C-reactive protein, and CD40L levels.


High resolution assembly and characterization of genomes of Canadian isolates of Salmonella Enteritidis.
There is a need to characterize genomes of the foodborne pathogen, Salmonella enterica serovar Enteritidis (SE) and identify genetic information that could be ultimately deployed for differentiating strains of the organism, a need that is yet to be addressed mainly because of the high degree of clonality of the organism. In an effort to achieve the first characterization of the genomes of SE of Canadian origin, we carried out massively parallel sequencing of the nucleotide sequence of 11 SE isolates obtained from poultry production environments (n = 9), a clam and a chicken, assembled finished genomes and investigated diversity of the SE genome. The median genome size was 4,678,683 bp. A total of 4,833 chromosomal genes defined the pan genome of our field SE isolates consisting of 4,600 genes present in all the genomes, i.e., core genome, and 233 genes absent in at least one genome (accessory genome). Genome diversity was demonstrable by the presence of 1,360 loci showing single nucleotide polymorphism (SNP) in the core genome which was used to portray the genetic distances by means of a phylogenetic tree for the SE isolates. The accessory genome consisted mostly of previously identified SE prophage sequences as well as two, apparently full-sized, novel prophages namely a 28 kb sequence provisionally designated as SE-OLF-10058 (3) prophage and a 43 kb sequence provisionally designated as SE-OLF-10012 prophage. The number of SNPs identified in the relatively large core genome of SE is a reflection of substantial diversity that could be exploited for strain differentiation as shown by the development of an informative phylogenetic tree. Prophage sequences can also be exploited for SE strain differentiation and lineage tracking. This work has laid the ground work for further studies to develop a readily adoptable laboratory test for the subtyping of SE.


De novo transcriptome assemblies of four accessions of the metal hyperaccumulator plant Noccaea caerulescens.
Noccaea caerulescens of the Brassicaceae family has become the key model plant among the metal hyperaccumulator plants. Populations/accessions of N. caerulescens from geographic locations with different soil metal concentrations differ in their ability to hyperaccumulate and hypertolerate metals. Comparison of transcriptomes in several accessions provides candidates for detailed exploration of the mechanisms of metal accumulation and tolerance and local adaptation. This can have implications in the development of plants for phytoremediation and improved mineral nutrition. Transcriptomes from root and shoot tissues of four N. caerulescens accessions with contrasting Zn, Cd and Ni hyperaccumulation and tolerance traits were sequenced with Illumina Hiseq2000. Transcriptomes were assembled using the Trinity de novo assembler and were annotated and the protein sequences predicted. The comparison against the BUSCO plant early release dataset indicated high-quality assemblies. The predicted protein sequences have been clustered into ortholog groups with closely related species. The data serve as important reference sequences in whole transcriptome studies, in analyses of genetic differences between the accessions and other species, and for primer design.


Effect of gestational age and position on peak expiratory flow rate: a longitudinal study.
We sought to study the effects of gestational age and maternal position on peak expiratory flow rates. Peak expiratory flow rates were measured in the standing, sitting, and supine positions in 38 healthy pregnant women at 4-week intervals starting at less than 10 weeks until delivery and again at 6 weeks postpartum. The highest reading of 3 consecutive peak expiratory flow rate measurements for each encounter and position was used in the analysis. Repeated measures analysis of covariance was performed with subjects, gestational age, position, and gestational age times position as the model effects. Least squares mean peak expiratory flow rates were compared among positions at different gestation ages using Bonferroni-adjusted least significant difference t tests. Peak expiratory flow rate declined significantly throughout gestation in all positions (P < .001) with mean rate of decline of 0.65 L/min per week). The slopes of linear trends were not statistically different between positions (P = .222). However, the rate of decline for the supine position was higher than for standing and sitting positions (0.86 compared with 0.46 and 0.57 L/min per week), respectively. On average, the postpartum peak expiratory flow rate returned to 71.9% of its measurement in early gestation. Nomograms depicting mean and the 5th and 95th percentiles of peak expiratory flow rates were constructed for each position. Peak expiratory flow rate measurements are affected by maternal position and advancing gestational age, especially in the supine position. Adjustment of patient's flow rate in relation to gestational age and maternal position is recommended, especially in pregnant women with asthma. III.


Dignity of Older Persons With Mental Health Conditions: Why Should Clinicians Care?
With a steady increase in population aging, the proportion of older people living with mental illness is on rise. This has a significant impact on their autonomy, rights, quality of life and functionality. The biomedical approach to mental healthcare has undergone a paradigm shift over the recent years to become more inclusive and rights-based. Dignity comprises of independence, social inclusion, justice, equality, respect and recognition of one's identity. It has both subjective and objective components and influences life-satisfaction, treatment response as well as compliance. The multi-dimensional framework of dignity forms the central anchor to person-centered mental healthcare for older adults. Mental health professionals are uniquely positioned to incorporate the strategies to promote dignity in their clinical care and research as well as advocate for related social/health policies based on a human rights approach. However, notwithstanding the growing body of research on the neurobiology of aging and old age mental health disorders, dignity-based mental healthcare is considered to be an abstract and hypothetical identity, often neglected in clinical practice. In this paper, we highlight the various components of dignity in older people, the impact of ageism and mental health interventions based on dignity, rights, respect, and equality (including dignity therapy). It hopes to serve as a framework for clinicians to incorporate dignity as a principle in mental health service delivery and research related to older people.


Sprouty: a common antagonist of FGF and EGF signaling pathways in Drosophila.
Extracellular factors such as FGF and EGF control various aspects of morphogenesis, patterning and cellular proliferation in both invertebrates and vertebrates. In most systems, it is primarily the distribution of these factors that controls the differential behavior of the responding cells. Here we describe the role of Sprouty in eye development. Sprouty is an extracellular protein that has been shown to antagonize FGF signaling during tracheal branching in Drosophila. It is a novel type of protein with a highly conserved cysteine-rich region. In addition to the embryonic tracheal system, sprouty is also expressed in other tissues including the developing eye imaginal disc, embryonic chordotonal organ precursors and the midline glia. In each of these tissues, EGF receptor signaling is known to participate in the control of the correct number of neurons or glia. We show that, in all three tissues, the loss of sprouty results in supernumerary neurons or glia, respectively. Furthermore, overexpression of sprouty in wing veins and ovarian follicle cells, two other tissues where EGF signaling is required for patterning, results in phenotypes that resemble the loss-of-function phenotypes of Egf receptor. These results suggest that Sprouty acts as an antagonist of EGF as well as FGF signaling pathways. These receptor tyrosine kinase-mediated pathways may share not only intracellular signaling components but also extracellular factors that modulate the strength of the signal.


Effectiveness and safety of 3 and 5 day courses of artemether-lumefantrine for the treatment of uncomplicated falciparum malaria in an area of emerging artemisinin resistance in Myanmar.
Artemisinin resistance in Plasmodium falciparum has emerged and spread in Southeast Asia. In areas where resistance is established longer courses of artemisinin-based combination therapy have improved cure rates. The standard 3-day course of artemether-lumefantrine (AL) was compared with an extended 5-day regimen for the treatment of uncomplicated falciparum malaria in Kayin state in South-East Myanmar, an area of emerging artemisinin resistance. Late parasite clearance dynamics were described by microscopy and quantitative ultra-sensitive PCR. Patients were followed up for 42 days. Of 154 patients recruited (105 adults and 49 children < 14 years) 78 were randomized to 3 days and 76 to 5 days AL. Mutations in the P. falciparum kelch13 propeller gene (k13) were found in 46% (70/152) of infections, with F446I the most prevalent propeller mutation (29%; 20/70). Both regimens were well-tolerated. Parasite clearance profiles were biphasic with a slower submicroscopic phase which was similar in k13 wild-type and mutant infections. The cure rates were 100% (70/70) and 97% (68/70) in the 3- and 5-day arms respectively. Genotyping of the two recurrences was unsuccessful. Despite a high prevalence of k13 mutations, the current first-line treatment, AL, was still highly effective in this area of South-East Myanmar. The extended 5 day regimen was very well tolerated, and would be an option to prolong the useful therapeutic life of AL. Trial registration NCT02020330. Registered 24 December 2013, https://clinicaltrials.gov/NCT02020330.


Observation of entanglement of a single photon with a trapped atom.
We report the observation of entanglement between a single trapped atom and a single photon at a wavelength suitable for low-loss communication over large distances, thereby achieving a crucial step towards long range quantum networks. To verify the entanglement, we introduce a single atom state analysis. This technique is used for full state tomography of the atom-photon qubit pair. The detection efficiency and the entanglement fidelity are high enough to allow in a next step the generation of entangled atoms at large distances, ready for a final loophole-free Bell experiment.


Short-Term Microbiota Manipulation and Forearm Substrate Metabolism in Obese Men: A Randomized, Double-Blind, Placebo-Controlled Trial.
To investigate the impact of gut microbiota manipulation on fasting and postprandial skeletal muscle metabolism in humans. 40 obese, insulin-resistant males were randomized to amoxicillin (broad-spectrum antibiotic), vancomycin (narrow-spectrum antibiotic), or placebo (7 days, 1,500 mg/day). Before and after treatment, forearm blood flow and metabolite fluxes across forearm muscle were measured under fasting and postprandial (high-fat mixed-meal) conditions. Vancomycin decreased bacterial diversity, reduced the abundance of Gram-positive Firmicutes, and increased the abundance of Gram-negative Proteobacteria, whereas amoxicillin did not affect microbial composition. Neither vancomycin nor amoxicillin treatment affected fasting and postprandial plasma glucose, free fatty acid (FFA), triacylglycerol (TAG), glycerol, lactate, and insulin concentrations or forearm blood flow. Fasting and postprandial net forearm muscle glucose uptake and the release of lactate were not significantly altered by antibiotic treatment as compared to placebo. Finally, antibiotic treatment did not change fasting and postprandial glycerol, FFA, and TAG fluxes across forearm muscle. The present study demonstrates that short-term antibiotic treatment has no effects on fasting and postprandial forearm substrate metabolism and blood flow in obese men with impaired glucose metabolism. These data suggest that short-term strategies targeting the gut microbiota to improve metabolic health may not be effective in obese humans.


Increased hand hygiene compliance in nursing homes after a multimodal intervention: A cluster randomized controlled trial (HANDSOME).
To assess the effect of a multimodal intervention on hand hygiene compliance (HHC) in nursing homes. HHC was evaluated using direct, unobtrusive observation in a cluster randomized controlled trial at publicly funded nursing homes in the Netherlands. In total, 103 nursing home organizations were invited to participate; 18 organizations comprising 33 nursing homes (n = 66 nursing home units) participated in the study. Nursing homes were randomized into a control group (no intervention, n = 30) or an intervention group (multimodal intervention, n = 36). The primary outcome measure was HHC of nurses. HHC was appraised at baseline and at 4, 7, and 12 months after baseline. Observers and nurses were blinded. Audits regarding hand hygiene (HH) materials and personal hygiene rules, 3 live lessons, an e-learning program, posters, and a photo contest. We used a new method to teach the nurses the WHO-defined 5 moments of HH: Room In, Room Out, Before Clean, and After Dirty. HHC increased in both arms. The increase after 12 months was larger for units in the intervention arm (from 12% to 36%) than for control units (from 13% to 21%) (odds ratio [OR], 2.10; confidence interval [CI], 1.35-3.28). The intervention arm exhibited a statistically significant increase in HHC at 4 of the 5 WHO-defined HH moments. At follow-up, HHC in the intervention arm remained statistically significantly higher (OR, 1.93; 95% CI, 1.59-2.34) for indications after an activity (from 37% to 39%) than for indications before an activity (from 14% to 27%). The HANDSOME intervention is successful in improving HHC in nursing homes.


Therapeutic advances in narcolepsy.
Narcolepsy treatment has changed dramatically over the last century. For the treatment of sleepiness in narcolepsy, we have progressed from the early use of caffeine. We have available a variety of different stimulants, and a wake-promoting agent, modafinil, which is widely regarded as the first-line medication for narcolepsy. Cataplexy is managed by medications whereas behavioral treatment, such as avoidance of emotion, was the only treatment available in the past. Following the widespread use of antidepressant medications for cataplexy, we now have sodium oxybate, which works by an unknown mechanism but is the only Food and Drug Administration (FDA)-approved medication for cataplexy. We also recognize that other sleep disorders can occur in narcolepsy, such as obstructive sleep apnea syndrome or rapid eye movement sleep behavior disorder, and new treatments allow these comorbid conditions to be effectively treated. However, although we cannot cure narcolepsy, the current treatments for excessive sleepiness and cataplexy can be effective for many patients. We are improving the quality of life for our patients without producing clinically significant adverse effects. We need new therapeutic advances and several medications that work, though different mechanisms are likely to be available in the near future.


Contribution of Indian psychiatry in the development of psychiatry in Nepal.
Psychiatric services remained virtually unknown in Nepal until 1961. The first psychiatric outpatient service was started in 1961, at Bir Hospital, Kathmandu. In 1984, the Psychiatry Department at Bir Hospital was separated and a mental hospital was created, which was later shifted to its current location at Lagankhel, Patan, in Kathmandu valley, in 1985. It is the only mental hospital in Nepal with a current bed strength of 50 beds. The new era in medical learning and teaching was ushered in Nepal with the establishment of the Institute of Medicine (IOM) under the Tribhuban University and the 400-bed Tribhuban University-Teaching Hospital (TU-Teaching Hospital), in the year 1983. BP Koirala Institute of Health Sciences (BPKIHS) at Dharan was established in 1993, as a part of the joint Indo-Nepal collaboration on developing an international standard teaching, training, and research-oriented medical institute similar to AIIMS, New Delhi. During the last one-and-half decades a number of privately run medical colleges have come up in Nepal. Outpatient and inpatient Psychiatry Departments have been established in most of these government as well as private medical institutes. At present, the postgraduate course (MD) in psychiatry has been running in two government-run institutes as well as three privately run medical colleges. Indian psychiatrists have played and are still playing significant roles in establishing as well as maintaining Psychiatry Departments, especially in the private sector medical colleges. They have also contributed to the growth of psychiatry research and postgraduate teaching in psychiatry, in Nepal.


Andersen-Tawil syndrome.
Andersen-Tawil syndrome (ATS) is a rare condition consisting of ventricular arrhythmias, periodic paralysis, and dysmorphic features. In 2001, mutations in KCNJ2, which encodes the a subunit of the potassium channel Kir2.1, were identified in patients with ATS. To date, KCNJ2 is the only gene implicated in ATS, accounting for approximately 60% of cases. ATS is a unique channelopathy, and represents the first link between cardiac and skeletal muscle excitability. The arrhythmias observed in ATS are distinctive; patients may be asymptomatic, or minimally symptomatic despite a high arrhythmia burden with frequent ventricular ectopy and bidirectional ventricular tachycardia. However, patients remain at risk for life-threatening arrhythmias, including torsades de pointes and ventricular fibrillation, albeit less commonly than observed in other genetic arrhythmia syndromes. The characteristic heterogeneity at both the genotypic and phenotypic levels contribute to the continued difficulties with appropriate diagnosis, risk stratification, and effective therapy. The initial recognition of a syndromic association of clinically diverse symptoms, and the subsequent identification of the underlying molecular genetic basis of ATS has enhanced both clinical care, and our understanding of the critical function of Kir2.1 on skeletal muscle excitability and cardiac action potential.


In vitro stability study of organophosphonic self assembled monolayers (SAMs) on cobalt chromium (Co-Cr) alloy.
Surface modification of cobalt chromium (Co-Cr) alloy is being investigated as a possible solution to the biomedical challenges arising from its usage. Self assembled monolayers (SAMs) of organophosphonic octadecylphosphonic acid (ODPA) were formed on the oxide surface of Co-Cr alloy by chemisorption using the solution deposition technique. High quality and well-ordered SAMs were formed which were characterized using Fourier transform infrared spectroscopy-attenuated total reflectance (FTIR-ATR), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM), contact angle measurements and ellipsometry. The resulting monolayers were then exposed to in vitro conditions using phosphate buffered saline (PBS) solution. The samples were analyzed for a period of 1, 3, 7 and 14 days. The resulting samples were characterized using XPS, AFM and Contact angle measurements. XPS atomic concentrations and detailed high energy elemental scans gave an insight into the trends of elemental concentrations over the duration of the study. SAMs were found to be strongly bound to the oxide surface after PBS exposure. AFM gave the topographic details of SAMs presence by island formation before and after SAM formation and also over the duration of the PBS exposure. Contact Angle Measurements confirmed the hydrophobicity of the surface after SAM formation and indicated a slight disorder of the SAM alkyl chain upon exposure to PBS. Thus, ODPA SAMs were successfully coated on Cobalt Chromium (Co-Cr) alloy surface and were found to be stable and strongly bound after PBS exposure.


Relationships, expertise, incentives, and governance: supporting care home residents' access to health care. An interview study from England.
To explore what commissioners of care, regulators, providers, and care home residents in England identify as the key mechanisms or components of different service delivery models that support the provision of National Health Service (NHS) provision to independent care homes. Qualitative, semistructured interviews with a purposive sample of people with direct experience of commissioning, providing, and regulating health care provision in care homes and care home residents. Data from interviews were augmented by a secondary analysis of previous interviews with care home residents on their personal experience of and priorities for access to health care. Analysis was framed by the assumptions of realist evaluation and drew on the constant comparative method to identify key themes about what is required to achieve quality health care provision to care homes and resident health. Participants identified 3 overlapping approaches to the provision of NHS that they believed supported access to health care for older people in care homes: (1) Investment in relational working that fostered continuity and shared learning between visiting NHS staff and care home staff, (2) the provision of age-appropriate clinical services, and (3) governance arrangements that used contractual and financial incentives to specify a minimum service that care homes should receive. The 3 approaches, and how they were typified as working, provide a rich picture of the stakeholder perspectives and the underlying assumptions about how service delivery models should work with care homes. The findings inform how evidence on effective working in care homes will be interrogated to identify how different approaches, or specifically key elements of those approaches, achieve different health-related outcomes in different situations for residents and associated health and social care organizations.


Structure-based assembly of protein complexes in yeast.
Images of entire cells are preceding atomic structures of the separate molecular machines that they contain. The resulting gap in knowledge can be partly bridged by protein-protein interactions, bioinformatics, and electron microscopy. Here we use interactions of known three-dimensional structure to model a large set of yeast complexes, which we also screen by electron microscopy. For 54 of 102 complexes, we obtain at least partial models of interacting subunits. For 29, including the exosome, the chaperonin containing TCP-1, a 3'-messenger RNA degradation complex, and RNA polymerase II, the process suggests atomic details not easily seen by homology, involving the combination of two or more known structures. We also consider interactions between complexes (cross-talk) and use these to construct a structure-based network of molecular machines in the cell.


Mesyl Phosphoramidate Oligonucleotides as Potential Splice-Switching Agents: Impact of Backbone Structure on Activity and Intracellular Localization.
A series of 2'-deoxy and novel 2'-O-methyl and 2'-O-(2-methoxyethyl) (2'-MOE) oligonucleotides with internucleotide methanesulfonyl (mesyl, μ) or 1-butanesulfonyl (busyl, β) phosphoramidate groups has been synthesized for evaluation as potential splice-switching oligonucleotides. Evaluation of their splice-switching activity in spinal muscular atrophy patient-derived fibroblasts revealed no significant difference in splice-switching efficacy between 2'-MOE mesyl oligonucleotide and the corresponding phosphorothioate (nusinersen). Yet, a survival study with model neonatal mice has shown the antisense 2'-MOE mesyl oligonucleotide to be inferior to nusinersen at the highest dose of 40 mg/kg. A reason for their lower activity in vivo as ascertained by cellular uptake study by fluorescent confocal microscopy in HEK293 cell line could possibly be ascribed to compromised endosomal release and/or nuclear uptake of the 2'-OMe or 2'-MOE μ- and β-oligonucleotides compared to their phosphorothioate analog.


[Variation in serum cholesterol level in the same person: what is the true value and when is it called a significant change?].
The blood cholesterol level is not constant over time. Changing values may be found when rechecking high values, especially in patients on a diet or receiving medication. Repeated measurements are necessary. It will then be possible to estimate the 'true mean value'. This is the theoretical average of a large number of measurements taken from one person. The study concentrated on the intra-individual variation of the serum cholesterol and the consequences for screening and follow-up. For this purpose, during a period of four weeks, cholesterol levels were measured 12 times in 33 men aged 25-40. The mean coefficient of variation was 5.7%, with wide differences between participants, ranging from 2.9% to 9.8%. The position of the 'true mean value' was estimated (with 90% confidence), after I resp. 3 determinations. These findings have consequences for the classification of subjects in the different risk categories as defined in the Dutch Cholesterol Consensus. It is also possible to determine if, after a period of intervention, there is a significant decline in the cholesterol level. Roughly, a decline of 10-12% indicates a significant difference.


RNA complementary to a herpesvirus alpha gene mRNA is prominent in latently infected neurons.
In initial attempts to define the molecular events responsible for the latent state of herpes simplex virus, in situ hybridization was utilized to search for virally encoded RNA transcripts in latently infected sensory neurons. The use of cloned probes representing the entire viral genome indicated that transcripts encoded within terminal repeats were present. When the alpha genes encoding ICP-0, ICP-4, and ICP-27 and the gamma 1 gene encoding VP-5 were employed, only RNA transcripts hybridizing to the ICP-0 probe were detected. In latently infected cells, the ICP-0--related transcripts were localized principally in the nucleus; this was not the case in acutely (productively) infected neurons or in neurons probed for RNA transcripts coding for actin. In Northern blotting experiments, an RNA of 2.6 kilobases was detected with the ICP-0 probe. When single-stranded DNAs from the ICP-0 region were used as probes, RNA from the strand complementary to that encoding ICP-0 messenger RNA (mRNA) was the major species detected. This RNA species may play a significant role in maintaining the latent infection.


Urelumab alone or in combination with rituximab in patients with relapsed or refractory B-cell lymphoma.
Urelumab, a fully human, non-ligand binding, CD137 agonist IgG4 monoclonal antibody, enhances T-cell and natural killer-cell antitumor activity in preclinical models, and may enhance cytotoxic activity of rituximab. Here we report results in patients with relapsed or refractory diffuse large B-cell lymphoma (DLBCL), follicular lymphoma (FL), and other B-cell lymphomas, in phase 1 studies evaluating urelumab alone (NCT01471210) or combined with rituximab (NCT01775631). Sixty patients received urelumab (0.3 mg/kg IV Q3W, 8 mg IV Q3W, or 8 mg IV Q6W); 46 received urelumab (0.1 mg/kg, 0.3 mg/kg, or 8 mg IV Q3W) plus rituximab 375 mg/m2 IV QW. The maximum tolerated dose (MTD) of urelumab was determined to be 0.1 mg/kg or 8 mg Q3W after a single event of potential drug-induced liver injury occurred with urelumab 0.3 mg/kg. Treatment-related AEs were reported in 52% (urelumab: grade 3/4, 15%) and 72% (urelumab + rituximab: grade 3/4, 28%); three led to discontinuation (grade 3 increased AST, grade 4 acute hepatitis [urelumab]; one death from sepsis syndrome [urelumab plus rituximab]). Objective response rates/disease control rates were 6%/19% (DLBCL, n = 31), 12%/35% (FL, n = 17), and 17%/42% (other B-cell lymphomas, n = 12) with urelumab and 10%/24% (DLBCL, n = 29) and 35%/71% (FL, n = 17) with urelumab plus rituximab. Durable remissions in heavily pretreated patients were achieved; however, many were observed at doses exceeding the MTD. These data show that urelumab alone or in combination with rituximab demonstrated manageable safety in B-cell lymphoma, but the combination did not enhance clinical activity relative to rituximab alone or other current standard of care.