Residue-specific mass signatures for the efficient detection of protein modifications by mass spectrometry.
Currently available mass spectrometric (MS) techniques lack specificity in identifying protein modifications because molecular mass is the only parameter used to characterize these changes. Consequently, the suspected modified peptides are subjected to tandem MS/MS sequencing that may demand more time and sample. We report the use of stable isotope-enriched amino acids as residue-specific "mass signatures" for the rapid and sensitive detection of protein modifications directly from the peptide mass map (PMM) without enrichment of the modified peptides. These mass signatures are easily recognized through their characteristic spectral patterns and provide fingerprints for peptides containing the same content of specific amino acid residue(s) in a PMM. Without the need for tandem MS/MS sequencing, a peptide and its modified form(s) can readily be identified through their identical fingerprints, regardless of the nature of modifications. In this report, we demonstrate this strategy for the detection of methionine oxidation and protein phosphorylation. More interestingly, the phosphorylation of a histone protein, H2A.X, obtained from human skin fibroblast cells, was effectively identified in response to low-dose radiation. In general, this strategy of residue-specific mass tagging should be applicable to other posttranslational modifications.


Copy-Number Variation Contributes to the Mutational Load of Bardet-Biedl Syndrome.
Bardet-Biedl syndrome (BBS) is a defining ciliopathy, notable for extensive allelic and genetic heterogeneity, almost all of which has been identified through sequencing. Recent data have suggested that copy-number variants (CNVs) also contribute to BBS. We used a custom oligonucleotide array comparative genomic hybridization (aCGH) covering 20 genes that encode intraflagellar transport (IFT) components and 74 ciliopathy loci to screen 92 unrelated individuals with BBS, irrespective of their known mutational burden. We identified 17 individuals with exon-disruptive CNVs (18.5%), including 13 different deletions in eight BBS genes (BBS1, BBS2, ARL6/BBS3, BBS4, BBS5, BBS7, BBS9, and NPHP1) and a deletion and a duplication in other ciliopathy-associated genes (ALMS1 and NPHP4, respectively). By contrast, we found a single heterozygous exon-disruptive event in a BBS-associated gene (BBS9) in 229 control subjects. Superimposing these data with resequencing revealed CNVs to (1) be sufficient to cause disease, (2) Mendelize heterozygous deleterious alleles, and (3) contribute oligogenic alleles by combining point mutations and exonic CNVs in multiple genes. Finally, we report a deletion and a splice site mutation in IFT74, inherited under a recessive paradigm, defining a candidate BBS locus. Our data suggest that CNVs contribute pathogenic alleles to a substantial fraction of BBS-affected individuals and highlight how either deletions or point mutations in discrete splice isoforms can induce hypomorphic mutations in genes otherwise intolerant to deleterious variation. Our data also suggest that CNV analyses and resequencing studies unbiased for previous mutational burden is necessary to delineate the complexity of disease architecture.


Modeling Decreased Resilience of Shallow Lake Ecosystems toward Eutrophication due to Microplastic Ingestion across the Food Web.
The discovery of microplastic (MP) being present in freshwaters has stimulated research on the impacts of MP on freshwater organisms. To date, research has focused on primary effects, leaving questions with respect to secondary effects at the level of freshwater food webs unanswered. Here, we use a theoretical modeling approach to investigate the hypothesis that MP imposes negative impacts on the level of freshwater shallow lake food webs. We find that increasing MP levels have the potential to affect the critical phosphorus loading (CPL), which is defined as the threshold for regime shifts between clear and turbid states of the water column. The possible occurrence of catastrophic cascades due to MP pollution is predominantly driven by the negative effects of MP on zooplankton. We explore the possible states of the food web by scenario analysis and show that the secondary effects of MP at current concentrations are likely to be negligible. However, at the current rate of MP production, a 20-40% reduction in the CPL would occur by the end of this century, suggesting a loss of resilience in shallow lakes that would be subject to abrupt changes in the food web under lower nutrient loading.


Prophylactic activity of atovaquone against Plasmodium falciparum in humans.
The prophylactic antimalarial activity of atovaquone was determined in a randomized, double-blind, placebo-controlled study of healthy volunteers who were challenged by the bite of Plasmodium falciparum-infected Anopheles stephensi. Subjects were randomly assigned to one of three groups: six received seven daily doses of 750 mg of atovaquone, starting the day before challenge; six received a single dose of 250 mg of atovaquone the day before challenge; and four received placebo. Polymerase chain reaction- and culture-confirmed parasitemia developed in all four placebo recipients, but in none of the drug recipients, indicating that either of the atovaquone regimens provides effective prophylaxis (P = 0.005). However, in low-dose recipients, the drug levels by day 6.5 were profoundly subtherapeutic, indicating that parasites were eliminated prior to the establishment of erythrocytic infection. Atovaquone thus protects non-immune subjects against mosquito-transmitted falciparum malaria, and has causal prophylactic activity.


Characterization of different water pools in solid-state NMR protein samples.
We observed and characterized two distinct signals originating from different pools of water protons in solid-state NMR protein samples, namely from crystal water which exchanges polarization with the protein (on the NMR timescale) and is located in the protein-rich fraction at the periphery of the magic-angle spinning (MAS) sample container, and supernatant water located close to the axis of the sample container. The polarization transfer between the water and the protein can be probed by two-dimensional exchange spectroscopy, and we show that the supernatant water does not interact with protein on the timescale of the experiments. The two water pools have different spectroscopic properties, including resonance frequency, longitudinal, transverse and rotating frame relaxation times. The supernatant water can be removed almost completely physically or can be frozen selectively. Both measures lead to an enhancement of the quality factor of the probe circuit, accompanied by an improvement of the experimental signal/noise, and greatly simplify solvent-suppression by substantially reducing the water signal. We also present a tool, which allows filling solid-state NMR sample containers in a more efficient manner, greatly reducing the amount of supernatant water and maximizing signal/noise.


An isovelocity dynamometer method to determine monoarticular and biarticular muscle parameters.
This study aimed to determine whether subject-specific individual muscle models for the ankle plantar flexors could be obtained from single joint isometric and isovelocity maximum torque measurements in combination with a model of plantar flexion. Maximum plantar flexion torque measurements were taken on one subject at six knee angles spanning full flexion to full extension. A planar three-segment (foot, shank and thigh), two-muscle (soleus and gastrocnemius) model of plantar flexion was developed. Seven parameters per muscle were determined by minimizing a weighted root mean square difference (wRMSD) between the model output and the experimental torque data. Valid individual muscle models were obtained using experimental data from only two knee angles giving a wRMSD score of 16 N m, with values ranging from 11 to 17 N m for each of the six knee angles. The robustness of the methodology was confirmed through repeating the optimization with perturbed experimental torques (± 20%) and segment lengths (± 10%) resulting in wRMSD scores of between 13 and 20 N m. Hence, good representations of maximum torque can be achieved from subject-specific individual muscle models determined from single joint maximum torque measurements. The proposed methodology could be applied to muscle-driven models of human movement with the potential to improve their validity.


Perioperative bridging anticoagulation during dabigatran or warfarin interruption among patients who had an elective surgery or procedure. Substudy of the RE-LY trial.
In patients with atrial fibrillation (AF) who require interruption of dabigatran or warfarin for an elective surgery/procedure, the risks and benefits of perioperative bridging anticoagulation is uncertain.We accessed the database from RE-LY, a randomised trial comparing dabigatran with warfarin for stroke prevention in AF, to assess the potential benefits and risks of bridging. In patients who had a first interruption of dabigatran or warfarin for an elective surgery/procedure, we compared the risk for major bleeding (MB), stroke or systemic embolism (SSE) and any thromboembolism (TE) in patients who were bridged or not bridged during the period of seven days before until 30 days after surgery/procedure. We used multivariable Cox regression to adjust for potential confounders.Bridging was used more during warfarin interruption than dabigatran interruption (27.5 % vs 15.4 %; p< 0.001). With dabigatran interruption, bridged patients had more MB (6.5 % vs 1.8 %, p< 0.001) than those not bridged but bridged and not bridged groups did not differ for any TE (1.2 % vs 0.6 %, p=0.16) and SSE (0.5 % vs 0.3 %, p=0.46). With warfarin interruption, bridged patients had more MB (6.8 % vs 1.6 %, p< 0.001) and any TE (1.8 % vs 0.3 %, p=0.007) than those not bridged but bridged and not bridged groups did not differ for SSE (0.5 % vs 0.2 %, p=0.321). In conclusion, in patients who interrupted dabigatran or warfarin for a surgery/ procedure in the RE-LY trial, use of bridging anticoagulation appeared to increase the risk for major bleeding irrespective of dabigatran or warfarin interruption.


Association Between Diabetes and Hippocampal Atrophy in Elderly Japanese: The Hisayama Study.
To investigate the association between diabetes and brain or hippocampal atrophy in an elderly population. A total of 1,238 community-dwelling Japanese subjects aged ≥65 years underwent brain MRI scans and a comprehensive health examination in 2012. Total brain volume (TBV), intracranial volume (ICV), and hippocampal volume (HV) were measured using MRI scans for each subject. We examined the associations between diabetes-related parameters and the ratios of TBV to ICV (an indicator of global brain atrophy), HV to ICV (an indicator of hippocampal atrophy), and HV to TBV (an indicator of hippocampal atrophy beyond global brain atrophy) after adjustment for other potential confounders. The multivariable-adjusted mean values of the TBV-to-ICV, HV-to-ICV, and HV-to-TBV ratios were significantly lower in the subjects with diabetes compared with those without diabetes (77.6% vs. 78.2% for the TBV-to-ICV ratio, 0.513% vs. 0.529% for the HV-to-ICV ratio, and 0.660% vs. 0.676% for the HV-to-TBV ratio; all P < 0.01). These three ratios decreased significantly with elevated 2-h postload glucose (PG) levels (all P for trend <0.05) but not fasting plasma glucose levels. Longer duration of diabetes was significantly associated with lower TBV-to-ICV, HV-to-ICV, and HV-to-TBV ratios. The subjects with diabetes diagnosed in midlife had significantly lower HV-to-ICV and HV-to-TBV ratios than those without and those diagnosed in late life. Our data suggest that a longer duration of diabetes and elevated 2-h PG levels, a marker of postprandial hyperglycemia, are risk factors for brain atrophy, particularly hippocampal atrophy.


Apple skin patterning is associated with differential expression of MYB10.
Some apple (Malus × domestica Borkh.) varieties have attractive striping patterns, a quality attribute that is important for determining apple fruit market acceptance. Most apple cultivars (e.g. 'Royal Gala') produce fruit with a defined fruit pigment pattern, but in the case of 'Honeycrisp' apple, trees can produce fruits of two different kinds: striped and blushed. The causes of this phenomenon are unknown. Here we show that striped areas of 'Honeycrisp' and 'Royal Gala' are due to sectorial increases in anthocyanin concentration. Transcript levels of the major biosynthetic genes and MYB10, a transcription factor that upregulates apple anthocyanin production, correlated with increased anthocyanin concentration in stripes. However, nucleotide changes in the promoter and coding sequence of MYB10 do not correlate with skin pattern in 'Honeycrisp' and other cultivars differing in peel pigmentation patterns. A survey of methylation levels throughout the coding region of MYB10 and a 2.5 Kb region 5' of the ATG translation start site indicated that an area 900 bp long, starting 1400 bp upstream of the translation start site, is highly methylated. Cytosine methylation was present in all three contexts, with higher methylation levels observed for CHH and CHG (where H is A, C or T) than for CG. Comparisons of methylation levels of the MYB10 promoter in 'Honeycrisp' red and green stripes indicated that they correlate with peel phenotypes, with an enrichment of methylation observed in green stripes. Differences in anthocyanin levels between red and green stripes can be explained by differential transcript accumulation of MYB10. Different levels of MYB10 transcript in red versus green stripes are inversely associated with methylation levels in the promoter region. Although observed methylation differences are modest, trends are consistent across years and differences are statistically significant. Methylation may be associated with the presence of a TRIM retrotransposon within the promoter region, but the presence of the TRIM element alone cannot explain the phenotypic variability observed in 'Honeycrisp'. We suggest that methylation in the MYB10 promoter is more variable in 'Honeycrisp' than in 'Royal Gala', leading to more variable color patterns in the peel of this cultivar.


Weissella soli sp. nov., a lactic acid bacterium isolated from soil.
Phylogenetic analysis of the 16S rRNA gene of bacterial isolates from garden soil showed relatedness to Weissella kandleri and Weissella confusa. However, the sequences had notable differences, and DNA-DNA hybridizations confirmed that the isolates are separate from these two species. The isolates could be further distinguished from all previously described Weissella species by electrophoretic analysis of whole-cell proteins, as well as by the results from different biochemical tests. The name Weissella soli is proposed for the new species, the type strain being Mi268T (= LMG 20113T = DSM 14420T).


Impact of barbecued meat consumed in pregnancy on birth outcomes accounting for personal prenatal exposure to airborne polycyclic aromatic hydrocarbons: Birth cohort study in Poland.
We previously reported an association between prenatal exposure to airborne polycyclic aromatic hydrocarbons (PAH) and lower birth weight, birth length, and head circumference. The main goal of the present analysis was to assess the possible impact of coexposure to PAH-containing barbecued meat consumed during pregnancy on birth outcomes. The birth cohort consisted of 432 pregnant women who gave birth at term (>36 wk of gestation). Only non-smoking women with singleton pregnancies, 18-35 y of age, and who were free from chronic diseases such as diabetes and hypertension, were included in the study. Detailed information on diet over pregnancy was collected through interviews and the measurement of exposure to airborne PAHs was carried out by personal air monitoring during the second trimester of pregnancy. The effect of barbecued meat consumption on birth outcomes (birth weight, length, and head circumference at birth) was adjusted in multiple linear regression models for potential confounding factors such as prenatal exposure to airborne PAHs, child's sex, gestational age, parity, size of mother (maternal prepregnancy weight, weight gain in pregnancy), and prenatal environmental tobacco smoke. The multivariable regression model showed a significant deficit in birth weight associated with barbecued meat consumption in pregnancy (coeff = -106.0 g; 95%CI: -293.3, -35.8). The effect of exposure to airborne PAHs was about the same magnitude order (coeff. = -164.6 g; 95%CI: -172.3, -34.7). Combined effect of both sources of exposure amounted to birth weight deficit of 214.3 g (95%CI: -419.0, -9.6). Regression models performed for birth length and head circumference showed similar trends but the estimated effects were of borderline significance level. As the intake of barbecued meat did not affect the duration of pregnancy, the reduced birth weight could not have been mediated by a shortened gestation period. In conclusion, the study results provided epidemiologic evidence that prenatal PAH exposure from diet including grilled meat might be hazardous for fetal development.


Bisphenol A, bisphenol F and bisphenol S affect differently 5α-reductase expression and dopamine-serotonin systems in the prefrontal cortex of juvenile female rats.
Early-life exposure to the endocrine disruptor bisphenol A (BPA) affects brain function and behavior, which might be attributed to its interference with hormonal steroid signaling and/or neurotransmitter systems. Alternatively, the use of structural analogs of BPA, mainly bisphenol F (BPF) and bisphenol S (BPS), has increased recently. However, limited in vivo toxicity data exist. We investigated the effects of BPA, BPF and BPS on 5α-reductase (5α-R), a key enzyme involved in neurosteroidogenesis, as well as on dopamine (DA)- and serotonin (5-HT)-related genes, in the prefrontal cortex (PFC) of juvenile female rats. Gestating Wistar rats were treated with either vehicle or 10 μg/kg/day of BPA, BPF or BPS from gestational day 12 to parturition. Then, female pups were exposed from postnatal day 1 through day 21 (PND21), when they were euthanized and RT-PCR, western blot and quantitative PCR-array experiments were performed. BPA decreased 5α-R2 and 5α-R3 mRNA and protein levels, while both BPF and BPS decreased 5α-R3 mRNA levels in PFC at PND21. Further, BPA, BPF and BPS significantly altered, respectively, the transcription of 25, 56 and 24 genes out of the 84 DA and 5-HT-related genes assayed. Of particular interest was the strong induction by all these bisphenols of Cyp2d4, implicated in corticosteroids synthesis. Our results demonstrate for the first time that BPA, BPF and BPS differentially affect 5α-R and genes related to DA/5-HT systems in the female PFC. In vivo evidence of the potential adverse effects of BPF and BPS in the brain of mammals is provided in this work, raising questions about the safety of these chemicals as substitutes for BPA.


Norms and the Flexibility of Moral Action.
A complex web of social and moral norms governs many everyday human behaviors, acting as the glue for social harmony. The existence of moral norms helps elucidate the psychological motivations underlying a wide variety of seemingly puzzling behavior, including why humans help or trust total strangers. In this review, we examine four widespread moral norms: Fairness, altruism, trust, and cooperation, and consider how a single social instrument-reciprocity-underpins compliance to these norms. Using a game theoretic framework, we examine how both context and emotions moderate moral standards, and by extension, moral behavior. We additionally discuss how a mechanism of reciprocity facilitates the adherence to, and enforcement of, these moral norms through a core network of brain regions involved in processing reward. In contrast, violating this set of moral norms elicits neural activation in regions involved in resolving decision conflict and exerting cognitive control. Finally, we review how a reinforcement mechanism likely governs learning about morally normative behavior. Together, this review aims to explain how moral norms are deployed in ways that facilitate flexible moral choices.


Two languages, two minds: flexible cognitive processing driven by language of operation.
People make sense of objects and events around them by classifying them into identifiable categories. The extent to which language affects this process has been the focus of a long-standing debate: Do different languages cause their speakers to behave differently? Here, we show that fluent German-English bilinguals categorize motion events according to the grammatical constraints of the language in which they operate. First, as predicted from cross-linguistic differences in motion encoding, bilingual participants functioning in a German testing context prefer to match events on the basis of motion completion to a greater extent than do bilingual participants in an English context. Second, when bilingual participants experience verbal interference in English, their categorization behavior is congruent with that predicted for German; when bilingual participants experience verbal interference in German, their categorization becomes congruent with that predicted for English. These findings show that language effects on cognition are context-bound and transient, revealing unprecedented levels of malleability in human cognition.


Interferon-gamma modulates prolactin and tissue factor expression in differentiating human endometrial stromal cells.
Cytokines such as interferon-gamma (IFNgamma) released by resident uterine immune cells are thought to influence the expression of differentiated function in the human endometrium. Decidualization of the stromal cell compartment is confined to the superficial endometrial layer in the nonpregnant uterus. To explore the molecular mechanism underlying the spatial expression of the decidual phenotype, the effect of IFNgamma on the induction of two well characterized markers of endometrial stromal (ES) cell differentiation, PRL and tissue factor (TF), has been investigated. IFNgamma antagonizes cAMP-mediated PRL protein and messenger RNA expression in primary ES cell cultures through inhibition of decidual PRL promoter activity. In parallel, IFNgamma stimulates Stat-1 (signal transducer and activator of transcription-1) expression, phosphorylation, and translocation to the nucleus. Exogenously expressed Stat-1 potently represses decidual PRL promoter activation, indicating the potential for the inhibitory effects of IFNgamma to be mediated by Stat-1. We demonstrate that although the coactivator CREB-binding protein/p300 is essential for decidual PRL transcription, this coactivator does not appear to be the target for IFNgamma-mediated repression. By contrast, IFNgamma has little effect on cAMP-mediated TF expression, but induces TF in ES cells not exposed to a decidualizing stimulus. This suggested that in vivo TF expression may not be restricted to decidualizing cells of the superficial layer and was confirmed by immunohistochemical analysis demonstrating intense TF staining in the basal stromal compartment during the regeneration phase of the cycle. The differential sensitivity of decidualization-associated genes to IFNgamma illustrates its potential role as a selective biological response modifier that influences regional function within the endometrium.


Arrhythmogenic right ventricular dysplasia, a cell adhesion cardiomyopathy: insights into disease pathogenesis from preliminary genotype--phenotype assessment.
Arrhythmogenic right ventricular dysplasia/cardiomyopathy (ARVC) is a genetically determined heart muscle disorder presenting clinically with even lethal ventricular arrhythmias, particularly in the young and athletes. It is reported familial with recessive and most commonly dominant inheritance. Disease-causing genes are increasingly recognised among desmosomal proteins plakoglobin, desmoplakin, plakophilin2, and desmoglein2 displaying phenotypic heterogeneity. Mutations in the plakoglobin and desmoplakin genes have been identified to underlie recessive ARVC associated with woolly hair and palmoplantar keratoderma (Naxos disease), while mutations in plakophilin2, desmoglein2 as well as desmoplakin have been identified to underlie the dominant non-syndromic form. Preliminary genotype-phenotype assessment indicates that mutations affecting the outer dense plaque of desmosome (desmoglein2, plakoglobin, plakophilin2 and the N-terminal of desmoplakin) result in ARVC with the ordinary described phenotype. However, mutations at the inner dense plaque, particularly affecting the desmin-binding site of desmoplakin, may result in ARVC with predominantly left ventricular involvement and clinical overlapping with dilated cardiomyopathy. The interesting finding of abnormal distribution of plakoglobin, independently of the primarily affected protein, might suggest a common pathway for plakoglobin in ARVC pathogenesis.


Implant prosthodontic rehabilitation of patients with rheumatic disorders: a case series report.
This retrospective study assessed implant and prosthodontic treatment outcomes of patients suffering from rheumatic disorders such as rheumatoid arthritis (RA) and connective tissue diseases (CTDs). This study included 22 patients (all women) suffering from autoimmune rheumatic disorders such as isolated RA (n = 16), RA with concomitant CTDs (n = 5), or isolated CTDs (n = 1). Overall, 89 implants were placed for rehabilitations such as single-tooth replacement (n = 8), fixed partial dentures (n = 14), complete dentures (n = 5), and overdentures (n = 2), and were evaluated after a mean of 42.6 +/- 25.2 months. The cumulative implant survival and success rates and peri-implant conditions (marginal bone loss, pocket depth, Plaque Index, Gingival Index, Bleeding Index, and Calculus Index) were evaluated with a special focus on RA and CTDs. In addition, incidence and type of prosthodontic maintenance were evaluated. A high implant survival rate was noted during follow-up with a cumulative 3-year implant success rate of 96.1%. Patients with RA demonstrated acceptable marginal bone resorption (mean: 2.1 +/- 0.5 mm) and good soft tissue conditions, while CTD patients showed increased bone resorption (mean: 3.1 +/- 0.7 mm). This was especially noted in scleroderma patients, as were major peri-implant soft tissue alterations (Bleeding Index) in patients suffering from Sjogren syndrome. A high implant and prothodontic success rate can be anticipated even for patients suffering from autoimmune rheumatic disorders such as RA and CTDs. A scrupulous maintenance program that includes optimal oral hygiene could assist in ensuring stable long-term results for CTD patients with more vulnerable soft tissue conditions. Int J Prosthodont 2010;23:22-28.


Structures and chiroptical properties of the BINAS-monosubstituted Au38(SCH3)24 cluster.
The structure and optical properties of a set of R-1,1'-binaphthyl-2,2'-dithiol (R-BINAS) monosubstituted A-Au38(SCH3)24 clusters are studied by means of time dependent density functional theory (TD-DFT). While it was proposed earlier that BINAS selectively binds to monomer motifs (SR-Au-SR) covering the Au23 core, our calculations suggest a binding mode that bridges two dimer (SR-Au-SR-Au-RS) motifs. The more stable isomers show a negligible distortion induced by BINAS adsorption on the Au38(SCH3)24 cluster which is reflected by similar optical and Circular Dichroism (CD) spectra to those found for the parent cluster. The results furthermore show that BINAS adsorption does not enhance the CD signals of the Au38(SCH3)24 cluster.


A phenology of the evolution of endothermy in birds and mammals.
Recent palaeontological data and novel physiological hypotheses now allow a timescaled reconstruction of the evolution of endothermy in birds and mammals. A three-phase iterative model describing how endothermy evolved from Permian ectothermic ancestors is presented. In Phase One I propose that the elevation of endothermy - increased metabolism and body temperature (Tb ) - complemented large-body-size homeothermy during the Permian and Triassic in response to the fitness benefits of enhanced embryo development (parental care) and the activity demands of conquering dry land. I propose that Phase Two commenced in the Late Triassic and Jurassic and was marked by extreme body-size miniaturization, the evolution of enhanced body insulation (fur and feathers), increased brain size, thermoregulatory control, and increased ecomorphological diversity. I suggest that Phase Three occurred during the Cretaceous and Cenozoic and involved endothermic pulses associated with the evolution of muscle-powered flapping flight in birds, terrestrial cursoriality in mammals, and climate adaptation in response to Late Cenozoic cooling in both birds and mammals. Although the triphasic model argues for an iterative evolution of endothermy in pulses throughout the Mesozoic and Cenozoic, it is also argued that endothermy was potentially abandoned at any time that a bird or mammal did not rely upon its thermal benefits for parental care or breeding success. The abandonment would have taken the form of either hibernation or daily torpor as observed in extant endotherms. Thus torpor and hibernation are argued to be as ancient as the origins of endothermy itself, a plesiomorphic characteristic observed today in many small birds and mammals.


β-alanine supplementation improves tactical performance but not cognitive function in combat soldiers.
There are no known studies that have examined β-alanine supplementation in military personnel. Considering the physiological and potential neurological effects that have been reported during sustained military operations, it appears that β-alanine supplementation may have a potential benefit in maintaining physical and cognitive performance during high-intensity military activity under stressful conditions. The purpose of this study was to examine the effect of 28 days of β-alanine ingestion in military personnel while fatigued on physical and cognitive performance. Twenty soldiers (20.1 ± 0.9 years) from an elite combat unit were randomly assigned to either a β-alanine (BA) or placebo (PL) group. Soldiers were involved in advanced military training, including combat skill development, navigational training, self-defense/hand-to-hand combat and conditioning. All participants performed a 4-km run, 5-countermovement jumps using a linear position transducer, 120-m sprint, a 10-shot shooting protocol with assault rifle, including overcoming a misfire, and a 2-min serial subtraction test to assess cognitive function before (Pre) and after (Post) 28 days of supplementation. The training routine resulted in significant increases in 4-km run time for both groups, but no between group differences were seen (p = 0.597). Peak jump power at Post was greater for BA than PL (p = 0.034), while mean jump power for BA at Post was 10.2% greater (p = 0.139) than PL. BA had a significantly greater (p = 0.012) number of shots on target at Post (8.2 ± 1.0) than PL (6.5 ± 2.1), and their target engagement speed at Post was also significantly faster (p = 0.039). No difference in serial subtraction performance was seen between the groups (p = 0.844). Results of this study indicate that 4-weeks of β-alanine ingestion in young, healthy soldiers did not impact cognitive performance, but did enhance power performance, marksmanship and target engagement speed from pre-ingestion levels.


Reliable Model Selection without Reference Values by Utilizing Model Diversity with Prediction Similarity.
Predictive modeling (calibration or training) with various data formats, such as near-infrared (NIR) spectra and quantitative structure-activity relationship (QSAR) data, provides essential information if a proper model is selected. Similarly, with a general model selection approach, spectral model maintenance (updating) from original modeling conditions to new conditions can be performed for dynamic modeling. Fundamental modeling (partial least-squares (PLS) and others) and maintenance processes (domain adaptation or transfer learning and others) require selection of tuning parameter(s) values to isolate models that can accurately predict new samples or molecules, e.g., number of PLS latent variables to predict analyte concentration. Regardless of the modeling task, model selection is complex and without a reliable protocol. Tuning parameter selection typically depends on only one model quality measure assessing model bias using prediction accuracy. Developed in this paper is a generic model selection process using concepts from consensus modeling and QSAR activity landscapes. It is a consensus filtering approach that prioritizes model diversity (MD) while conserving prediction similarity (PS) fused with a common bias-variance trade-off measure. A significant feature of MDPS is that a cross-validation scheme is not needed because models are selected relative to predicting new samples or molecules, i.e., model selection uses unlabeled samples (without reference values) for active predictions. The versatility and reliability of MDPS model selection is shown using four NIR data sets and a QSAR data set. The study also substantiates the Rashomon effect where there is not one best model tuning parameter value that provides accurate predictions.


Effectiveness and cost-effectiveness of Improving clinicians' diagnostic and communication Skills on Antibiotic prescribing Appropriateness in patients with acute Cough in primary care in CATalonia (the ISAAC-CAT study): study protocol for a cluster randomised controlled trial.
Despite their marginal benefit, about 60% of acute lower respiratory tract infections (ALRTIs) are currently treated with antibiotics in Catalonia. This study aims to evaluate the effectiveness and efficiency of a continuous disease-focused intervention (C-reactive protein [CRP]) and an illness-focused intervention (enhancement of communication skills to optimise doctor-patient consultations) on antibiotic prescribing in patients with ALRTIs in Catalan primary care centres. A cluster randomised, factorial, controlled trial aimed at including 20 primary care centres (N = 2940 patients) with patients older than 18 years of age presenting for a first consultation with an ALRTI will be included in the study. Primary care centres will be identified on the basis of socioeconomic data and antibiotic consumption. Centres will be randomly assigned according to hierarchical clustering to any of four trial arms: usual care, CRP testing, enhanced communication skills backed up with patient leaflets, or combined interventions. A cost-effectiveness and cost-utility analysis will be performed from the societal and national healthcare system perspectives, and the time horizon of the analysis will be 1 year. Two qualitative studies (pre- and post-clinical trial) aimed to identify the expectations and concerns of patients with ALRTIs and the barriers and facilitators of each intervention arm will be run. Family doctors and nurses assigned to the interventions will participate in a 2-h training workshop before the inception of the trial and will receive a monthly intervention-tailored training module during the year of the trial period. Primary outcomes will be antibiotic use within the first 6 weeks, duration of moderate to severe cough, and the quality-adjusted life-years. Secondary outcomes will be duration of illness and severity of cough measured using a symptom diary, healthcare re-consultations, hospital admissions, and complications. Healthcare costs will be considered and expressed in 2021 euros (year foreseen to finalise the study) of the current year of the analysis. Univariate and multivariate sensitivity analyses will be carried out. The ISAAC-CAT project will contribute to evaluate the effectiveness and efficiency of different strategies for more appropriate antibiotic prescribing that are currently out of the scope of the actual clinical guidelines. ClinicalTrials.gov, NCT03931577.


Breast cancer detection with short-interval follow-up compared with return to annual screening in patients with benign stereotactic or US-guided breast biopsy results.
To compare the cancer detection rate and stage after benign stereotactic or ultrasonography (US)-guided core breast biopsy between patients with short-interval follow-up (SIFU) and those who return to annual screening. The Breast Cancer Surveillance Consortium (BCSC) registry and the BCSC Statistical Coordinating Center received institutional review board approval for active and passive consent processes and a waiver of consent. All procedures were HIPAA compliant. BCSC data for 1994-2010 were used to compare ipsilateral breast cancer detection rates and tumor characteristics for diagnoses within 3 months after SIFU (3-8 months) versus return to annual screening (RTAS) mammography (9-18 months) after receiving a benign pathology result from image-guided breast biopsy. In total, 17 631 biopsies with benign findings were identified with SIFU or RTAS imaging. In the SIFU group, 27 ipsilateral breast cancers were diagnosed in 10 715 mammographic examinations (2.5 cancers per 1000 examinations) compared with 16 cancers in 6916 mammographic examinations in the RTAS group (2.3 cancers per 1000 examinations) (P = .88). Sixteen cancers after SIFU (59%; 95% confidence interval [CI]: 39%, 78%) were invasive versus 12 after RTAS (75%; 95% CI: 48%, 93%). The invasive cancer rate was 1.5 per 1000 examinations after SIFU (95% CI: 0.9, 2.4) and 1.7 per 1000 examinations (95% CI: 0.9, 3.0) after RTAS (P = .70). Among invasive cancers, 25% were late stage (stage 2B, 3, or 4) in the SIFU group (95% CI: 7%, 52%) versus 27% in the RTAS group (95% CI: 6%, 61%). Positive lymph nodes were found in seven (44%; 95% CI: 20%, 70%) invasive cancers after SIFU and in three (25%; 95% CI: 5%, 57%) invasive cancers after RTAS. Similar rates of cancer detection were found between SIFU and RTAS after benign breast biopsy with no significant differences in stage, tumor size, or nodal status, although the present study was limited by sample size. These findings suggest that patients with benign radiologic-pathologic-concordant percutaneous breast biopsy results could return to annual screening.


Classical conditioning in the vegetative and minimally conscious state.
Pavlovian trace conditioning depends on the temporal gap between the conditioned and unconditioned stimuli. It requires, in mammals, functional medial temporal lobe structures and, in humans, explicit knowledge of the temporal contingency. It is therefore considered to be a plausible objective test to assess awareness without relying on explicit reports. We found that individuals with disorders of consciousness (DOCs), despite being unable to report awareness explicitly, were able to learn this procedure. Learning was specific and showed an anticipatory electromyographic response to the aversive conditioning stimulus, which was substantially stronger than to the control stimulus and was augmented as the aversive stimulus approached. The amount of learning correlated with the degree of cortical atrophy and was a good indicator of recovery. None of these effects were observed in control subjects under the effect of anesthesia (propofol). Our results suggest that individuals with DOCs might have partially preserved conscious processing, which cannot be mediated by explicit reports and is not detected by behavioral assessment.


Outcomes of Gamma Knife surgery for trigeminal neuralgia secondary to vertebrobasilar ectasia.
Vertebrobasilar ectasia (VBE) is an unusual cause of trigeminal neuralgia (TN). The surgical options for patients with medically refractory pain include percutaneous or microsurgical rhizotomy and microvascular decompression (MVD). All such procedures can be technically challenging. This report evaluates the response to a minimally invasive procedure, Gamma Knife surgery (GKS), in patients with TN associated with severe vascular compression caused by VBE. Twenty patients underwent GKS for medically refractory TN associated with VBE. The median patient age was 74 years (range 48-95 years). Prior surgical procedures had failed in 11 patients (55%). In 9 patients (45%), GKS was the first procedure they had undergone. The median target dose for GKS was 80 Gy (range 75-85 Gy). The median follow-up was 29 months (range 8-123 months) after GKS. The treatment outcomes were compared with 80 case-matched controls who underwent GKS for TN not associated with VBE. Intraoperative MR imaging or CT scanning revealed VBE that deformed the brainstem in 50% of patients. The trigeminal nerve was displaced in cephalad or lateral planes in 60%. In 4 patients (20%), the authors could identify only the distal cisternal component of the trigeminal nerve as it entered into the Meckel cave. After GKS, 15 patients (75%) achieved initial pain relief that was adequate or better, with or without medication (Barrow Neurological Institute [BNI] pain scale, Grades I-IIIb). The median time until pain relief was 5 weeks (range 1 day-6 months). Twelve patients (60%) with initial pain relief reported recurrent pain between 3 and 43 months after GKS (median 12 months). Pain relief was maintained in 53% at 1 year, 38% at 2 years, and 10% at 5 years. Some degree of facial sensory dysfunction occurred in 10% of patients. Eventually, 14 (70%) of the 20 patients underwent an additional surgical procedure including repeat GKS, percutaneous procedure, or MVD at a median of 14 months (range 5-50 months) after the initial GKS. At the last follow-up, 15 patients (75%) had satisfactory pain control (BNI Grades I-IIIb), but 5 patients (25%) continued to have unsatisfactory pain control (BNI Grade IV or V). Compared with patients without VBE, patients with VBE were much less likely to have initial (p = 0.025) or lasting (p = 0.006) pain relief. Pain control rates of GKS in patients with TN associated with VBE were inferior to those of patients without VBE. Multimodality surgical or medical management strategies were required in most patients with VBE.


Quality of care among obese patients.
Clinicians often have negative attitudes toward obesity and express dissatisfaction in caring for obese patients. Moreover, obese patients often feel that clinicians are biased or disrespectful because of their weight. These observations raise the concern that obese patients may receive lower quality of care. To determine whether performance on common outpatient quality measures differs by patient weight status. Eight different performance measures were examined in 2 national-level patient populations: (1) Medicare beneficiaries (n = 36 122) using data from the Medicare Beneficiary Survey (1994-2006); and (2) recipients of care from the Veterans Health Administration (VHA) (n = 33 550) using data from an ongoing performance-evaluation program (2003-2004). Performance measures among eligible patients for diabetes care (eye examination, glycated hemoglobin [HbA(1c)] testing, and lipid screening), pneumococcal vaccination, influenza vaccination, screening mammography, colorectal cancer screening, and cervical cancer screening. Measures were based on a combination of administrative claims, survey, and chart review data. We found no evidence that obese or overweight patients were less likely to receive recommended care relative to normal-weight patients. Moreover, success rates were marginally higher for obese and/or overweight patients on several measures. The most notable differentials were observed for recommended diabetes care among Medicare beneficiaries: comparing obese vs normal-weight patients with diabetes, obese patients were more likely to receive recommended care on lipid screening (72% vs 65%; odds ratio, 1.37 [95% confidence interval, 1.09-1.73]) and HbA(1c) testing (74% vs 62%; odds ratio, 1.73 [95% confidence interval, 1.41-2.11]). All analyses were adjusted for sociodemographic factors, health status, clinical complexity, and visit frequency. Among samples of patients from the Medicare and VHA populations, there was no evidence across 8 performance measures that obese or overweight patients received inferior care when compared with normal-weight patients. Being obese or overweight was associated with a marginally higher rate of recommended care on several measures.


The effect of developmental temperature on the genetic architecture underlying size and thermal clines in Drosophila melanogaster and D. simulans from the east coast of Australia.
Body size and thermal tolerance clines in Drosophila melanogaster occur along the east coast of Australia. However the extent to which temperature affects the genetic architecture underlying the observed clinal divergence remains unknown. Clinal variation in these traits is associated with cosmopolitan chromosome inversions that cline in D. melanogaster. Whether this association influences the genetic architecture for these traits in D. melanogaster is unclear. Drosophila simulans shows linear clines in body size, but nonlinear clines in cold resistance. Clinally varying inversions are absent in D. simulans. Line-cross and clinal analyses were performed between tropical and temperate populations of D. melanogaster and D. simulans from the east coast of Australia to investigate whether clinal patterns and genetic effects contributing to clinal divergence in wing centroid size, thorax length, wing-to-thorax ratio, cold and heat resistance differed under different developmental temperatures (18 °C, 25 °C, and 29 °C). Developmental temperature influenced the genetic architecture in both species. Similarities between D. melanogaster and D. simulans suggest clinally varying inversion polymorphisms have little influence on the genetic architecture underlying clinal divergence in size in D. melanogaster. Differing genetic architectures across different temperatures highlight the need to consider different environments in future evolutionary and molecular studies of phenotypic divergence.


Optimal control-based bayesian detection of clinical and behavioral state transitions.
Accurately detecting hidden clinical or behavioral states from sequential measurements is an emerging topic in neuroscience and medicine, which may dramatically impact neural prosthetics, brain-computer interface and drug delivery. For example, early detection of an epileptic seizure from sequential electroencephalographic (EEG) measurements would allow timely administration of anticonvulsant drugs or neurostimulation, thus reducing physical impairment and risks of overtreatment. We develop a Bayesian paradigm for state transition detection that combines optimal control and Markov processes. We define a hidden Markov model of the state evolution and develop a detection policy that minimizes a loss function of both probability of false positives and accuracy (i.e., lag between estimated and actual transition time). Our strategy automatically adapts to each newly acquired measurement based on the state evolution model and the relative loss for false positives and accuracy, thus resulting in a time varying threshold policy. The paradigm was used in two applications: 1) detection of movement onset (behavioral state) from subthalamic single unit recordings in Parkinson's disease patients performing a motor task; 2) early detection of an approaching seizure (clinical state) from multichannel intracranial EEG recordings in rodents treated with pentylenetetrazol chemoconvulsant. Our paradigm performs significantly better than chance and improves over widely used detection algorithms.


Automated SNP detection from a large collection of white spruce expressed sequences: contributing factors and approaches for the categorization of SNPs.
High-throughput genotyping technologies represent a highly efficient way to accelerate genetic mapping and enable association studies. As a first step toward this goal, we aimed to develop a resource of candidate Single Nucleotide Polymorphisms (SNP) in white spruce (Picea glauca [Moench] Voss), a softwood tree of major economic importance. A white spruce SNP resource encompassing 12,264 SNPs was constructed from a set of 6,459 contigs derived from Expressed Sequence Tags (EST) and by using the bayesian-based statistical software PolyBayes. Several parameters influencing the SNP prediction were analysed including the a priori expected polymorphism, the probability score (PSNP), and the contig depth and length. SNP detection in 3' and 5' reads from the same clones revealed a level of inconsistency between overlapping sequences as low as 1%. A subset of 245 predicted SNPs were verified through the independent resequencing of genomic DNA of a genotype also used to prepare cDNA libraries. The validation rate reached a maximum of 85% for SNPs predicted with either PSNP > or = 0.95 or > or = 0.99. A total of 9,310 SNPs were detected by using PSNP > or = 0.95 as a criterion. The SNPs were distributed among 3,590 contigs encompassing an array of broad functional categories, with an overall frequency of 1 SNP per 700 nucleotide sites. Experimental and statistical approaches were used to evaluate the proportion of paralogous SNPs, with estimates in the range of 8 to 12%. The 3,789 coding SNPs identified through coding region annotation and ORF prediction, were distributed into 39% nonsynonymous and 61% synonymous substitutions. Overall, there were 0.9 SNP per 1,000 nonsynonymous sites and 5.2 SNPs per 1,000 synonymous sites, for a genome-wide nonsynonymous to synonymous substitution rate ratio (Ka/Ks) of 0.17. We integrated the SNP data in the ForestTreeDB database along with functional annotations to provide a tool facilitating the choice of candidate genes for mapping purposes or association studies.


Noiseless independent signal and power amplification.
We present a noiseless optical amplifier comprising a signal-amplifying feed-forward loop and a power-amplifying injection-locked laser. We demonstrate that the signal amplifier can attain a signal-transfer coefficient limited solely by the quantum efficiency of our in-loop photodetector and that we can independently amplify the optical power while leaving the normalized intensity-noise spectral density of the input field unchanged.


Molecular Characterization of Vitellogenin and Vitellogenin Receptor of Bemisia tabaci.
Vitellogenin (Vg) plays vital role in oocytes and embryo development in insects. Vg is synthesized in the fat body, moves through haemolymph and accumulates in oocytes. Vitellogenin receptors (VgR) present on the surface of oocytes, are responsible for Vg transportation from haemolymph to oocytes. Here, we cloned and characterized these genes from Bemisia tabaci Asia1 (BtA1) species. The cloned BtA1Vg and BtA1VgR genes consisted of 6,330 and 5,430 bp long open reading frames, which encoded 2,109 and 1,809 amino acid (AA) residues long protein. The BtA1Vg protein comprised LPD_N, DUF1943 and VWFD domains, typical R/KXXR/K, DGXR and GL/ICG motifs, and polyserine tracts. BtA1VgR protein contained 12 LDLa, 10 LDLb and 7 EGF domains, and a trans-membrane and cytoplasmic region at C-terminus. Phylogenetic analyses indicated evolutionary association of BtA1Vg and BtA1VgR with the homologous proteins from various insect species. Silencing of BtA1VgR by siRNA did not affect the transcript level of BtA1Vg. However, BtA1Vg protein accumulation in oocytes was directly influenced with the expression level of BtA1VgR. Further, BtA1VgR silencing caused significant mortality and reduced fecundity in adult whiteflies. The results established the role of BtA1VgR in transportation of BtA1Vg in oocytes. Further, these proteins are essential for fecundity, and therefore these can be potential RNAi targets for insect control in crop plants.


Cyclomaltodextrinase, neopullulanase, and maltogenic amylase are nearly indistinguishable from each other.
Over 20 enzymes denoted as cyclomaltodextrinase, maltogenic amylase, or neopullulanase that share 40-86% sequence identity with each other are found in public data bases. These enzymes are distinguished from typical alpha-amylases by containing a novel N-terminal domain and exhibiting preferential substrate specificities for cyclomaltodextrins (CDs) over starch. In this research field, a great deal of confusion exists regarding the features distinguishing the three groups of enzymes from one another. Although a different enzyme code has been assigned to each of the three different enzyme names, even a single differentiating enzymatic property has not been documented in the literature. On the other hand, an outstanding question related to this issue concerns the structural basis for the preference of these enzymes for CDs. To clarify the confusion and to address this question, we have determined the structures of two enzymes, one from alkalophilic Bacillus sp. I-5 and named cyclomaltodextrinase and the other from a Thermus species and named maltogenic amylase. The structure of the Bacillus enzyme reveals a dodecameric assembly composed of six copies of the dimer, which is the structural and functional unit of the Thermus enzyme and an enzyme named neopullulanase. The structure of the Thermus enzyme in complex with beta-CD led to the conclusion that Trp47, a well conserved N-terminal domain residue, contributes greatly to the preference for beta-CD. The common dimer formation through the novel N-terminal domain, which contributes to the preference for CDs by lining the active-site cavity, convincingly indicates that the three groups of enzymes are not different enough to preserve the different names and enzyme codes.


The histology of brain tumors for 67 331 children and 671 085 adults diagnosed in 60 countries during 2000-2014: a global, population-based study (CONCORD-3).
Global variations in survival for brain tumors are very wide when all histological types are considered together. Appraisal of international differences should be informed by the distribution of histology, but little is known beyond Europe and North America. The source for the analysis was the CONCORD database, a program of global surveillance of cancer survival trends, which includes the tumor records of individual patients from more than 300 population-based cancer registries. We considered all patients aged 0-99 years who were diagnosed with a primary brain tumor during 2000-2014, whether malignant or nonmalignant. We presented the histology distribution of these tumors, for patients diagnosed during 2000-2004, 2005-2009, and 2010-2014. Records were submitted from 60 countries on 5 continents, 67 331 for children and 671 085 for adults. After exclusion of irrelevant morphology codes, the final study population comprised 60 783 children and 602 112 adults. Only 59 of 60 countries covered in CONCORD-3 were included because none of the Mexican records were eligible. We defined 12 histology groups for children, and 11 for adults. In children (0-14 years), the proportion of low-grade astrocytomas ranged between 6% and 50%. Medulloblastoma was the most common subtype in countries where low-grade astrocytoma was less commonly reported. In adults (15-99 years), the proportion of glioblastomas varied between 9% and 69%. International comparisons were made difficult by wide differences in the proportion of tumors with unspecified histology, which accounted for up to 52% of diagnoses in children and up to 65% in adults. To our knowledge, this is the first account of the global histology distribution of brain tumors, in children and adults. Our findings provide insights into the practices and the quality of cancer registration worldwide.


Venereal transmission of La Crosse virus from male to female Aedes triseriatus.
Horizontal transmission of La Crosse (LAC) arbovirus in colonized Aedes triseriatus was first observed in mating cages. LAC antigen was visualized by fluorescent antibody technique in multiple organs of 5 (2.5% of 198 females dissected after contact in mating cages with males, with a transovarial infection rate of 31%. Venereal transmission of LAC virus was demonstrated by limited contact induced mating. Antigen was detected in bursal contents of 35 (53%) of 66 females dissected within 24 h after induced mating by males previously infected by intrathoracic inoculation. Antigen was observed in ovaries, heart, gut, salivary glands, or other organs in 13 (3.4%) of 379 females dissected 1 to 14 days post-mating. LAC virus was isolated from infected males, from contents of the bursa copulatrix of females following mating, and from dissection remnants of 6 of the 13 females in which disseminated antigen had been observed. Venereally infected females were shown to be capable of transmission of LAC virus by bite to mice and to eggs.


The SGLT2 inhibitor dapagliflozin in heart failure with preserved ejection fraction: a multicenter randomized trial.
Patients with heart failure and preserved ejection fraction (HFpEF) have a high burden of symptoms and functional limitations, and have a poor quality of life. By targeting cardiometabolic abmormalities, sodium glucose cotransporter 2 (SGLT2) inhibitors may improve these impairments. In this multicenter, randomized trial of patients with HFpEF (NCT03030235), we evaluated whether the SGLT2 inhibitor dapagliflozin improves the primary endpoint of Kansas City Cardiomyopathy Questionnaire Clinical Summary Score (KCCQ-CS), a measure of heart failure-related health status, at 12 weeks after treatment initiation. Secondary endpoints included the 6-minute walk test (6MWT), KCCQ Overall Summary Score (KCCQ-OS), clinically meaningful changes in KCCQ-CS and -OS, and changes in weight, natriuretic peptides, glycated hemoglobin and systolic blood pressure. In total, 324 patients were randomized to dapagliflozin or placebo. Dapagliflozin improved KCCQ-CS (effect size, 5.8 points (95% confidence interval (CI) 2.3-9.2, P = 0.001), meeting the predefined primary endpoint, due to improvements in both KCCQ total symptom score (KCCQ-TS) (5.8 points (95% CI 2.0-9.6, P = 0.003)) and physical limitations scores (5.3 points (95% CI 0.7-10.0, P = 0.026)). Dapagliflozin also improved 6MWT (mean effect size of 20.1 m (95% CI 5.6-34.7, P = 0.007)), KCCQ-OS (4.5 points (95% CI 1.1-7.8, P = 0.009)), proportion of participants with 5-point or greater improvements in KCCQ-OS (odds ratio (OR) = 1.73 (95% CI 1.05-2.85, P = 0.03)) and reduced weight (mean effect size, 0.72 kg (95% CI 0.01-1.42, P = 0.046)). There were no significant differences in other secondary endpoints. Adverse events were similar between dapagliflozin and placebo (44 (27.2%) versus 38 (23.5%) patients, respectively). These results indicate that 12 weeks of dapagliflozin treatment significantly improved patient-reported symptoms, physical limitations and exercise function and was well tolerated in chronic HFpEF.


Reducing the silent burden of impaired mental health.
Mental and behavioral disorders account for about one third of the world's disability caused by all ill health among adults, with unipolar depressive disorders set to be the world's number one cause of illhealth and premature death in 2030, affecting high- and low-income countries. There is a range of evidence-based cost-effective interventions that can be implemented in parenting, at schools, at the workplace, and in older age that can promote health and well-being, reduce mental disorders, lead to improved productivity, and increase resilience to cope with many of the stressors in the world. These facts need to be better communicated to policymakers to ensure that the silent burden of impaired mental health is adequately heard and reduced.


A multi-layered and dynamic apical extracellular matrix shapes the vulva lumen in Caenorhabditis elegans.
Biological tubes must develop and maintain their proper diameter to transport materials efficiently. These tubes are molded and protected in part by apical extracellular matrices (aECMs) that line their lumens. Despite their importance, aECMs are difficult to image in vivo and therefore poorly understood. The Caenorhabditis elegans vulva has been a paradigm for understanding many aspects of organogenesis. Here we describe the vulva luminal matrix, which contains chondroitin proteoglycans, Zona Pellucida (ZP) domain proteins, and other glycoproteins and lipid transporters related to those in mammals. Confocal and transmission electron microscopy revealed, with unprecedented detail, a complex and dynamic aECM. Different matrix factors assemble on the apical surfaces of each vulva cell type, with clear distinctions seen between Ras-dependent (1°) and Notch-dependent (2°) cell types. Genetic perturbations suggest that chondroitin and other aECM factors together generate a structured scaffold that both expands and constricts lumen shape.


Compression behavior of single-layer graphenes.
Central to most applications involving monolayer graphenes is its mechanical response under various stress states. To date most of the work reported is of theoretical nature and refers to tension and compression loading of model graphenes. Most of the experimental work is indeed limited to the bending of single flakes in air and the stretching of flakes up to typically approximately 1% using plastic substrates. Recently we have shown that by employing a cantilever beam we can subject single graphenes to various degrees of axial compression. Here we extend this work much further by measuring in detail both stress uptake and compression buckling strain in single flakes of different geometries. In all cases the mechanical response is monitored by simultaneous Raman measurements through the shift of either the G or 2D phonons of graphene. Despite the infinitely small thickness of the monolayers, the results show that graphenes embedded in plastic beams exhibit remarkable compression buckling strains. For large length (l)-to-width (w) ratios (> or =0.2) the buckling strain is of the order of -0.5% to -0.6%. However, for l/w < 0.2 no failure is observed for strains even higher than -1%. Calculations based on classical Euler analysis show that the buckling strain enhancement provided by the polymer lateral support is more than 6 orders of magnitude compared to that of suspended graphene in air.


Flood or drought: how do aerosols affect precipitation?
Aerosols serve as cloud condensation nuclei (CCN) and thus have a substantial effect on cloud properties and the initiation of precipitation. Large concentrations of human-made aerosols have been reported to both decrease and increase rainfall as a result of their radiative and CCN activities. At one extreme, pristine tropical clouds with low CCN concentrations rain out too quickly to mature into long-lived clouds. On the other hand, heavily polluted clouds evaporate much of their water before precipitation can occur, if they can form at all given the reduced surface heating resulting from the aerosol haze layer. We propose a conceptual model that explains this apparent dichotomy.


Challenging homophobia and heterosexism through storytelling and critical dialogue among Hong Kong Chinese immigrant parents in Toronto.
Homophobia and heterosexism are ubiquitous in Canadian society. They contribute to significant health and mental health disparities for lesbian, gay and bisexual youth and their families. Anti-homophobia efforts tend to focus on students and teachers at school. While these efforts are important, they do not reach parents, who play an important role in shaping young people's attitudes towards gender and sexuality. To eliminate bullying and victimisation associated with homophobia at school and in the community, concerted efforts are urgently needed to mobilise parents to become champions against homophobia and heterosexism. In this paper, we report on our use of storytelling and critical dialogue to engage a group of Hong Kong Chinese immigrant parents in Toronto to interrogate their values and assumptions about homosexuality. In particular, we illustrate how we use storytelling to create a liminal space whereby the narrators and listeners collaborate to create counter-discourses that challenge social domination and exclusion. We then discuss the implications of using a critical dialogical approach to integrate anti-homophobia efforts in community parenting programmes.


Maximum speed and mechanical power output in lizards.
The goal of the present study was to test the hypothesis that maximum running speed is limited by how much mechanical power the muscular system can produce. To test this hypothesis, two species of lizards, Coleonyx variegatus and Eumeces skiltonianus, sprinted on hills of different slopes. According to the hypothesis, maximum speed should decrease on steeper uphill slopes but mechanical power output at maximum speed should be independent of slope. For level sprinting, the external mechanical power output was determined from force platform data. For uphill sprinting, the mechanical power output was approximated as the power required to lift the center of mass vertically. When the slope increased from level to 40 degrees uphill, maximum speed decreased by 28% in C. variegatus and by 16% in E. skiltonianus. At maximum speed on a 40 degrees uphill slope in both species, the mechanical power required to lift the body vertically was approximately 3.9 times greater than the external mechanical power output at maximum speed on the level. Because total limb mass is small in both species (6-16% of body mass) and stride frequency is similar at maximum speed on all slopes, the internal mechanical power output is likely to be small and similar in magnitude on all slopes. I conclude that the muscular system is capable of producing substantially more power during locomotion than it actually produces during level sprinting. Thus, the capacity of the muscular system to produce power does not limit maximum running speed.


Fabrication of 150 nm half-pitch grating templates for nanoimprint lithography.
We present the fabrication of 150 nm half-pitch Si grating templates by reactive ion etch (RIE), which are used in nanoimprint lithography (NIL) for high groove density gratings in SU-8 plastic. The etch properties such as the etch rate, profile and etching selectivity of Si over Cr as etch mask were carefully studied. Under the optimum condition Si gratings with 150 nm in linewidth, 480 nm in height and nearly 90 degree in verticality of the sidewall have been achieved. 150 nm half-pitch gratings on SU-8 were then successfully imprinted using the fabricated templates. The diffraction pattern of zeroth and first order from the SU-8 gratings was observed using a 266 nm laser beam. The developed nanofabrication technique in this work is applicable not only for templates but also for other nanostructures in silicon.


Safety Assessment of Acyl Glucuronides-A Simplified Paradigm.
While simple O- (ether-linked) and N-glucuronide drug conjugates generally are unreactive and considered benign from a safety perspective, the acyl glucuronides that derive from metabolism of carboxylic acid-containing xenobiotics can exhibit a degree of chemical reactivity that is dependent upon their molecular structure. As a result, concerns have arisen over the safety of acyl glucuronides as a class, several members of which have been implicated in the toxicity of their respective parent drugs. However, direct evidence in support of these claims remains sparse, and due to frequently encountered species differences in the systemic exposure to acyl glucuronides (both of the parent drug and oxidized derivatives thereof), coupled with their instability in aqueous media and potential to undergo chemical rearrangement (acyl migration), qualification of these conjugates by traditional safety assessment methods can be very challenging. In this Commentary, we discuss alternative (non-acyl glucuronide) mechanisms by which carboxylic acids may cause serious adverse reactions, and propose a novel, practical approach to compare systemic exposure to acyl glucuronide metabolites in humans to that in animal species used in preclinical safety assessment based on relative estimates of the total body burden of these circulating conjugates.


Tobacco and electronic cigarette cues for smoking and vaping: an online experimental study.
This study examined whether exposure to smoking and vaping cues the urge to smoke or vape. It extends previous studies on first-generation cigalikes (visually similar to cigarettes) and second-generation devices (visually similar to pens) by including third-generation tank system devices (larger bulky units). In an online experiment, participants were randomly assigned to view one of four videos, which included smoking, vaping (cigalike or tank system), or neutral cues. The primary outcome was urge to smoke. Secondary outcomes were urge to vape, desire to smoke and vape, and intention to quit or remain abstinent from smoking. UK adults varying in smoking (current or former) and vaping (user or non-user) status (n = 1120) completed the study: 184 (16%) failed study attention checks meaning 936 were included in the final analysis. Urges to smoke were similar across cue groups. Urges to vape were higher following exposure to vaping compared to neutral cues. There was no clear evidence of an interaction between cue group and smoking or vaping status. The lack of cueing effects on smoking urges is inconsistent with previous research, raising questions about the ability to assess craving in online settings.


Mature murine B and T cells transferred to SCID mice can survive indefinitely and many maintain a virgin phenotype.
To seek information on the potential lifespan of normal B and T lymphocytes, lymph node (LN) cells from unprimed mice were transferred to H-2-identical severe combined immunodeficiency (SCID) hosts. At a population level, the donor B and T cells survived for at least 10 mo post-transfer with no reduction in their numbers. In terms of antibody production, LN-injected SCID mice remained responsive to several different antigens and contained unprimed precursors of memory cells for greater than or equal to 6 mo post-transfer. Most of the B and T cells recovered from LN-injected SCID mice expressed the typical virgin phenotype of mature lymphocytes from young mice. These findings suggest that many of the transferred lymphocytes might have remained in interphase as virgin cells from the time of injection. This did not apply to all of the transferred cells, however, because 20-40% of CD4+ cells from long-term SCID hosts displayed a memory phenotype, 7% incorporated 2-bromodeoxyuridine over 5 d, and total numbers of B and T cells increased gradually (twofold) over a 10-mo period. Collectively, the data favor the view that the pool of mature B and T cells in adult mice is largely self sufficient: some of the cells proliferate, presumably in response to environmental antigens, but many mature cells can remain quiescent for prolonged periods. Input of new cells from the primary lymphoid organs continues, but at a much reduced rate relative to young life.


Laser welding method for removal of instruments debris from root canals.
The purpose of this study was to clarify the viability of a novel method for removing debris from broken instruments from root canals using a laser apparatus. Laser welding was performed on stainless steel or nickel titanium files using an Nd:YAG laser. Retention force between the files and extractors was measured. Increase in temperature on the root surface during laser irradiation was recorded and the irradiated areas evaluated with a scanning electron microscope. Retention force on stainless steel was significantly greater than that on nickel titanium. The maximum temperature increase was 4.1°C. The temperature increase on the root surface was greater in the vicinity of the welded area than that at the apical area. Scanning electron microscopy revealed that the files and extractors were welded together. These results indicate that the laser welding method is effective in removing debris from broken instruments from root canals.


Effects of sounds of locomotion on speech perception.
Human locomotion typically creates noise, a possible consequence of which is the masking of sound signals originating in the surroundings. When walking side by side, people often subconsciously synchronize their steps. The neurophysiological and evolutionary background of this behavior is unclear. The present study investigated the potential of sound created by walking to mask perception of speech and compared the masking produced by walking in step with that produced by unsynchronized walking. The masking sound (footsteps on gravel) and the target sound (speech) were presented through the same speaker to 15 normal-hearing subjects. The original recorded walking sound was modified to mimic the sound of two individuals walking in pace or walking out of synchrony. The participants were instructed to adjust the sound level of the target sound until they could just comprehend the speech signal ("just follow conversation" or JFC level) when presented simultaneously with synchronized or unsynchronized walking sound at 40 dBA, 50 dBA, 60 dBA, or 70 dBA. Synchronized walking sounds produced slightly less masking of speech than did unsynchronized sound. The median JFC threshold in the synchronized condition was 38.5 dBA, while the corresponding value for the unsynchronized condition was 41.2 dBA. Combined results at all sound pressure levels showed an improvement in the signal-to-noise ratio (SNR) for synchronized footsteps; the median difference was 2.7 dB and the mean difference was 1.2 dB [P < 0.001, repeated-measures analysis of variance (RM-ANOVA)]. The difference was significant for masker levels of 50 dBA and 60 dBA, but not for 40 dBA or 70 dBA. This study provides evidence that synchronized walking may reduce the masking potential of footsteps.


Mutual interference between genomic RNA replication and subgenomic mRNA transcription in brome mosaic virus.
Replication by many positive-strand RNA viruses includes genomic RNA amplification and subgenomic mRNA (sgRNA) transcription. For brome mosaic virus (BMV), both processes occur in virus-induced, membrane-associated compartments, require BMV replication factors 1a and 2a, and use negative-strand RNA3 as a template for genomic RNA3 and sgRNA syntheses. To begin elucidating their relations, we examined the interaction of RNA3 replication and sgRNA transcription in Saccharomyces cerevisiae expressing 1a and 2a, which support the full RNA3 replication cycle. Blocking sgRNA transcription stimulated RNA3 replication by up to 350%, implying that sgRNA transcription inhibits RNA3 replication. Such inhibition was independent of the sgRNA-encoded coat protein and operated in cis. We further found that sgRNA transcription inhibited RNA3 replication at a step or steps after negative-strand RNA3 synthesis, implying competition with positive-strand RNA3 synthesis for negative-strand RNA3 templates, viral replication factors, or common host components. Consistent with this, sgRNA transcription was stimulated by up to 400% when mutations inhibiting positive-strand RNA3 synthesis were introduced into the RNA3 5'-untranslated region. Thus, BMV subgenomic and genomic RNA syntheses mutually interfered with each other, apparently by competition for one or more common factors. In plant protoplasts replicating all three BMV genomic RNAs, mutations blocking sgRNA transcription often had lesser effects on RNA3 accumulation, possibly because RNA3 also competed with RNA1 and RNA2 replication templates and because any increase in RNA3 replication at the expense of RNA1 and RNA2 would be self-limited by decreased 1a and 2a expression from RNA1 and RNA2.


Morphological and cytochemical identification of the Golgi apparatus.
1. A modification of Elftman's direct silver method reveals both the lipochondria of Baker and the network of Golgi in the same cell. For purpose of distinction, it is proposed to call Baker's lipochondria the nucleopetal fraction, the Golgi network the nucleofugal fraction of the Golgi apparatus. 2. The nucleopetal fraction is located closer to the nucleus. It is spherical in shape and appears black in color. The nucleofugal fraction is located farther away from the nucleus. It is reticular in form and appears brown in color after silver impregnation by the modified Elftman's method. 3. These two fractions are separate entities. The network of Golgi is not due to deposition of silver on lipochondria. Lipochondria do not represent Golgi apparatus in living cells. 4. Aldehydes facilitate the demonstration of the nucleofugal fraction. Based on the circumstantial evidence presented, it appears that aldehyde dehydrogenase composed of a specific protein bound to a prosthetic group of flavin-adenine dinucleotide may be concentrated in this fraction. 5. Aldehyde dehydrogenase also functions as xanthine oxidase. It is suggested as a working hypothesis that under physiological condition, one of the functions of the nucleofugal fraction (Golgi network) is concerned with purine metabolism of nucleoproteins.


Molecular markers to predict clinical outcome and radiation induced toxicity in lung cancer.
The elucidation of driver mutations involved in the molecular pathogenesis of cancer has led to a surge in the application of novel targeted therapeutics in lung cancer. Novel oncologic research continues to lead investigators towards targeting personalized tumor characteristics rather than applying targeted therapy to broad patient populations. Several driver genes, in particular epidermal growth factor receptor (EGFR) and ALK fusions, are the earliest to have made their way into clinical trials. The avant-garde role of genomic profiling has led to important clinical challenges when adapting current standard treatments to personalized oncologic care. This new frontier of medicine requires newer biomarkers for toxicity that will identify patients at risk, as well as, new molecular markers to predict and assess clinical outcomes. Thus far, several signature genes have been developed to predict outcome as well as genetic factors related to inflammation to predict toxicity.


Process-Structure-Quality Relationships of Three-Dimensional Printed Poly(Caprolactone)-Hydroxyapatite Scaffolds.
Bone defects are common and, in many cases, challenging to treat. Tissue engineering is an interdisciplinary approach with promising potential for treating bone defects. Within tissue engineering, three-dimensional (3D) printing strategies have emerged as potent tools for scaffold fabrication. However, reproducibility and quality control are critical aspects limiting the translation of 3D printed scaffolds to clinical use, which remain to be addressed. To elucidate the factors that yield to the generation of defects in bioprinting and to achieve reproducible biomaterial printing, the objective of this article is to frame a systematic approach for optimizing and validating 3D printing of poly(caprolactone) (PCL)-hydroxyapatite (HAp) composite scaffolds. We delineate the effect of PCL-to-HAp ratio, print velocity, print temperature, and extrusion pressure on the architectural and mechanical properties of the 3D printed scaffold. Furthermore, we present an in situ image-based monitoring approach to quantify key quality-related aspects of constructs, such as the ability to deposit material consistently and print elementary shapes with fewer flaws. Our results show that small defects generated during the printing process have a significant role in lowering the mechanical properties of 3D printed polymeric scaffolds. In addition, the in vitro osteoinductivity of the fabricated scaffolds is demonstrated. Impact statement Identifying quality control measures is essential in the translation of three-dimensional (3D) printed scaffolds into clinical practice. In this article, we highlighted the importance of selected printing parameters on the quality of the 3D printed scaffolds. We also demonstrated that flaws, such as voids, significantly lower the mechanical properties (compressive modulus) of 3D printed polymeric scaffolds.


Response Coordination Emerges in Cooperative but Not Competitive Joint Task.
Effective social interactions rely on humans' ability to attune to others within social contexts. Recently, it has been proposed that the emergence of shared representations, as indexed by the Joint Simon effect (JSE), might result from interpersonal coordination (Malone et al., 2014). The present study aimed at examining interpersonal coordination in cooperative and competitive joint tasks. To this end, in two experiments we investigated response coordination, as reflected in instantaneous cross-correlation, when co-agents cooperate (Experiment 1) or compete against each other (Experiment 2). In both experiments, participants performed a go/no-go Simon task alone and together with another agent in two consecutive sessions. In line with previous studies, we found that social presence differently affected the JSE under cooperative and competitive instructions. Similarly, cooperation and competition were reflected in co-agents response coordination. For the cooperative session (Experiment 1), results showed higher percentage of interpersonal coordination for the joint condition, relative to when participants performed the task alone. No difference in the coordination of responses occurred between the individual and the joint conditions when co-agents were in competition (Experiment 2). Finally, results showed that interpersonal coordination between co-agents implies the emergence of the JSE. Taken together, our results suggest that shared representations seem to be a necessary, but not sufficient, condition for interpersonal coordination.


Differential regulation of the period genes in striatal regions following cocaine exposure.
Several studies have suggested that disruptions in circadian rhythms contribute to the pathophysiology of multiple psychiatric diseases, including drug addiction. In fact, a number of the genes involved in the regulation of circadian rhythms are also involved in modulating the reward value for drugs of abuse, like cocaine. Thus, we wanted to determine the effects of chronic cocaine on the expression of several circadian genes in the Nucleus Accumbens (NAc) and Caudate Putamen (CP), regions of the brain known to be involved in the behavioral responses to drugs of abuse. Moreover, we wanted to explore the mechanism by which these genes are regulated following cocaine exposure. Here we find that after repeated cocaine exposure, expression of the Period (Per) genes and Neuronal PAS Domain Protein 2 (Npas2) are elevated, in a somewhat regionally selective fashion. Moreover, NPAS2 (but not CLOCK (Circadian Locomotor Output Cycles Kaput)) protein binding at Per gene promoters was enhanced following cocaine treatment. Mice lacking a functional Npas2 gene failed to exhibit any induction of Per gene expression after cocaine, suggesting that NPAS2 is necessary for this cocaine-induced regulation. Examination of Per gene and Npas2 expression over twenty-four hours identified changes in diurnal rhythmicity of these genes following chronic cocaine, which were regionally specific. Taken together, these studies point to selective disruptions in Per gene rhythmicity in striatial regions following chronic cocaine treatment, which are mediated primarily by NPAS2.


A hierarchical Ca/TiO2/NH2-MIL-125 nanocomposite photocatalyst for solar visible light induced photodegradation of organic dye pollutants in water.
In this study, for the first time, the Ca/TiO2/NH2-MIL-125 nanocomposite photocatalyst was synthesized for the purpose of photodegradation of Methyl Orange (MO) and Rhodamine B (RhB) dyes under visible light irradiation. The structural and chemical properties of the nanocomposite photocatalyst were characterized through FTIR, XRD, TGA, PL, XPS, ICP-OES and UV-DRS. For the photodegradation efficiency analysis, the effect of pH (3, 5, 7, 9, and 11), photocatalyst dosage (0.1, 0.2, 0.4, 0.6, and 0.8 g L-1), dye concentration (1-40 mg L-1), and contact time (10-120 min) was precisely evaluated. The largest photodegradation efficiency for RhB and MO dye models was 82.87% and 86.22%, respectively, that was obtained under optimal conditions in terms of pH and photocatalyst dosage and for Ca(30%)/TiO2/NH2-MIL-125. The photodegradation process of the dyes complied well with the first-order kinetic model. Moreover, the nanocomposite photocatalyst showed consistent photodegradation efficiency and after 6 successive cycles with fresh dye solutions, it could still perform comparably well. Taken together, Ca/TiO2/NH2-MIL-125 photocatalyst is able to show a high photodegradation efficiency for dye pollutants and optimum stability and reusability.


Drug Interactions between Dolutegravir and Artemether-Lumefantrine or Artesunate-Amodiaquine.
Across sub-Saharan Africa, patients with HIV on antiretrovirals often get malaria and need cotreatment with artemisinin-containing therapies. We undertook two pharmacokinetic studies in healthy volunteers, using standard adult doses of artemether-lumefantrine or artesunate-amodiaquine given with 50 mg once daily dolutegravir (DTG) to investigate the drug-drug interaction between artemether-lumefantrine or artesunate-amodiaquine and dolutegravir. The dolutegravir/artemether-lumefantrine interaction was evaluated in a two-way crossover study and measured artemether, dihydroartemisinin, lumefantrine, and desbutyl-lumefantrine over 264 h. The dolutegravir/artesunate-amodiaquine interaction was investigated using a parallel study design due to long half-life of the amodiaquine metabolite, desethylamodiaquine and measured artesunate, amodiaquine, and desethylamodiaquine over 624 h. Noncompartmental analysis was performed, and geometric mean ratios and 90% confidence intervals were generated for evaluation of both interactions. Dolutegravir did not significantly change the maximum concentration in plasma, the time to maximum concentration, and the area under the concentration-time curve (AUC) for artemether, dihydroartemisinin, lumefantrine, and desbutyl-lumefantrine, nor did it significantly alter the AUC for artesunate, dihydroartemisinin, amodiaquine, and desethylamodiaquine. Coadministration of dolutegravir with artemether-lumefantrine resulted in a 37% decrease in DTG trough concentrations. Coadministration of dolutegravir with artesunate-amodiaquine resulted in 42 and 24% approximate decreases in the DTG trough concentrations and the AUC, respectively. The significant decreases in DTG trough concentrations with artemether-lumefantrine and artesunate-amodiaquine and dolutegravir exposure with artesunate-amodiaquine are unlikely to be of clinical significance since the DTG trough concentrations were above dolutegravir target concentrations of 300 ng/ml. Study drugs were well tolerated with no serious adverse events. Standard doses of artemether-lumefantrine and artesunate-amodiaquine should be used in patients receiving dolutegravir. (This study has been registered at ClinicalTrials.gov under identifier NCT02242799.).


Chemical Signals of Vector Beetle Facilitate the Prevalence of a Native Fungus and the Invasive Pinewood Nematode.
In China, the invasive Bursaphelenchus xylophilus, the vector Monochamus alternatus beetle, and associated fungi exhibit a symbiotic relationship causing serious losses to pine forests. Although this complex system has been intensively investigated, the role of vector beetles on the development of associated fungi and their indirect contribution to the prevalence of pinewood nematode (PWN) is yet unknown. Here, three of the highly prevalent fungal species, viz., Sporothrix sp. 1, Ophiostoma ips, and Sporothrix sp. 2 were isolated from beetle chambers in diseased trees in Guangdong province, southeast China. Pairwise cultivation of isolated fungi demonstrated the dominance of Sporothrix sp. 1 over O. ips and Sporothrix sp. 2. On the other hand, two fatty acid ethyl esters (FAEE), ethyl palmitate (EP) and ethyl linoleate (EL), isolated from the body surface of the vector beetle enhanced the growth of Sporothrix sp. 1. When PWN were cultured on Sporothrix sp. 1, the fecundity and the body length were increased significantly as compared with when cultured on O. ips and Sporothrix sp. 1. Our results suggest that the vector beetles promote Sporothrix sp. 1 to occupy more niches by rapid growth and spread, which in turn better support PWN population, hence facilitate PWN pathogenicity in the invasive regions.


A Comparative Study of the Metabolic and Skeletal Response of C57BL/6J and C57BL/6N Mice in a Diet-Induced Model of Type 2 Diabetes.
Type 2 diabetes mellitus (T2DM) represents a complex clinical scenario of altered energy metabolism and increased fracture incidence. The C57BL/6 mouse model of diet-induced obesity has been used to study the mechanisms by which altered glucose homeostasis affects bone mass and quality, but genetic variations in substrains of C57BL/6 may have confounded data interpretation. This study investigated the long-term metabolic and skeletal consequences of two commonly used C57BL/6 substrains to a high fat (HF) diet. Male C57BL/6J, C57BL/6N, and the negative control strain, C3H/HeJ, mice were fed a control or HF diet for 24 wks. C57BL/6N mice on a HF diet demonstrated an increase in plasma insulin and blood glucose as early as 4 wk, whereas these responses were delayed in the C57BL/6J mice. The C57BL/6N mice exhibited more severe hepatic steatosis and inflammation. Only the C57BL/6N mice lost significant trabecular bone in response to the high fat diet. The C3H/HeJ mice were protected from bone loss. The data show that C57BL/6J and C57BL/6N mice differ in their metabolic and skeletal response when fed a HF diet. These substrain differences should be considered when designing experiments and are likely to have implications on data interpretation and reproducibility.


Changes in drug use are associated with health-related quality of life improvements among methadone maintenance patients with HIV/AIDS.
This longitudinal study assessed the changes in drug use patterns and health-related quality of life (HRQL) among HIV-positive drug users in the first methadone maintenance treatment (MMT) cohort in Vietnam. A secondary analysis was conducted on 370 HIV-positive drug users (age: mean ± SD: 29.5 ± 5.9 years; 95.7% men). Modified WHOQOL-BREF, self-report, and opioid confirmatory urine tests were used to assess HRQL and drug use behaviours at baseline, 3, 6, and 9 months. Generalized estimating equations (GEE) models were constructed to adjust for intra-individual correlations. MMT response rate after 9 months was 89.9%. Rates of positive heroin urine tests rapidly decreased at the first trimester (18.1%) and then stabilized during the next 2 trimesters (11.8 and 14.4%). Among patients with continued drug use, frequency of use decreased from 3.4 to 0.7 time/day. Improvements in HRQL were large over the course of the study and highest in the psychological domain. Adjusting for propensity score in GEE models, ongoing heroin use during MMT resulted in large decrements in all HRQL domains. MMT improved the outcomes of treatment for drug users in ways that might facilitate success of antiretroviral therapy. Integrating MMT to HIV care and treatment services could be beneficial in injection-driven HIV epidemics in resource-scare settings.


Comparison of Salvage Total Pharyngolaryngectomy and Cervical Esophagectomy Between Hypopharyngeal Cancer and Cervical Esophageal Cancer.
Total pharyngolaryngectomy and cervical esophagectomy (TPLCE) after chemoradiotherapy remains a challenge because of the high rate of complications and few available data on outcomes and safety. The purpose of this study was to evaluate the clinical significance of salvage TPLCE and to compare treatment outcomes between hypopharyngeal cancer and cervical esophageal cancer. Data from 37 consecutive patients who were diagnosed with potentially resectable hypopharyngeal and cervical esophageal cancer after chemoradiotherapy were retrospectively analyzed. The survival and surgical outcomes were investigated between the hypopharyngeal cancer and cervical esophageal cancer groups. Twenty-six patients were included in hypopharyngeal cancer group and 11 patients were included in cervical esophageal cancer group. The baseline characteristics were balanced between the two groups. Compared to the hypopharyngeal cancer group, the cervical esophageal cancer group had significantly more frequent tracheal-related complications (p < 0.05) and stronger association of distal margin of the cervical esophagus and radiation field with tracheal ischemia after salvage surgery. Salvage TPLCE can offer the exclusive chance of prolonged survival. Association of tracheal ischemia with salvage TPLCE was seen more frequently for cervical esophageal cancer. Therefore, the indication for salvage TPLCE must be carefully considered to maintain the balance between curability and safety.


A proinflammatory activity of interleukin 8 in human skin: expression of the inducible nitric oxide synthase in psoriatic lesions and cultured keratinocytes.
Psoriasis is a common chronic skin disease mediated by cellular immune mechanisms and characterized by an intense neutrophil cell infiltrate and proliferative activation of epidermal keratinocytes. We have previously described the expression of the inducible nitric oxide synthase (iNOS) in epidermal keratinocytes of psoriatic skin lesions. In this study, the role of iNOS in psoriatic inflammation was explored ex vivo in psoriatic skin biopsies and in vitro in primary cultures of human keratinocytes. Messenger RNA for the iNOS enzyme (iNOS mRNA) was detected by reverse transcriptase polymerase chain reaction in skin biopsies from patients with psoriasis, but not in skin specimens from patients with atopic eczema or from healthy volunteers. As demonstrated by in situ hybridization and immunohistochemistry, expression of iNOS mRNA and its gene product was localized to the epidermal keratinocytes of psoriatic skin lesions. In situ hybridization further revealed a complete colocalization of mRNA expression for iNOS with interleukin (IL) 8 receptor-specific mRNA either in the basal germinative cell layer or at focal sites of ongoing neutrophil inflammation in suprabasal cell layers. Because psoriatic keratinocytes have previously been shown to express mRNA transcripts for IL-8, it seemed reasonable to hypothesize that iNOS expression could be induced in an autocrine loop by IL-8. This hypothesis was substantiated by our in vitro experiments showing that a combination of IL-8 and interferon gamma induces the expression of iNOS-specific mRNA and of the functional enzyme in cultured human keratinocytes. These results suggest an important role for iNOS in concert with IL-8 and its receptor early during the formation of psoriatic lesions.


Biotic potential, fertility and life table of Spodoptera albula (Walker) (Lepidoptera: Noctuidae), under controlled conditions.
This study aimed to evaluate the biotic potential, life table parameters and fertility of Spodoptera albula (Walker, 1857) under controlled conditions (25 ± 1°C, 70 ± 10% RH and 14 hour photo phase). The longevity, pre, post and oviposition periods, fecundity and fertility of 13 couples were evaluated. The longevity of females (13.500 days) was significantly higher than those of males (11.154 days). The mean durations of the pre, post and oviposition periods were 2.615, 1.769 and 9.385 days, respectively. The mean fecundity was 1.417.69 eggs and mean fertility was 1.340.401 larvae, per female. On average, females copulated 1, 231 times. A strong positive correlation was observed between the number of copulations and fecundity (r = 0.847, p <0.001), as well as a strong negative correlation between the number of copulations and the duration of the pre-oviposition period (r = -0.762, p = 0.002), and longevity (r = -0.788, p = 0.001). The biotic potential of S. albula was estimated at 8.768 x 1022 individuals / female / year. The net reproductive rate (Ro) was 353,904 times per generation and the mean generation time (T) was 37.187 days. The intrinsic rate of increase (rm) was 1,105, with a finite rate of increase (λ) of 3,019.


Queen Nefertari, the Royal Spouse of Pharaoh Ramses II: A Multidisciplinary Investigation of the Mummified Remains Found in Her Tomb (QV66).
Queen Nefertari, the favourite Royal Consort of Pharaoh Ramses II (Ancient Egypt, New Kingdom, 19th Dynasty c. 1250 BC) is famous for her beautifully decorated tomb in the Valley of the Queens. Her burial was plundered in ancient times yet still many objects were found broken in the debris when the tomb was excavated. Amongst the found objects was a pair of mummified legs. They came to the Egyptian Museum in Turin and are henceforth regarded as the remains of this famous Queen, although they were never scientifically investigated. The following multidisciplinary investigation is the first ever performed on those remains. The results (radiocarbon dating, anthropology, paleopathology, genetics, chemistry and Egyptology) all strongly speak in favour of an identification of the remains as Nefertari's, although different explanations-albeit less likely-are considered and discussed. The legs probably belong to a lady, a fully adult individual, of about 40 years of age. The materials used for embalming are consistent with Ramesside mummification traditions and indeed all objects within the tomb robustly support the burial as of Queen Nefertari.