Light rescues circadian behavior and brain dopamine abnormalities in diurnal rodents exposed to a winter-like photoperiod.
Seasonal affective disorder (SAD), beyond mood changes, is characterized by alterations in daily rhythms of behavior and physiology. The pathophysiological conditions of SAD involve changes in day length and its first-line treatment is bright light therapy. Animal models using nocturnal rodents have been studied to elucidate the neurobiological mechanisms of depression, but might be ill suited to study the therapeutic effects of light in SAD since they exhibit light-aversive responses. Here Arvicanthis ansorgei, a diurnal rodent, was used to determine behavioral, molecular and brain dopamine changes in response to exposure to a winter-like photoperiod consisting of a light-dark cycle with 8 h of light, under diminished light intensity, and 16 h of darkness. Furthermore, we evaluated whether timed-daily bright light exposure has an effect on behavior and brain physiology of winter-like exposed animals. Arvicanthis under a winter-like condition showed alterations in the synchronization of the locomotor activity rhythm to the light-dark cycle. Moreover, alterations in day-night activity of dopaminergic neurotransmission were revealed in the nucleus accumbens and the dorsal striatum, and in the day-night clock gene expression in the suprachiasmatic nucleus. Interestingly, whereas dopamine disturbances were reversed in animals exposed to daily light at early or late day, altered phase of the daily rhythm of locomotion was reverted only in animals exposed to light at the late day. Moreover, Per2 gene expression in the SCN was also affected by light exposure at late day in winter-like exposed animals. These findings suggest that light induces effects on behavior by mechanisms that rely on both circadian and rhythm-independent pathways influencing the dopaminergic circuitry. This last point might be crucial for understanding the mechanisms of non-pharmacological treatment in SAD.


Comparing the climate effect of emissions of short- and long-lived climate agents.
Multi-gas climate agreements require a metric by which emissions of gases with different lifetimes and radiative properties can be placed on a common scale. The Kyoto Protocol to the United Nations Framework Convention on Climate Change uses the global warming potential (GWP) as such a metric. The GWP has attracted particular criticism as being inappropriate in the context of climate policy which seeks to restrict warming below a given target, because it gives equal weight to emissions irrespective of the target and the proximity to the target. The use of an alternative metric, the time-dependent global temperature change potential (GTP), is examined for its suitability and the prospects for it including very short-lived species. It retains the transparency and relative ease of use, which are attractive features of the GWP, but explicitly includes a dependence on the target of climate policy. The weighting of emissions using the GTP is found to be significantly dependent on the scenarios of future emissions and the sensitivity of the climate system. This may indicate that the use of any GTP-based weighting in future policymaking would necessitate regular revisions, as the global-mean temperature moves towards a specified target.


Big data challenges in bone research: genome-wide association studies and next-generation sequencing.
Genome-wide association studies (GWAS) have been developed as a practical method to identify genetic loci associated with disease by scanning multiple markers across the genome. Significant advances in the genetics of complex diseases have been made owing to advances in genotyping technologies, the progress of projects such as HapMap and 1000G and the emergence of genetics as a collaborative discipline. Because of its great potential to be used in parallel by multiple collaborators, it is important to adhere to strict protocols assuring data quality and analyses. Quality control analyses must be applied to each sample and each single-nucleotide polymorphism (SNP). The software package PLINK is capable of performing the whole range of necessary quality control tests. Genotype imputation has also been developed to substantially increase the power of GWAS methodology. Imputation permits the investigation of associations at genetic markers that are not directly genotyped. Results of individual GWAS reports can be combined through meta-analysis. Finally, next-generation sequencing (NGS) has gained popularity in recent years through its capacity to analyse a much greater number of markers across the genome. Although NGS platforms are capable of examining a higher number of SNPs compared with GWA studies, the results obtained by NGS require careful interpretation, as their biological correlation is incompletely understood. In this article, we will discuss the basic features of such protocols.


Cooperative and redundant roles of VEGFR-2 and VEGFR-3 signaling in adult lymphangiogenesis.
Activation of vascular endothelial growth factor (VEGF) receptor-3 (VEGFR-3) by VEGF-C initiates lymphangiogenesis by promoting lymphatic proliferation and migration. However, it is unclear whether VEGFR-3 signaling is required beyond these initial stages, namely during the organization of new lymphatic endothelial cells (LECs) into functional capillaries. Furthermore, the role of VEGFR-2, which is also expressed on LECs and binds VEGF-C, is unclear. We addressed these questions by selectively neutralizing VEGFR-3 and/or VEGFR-2 for various time periods in an adult model of lymphangiogenesis in regenerating skin. While blocking either VEGFR-2 or VEGFR-3 with specific antagonist mAbs (DC101 and mF4-31C1, respectively) prior to lymphatic migration prevented lymphangiogenesis, blocking VEGFR-3 subsequent to migration did not affect organization into functional capillaries, and VEGFR-2 blocking had only a small hindrance on organization. These findings were confirmed in vitro using human LECs and anti-human antagonist mAbs (IMC-1121a and hF4-3C5): both VEGFR-2 and -3 signaling were required for migration and proliferation, but tubulogenesis in 3D cultures was unaffected by VEGFR-3 blocking and partially hindered by VEGFR-2 blocking. Furthermore, both in vitro and in vivo, while VEGFR-3 blocking had no effect on LEC organization, coneutralization of VEGFR-2, and VEGFR-3 completely prevented lymphatic organization. Our findings demonstrate that cooperative signaling of VEGFR-2 and -3 is necessary for lymphatic migration and proliferation, but VEGFR-3 is redundant with VEGFR-2 for LEC organization into functional capillaries.


The relative importance of dispositional optimism and control appraisals in quality of life after coronary artery bypass surgery.
Similar mechanisms have been proposed to explain the stress-buffering effects of both dispositional optimism and perceived control. Yet dispositional optimism as a personal resource should function independently of situational control appraisals. To evaluate the unique and additive contributions to adaptation of control appraisals and optimism, we followed 49 individuals scheduled for coronary artery bypass surgery. One month before surgery dispositional optimism was associated with neither health locus of control nor specific expectancies about the outcomes of surgery. Dispositional optimism, however, was associated with perceived control over the course of the illness and with quality of life appraisals. Although presurgery optimism predicted life quality 8 months after surgery, this was not the case when general and specific control appraisals and specific expectancies were included in the prediction. These findings are discussed as they relate to current conceptions of trait optimism.


Mechanism of benzylselenocyanate inhibition of azoxymethane-induced colon carcinogenesis in F344 rats.
Benzylselenocyanate (BSC), a novel organoselenium compound, has been found to inhibit azoxymethane (AOM)-induced colon carcinogenesis in rats during initiation. To investigate its mechanism of action, we examined the effects of BSC feeding on the following parameters: (a) metabolism of [14C]AOM to 14CO2 in vivo; (b) metabolic activation of AOM to MAM and of MAM to formic acid and methanol by rat liver microsomes in vitro; and (c) AOM-induced DNA methylation in rat livers and colons. Five-week-old male F344 rats were fed modified (23% corn oil) AIN-76A diets containing 0 (control), 25, or 50 ppm of BSC or benzylthiocyanate (BTC), a sulfur analogue of BSC which does not inhibit the colon carcinogenicity of AOM. After 3 weeks, rats were either sacrificed for the isolation of liver microsomes or were given 15 mg/kg of [14C]AOM s.c. to determine the rate of carcinogen metabolism in vivo. No difference in [14C]AOM metabolism was found between rats fed the BTC diets and those fed the control diet. In contrast, the rate of [14C]AOM metabolism, as determined by exhaled radioactivity, was 2-3 times higher in rats fed the BSC diets. While liver microsomes from rats fed the BTC diets metabolized AOM and MAM at rates not significantly different from those obtained with control liver microsomes, the metabolic activation of AOM as well as of MAM was stimulated severalfold when assayed with liver microsomes from rats fed the BSC diets. An increase in total liver cytochrome P-450 was also observed in the BSC-fed rats. Following the administration of 15 mg/kg AOM, significantly less O6-methylguanine and 7-methylguanine was present in the colon DNA from rats consuming the BSC diets than in rats fed the BTC or control diets. The body weight gains of rats fed the 25- and 50-ppm BSC-containing diets for 3 weeks were less (27 and 43%, respectively) than those of rats fed either the control or BTC-containing diets. These results indicate that dietary BSC significantly induces the hydroxylation of AOM and the oxidation of MAM in rat liver. An increase in the rates of AOM and MAM metabolism in the liver due to enzyme induction by BSC will result in decreased delivery of MAM to the colon via the bloodstream. This will be reflected in decreased DNA alkylation, as observed, and is likely to be a major factor in the inhibition of AOM-induced colon carcinogenesis by BSC.


Single-molecule decoding of combinatorially modified nucleosomes.
Different combinations of histone modifications have been proposed to signal distinct gene regulatory functions, but this area is poorly addressed by existing technologies. We applied high-throughput single-molecule imaging to decode combinatorial modifications on millions of individual nucleosomes from pluripotent stem cells and lineage-committed cells. We identified definitively bivalent nucleosomes with concomitant repressive and activating marks, as well as other combinatorial modification states whose prevalence varies with developmental potency. We showed that genetic and chemical perturbations of chromatin enzymes preferentially affect nucleosomes harboring specific modification states. Last, we combined this proteomic platform with single-molecule DNA sequencing technology to simultaneously determine the modification states and genomic positions of individual nucleosomes. This single-molecule technology has the potential to address fundamental questions in chromatin biology and epigenetic regulation.


HapCUT2: robust and accurate haplotype assembly for diverse sequencing technologies.
Many tools have been developed for haplotype assembly-the reconstruction of individual haplotypes using reads mapped to a reference genome sequence. Due to increasing interest in obtaining haplotype-resolved human genomes, a range of new sequencing protocols and technologies have been developed to enable the reconstruction of whole-genome haplotypes. However, existing computational methods designed to handle specific technologies do not scale well on data from different protocols. We describe a new algorithm, HapCUT2, that extends our previous method (HapCUT) to handle multiple sequencing technologies. Using simulations and whole-genome sequencing (WGS) data from multiple different data types-dilution pool sequencing, linked-read sequencing, single molecule real-time (SMRT) sequencing, and proximity ligation (Hi-C) sequencing-we show that HapCUT2 rapidly assembles haplotypes with best-in-class accuracy for all data types. In particular, HapCUT2 scales well for high sequencing coverage and rapidly assembled haplotypes for two long-read WGS data sets on which other methods struggled. Further, HapCUT2 directly models Hi-C specific error modalities, resulting in significant improvements in error rates compared to HapCUT, the only other method that could assemble haplotypes from Hi-C data. Using HapCUT2, haplotype assembly from a 90× coverage whole-genome Hi-C data set yielded high-resolution haplotypes (78.6% of variants phased in a single block) with high pairwise phasing accuracy (∼98% across chromosomes). Our results demonstrate that HapCUT2 is a robust tool for haplotype assembly applicable to data from diverse sequencing technologies.


Tadalafil Preserves Penile Nitric Oxide Synthase from Detrimental Effect of Paroxetine in Rats.
Paroxetine is a commonly prescribed SSRI that can impair erectile function in animal models via inhibition of nitric oxide synthase (NOS). Tadalafil potentiates nitric oxide (NO)-mediated responses in isolated trabecular smooth muscle and penile erection. The purpose of this study was to evaluate the impact of co-administering tadalafil with paroxetine on penile NOS levels in rats. A total of 30 male Sprague-Dawley rats were divided into 3 groups as control (Group-C), paroxetine (Group-P) and paroxetine plus tadalafil (Group-P+T). After 28 days of treatment, rats were sacrificed and their penile tissues were harvested for analysis. NOS isoform protein levels and immunoreactivity scores of NOS were assessed. Statistical significance level was set at p<0.05. Neuronal NOS (nNOS) levels were significantly decreased in group-P, compared with group-C (p<0.001). In comparison, rats in group-P+T had significantly higher nNOS levels compared to group-P (p<0.001). Endothelial NOS (eNOS) and inducible NOS (iNOS) levels were significantly higher in group-P compared with group-C (p<0.01). The levels of eNOS and iNOS in group-P+T were similar to group-C. Daily treatment with tadalafil prevented chronic paroxetine-induced changes in all three NOS isoform levels. Tadalafil treatment may therefore be a useful therapy in men with paroxetine-associated erectile dysfunction.


The behaviour of inositol 1,3,4,5,6-pentakisphosphate in the presence of the major biological metal cations.
The inositol phosphates are ubiquitous metabolites in eukaryotes, of which the most abundant are inositol hexakisphosphate (InsP 6) and inositol 1,3,4,5,6-pentakisphosphate [Ins(1,3,4,5,6)P5)]. These two compounds, poorly understood functionally, have complicated complexation and solid formation behaviours with multivalent cations. For InsP 6, we have previously described this chemistry and its biological implications (Veiga et al. in J Inorg Biochem 100:1800, 2006; Torres et al. in J Inorg Biochem 99:828, 2005). We now cover similar ground for Ins(1,3,4,5,6)P5, describing its interactions in solution with Na+, K+, Mg2+, Ca2+, Cu2+, Fe2+ and Fe3+, and its solid-formation equilibria with Ca2+ and Mg2+. Ins(1,3,4,5,6)P5 forms soluble complexes of 1:1 stoichiometry with all multivalent cations studied. The affinity for Fe3+ is similar to that of InsP6 and inositol 1,2,3-trisphosphate, indicating that the 1,2,3-trisphosphate motif, which Ins(1,3,4,5,6)P5 lacks, is not absolutely necessary for high-affinity Fe3+ complexation by inositol phosphates, even if it is necessary for their prevention of the Fenton reaction. With excess Ca2+ and Mg2+, Ins(1,3,4,5,6)P5 also forms the polymetallic complexes [M4(H2L)] [where L is fully deprotonated Ins(1,3,4,5,6)P5]. However, unlike InsP6, Ins(1,3,4,5,6)P5 is predicted not to be fully associated with Mg2+ under simulated cytosolic/nuclear conditions. The neutral Mg2+ and Ca2+ complexes have significant windows of solubility, but they precipitate as [Mg4(H2L)] x 23H2O or [Ca4(H2L)] x 16H2O whenever they exceed 135 and 56 microM in concentration, respectively. Nonetheless, the low stability of the [M4(H2L)] complexes means that the 1:1 species contribute to the overall solubility of Ins(1,3,4,5,6)P 5 even under significant Mg2+ or Ca2+ excesses. We summarize the solubility behaviour of Ins(1,3,4,5,6)P5 in straightforward plots.


Murine isograft studies of gut immunity: recirculation and homing of mononuclear cells.
Recirculation of lymphocytes requires appropriate signals on the lymphocytes as well as the vascular endothelium. The aim of this report was to study the expression of the mucosal addressin required for Peyer's patch-selective lymphocyte homing during ontogeny of the murine intestine and investigate the role of this addressin in recirculation of mononuclear cells. Immunohistochemistry and murine intestinal isografts were used in which the small intestine from neonatal mice is implanted subcutaneously in mice with severe combined immunodeficiency disease. Expression of the mucosal addressin cell adhesion molecule 1 during murine intestinal ontogeny was detected from embryonic day 15 in developing gut and in adult Peyer's patches and lamina propria. Isografted intestine developed normally; it was not exposed to luminal contents but expressed the mucosal addressin cell adhesion molecule 1. Mononuclear cells recirculated to native and isografted intestine and recirculation was inhibited by monoclonal antibody to mucosal addressin in a regionally specific manner with greatest inhibition in the distal intestine. The mucosal vascular addressin is expressed early in ontogeny and can be detected in lamina propria in addition to Peyer's patches of the gut. The isograft not only developed into a morphologically normal intestine but also expressed differentiation antigens required for normal lymphoid homing to gut without exposure to luminal contents or lymphocytes.


Joint effects of physical activity and BMI on risk of hypertension in women: a longitudinal study.
There is debate as to whether physical activity counteracts the adverse effect of weight on health outcomes. We investigated how physical activity modifies the effect of body mass index (BMI) on hypertension risk. BMI, physical activity, and hypertension were measured at baseline and at three-year interval for 14 years (from 1996 to 2010), in 10,339 participants in the Australian Longitudinal Study on Women's Health. Generalised estimating equation models for binary repeated measures were performed to determine the individual and joint effects of BMI and physical activity on incident hypertension. At baseline (mean age 47.6 ± 1.5 SD), 57% were healthy weight, 28% overweight, and 14% obese. Increasing BMI and decreasing physical activity were associated with increased risk of hypertension. Physical activity attenuated the positive association between weight and risk of hypertension, especially for obese women. Compared to healthy weight high active women, risk of hypertension in obese high active women was 3.4 times greater (OR 3.43, 95% CI 2.68, 4.39) and in obese inactive women 4.9 times greater (OR 4.91, 95% CI 3.92, 6.13). Both physical activity and maintenance of a healthy body weight are associated with lower risk of hypertension. Physical activity reduced but did not remove the effect of obesity on hypertension risk.


CLEVER: clique-enumerating variant finder.
Next-generation sequencing techniques have facilitated a large-scale analysis of human genetic variation. Despite the advances in sequencing speed, the computational discovery of structural variants is not yet standard. It is likely that many variants have remained undiscovered in most sequenced individuals. Here, we present a novel internal segment size based approach, which organizes all, including concordant, reads into a read alignment graph, where max-cliques represent maximal contradiction-free groups of alignments. A novel algorithm then enumerates all max-cliques and statistically evaluates them for their potential to reflect insertions or deletions. For the first time in the literature, we compare a large range of state-of-the-art approaches using simulated Illumina reads from a fully annotated genome and present relevant performance statistics. We achieve superior performance, in particular, for deletions or insertions (indels) of length 20-100 nt. This has been previously identified as a remaining major challenge in structural variation discovery, in particular, for insert size based approaches. In this size range, we even outperform split-read aligners. We achieve competitive results also on biological data, where our method is the only one to make a substantial amount of correct predictions, which, additionally, are disjoint from those by split-read aligners. CLEVER is open source (GPL) and available from http://clever-sv.googlecode.com. firstname.lastname@example.com or firstname.lastname@example.com. Supplementary data are available at Bioinformatics online.


Linear, self-referenced technique for single-shot and real-time full characterization of (sub-)picosecond optical pulses.
We propose and experimentally demonstrate single-shot, real-time ultrashort pulse intensity and phase characterization using a self-referenced and highly sensitive (linear) technique. The proposed method is based on a direct reconstruction of the spectral phase of the pulse-under-test (PUT) from three different measured spectra, two of which are obtained by a suitable time-synchronized electro-optic intensity modulation of the PUT. The required set of spectra are temporally interleaved and mapped along the time domain by linear dispersion for single-shot acquisition using a real-time scope. A dynamic nonlinear compression experiment of a picosecond pulse is fully monitored in real time using the proposed method.


Influence of cardiovascular absolute risk assessment on prescribing of antihypertensive and lipid-lowering medications: a cluster randomized controlled trial.
Guidelines for management of hypertension and lipids recommend using cardiovascular absolute risk (CVAR) to manage patients. This randomized controlled trial investigated the impact of CVAR assessment in family practice on management of cardiovascular risk, including prescription of antihypertensive and lipid-lowering medication. A cluster randomized controlled trial was conducted from 2008 to 2010 in Sydney, Australia. Family practices were randomized, and patients aged 45 to 69 years were invited to participate. Intervention family physicians (FP) were trained in use of CVAR, provided with an electronic CVAR calculator, and assessed their patients' absolute risk in a dedicated consultation. Control practice patients received a general health check. Primary outcome analyzed was the proportion of patients in each group on antihypertensive and/or lipid-lowering medication at 12 months. Multilevel logistic regression was performed to explore variables influencing changes in pharmacologic therapy. The study recruited 36 FPs from 34 practices and 1,074 patients, of which 906 (84.4%) completed 12-month follow-up. At 12 months, there was no significant difference between the intervention and control groups in proportion of patients on antihypertensives (31.2% vs 34.3%, P = .31), but control group patients were more likely to be on lipid-lowering medications (30.2% vs 22.7%, P = .01). After multilevel analysis, this difference was not present. Intensification or reduction of pharmacologic therapy was associated with meeting treatment targets for blood pressure and lipids but not with the CVAR or intervention group. Single-risk factor management remains a strong influence on FP prescribing practices. Shifting to an approach based on CVAR will require more intensive intervention.


Human serum-derived protein removes the need for coating in defined human pluripotent stem cell culture.
Reliable, scalable and time-efficient culture methods are required to fully realize the clinical and industrial applications of human pluripotent stem (hPS) cells. Here we present a completely defined, xeno-free medium that supports long-term propagation of hPS cells on uncoated tissue culture plastic. The medium consists of the Essential 8 (E8) formulation supplemented with inter-α-inhibitor (IαI), a human serum-derived protein, recently demonstrated to activate key pluripotency pathways in mouse PS cells. IαI efficiently induces attachment and long-term growth of both embryonic and induced hPS cell lines when added as a soluble protein to the medium at seeding. IαI supplementation efficiently supports adaptation of feeder-dependent hPS cells to xeno-free conditions, clonal growth as well as single-cell survival in the absence of Rho-associated kinase inhibitor (ROCKi). This time-efficient and simplified culture method paves the way for large-scale, high-throughput hPS cell culture, and will be valuable for both basic research and commercial applications.


Cellular pathways of galactose-terminal ligand movement in a cloned human hepatoma cell line.
The intracellular pathways taken by galactose-terminal glycoproteins were examined following endocytosis by the asialoglycoprotein receptor in monolayers of the human hepatoma cell line, Hep G2. In addition to a pathway leading to lysosomal degradation, single cohort kinetics revealed that up to 28% of surface-bound and internalized 125I-asialoorosomucoid (ASOR) eventually returned undegraded to the extracellular medium over 6 hr in the presence or absence of free ASOR in the exocytosis medium. This reappearance of ligand in the exocytosis medium represented a constant fraction of surface bound and internalized 125I-ASOR, and followed pseudo-first order kinetics with t1/2 = 84 min (long transit pool). Under conditions of enhanced ligand-receptor dissociation (incubation with 100 mM N-acetylgalactosamine (GalNAc), at least 50% of initially internalized 125I-ASOR returned to the cell surface as ligand-receptor complexes, followed by dissociation of free ligand into the exocytosis medium. This rapid transit pool of ligand also displayed pseudo-first order kinetics with t1/2 = 24 min. Exocytosis of 125I-Gal-cytochrome c, a synthesized ligand displaying rapid dissociation from the asialoglycoprotein receptor (ASGP-R), paralleled the kinetics of the rapid transit pool of 125I-ASOR (t1/2 = 28 min). Furthermore, in addition to spontaneous dissociation from ASPG-R following return to the cell surface, studies conducted in saponin-permeabilized monolayers support the return of free intracellular 125I-Gal-cytochrome c to the cell surface during exocytosis. The rapid transit pool of ligand was insensitive to inhibition by 10 mM sodium azide or 0.1 mM primaquine. In contrast, the long transit pool destined for exocytosis was inhibited 50% by 10 mM sodium azide, but insensitive to inhibition by 0.1 mM primaquine. These data suggest that, following internalization by the ASGP-R, a major pathway of ligand movement includes the rapid return of ligand-receptor complexes and/or free ligand to the cell surface. Return of ligand-receptor complexes or free ligand to the cell surface occurs prior to an acidic sorting compartment, can involve multiple cycles of return to the cell surface, and may involve passage through other nonlysosomal intracellular organelles.


Patterned gene expression directs bipolar planar polarity in Drosophila.
During convergent extension in Drosophila, polarized cell movements cause the germband to narrow along the dorsal-ventral (D-V) axis and more than double in length along the anterior-posterior (A-P) axis. This tissue remodeling requires the correct patterning of gene expression along the A-P axis, perpendicular to the direction of cell movement. Here, we demonstrate that A-P patterning information results in the polarized localization of cortical proteins in intercalating cells. In particular, cell fate differences conferred by striped expression of the even-skipped and runt pair-rule genes are both necessary and sufficient to orient planar polarity. This polarity consists of an enrichment of nonmuscle myosin II at A-P cell borders and Bazooka/PAR-3 protein at the reciprocal D-V cell borders. Moreover, bazooka mutants are defective for germband extension. These results indicate that spatial patterns of gene expression coordinate planar polarity across a multicellular population through the localized distribution of proteins required for cell movement.


The use of alternative medicine in pediatric patients with atopic dermatitis.
Alternative medicine has been defined as forms of therapy or examination that have no scientific basis and for which no effective or diagnostic reliability has been demonstrated by scientific methods. The use of complementary or alternative medicine is increasing and controlled clinical trials on the subject are few. We performed a questionnaire-based study of 80 pediatric patients with atopic dermatitis. This questionnaire assessed the duration of treatment, the reason(s) for trying alternative therapy, the approximate cost and the success of the treatment, the duration of the childhood eczema, and whether the child had ever required hospital admission for eczema. Of the total, 34 (42.5%) patients had used alternative medicine. Herbal remedies and homeopathy were used most often. Most treatments were reported to show no benefit and in three instances deterioration was reported. This study has prompted us to enquire routinely regarding alternative medicine use. Alternative therapies are subject to minimal regulation and have been associated with serious side effects. We would recommend enquiries regarding alternative medicine use in all pediatric dermatology patients.


Shear flow affects selective monocyte recruitment into MCP-1-loaded scaffolds.
Novel cardiovascular replacements are being developed by using degradable synthetic scaffolds, which function as a temporary guide to induce neotissue formation directly in situ. Priming of such scaffolds with fast-releasing monocyte chemoattractant protein-1 (MCP-1) was shown to improve the formation of functional neoarteries in rats. However, the underlying mechanism has not been clarified. Therefore, the goal of this study was to investigate the effect of a burst-release of MCP-1 from a synthetic scaffold on the local recruitment of circulating leucocytes under haemodynamic conditions. Herein, we hypothesized that MCP-1 initiates a desired healing cascade by recruiting favourable monocyte subpopulations into the implanted scaffold. Electrospun poly(ε-caprolactone) scaffolds were loaded with fibrin gel containing various doses of MCP-1 and exposed to a suspension of human peripheral blood mononuclear cells in static or dynamic conditions. In standard migration assay, a dose-dependent migration of specific CD14(+) monocyte subsets was observed, as measured by flow cytometry. In conditions of pulsatile flow, on the other hand, a marked increase in immediate monocyte recruitment was observed, but without evident selectivity in monocyte subsets. This suggests that the selectivity was dependent on the release kinetics of the MCP-1, as it was overruled by the effect of shear stress after the initial burst-release. Furthermore, these findings demonstrate that local recruitment of specific MCP-1-responsive monocytes is not the fundamental principle behind the improved neotissue formation observed in long-term in vivo studies, and mobilization of MCP-1-responsive cells from the bone marrow into the bloodstream is suggested to play a predominant role in vivo.


Emergence of patterned stem cell differentiation within multicellular structures.
The ability of stem cells to differentiate into specified lineages in the appropriate locations is vital to morphogenesis and adult tissue regeneration. Although soluble signals are important regulators of patterned differentiation, here we show that gradients of mechanical forces can also drive patterning of lineages. In the presence of soluble factors permitting osteogenic and adipogenic differentiation, human mesenchymal stem cells at the edge of multicellular islands differentiate into the osteogenic lineage, whereas those in the center became adipocytes. Interestingly, changing the shape of the multicellular sheet modulated the locations of osteogenic versus adipogenic differentiation. Measuring traction forces revealed gradients of stress that preceded and mirrored the patterns of differentiation, where regions of high stress resulted in osteogenesis, whereas stem cells in regions of low stress differentiated to adipocytes. Inhibiting cytoskeletal tension suppressed the relative degree of osteogenesis versus adipogenesis, and this spatial patterning of differentiation was also present in three-dimensional multicellular clusters. These findings demonstrate a role for mechanical forces in linking multicellular organization to spatial differentials of cell differentiation, and they represent an important guiding principle in tissue patterning that could be exploited in stem cell-based therapies. Disclosure of potential conflicts of interest is found at the end of this article.


A psychiatric study of patients with diseases of the small intestine.
A group of 80 patients with diseases of the small intestine were given a standardized psychiatric assessment every time they attended the outpatients' department over a period of a year. Forty-six patients with idiopathic steatorrhoea, 23 patients with Crohn's disease, and 11 patients with alactasia were interviewed on 158 occasions. During this time 27 of them (34%) were found to be psychiatrically ill, their illnesses being minor affective disorders. Psychiatric illness was not related either to generalized malabsorption or to deficiencies of specific substances such as folic acid. Although a clear relationship was demonstrated between emotional disturbances and bowel action, diarrhoea was neither sufficient nor necessary for psychiatric illness, and distress associated with diarrhoea was unusual. No single characteristic type of personality was found in any of the patients with the three diseases, but some traits were considerably more commonly associated with some of the diseases than others, and patients with psychiatric illness were shown to attend the outpatients' department more frequently than stable patients. Patients with a family history or a previous history of psychiatric illness, or with depressive traits in their previous personality, were much more likely to fall ill in the survey year. A positive family history of psychiatric illness was significantly more common in idiopathic steatorrhoea. Three patients suffered relapses in the survey year, and in all of these the relapse appeared to be related to emotional factors.


Efficacy and safety of treatments for refractory generalized anxiety disorder: a systematic review.
This study systematically collated clinical evidence on refractory generalized anxiety disorder (GAD). Refractory GAD patients are those who have failed to respond adequately to at least one earlier treatment for GAD. MEDLINE, EMBASE, The Cochrane Library and conference proceedings were searched to identify trials. Four placebo-controlled trials (pregabalin, olanzapine, quetiapine, risperidone) and four single-arm studies (aripiprazole, risperidone, quetiapine, ziprasidone) evaluated the add-ons to initial treatment(s) or switch of treatment(s) because of inadequate efficacy. The most robust trial was the pregabalin study, with a study duration of 8 weeks and a largest sample size that consists of 356 patients. A significant reduction in the Hamilton Anxiety Scale (HAM-A) score was found for pregabalin and risperidone augmentation compared with placebo. Olanzapine augmentation resulted in a significantly higher proportion of responders (using HAM-A scores) compared with placebo. Quetiapine augmentation did not result in significantly greater mean reductions in the HAM-Ascores compared with placebo. There is a need for effective and safe augmentation treatments for patient's refractory to initial treatments for GAD. This study has located one large robust trial assessing the add-on to pregabalin. All other trials were small and unpowered studies with less than 50 patients. Further high-quality trials of augmentation treatment on refractory GAD are required.


Preparation, characterisation, and in vitro evaluation of electrically conducting poly(ɛ-caprolactone)-based nanocomposite scaffolds using PC12 cells.
In the current study, we describe the synthesis, material characteristics, and cytocompatibility of conducting poly (ɛ-caprolactone) (PCL)-based nano-composite films. Electrically conducting carbon nano-fillers (carbon nano-fiber (CNF), nano-graphite (NG), and liquid exfoliated graphite (G)) were used to prepare porous film type scaffolds using modified solvent casting methods. The electrical conductivity of the nano-composite films was increased when carbon nano-fillers were incorporated in the PCL matrix. CNF-based nano-composite films showed the highest increase in electrical conductivity. The presence of an ionic solution significantly improved the conductivity of some of the polymers, however at least 24 h was required to absorb the simulated ion solutions. CNF-based nano-composite films were found to have good thermo-mechanical properties compared to other conducting polymer films due to better dispersion and alignment in the critical direction. Increased nano-filler content increased the crystallisation temperature. Analysis of cell viability revealed no increase in cell death on any of the polymers compared to tissue culture plastic controls, or compared to PCL polymer without nano-composites. The scaffolds showed some variation when tested for PC12 cell attachment and proliferation, however all the polymers supported PC12 attachment and differentiation in the absence of cell adhesion molecules. In general, CNF-based nano-composite films with highest electrical conductivity and moderate roughness showed highest cell attachment and proliferation. These polymers are promising candidates for use in neural applications in the area of bionics and tissue engineering due to their unique properties.


MicroRNA-181b regulates NF-κB-mediated vascular inflammation.
EC activation and dysfunction have been linked to a variety of vascular inflammatory disease states. The function of microRNAs (miRNAs) in vascular EC activation and inflammation remains poorly understood. Herein, we report that microRNA-181b (miR-181b) serves as a potent regulator of downstream NF-κB signaling in the vascular endothelium by targeting importin-α3, a protein that is required for nuclear translocation of NF-κB. Overexpression of miR-181b inhibited importin-α3 expression and an enriched set of NF-κB-responsive genes such as adhesion molecules VCAM-1 and E-selectin in ECs in vitro and in vivo. In addition, treatment of mice with proinflammatory stimuli reduced miR-181b expression. Rescue of miR-181b levels by systemic administration of miR-181b "mimics" reduced downstream NF-κB signaling and leukocyte influx in the vascular endothelium and decreased lung injury and mortality in endotoxemic mice. In contrast, miR-181b inhibition exacerbated endotoxin-induced NF-κB activity, leukocyte influx, and lung injury. Finally, we observed that critically ill patients with sepsis had reduced levels of miR-181b compared with control intensive care unit (ICU) subjects. Collectively, these findings demonstrate that miR-181b regulates NF-κB-mediated EC activation and vascular inflammation in response to proinflammatory stimuli and that rescue of miR-181b expression could provide a new target for antiinflammatory therapy and critical illness.


Cloning of the eukaryotic expression vector with nerve growth factor in rats and its effects on proliferation and differentiation of mesencephal neural stem cells of fetal rats.
The eukaryotic expression vector containing full-length cDNA sequence of rate nerve growth factor (NGF) beta subunit was constructed and its effects on proliferation and differentiation of neural stem cells were observed. By using PCR, full-length cDNA sequence of NGF beta subunit in rats was cloned and ligated into the eukaryotic expression vector pEGFP-N1-NGF. The recombinant plasmid pEGFP-N1-NGF was transfected into the mesencephal neural stem cells of embryonic rats by Lipofectamin and transiently expressed. MTT method was used to determine the effects of NGF on proliferation of neural stem cells, and under phase-contrast microscopy, the effects of NGF on growth of nervous processes following differentiation of neural stem cells were observed. Sequence analysis indicated that the cloned full-length cDNA sequence of rat NGF beta was identical to that of published sequence encoding NGF in gene GeneBank. The transfection of recombinant plasmid pEGFP-N1-NGF into mesencephal neural stem cells of embryonic rats could obviously promote proliferation of neural stem cells and facilitate the growth of neural stem cells-derived nerve cells. It was suggested that neural stem cells could be used as a vehicle of gene transfer, and the expression of NGF beta subunit in the neural stem cells could promote the growth of nerve cells derived from neural stem cells.


Coding of object location in the vibrissal thalamocortical system.
In whisking rodents, object location is encoded at the receptor level by a combination of motor and sensory related signals. Recoding of the encoded signals can result in various forms of internal representations. Here, we examined the coding schemes occurring at the first forebrain level that receives inputs necessary for generating such internal representations--the thalamocortical network. Single units were recorded in 8 thalamic and cortical stations in artificially whisking anesthetized rats. Neuronal representations of object location generated across these stations and expressed in response latency and magnitude were classified based on graded and binary coding schemes. Both graded and binary coding schemes occurred across the entire thalamocortical network, with a general tendency of graded-to-binary transformation from thalamus to cortex. Overall, 63% of the neurons of the thalamocortical network coded object position in their firing. Thalamocortical responses exhibited a slow dynamics during which the amount of coded information increased across 4-5 whisking cycles and then stabilized. Taken together, the results indicate that the thalamocortical network contains dynamic mechanisms that can converge over time on multiple coding schemes of object location, schemes which essentially transform temporal coding to rate coding and gradual to labeled-line coding.


Can serial exercise testing improve the prediction of coronary events in asymptomatic individuals?
An abnormal ST segment response to treadmill exercise has a low predictive value for future coronary events (angina pectoris, nonfatal myocardial infarction, or cardiac death) in apparently healthy individuals. To determine whether the conversion from a normal to an abnormal ST segment response might identify individuals at high risk for a future coronary event, we analyzed the results of serial exercise tests performed at 2-4-year intervals in 726 male and female volunteers, aged 22-84 years (mean, 55.1 years), from the Baltimore Longitudinal Study of Aging (BLSA). All subjects were free of cardiovascular disease at entry by history, physical examination, and resting 12-lead electrocardiogram. Over a mean overall follow-up of 7.4 years, coronary events occurred in 34 of 178 (19.1%) of those with an abnormal ST response to exercise versus 30 of 548 (5.5%) in those with a normal response (p = 0.001). Angina pectoris was the most common presenting coronary event regardless of ST segment exercise response. Among individuals with an abnormal ST segment response, the incidence of events was virtually identical between those with an initially abnormal response (group 1) and those who converted from a normal to an abnormal response (group 2), 19.8% versus 18.5%. After adjustment for standard coronary risk factors by proportional hazards regression analysis, the risk of a coronary event relative to subjects with persistently normal ST segment responses (group 3) remained nearly identical in the two groups, 2.72 in group 1 (p less than 0.003) and 2.80 in group 2 (p less than 0.002).(ABSTRACT TRUNCATED AT 250 WORDS)


Sweat rate, salt loss, and fluid intake during an intense on-ice practice in elite Canadian male junior hockey players.
Previous research in many sports suggests that losing ~1%-2% body mass through sweating impairs athletic performance. Elite-level hockey involves high-intensity bursts of skating, arena temperatures are >10 degrees C, and players wear protective equipment, all of which promote sweating. This study examined the pre-practice hydration, on-ice fluid intake, and sweat and sodium losses of 44 candidates for Canada's junior men's hockey team (mean +/- SE age, 18.4 +/- 0.1 y; height, 184.8 +/- 0.9 cm; mass, 89.9 +/- 1.1 kg). Players were studied in groups of 10-12 during 4 intense 1 h practices (13.9 degrees C, 66% relative humidity) on 1 day. Hydration status was estimated by measuring urine specific gravity (USG). Sweat rate was calculated from body mass changes and fluid intake. Sweat sodium concentration ([Na]) was analyzed in forehead sweat patch samples and used with sweat rate to estimate sodium loss. Over 50% of players began practice mildly hypohydrated (USG > 1.020). Sweat rate during practice was 1.8 +/- 0.1 L.h(-1) and players replaced 58% (1.0 +/- 0.1 L.h(-1)) of the sweat lost. Body mass loss averaged 0.8% +/- 0.1%, but 1/3 of players lost more than 1%. Sweat [Na] was 54.2 +/- 2.4 mmol.L(-1) and sodium loss averaged 2.26 +/- 0.17 g during practice. Players drank only water during practice and replaced no sodium. In summary, elite junior hockey players incurred large sweat and sodium losses during an intense practice, but 2/3 of players drank enough to minimize body mass loss. However, 1/3 of players lost more than 1% body mass despite ready access to fluid and numerous drinking opportunities from the coaches.


Autoamputation of Dermatofibrosarcoma Protuberans: A Novel and Rare Presentation of a Familiar Entity.
Dermatofibrosarcoma protuberans (DFSP) is categorized as a fibrohistiocytic tumor of intermediate malignant potential. It has significant risk for local recurrence and, less commonly, local or distant metastasis. Initially, these tumors typically arise as a firm plaque on the skin that slowly progresses to a nodular and protuberant dermal lesion. DFSP can also exhibit ulceration, hemorrhage, and accelerated growth, but autoamputation has not been described in the English literature. In this article, we report a case of an asymptomatic classical DFSP on the upper back in which the protuberant portion spontaneously autoamputated. In this case, the residual lesion was treated with Mohs micrographic surgery. The presentation, features, and implications of this interesting mode of presentation are discussed.


Multi-omics Quantification of Species Variation of Escherichia coli Links Molecular Features with Strain Phenotypes.
Escherichia coli strains are widely used in academic research and biotechnology. New technologies for quantifying strain-specific differences and their underlying contributing factors promise greater understanding of how these differences significantly impact physiology, synthetic biology, metabolic engineering, and process design. Here, we quantified strain-specific differences in seven widely used strains of E. coli (BL21, C, Crooks, DH5a, K-12 MG1655, K-12 W3110, and W) using genomics, phenomics, transcriptomics, and genome-scale modeling. Metabolic physiology and gene expression varied widely with downstream implications for productivity, product yield, and titer. These differences could be linked to differential regulatory structure. Analyzing high-flux reactions and expression of encoding genes resulted in a correlated and quantitative link between these sets, with strain-specific caveats. Integrated modeling revealed that certain strains are better suited to produce given compounds or express desired constructs considering native expression states of pathways that enable high-production phenotypes. This study yields a framework for quantitatively comparing strains in a species with implications for strain selection.


The epigenetic mechanism of mechanically induced osteogenic differentiation.
Epigenetic regulation of gene expression occurs due to alterations in chromatin proteins that do not change DNA sequence, but alter the chromatin architecture and the accessibility of genes, resulting in changes to gene expression that are preserved during cell division. Through this process genes are switched on or off in a more durable fashion than other transient mechanisms of gene regulation, such as transcription factors. Thus, epigenetics is central to cellular differentiation and stem cell linage commitment. One such mechanism is DNA methylation, which is associated with gene silencing and is involved in a cell's progression towards a specific fate. Mechanical signals are a crucial regulator of stem cell behavior and important in tissue differentiation; however, there has been no demonstration of a mechanism whereby mechanics can affect gene regulation at the epigenetic level. In this study, we identified candidate DNA methylation sites in the promoter regions of three osteogenic genes from bone marrow derived mesenchymal stem cells (MSCs). We demonstrate that mechanical stimulation alters their epigenetic state by reducing DNA methylation and show an associated increase in expression. We contrast these results with biochemically induced differentiation and distinguish expression changes associated with durable epigenetic regulation from those likely to be due to transient changes in regulation. This is an important advance in stem cell mechanobiology as it is the first demonstration of a mechanism by which the mechanical micro-environment is able to induce epigenetic changes that control osteogenic cell fate, and that can be passed to daughter cells. This is a first step to understanding that will be vital to successful bone tissue engineering and regenerative medicine, where continued expression of a desired long-term phenotype is crucial.


A renal-like organic anion transport system in the ciliary epithelium of the bovine and human eye.
The purpose of this study was to determine the direction of organic anion (OA) transport across the ciliary body and the transport proteins that may contribute. Transport of several OAs across the bovine ciliary body was examined using ciliary body sections mounted in Ussing chambers and a perfused eye preparation. Microarray, reverse-transcription polymerase chain reaction (RT-PCR), immunoblotting, and immunohistochemistry were used to examine OA transporter expression in human ocular tissues. Microarray analysis showed that many OA transporters common to other barrier epithelia are expressed in ocular tissues. mRNA (RT-PCR) and protein (immunoblotting) for OAT1, OAT3, NaDC3, and MRP4 were detected in extracts of the human ciliary body from several donors. OAT1 and OAT3 localized to basolateral membranes of nonpigmented epithelial cells and MRP4 to basolateral membranes of pigmented cells in the human eye. Para-aminohippurate (PAH) and estrone-3-sulfate transport across the bovine ciliary body in the Ussing chambers was greater in the aqueous humor-to-blood direction than in the blood-to-aqueous humor direction, and active. There was little net directional movement of cidofovir. Probenecid (0.1 mM) or novobiocin (0.1 mM) added to the aqueous humor side of the tissue, or MK571 (5-(3-(2-(7-chloroquinolin-2-yl)ethenyl)phenyl)-8-dimethylcarbamyl-4,6-dithiaoctanoic acid; 0.1 mM) added to the blood side significantly reduced net active PAH transport. The rate of 6-carboxyfluorescein elimination from the aqueous humor of the perfused eye was reduced 80% when novobiocin (0.1 mM) was present in the aqueous humor. These data indicate that the ciliary body expresses a variety of OA transporters, including those common to the kidney. They are likely involved in clearing potentially harmful endobiotic and xenobiotic OAs from the eye.


Prevalence of Abuse Among Young Children With Rib Fractures: A Systematic Review.
We aimed to estimate the prevalence of abuse in young children presenting with rib fractures and to identify demographic, injury, and presentation-related characteristics that affect the probability that rib fractures are secondary to abuse. We searched PubMed/MEDLINE and CINAHL databases for articles published in English between January 1, 1990, and June 30, 2014 on rib fracture etiology in children 5 years or younger. Two reviewers independently extracted predefined data elements and assigned quality ratings to included studies. Study-specific abuse prevalences and the sensitivities, specificities, and positive and negative likelihood ratios of patients' demographic and clinical characteristics for abuse were calculated with 95% confidence intervals. Data for 1396 children 48 months or younger with rib fractures were abstracted from 10 articles. Among infants younger than 12 months, abuse prevalence ranged from 67% to 82%, whereas children 12 to 23 and 24 to 35 months old had study-specific abuse prevalences of 29% and 28%, respectively. Age younger than 12 months was the only characteristic significantly associated with increased likelihood of abuse across multiple studies. Rib fracture location was not associated with likelihood of abuse. The retrospective design of the included studies and variations in ascertainment of cases, inclusion/exclusion criteria, and child abuse assessments prevented further meta-analysis. Abuse is the most common cause of rib fractures in infants younger than 12 months. Prospective studies with standardized methods are needed to improve accuracy in determining abuse prevalence among children with rib fractures and characteristics associated with abusive rib fractures.


Pericardial Effusion in Obstructive Sleep Apnea without Pulmonary Arterial Hypertension and Daily Hypoxemia - is it Unusual?
Pericardial effusion in chronic hypoxemic lung diseases, such as Obstructive Sleep Apnea syndrome, usually occurs after the development of severe pulmonary arterial hypertension. However, data about the frequency of pericardial effusions in Obstructive Sleep Apnea syndrome without pulmonary arterial hypertension and/or daytime hypoxemia are still scarce, and their pathogenesis is unclear. To assess the prevalence of pericardial effusions and their volume and location in patients with obesity and Obstructive Sleep Apnea syndrome without pulmonary arterial hypertension and/or hypoxemia. Cross-sectional study. We included 279 consecutive patients (162 males) with newly diagnosed Obstructive Sleep Apnea syndrome having a mean age of 42.8±12.4 years and a mean body mass index of 37.3±7.8 kg/m2. Obstructive Sleep Apnea syndrome was confirmed by polysomnography. Main exclusion criteria were concomitant inflammatory diseases, thyroid dysfunction, daytime hypoxemia, nephrotic syndrome, left ventricular systolic dysfunction and pulmonary arterial hypertension. Pericardial effusion was found in 102 (36.56%) -all of them with moderate to severe obstructive Sleep Apnea syndrome. The mean effusion volume was mild to moderate (up to 250 mL). In 36 patients (35.3%) the pericardial effusion was diffuse, in 42 (41.2%), the pericardial effusion was located in front of the right atrium and the right ventricle, and in 24 (23.5%) the pericardial effusion was situated in front of the right cardiac cavities and the left atrium. We found a significant positive correlation between the presence of pericardial effusion and apnea-hypopnea index (r=0.374, p<0.001), body mass index (r=0.473, p<0.001), and desaturation time during sleep (r=0.289, p<0.001). Pericardial effusion in patients with obesity and moderate to severe Obstructive Sleep Apnea syndrome without daily hypoxemia and/or pulmonary hypertension is a relatively common finding. The occurrence of pericardial effusions is dependent mostly on the grade of Obstructive Sleep Apnea syndrome, degree of obesity, and duration of sleep desaturation.


CYTOCHROME CONTENT OF MITOCHONDRIA STRIPPED OF INNER MEMBRANE STRUCTURE.
The cytochrome composition of mitochondrial fractions which have been stripped of inner membrane subunits by exposure to high-frequency sound have been examined by lowtemperature spectroscopy. The ratio of cytochrome c to cytochrome a is not changed by the treatment, but the concentration of cytochrome per milligram of protein is increased and the concentrations of cytochromes c(1) and b change slightly. These cytochromes (c(1) and b) may be at least partly located in the subunits of the inner membrane, but the idea that all the respiratory components are located in single subunits of the mitochondrial cristae may be considered to be disproved.


The effect of yoga practice on glycemic control and other health parameters in the prediabetic state: A systematic review and meta-analysis.
A systematic review and meta-analysis was conducted to investigate the effects of yoga on glycemic control, lipid profiles, body composition and blood pressure in people in the pre-diabetic state. Studies on the effectiveness of yoga on population groups under high risk for diabetes, called prediabetic or suffering from metabolic syndromes were extracted from a thorough search of PubMed, Scopus, Cochrane Library, EBSCO and IndMED databases. Both Randomised Controlled Trial (RCT) and non-RCT studies were included in the systematic review and meta-analysis. Studies published between Jan 2002 and Dec 2018 were included. Studies were considered for evaluation if they investigated a yoga intervention to prevent T2DM, against a control group, while also reporting glycemic control and other health parameters of T2DM management. Summary effect sizes and 95% confidence intervals (CI) were calculated using the Comprehensive Meta-Analysis software in addition to publication bias. Of the 46,500 identified studies, 14 studies with 834 participants of whom were 50% women, were found to be eligible for inclusion in our systematic review. Our quantitative synthesis included 12 randomized control trials and 2 non-randomized control trials, with the follow-up period ranging from 4 to 52 weeks. Compared to controls, yoga intervention improved fasting blood glucose (FBG) [Standard Mean Difference (SMD -0.064 mg/dL (95% CI -0.201 to 0.074)]; low density lipoprotein (LDL) [SMD-0.090 mg/dL (95% CI -0.270 to 0.090)]; triglycerides [SMD -0.148 mg/dL (95% CI -0.285 to -0.012)]; total cholesterol [SMD -0.058 mg/dL (95% CI -0.220 to 0.104)] and systolic blood pressure [SMD -0.058 mm Hg (95% CI -0.168 to 0.053)]. This meta-analysis uncovered clinically improved effects of yoga intervention on glycemic control, lipid profiles and other parameters of T2DM management in prediabetic population. These results suggest that yoga intervention may be considered as a comprehensive and alternative approach to preventing T2DM. Further adequately powered, well designed RCTs are needed to support our findings and investigate the long-term effects of yoga in T2DM patients.


Applicability and importance of Caraka's concept of Aaturaparijnana Hetawah in understanding a patient.
Many concepts in Ayurveda are untouched or unexplored till date. One of such concepts to understand the healthy and unhealthy persons is that of the Aaturaparijnana Hetawah (APH), which is only mentioned in Caraka Samhita. The descriptions about Dushya, Desha, Bala, Kala, Prakriti, etc. are mentioned very briefly in all the classics. But no standard figure or pattern is maintained while considering them. It is only in the description of Aaturaparijnana Hetawah mentioned in Vimana Sthana that all the factors are pre-arranged in a chronological order to understand an individual according to one's Desha as from birth till grown up stage, and also to understand about the illness and most common disorders. In this article, it is aimed at presenting an in-depth evaluation about description of Aaturaparijnana Hetawah in Ayurveda. For this study, the references of Aturaparijnana Hetu have been collected through the Ayurvedic texts along with their commentaries. By making use of 14 factors described in Aaturaparijnana Hetawah, it is possible to have an in depth understanding about the patient's nature and factors that may be contributing to his health and illness. Aaturaparijnana Hetawah helps in an in-depth understanding of a patient's disease and heath status. More in-depth evaluation studies and application in the modern medical science should be done in order to avail the benefits of this approach.


Identification of the DNA binding site for NGFI-B by genetic selection in yeast.
An in vivo selection system for isolating targets of DNA binding proteins in yeast was developed and used to identify the DNA binding site for the NGFI-B protein, a member of the steroid-thyroid hormone receptor superfamily. The feasibility of the technique was verified by selecting DNA fragments that contained binding sites for GCN4, a well-characterized yeast transcriptional activator. The DNA binding domain of NGFI-B, expressed as part of a LexA-NGFI-B-GAL4 chimeric activator, was then used to isolate a rat genomic DNA fragment that contained an NGFI-B binding site. The NGFI-B response element (NBRE) is similar to but functionally distinct from elements recognized by the estrogen and thyroid hormone receptors and the hormone receptor-like proteins COUP-TF, CF1, and H-2RIIBP. Cotransfection experiments in mammalian cells demonstrated that NGFI-B can activate transcription from the NBRE with or without its putative ligand binding domain.


Self-regulation of amygdala activation using real-time FMRI neurofeedback.
Real-time functional magnetic resonance imaging (rtfMRI) with neurofeedback allows investigation of human brain neuroplastic changes that arise as subjects learn to modulate neurophysiological function using real-time feedback regarding their own hemodynamic responses to stimuli. We investigated the feasibility of training healthy humans to self-regulate the hemodynamic activity of the amygdala, which plays major roles in emotional processing. Participants in the experimental group were provided with ongoing information about the blood oxygen level dependent (BOLD) activity in the left amygdala (LA) and were instructed to raise the BOLD rtfMRI signal by contemplating positive autobiographical memories. A control group was assigned the same task but was instead provided with sham feedback from the left horizontal segment of the intraparietal sulcus (HIPS) region. In the LA, we found a significant BOLD signal increase due to rtfMRI neurofeedback training in the experimental group versus the control group. This effect persisted during the Transfer run without neurofeedback. For the individual subjects in the experimental group the training effect on the LA BOLD activity correlated inversely with scores on the Difficulty Identifying Feelings subscale of the Toronto Alexithymia Scale. The whole brain data analysis revealed significant differences for Happy Memories versus Rest condition between the experimental and control groups. Functional connectivity analysis of the amygdala network revealed significant widespread correlations in a fronto-temporo-limbic network. Additionally, we identified six regions--right medial frontal polar cortex, bilateral dorsomedial prefrontal cortex, left anterior cingulate cortex, and bilateral superior frontal gyrus--where the functional connectivity with the LA increased significantly across the rtfMRI neurofeedback runs and the Transfer run. The findings demonstrate that healthy subjects can learn to regulate their amygdala activation using rtfMRI neurofeedback, suggesting possible applications of rtfMRI neurofeedback training in the treatment of patients with neuropsychiatric disorders.


Impact of treatment with bevacizumab beyond disease progression: a randomized phase II study of docetaxel with or without bevacizumab after platinum-based chemotherapy plus bevacizumab in patients with advanced nonsquamous non-small cell lung cancer (WJOG 5910L).
Bevacizumab, a humanized antibody to vascular endothelial growth factor (VEGF), shows clinical activity against human cancer, with its addition to standard chemotherapy having been found to improve outcome in patients with advanced nonsquamous non-small cell lung cancer (NSCLC). However, there have been no evidence-based studies to support the continued use of bevacizumab beyond disease progression in such patients treated with the drug in first-line therapy. We have now designed a randomized phase II trial to examine the clinical benefit and safety of continued bevacizumab treatment in patients with advanced nonsquamous NSCLC whose disease has progressed after first-line treatment with bevacizumab plus a platinum-based doublet. WJOG 5910L was designed as a multicenter, open-label, randomized, phase II trial by the West Japan Oncology Group of docetaxel (arm A) versus docetaxel plus bevacizumab (arm B) in patients with recurrent or metatstatic nonsquamous NSCLC whose disease has progressed after first-line treatment with bevacizumab plus a platinum-based doublet. Patients in arm A will receive docetaxel at 60 mg/m2 and those in arm B will receive docetaxel at 60 mg/m2 plus bevacizumab at 15 mg/kg, with each drug administered on day 1 every 21 days until progression or unacceptable toxicity. The primary endpoint of the study is progression-free survival, with secondary endpoints including response rate, overall survival, and safety, for patients treated in either arm. UMIN (University Hospital Medical Information Network in Japan) 000004715.


Cervical spine injuries and helmet laws: a population-based study.
To assess the incidence of cervical spine (C-spine) injuries in patients admitted after motorcycle crash in states with mandatory helmet laws (MHL) compared with states without helmet laws or selective helmet laws. The Nationwide Inpatient Sample from the Healthcare and Utilization Project for the year 2008 was analyzed. International Classification of Diseases and Health Related Problems, Ninth Edition codes were used to identify patients with a diagnosis of motorcycle crash and C-spine injuries. National estimates were generated based on weighted analysis of the data. Outcome variables investigated were as follows: length of stay (LOS), in-hospital mortality, hospital teaching status, and discharge disposition. States were then stratified into states with MHL or selective helmet laws. A total of 30,117 discharges were identified. Of these, 2,041 (6.7%) patients had a C-spine injury. Patients in MHL states had a lower incidence of C-spine injuries (5.6 vs. 6.4%; p = 0.003) and less in-hospital mortality (1.8 vs. 2.6%; p = 0.0001). Patients older than 55 years were less likely to be discharged home (57.5% vs. 72.5%; p = 0.0001), more likely to die in-hospital (3.0% vs. 2.1%; p = 0.0001), and more likely to have a hospital LOS more than 21 days (7.7% vs. 6.2%; p = 0.0001). Patients admitted to the hospital in states with MHLs have decreased rate of C-spine injuries than those patients admitted in states with more flexible helmet laws. Patients older than 55 years are more likely to die in the hospital, have a prolonged LOS, and require services after discharge. III.


Ambulatory blood pressure monitoring using telemedicine: proof-of-concept cohort and failure modes and effects analyses.
Background: The COVID-19 pandemic has accelerated adoption of remote consulting in healthcare. Despite opportunities posed by telemedicine, most hypertension services in Europe have suspended ambulatory blood pressure monitoring (ABPM). Methods: We examined the process and performance of remotely delivered ABPM using two methodologies: firstly, a Failure Modes and Effects Analysis (FMEA) and secondly, a quantitative analysis comparing ABPM data from a subgroup of 65 participants of the Screening for Hypertension in the INpatient Environment (SHINE) diagnostic accuracy study. The FMEA was performed over seven sessions from February to March 2021, with a multidisciplinary team comprising a patient representative, a research coordinator with technical expertise and four research clinicians. Results: The FMEA identified a single high-risk step in the remote ABPM process. This was cleaning of monitoring equipment in the context of the COVID-19 pandemic, unrelated to the remote setting. A total of 14 participants were scheduled for face-to-face ABPM appointments, before the UK March 2020 COVID-19 lockdown; 62 were scheduled for remote ABPM appointments since emergence of the COVID-19 pandemic between November 2020 and August 2021. A total of 65 (88%) participants completed ABPMs; all obtained sufficient successful measurements for interpretation. For the 10 participants who completed face-to-face ABPM, there were 402 attempted ABPM measurements and 361 (89%) were successful. For the 55 participants who completed remote ABPM, there were 2516 attempted measurements and 2114 (88%) were successful. There was no significant difference in the mean per-participant error rate between face-to-face (0.100, SD 0.009) and remote (0.143, SD 0.132) cohorts (95% CI for the difference -0.125 to 0.045 and two-tailed P-value 0.353). Conclusions: We have demonstrated that ABPM can be safely and appropriately provided in the community remotely and without face-to-face contact, using video technology for remote fitting appointments, alongside courier services for delivery of equipment to participants.


Tissue distribution and quantification of the expression of mRNAs of peroxisome proliferator-activated receptors and liver X receptor-alpha in humans: no alteration in adipose tissue of obese and NIDDM patients.
Members of the peroxisome proliferator-activated receptor (PPAR) family might be involved in pathologies with altered lipid metabolism. They participate in the control of the expression of genes involved in lipid metabolism and adipocyte differentiation. In addition, thiazolidinediones improve insulin resistance in vivo by activating PPAR gamma. However, little is known regarding their tissue distribution and relative expression in humans. Using a quantitative and sensitive reverse transcription (RT)-competitive polymerase chain reaction (PCR) assay, we determined the distribution and relative mRNA expression of the four PPARs (alpha,beta, gamma1, and gamma2) and liver X receptor-alpha (LXR alpha) in the main tissues implicated in lipid metabolism. PPAR alpha and LXR alpha were mainly expressed in liver, while PPAR gamma1 predominated in adipose tissue and large intestine. We found that PPAR gamma2 mRNA was a minor isoform, even in adipose tissue, thus causing question of its role in humans. PPAR beta mRNA was present in all the tissues tested at low levels. In addition, PPAR gamma mRNA was barely detectable in skeletal muscle, suggesting that improvement of insulin resistance with thiazolidinediones may not result from a direct effect of these agents on PPAR gamma in muscle. Obesity and NIDDM were not associated with change in PPARs and LXR alpha expression in adipose tissue. The mRNA levels of PPAR gamma1, the predominant form in adipocytes, did not correlate with BMI, leptin mRNA levels, or fasting insulinemia in 29 subjects with various degrees of obesity. These results indicated that obesity is not associated with alteration in PPAR gene expression in abdominal subcutaneous adipose tissue in humans.


Defining the impact of melanopsin missense polymorphisms using in vivo functional rescue.
Melanopsin (OPN4) is an opsin photopigment expressed within intrinsically photosensitive retinal ganglion cells (ipRGCs) that mediate non-image forming (NIF) responses to light. Two single-nucleotide polymorphisms (SNPs) in human melanopsin (hOPN4), Pro10Leu and Thr394Ile, have recently been associated with abnormal NIF responses to light, including seasonal affective disorder. It has been suggested these behavioural changes are due to altered melanopsin signalling. However, there is currently no direct evidence to support this. Here we have used ipRGC-specific delivery of hOPN4 wild-type (WT), Pro10Leu or Thr394Ile adeno-associated viruses (AAV) to determine the functional consequences of hOPN4 SNPs on melanopsin-driven light responses and associated behaviours. Immunohistochemistry confirmed hOPN4 AAVs exclusively transduced mouse ipRGCs. Behavioural phenotyping performed before and after AAV injection demonstrated that both hOPN4 Pro10Leu and Thr394Ile could functionally rescue pupillary light responses and circadian photoentrainment in Opn4-/- mice, with no differences in NIF behaviours detected for animals expressing either SNP compared to hOPN4 WT. Multi-electrode array recordings revealed that ipRGCs expressing hOPN4 Thr394Ile exhibit melanopsin-driven light responses with significantly attenuated response amplitude, decreased sensitivity and faster offset kinetics compared to hOPN4 WT. IpRGCs expressing hOpn4 Pro10Leu also showed reduced response amplitude. Collectively these data suggest Thr394Ile and Pro10Leu may be functionally significant SNPs, which result in altered melanopsin signalling. To our knowledge, this study provides the first direct evidence for the effects of hOPN4 polymorphisms on melanopsin-driven light responses and NIF behaviours in vivo, providing further insight into the role of these SNPs in melanopsin function and human physiology.


Effect of training on muscle triacylglycerol and structural lipids: a relation to insulin sensitivity?
We studied whether endurance training impacts insulin sensitivity by affecting the structural and storage lipids in humans. Eight male subjects participated (age 25 +/- 1 years, height 178 +/- 3 cm, weight 76 +/- 4 kg [mean +/- SE]). Single-leg training was performed for 30 min/day for 4 weeks at approximately 70% of single-leg maximal oxygen uptake. After 8, 14, and 30 days, a two-step hyperinsulinemic-euglycemic glucose clamp, combined with catheterization of an artery and both femoral veins, was performed. In addition, a muscle biopsy was obtained from vastus lateralis of both legs. Maximal oxygen uptake increased by 7% in the trained leg (T), and training workload increased (P < 0.05) from 79 +/- 12 to 160 +/- 15 W. At day 8, glucose uptake was higher (P < 0.01) in the trained (0.8 +/- 0.2, 6.0 +/- 0.8, 13.4 +/- 1.2 mg x min(-1) x kg(-1) leg wt) than the untrained leg (0.5 +/- 0.2, 3.7 +/- 0.6, 10.5 +/- 1.5 mg x min(-1) x kg(-1) leg wt) at basal and the two succeeding clamp steps, respectively. After day 8, training did not further increase leg glucose uptake. Individual muscle triacylglycerol fatty acid composition and total triacylglycerol content were not significantly affected by training and thus showed no relation to leg glucose uptake. Individual muscle phospholipid fatty acids were not affected by training, but the content of phospholipid polyunsaturated fatty acids was higher (P < 0.06) after 30 than 8 days in T. Furthermore, after 30 days of training, the sum of phospholipid long-chain polyunsaturates was correlated to leg glucose uptake (r = 0.574, P < 0.04). Endurance training did not influence muscle triacylglycerol content or total triacylglycerol fatty acid composition. In contrast, training induced a minor increase in the content of phospholipid fatty acid membrane polyunsaturates, which may indicate that membrane lipids may have a role in the training-induced increase in insulin sensitivity.


Getting Recovery Right After Neck Dissection (GRRAND-F): Mixed-methods feasibility study to design a pragmatic randomised controlled trial.
To determine the feasibility of a randomised controlled trial to estimate the effectiveness and cost-effectiveness of a rehabilitation intervention following neck dissection (ND) after head and neck cancer (HNC). Two-arm, open, pragmatic, parallel, multicentre, randomised controlled feasibility trial. Two UK NHS hospitals. People who had HNC in whom a ND was part of their care. We excluded those with a life expectancy of six months or less, pre-existing, long-term neurological disease affecting the shoulder and cognitive impairment. Usual care (standard care supplemented with a booklet on postoperative self-management) was received by all participants. The GRRAND intervention programme consisted of usual care plus up to six individual physiotherapy sessions including neck and shoulder range of motion and progressive resistance exercises, advice and education. Between sessions, participants were advised to complete a home exercise programme. 1:1 randomisation. Allocation was based on minimisation, stratified by hospital site and spinal accessory nerve sacrifice. It was not possible to mask treatment received. Primary: Participant recruitment, retention and fidelity to the study protocol and interventions from study participants and staff at six months post-randomisation (and 12 months for those reaching that time-point). Secondary: clinical measures of pain, function, physical performance, health-related quality of life, health utilisation and adverse events. 36 participants were recruited and enrolled. The study achieved five of its six feasibility targets. These included consent - 70% of eligible participants were consented; intervention fidelity - 78% participants discharged completed the intervention sessions; contamination - none - no participants in the control arm received the GRRAND-F intervention and retention - 8% of participants were lost to follow-up. The only feasibility target that was not achieved was the recruitment target where only 36 of the planned 60 participants were recruited over 18 months. This was principally due to the COVID-19 pandemic which caused all research activity to be paused or reduced, with a subsequent reduction in. Based on the findings a full-trial can now be designed to better understand whether this proposed intervention is effective. https://www.isrctn.com/ISRCTN1197999, identifier ISRCTN11979997.


Training and development for radiographers' extended roles: a case of ad hoc implementation.
This paper examines the quality and consistency of postregistration training and development for extended role activities undertaken by radiographers. Although the undergraduate curriculum has changed to some extent to accommodate expanded role requirements, much of the training does not, and cannot, take place until radiographers are qualified and are in post. While undergraduate programmes in radiography must be approved by the Health Professions Council, and are normally accredited by the radiographers' professional body, the Society and College of Radiographers, much of the training provided for extended role activities is ad hoc and neither validated nor accredited. This paper reports the outcomes of a survey of imaging service managers and follow-up interviews with imaging service managers. Managers' views of the quality of training and development provided for extended role activities and their approaches to recruiting radiographers with extended role experience, were explored. The research identified concerns among managers regarding the training currently available and indicated potential career disadvantages for radiographers receiving unaccredited development. There is also some evidence of wastage arising from the duplication of training by trusts because of uncertainty about the standards of competence instilled by programmes.


Differential dynamics of alpha 5 integrin, paxillin, and alpha-actinin during formation and disassembly of adhesions in migrating cells.
To investigate the mechanisms by which adhesions form and disperse in migrating cells, we expressed alpha 5 integrin, alpha-actinin, and paxillin as green fluorescent protein (GFP) fusions. All localized with their endogenous counterparts and did not perturb migration when expressed at moderate levels. alpha 5-GFP also rescued the adhesive defects in CHO B2 cells, which are alpha 5 integrin deficient. In ruffling cells, alpha 5-GFP and alpha-actinin--GFP localized prominently at the leading edge in membrane protrusions. Of the three GFP fusion proteins that we examined, paxillin was the first component to appear visibly organized in protrusive regions of the cell. When a new protrusion formed, the paxillin appeared to remodel from older to newer adhesions at the leading edge. alpha-Actinin subsequently entered adhesions, which translocated toward the cell center, and inhibited paxillin turnover. The new adhesions formed from small foci of alpha-actinin--GFP and paxillin-GFP, which grew in size. Subsequently, alpha 5 integrin entered the adhesions to form visible complexes, which served to stabilize the adhesions. alpha 5-GFP also resided in endocytic vesicles that emanated from the leading edge of protrusions. Integrin vesicles at the cell rear moved toward the cell body. As cells migrated, alpha 5 vesicles also moved from a perinuclear region to the base of the lamellipodium. The alpha 5 vesicles colocalized with transferrin receptor and FM 4-64 dye. After adhesions broke down in the rear, alpha 5-GFP was found in fibrous structures behind the cell, whereas alpha-actinin--GFP and paxillin-GFP moved up the lateral edge of retracting cells as organized structures and then dissipated.


Laryngeal airway reconstruction indicates that rodent ultrasonic vocalizations are produced by an edge-tone mechanism.
Some rodents produce ultrasonic vocalizations (USVs) for social communication using an aerodynamic whistle, a unique vocal production mechanism not found in other animals. The functional anatomy and evolution of this sound production mechanism remains unclear. Using laryngeal airway reconstruction, we identified anatomical specializations critical for USV production. A robust laryngeal cartilaginous framework supports a narrow supraglottal airway. An intralaryngeal airsac-like cavity termed the ventral pouch was present in three muroid rodents (suborder Myomorpha), but was absent in a heteromyid rodent (suborder Castorimorpha) that produces a limited vocal repertoire and no documented USVs. Small lesions to the ventral pouch in laboratory rats caused dramatic changes in USV production, supporting the hypothesis that an interaction between a glottal exit jet and the alar edge generates ultrasonic signals in rodents. The resulting undulating airflow around the alar edge interacts with the resonance of the ventral pouch, which may function as a Helmholtz resonator. The proposed edge-tone mechanism requires control of intrinsic laryngeal muscles and sets the foundation for acoustic variation and diversification among rodents. Our work highlights the importance of anatomical innovations in the evolution of animal sound production mechanisms.


The effects of macrophage migratory inhibitory factor on acute-phase protein production in primary human hepatocytes.
Macrophage inhibitory factor (MIF) is a pituitary peptide released during the physiological stress response, a T-cell product secreted during the antigen-specific response and a pro-inflammatory macrophage cytokine secreted after LPS stimulation. It has become apparent that MIF is central to the regulation of the inflammatory response and is implicated in the pathogenesis of a variety of acute and chronic inflammatory conditions. This is, at least in part, due to the apparent counter-regulation of the anti-inflammatory actions of glucocorticoids, including the reversal of glucocorticoid-mediated IL-6 release inhibition. This study examines the effect of recombinant MIF on regulation of the acute phase response in isolated human hepatocytes. MIF alone increased C-reactive protein (CRP) release in a dose-dependent manner < or = 0.1 ng/ml after which the effects of MIF were attenuated. In combination with IL-6 both CRP and and alpha-1-antichymotrypsin (ACT) release were increased above levels found with either IL-6 or MIF treatment alone. Dexamethasone attenuated the effects of MIF upon CRP production but increased the MIF stimulated release of ACT. The study demonstrates that the effects of MIF upon the acute phase response are complex and can differentially modulate the production of acute phase proteins depending on the presence of other factors.


A membrane associated mCherry fluorescent reporter line for studying vascular remodeling and cardiac function during murine embryonic development.
The development of the cardiovascular system is a highly dynamic process dependent on multiple signaling pathways regulating proliferation, differentiation, migration, cell-cell and cell-matrix interactions. To characterize cell and tissue dynamics during the formation of the cardiovascular system in mice, we generated a novel transgenic mouse line, Tg(Flk1::myr-mCherry), in which endothelial cell membranes are brightly labeled with mCherry, a red fluorescent protein. Tg(Flk1::myr-mCherry) mice are viable, fertile, and do not exhibit any developmental abnormalities. High levels of mCherry are expressed in the embryonic endothelium and endocardium, and expression is also observed in capillaries in adult animals. Targeting of the fluorescent protein to the cell membrane allows for subcellular imaging and cell tracking. By acquiring confocal time lapses of live embryos cultured on the microscope stage, we demonstrate that the newly generated transgenic model beautifully highlights the sprouting behaviors of endothelial cells during vascular plexus formation. We have also used embryos from this line to imaging the endocardium in the beating embryonic mouse heart, showing that Tg(Flk1::myr-mCherry) mice are suitable for the characterization of cardio dynamics. Furthermore, when combined with the previously described Tg(Flk1::H2B-EYFP) line, cell number in addition to cell architecture is revealed, making it possible to determine how individual endothelial cells contribute to the structure of the vessel.


Sonic Hedgehog induces proliferation of committed skeletal muscle cells in the chick limb.
Myogenic Regulatory Factors (MRFs) are a family of transcription factors whose expression in a cell reflects the commitment of this cell to a myogenic fate before any cytological sign of muscle differentiation is detectable. Myogenic cells in limb skeletal muscles originate from the lateral half of the somites. Cells that migrate away from the lateral part of the somites to the limb bud do not initially express any member of the MRF family. Expression of MRFs in the muscle precursor cells starts after the migration process is completed. The extracellular signals involved in activating the myogenic programme in muscle precursor cells in the limb in vivo are not known. We wished to investigate whether Sonic Hedgehog (SHH) expressed in the posterior part of the limb bud could be involved in differentiation of the muscle precursor cells in the limb. We found that retrovirally overexpressed SHH in the limb bud induced the extension of the expression domain of the Pax-3 gene, then that of the MyoD gene and finally that of the myosin protein. This led to an hypertrophy of the muscles in vivo. Addition of SHH to primary cultures of myoblasts resulted in an increase in the proportion of myoblasts that incorporate bromodeoxyuridine, resulting in an increase of myotube number. These data show that SHH is able to activate myogenesis in vivo and in vitro in already committed myoblasts and suggest that the stimulation of the myogenic programme by SHH involves activation of cell proliferation.


Cognitive therapy for people with schizophrenia spectrum disorders not taking antipsychotic drugs: a single-blind randomised controlled trial.
Antipsychotic drugs are usually the first line of treatment for schizophrenia; however, many patients refuse or discontinue their pharmacological treatment. We aimed to establish whether cognitive therapy was effective in reducing psychiatric symptoms in people with schizophrenia spectrum disorders who had chosen not to take antipsychotic drugs. We did a single-blind randomised controlled trial at two UK centres between Feb 15, 2010, and May 30, 2013. Participants aged 16-65 years with schizophrenia spectrum disorders, who had chosen not to take antipsychotic drugs for psychosis, were randomly assigned (1:1), by a computerised system with permuted block sizes of four or six, to receive cognitive therapy plus treatment as usual, or treatment as usual alone. Randomisation was stratified by study site. Outcome assessors were masked to group allocation. Our primary outcome was total score on the positive and negative syndrome scale (PANSS), which we assessed at baseline, and at months 3, 6, 9, 12, 15, and 18. Analysis was by intention to treat, with an ANCOVA model adjusted for site, age, sex, and baseline symptoms. This study is registered as an International Standard Randomised Controlled Trial, number 29607432. 74 individuals were randomly assigned to receive either cognitive therapy plus treatment as usual (n=37), or treatment as usual alone (n=37). Mean PANSS total scores were consistently lower in the cognitive therapy group than in the treatment as usual group, with an estimated between-group effect size of -6.52 (95% CI -10.79 to -2.25; p=0.003). We recorded eight serious adverse events: two in patients in the cognitive therapy group (one attempted overdose and one patient presenting risk to others, both after therapy), and six in those in the treatment as usual group (two deaths, both of which were deemed unrelated to trial participation or mental health; three compulsory admissions to hospital for treatment under the mental health act; and one attempted overdose). Cognitive therapy significantly reduced psychiatric symptoms and seems to be a safe and acceptable alternative for people with schizophrenia spectrum disorders who have chosen not to take antipsychotic drugs. Evidence-based treatments should be available to these individuals. A larger, definitive trial is needed. National Institute for Health Research.


Vimentin Levels and Serine 71 Phosphorylation in the Control of Cell-Matrix Adhesions, Migration Speed, and Shape of Transformed Human Fibroblasts.
Metastasizing tumor cells show increased expression of the intermediate filament (IF) protein vimentin, which has been used to diagnose invasive tumors for decades. Recent observations indicate that vimentin is not only a passive marker for carcinoma, but may also induce tumor cell invasion. To clarify how vimentin IFs control cell adhesions and migration, we analyzed the nanoscale (30-50 nm) spatial organization of vimentin IFs and cell-matrix adhesions in metastatic fibroblast cells, using three-color stimulated emission depletion (STED) microscopy. We also studied whether wild-type and phospho-deficient or -mimicking mutants of vimentin changed the size and lifetime of focal adhesions (FAs), cell shape, and cell migration, using live-cell total internal reflection imaging and confocal microscopy. We observed that vimentin exists in fragments of different lengths. Short fragments were mostly the size of a unit-length filament and were mainly localized close to small cell-matrix adhesions. Long vimentin filaments were found in the proximity of large FAs. Vimentin expression in these cells caused a reduction in FAs size and an elongated cell shape, but did not affect FA lifetime, or the speed or directionality of cell migration. Expression of a phospho-mimicking mutant (S71D) of vimentin increased the speed of cell migration. Taken together, our results suggest that in highly migratory, transformed mesenchymal cells, vimentin levels control the cell shape and FA size, but not cell migration, which instead is linked to the phosphorylation status of S71 vimentin. These observations are consistent with the possibility that not only levels, but also the assembly status of vimentin control cell migration.


Cortical structure of hallucal metatarsals and locomotor adaptations in hominoids.
Diaphyseal morphology of long bones, in part, reflects in vivo loads experienced during the lifetime of an individual. The first metatarsal, as a cornerstone structure of the foot, presumably expresses diaphyseal morphology that reflects loading history of the foot during stance phase of gait. Human feet differ substantially from those of other apes in terms of loading histories when comparing the path of the center of pressure during stance phase, which reflects different weight transfer mechanisms. Here we use a novel approach for quantifying continuous thickness and cross-sectional geometric properties of long bones in order to test explicit hypotheses about loading histories and diaphyseal structure of adult chimpanzee, gorilla, and human first metatarsals. For each hallucal metatarsal, 17 cross sections were extracted at regularly-spaced intervals (2.5% length) between 25% and 65% length. Cortical thickness in cross sections was measured in one degree radially-arranged increments, while second moments of area were measured about neutral axes also in one degree radially-arranged increments. Standardized thicknesses and second moments of area were visualized using false color maps, while penalized discriminant analyses were used to evaluate quantitative species differences. Humans systematically exhibit the thinnest diaphyseal cortices, yet the greatest diaphyseal rigidities, particularly in dorsoplantar regions. Shifts in orientation of maximum second moments of area along the diaphysis also distinguish human hallucal metatarsals from those of chimpanzees and gorillas. Diaphyseal structure reflects different loading regimes, often in predictable ways, with human versus non-human differences probably resulting both from the use of arboreal substrates by non-human apes and by differing spatial relationships between hallux position and orientation of the substrate reaction resultant during stance. The novel morphological approach employed in this study offers the potential for transformative insights into form-function relationships in additional long bones, including those of extinct organisms (e.g., fossils).


Anthropometry, dietary intake, physical activity and sitting time patterns in adolescents aged 15-17 years: an international comparison in eight Latin American countries.
Although there is high prevalence of obesity and other cardiovascular risk factors among Latin American adolescents, there is limited evidence on dietary intake and physical activity (PA) patterns in this population. Therefore, we characterized anthropometry, dietary intake, PA and sitting time (ST) in adolescents aged 15-17 years from eight Latin American countries. Six hundred seventy-one adolescents (41.4% girls) from the Latin American Study of Nutrition and Health (ELANS) were included. Nutritional status was classified by four BMI (kg/m2) categories. Waist circumference (WC) was categorized as above or below thresholds. Dietary intake was assessed through two non-consecutive 24-h dietary recalls. PA and ST were measured using the International Physical Activity Questionnaire (IPAQ). We calculated overall and country-specific estimates by sex and tested for differences between boys and girls. Differences in the prevalence of overweightness (15.1 and 21.6%) and obesity (8.5 and 6.5%) between boys and girls, respectively, were statistically insignificant (p = 0.059). Average energy intake was 2289.7 kcal/day (95% CI: 2231-2350) for boys and 1904.2 kcal/day (95% CI: 1840-1963) for girls (p < 0.001). In relation to macronutrient intake for boys and girls, respectively, the average intake (expressed as percentage of total energy) was 15.0 and 14.9% for protein; 55.4 and 54.9% for carbohydrates; 14.1 and 14.5% for added sugar; 29.5 and 30.1% for total fat; and 9.6 and 9.9% for saturated fat (p > 0.05 for all outcomes). There was no statistically significant difference in the prevalence of total energy (TE) saturated fat and added sugar (>10% of TE) between girls and boys (49.6% versus 44.8 and 81.7% versus 76.1%, respectively). Prevalence of physical inactivity was 19% in boys and 43.7% in girls (p < 0.001). Median levels of vigorous-intensity PA and total PA were significantly higher for boys than for girls (p < 0.05 for both outcomes); whereas levels of ST were similar (273.7 versus 220.0 min/day for boys and girls, respectively; p > 0.05). These findings highlight the high prevalence of poor dietary intake and physical inactivity in adolescents from Latin American countries. Therefore, effective and sustainable strategies and programmes are needed that promote healthier diets, regular PA and reduce ST among Latin American adolescents. Clinical Trials NCT02226627. Retrospectively registered on August 27, 2014.


Impacts of river water consumption on aquatic biodiversity in life cycle assessment--a proposed method, and a case study for Europe.
In the context of climate change and food provisioning for a growing global population, the impacts of water consumption on aquatic biodiversity (e.g., river water consumption for irrigation) should be considered in Life Cycle Impact Assessment (LCIA). A previous LCIA method quantifying the potential impacts of river water consumption on fish biodiversity, using a species-discharge relationship (SDR), constituted an essential first step. This method is however limited in terms of regionalization and taxa considered, and predicts the potential risk of local species loss only. Here, we address these shortcomings by developing region-specific SDRs for Europe at various scales (continent, country, and eco-region), and including macro-invertebrate biodiversity. SDR exponents vary from 0.06 to 0.45 between regions, underlining the importance of such regionalization. Furthermore, we provide a new regionalized method which considers the location of water consumption within a river basin, by integrating the concept of longitudinal river zonation. This involves the use of a novel measure of potential loss of species richness, standardizing local species loss to an equivalent of global extinction and reflecting species vulnerability. The new method is applied in a Swiss case-study. The consideration of the location of water consumption within a basin was found to be of high importance in the assessment: potential species loss varied between 4.22 × 10(-3) and 3.95 × 10(-1) species (2 orders of magnitude) depending on location. This work thus provides enhancements in the assessment of potential impacts of river water consumption on aquatic biodiversity and contributes to the ecological relevance of the method.