Preparation of Highly Reactive Lithium Metal Dendrites for the Synthesis of Organolithium Reagents.
A long-standing problem in the area of organolithium chemistry has been the need for a highly reactive Li-metal source that mimics Li-powders but has the advantage of being freshly prepared from inexpensive and readily available Li-sources. Here, we report a simple and convenient activation method using liquid ammonia that furnishes a new Li-metal source in the form of crystalline Li-dendrites. The Li-dendrites were shown to have ca. 100 times greater surface area than conventional Li-sources created by prototypical mechanical activation methods. Concomitant with the surface area increase, the Li-dendrites were shown to exhibit significant rate enhancements over Li-powders, which are currently the industry standard for the preparation of organolithium compounds. These features were leveraged for the reproducible synthesis of organolithium reagents over a range of common laboratory scales.


The role of dimer asymmetry and protomer dynamics in enzyme catalysis.
Freeze-trapping x-ray crystallography, nuclear magnetic resonance, and computational techniques reveal the distribution of states and their interconversion rates along the reaction pathway of a bacterial homodimeric enzyme, fluoroacetate dehalogenase (FAcD). The crystal structure of apo-FAcD exhibits asymmetry around the dimer interface and cap domain, priming one protomer for substrate binding. This asymmetry is dynamically averaged through conformational exchange on a millisecond time scale. During catalysis, the protomer conformational exchange rate becomes enhanced, the empty protomer exhibits increased local disorder, and water egresses. Computational studies identify allosteric pathways between protomers. Water release and enhanced dynamics associated with catalysis compensate for entropic losses from substrate binding while facilitating sampling of the transition state. The studies provide insights into how substrate-coupled allosteric modulation of structure and dynamics facilitates catalysis in a homodimeric enzyme.


Fear Avoidance Beliefs in College Athletes with a History of Ankle Sprain.
Ankle sprains are the most prevalent injuries, and elevated fear avoidance beliefs after ankle sprain episodes could inhibit athletic performance and contribute to residual symptoms, such as functional and/or mechanical instability. However, it remains unclear how fear avoidance beliefs differ according to conditions of posttraumatic sequelae. The purpose of this study was to determine whether fear of movement/reinjury differed between individuals with and without functional ankle instability (FI, NFI) and healthy controls (CON) and to examine the relationship between fear and ankle joint laxity by sex. Participants (115 male athletes, 105 female athletes) completed the Identification of Functional Ankle Instability, Athlete Fear Avoidance Questionnaire (AFAQ), Tampa Scale for Kinesiophobia (TSK), and ankle joint laxity test. Total 168 athletes (79 males, 89 females) data were eligible for analysis. The results demonstrated that fear of movement/reinjury was lower in individuals in the absence of functional ankle instability although they experienced ankle sprain (FI; TSK=38.6±4.5, AFAQ=27.4±6.2, NFI; TSK=35.7±5.6, AFAQ=24.5±6.6). The fear of movement/reinjury had correlation with ankle joint laxity only in female athletes (TSK; r=0.285, p=0.013, AFAQ; r 0=0.322, p=0.045).


Inferring the Effects of Honokiol on the Notch Signaling Pathway in SW480 Colon Cancer Cells.
In a tumor cell, the development of acquired therapeutic resistance and the ability to survive in extracellular environments that differ from the primary site are the result of molecular adaptations in potentially highly plastic molecular networks. The accurate prediction of intracellular networks in a tumor remains a difficult problem in cancer informatics. In order to make truly rational patient-driven therapeutic decisions, it will be critical to develop methodologies that can accurately infer the molecular circuitry in the cells of a specific tumor. Despite enormous heterogeneity, cellular networks elicit deterministic digital-like responses. We discuss the use and limitations of methodologies that model molecular networks in cancer cells as a digital circuit. We also develop a network model of Notch signaling in colon cancer using a novel reverse engineering logic-based method and published western blot data to elucidate the interactions likely present in the circuits of the SW480 colon cancer cell line. Within this framework, we make predictions related to the role that honokiol may be playing as an anti-cancer drug.


Prediction of EGFR and KRAS mutation in non-small cell lung cancer using quantitative 18F FDG-PET/CT metrics.
This study investigated the relationship between epidermal growth factor receptor (EGFR) and Kirsten rat sarcoma viral oncogene homolog (KRAS) mutations in non-small-cell lung cancer (NSCLC) and quantitative FDG-PET/CT parameters including tumor heterogeneity. 131 patients with NSCLC underwent staging FDG-PET/CT followed by tumor resection and histopathological analysis that included testing for the EGFR and KRAS gene mutations. Patient and lesion characteristics, including smoking habits and FDG uptake parameters, were correlated to each gene mutation. Never-smoker (P < 0.001) or low pack-year smoking history (p = 0.002) and female gender (p = 0.047) were predictive factors for the presence of the EGFR mutations. Being a current or former smoker was a predictive factor for the KRAS mutations (p = 0.018). The maximum standardized uptake value (SUVmax) of FDG uptake in lung lesions was a predictive factor of the EGFR mutations (p = 0.029), while metabolic tumor volume and total lesion glycolysis were not predictive. Amongst several tumor heterogeneity metrics included in our analysis, inverse coefficient of variation (1/COV) was a predictive factor (p < 0.02) of EGFR mutations status, independent of metabolic tumor diameter. Multivariate analysis showed that being a never-smoker was the most significant factor (p < 0.001) for the EGFR mutations in lung cancer overall. The tumor heterogeneity metric 1/COV and SUVmax were both predictive for the EGFR mutations in NSCLC in a univariate analysis. Overall, smoking status was the most significant factor for the presence of the EGFR and KRAS mutations in lung cancer.


Sleep-Dependent Modulation of Metabolic Rate in Drosophila.
Dysregulation of sleep is associated with metabolic diseases, and metabolic rate (MR) is acutely regulated by sleep-wake behavior. In humans and rodent models, sleep loss is associated with obesity, reduced metabolic rate, and negative energy balance, yet little is known about the neural mechanisms governing interactions between sleep and metabolism. We have developed a system to simultaneously measure sleep and MR in individual Drosophila, allowing for interrogation of neural systems governing interactions between sleep and metabolic rate. Like mammals, MR in flies is reduced during sleep and increased during sleep deprivation suggesting sleep-dependent regulation of MR is conserved across phyla. The reduction of MR during sleep is not simply a consequence of inactivity because MR is reduced ~30 minutes following the onset of sleep, raising the possibility that CO2 production provides a metric to distinguish different sleep states in the fruit fly. To examine the relationship between sleep and metabolism, we determined basal and sleep-dependent changes in MR is reduced in starved flies, suggesting that starvation inhibits normal sleep-associated effects on metabolic rate. Further, translin mutant flies that fail to suppress sleep during starvation demonstrate a lower basal metabolic rate, but this rate was further reduced in response to starvation, revealing that regulation of starvation-induced changes in MR and sleep duration are genetically distinct. Therefore, this system provides the unique ability to simultaneously measure sleep and oxidative metabolism, providing novel insight into the physiological changes associated with sleep and wakefulness in the fruit fly.


Spatio-temporal transcript profiling of rice roots and shoots in response to phosphate starvation and recovery.
Using rice (Oryza sativa) as a model crop species, we performed an in-depth temporal transcriptome analysis, covering the early and late stages of Pi deprivation as well as Pi recovery in roots and shoots, using next-generation sequencing. Analyses of 126 paired-end RNA sequencing libraries, spanning nine time points, provided a comprehensive overview of the dynamic responses of rice to Pi stress. Differentially expressed genes were grouped into eight sets based on their responses to Pi starvation and recovery, enabling the complex signaling pathways involved in Pi homeostasis to be untangled. A reference annotation-based transcript assembly was also generated, identifying 438 unannotated loci that were differentially expressed under Pi starvation. Several genes also showed induction of unannotated splice isoforms under Pi starvation. Among these, PHOSPHATE2 (PHO2), a key regulator of Pi homeostasis, displayed a Pi starvation-induced isoform, which was associated with increased translation activity. In addition, microRNA (miRNA) expression profiles after long-term Pi starvation in roots and shoots were assessed, identifying 20 miRNA families that were not previously associated with Pi starvation, such as miR6250. In this article, we present a comprehensive spatio-temporal transcriptome analysis of plant responses to Pi stress, revealing a large number of potential key regulators of Pi homeostasis in plants.


Mechanism and evolution of hypoxia-tolerance in humans.
To physiologists, the term 'adaptation' usually refers to any trait that is considered advantageous; evolutionary biologists require a more rigorous definition (restricting it to traits arising and maintained under selection). By their definition, many physiological traits may merely reflect inheritance passed on through lineage. In considering the evolution of tolerance to reduced oxygen availability, we examined the issue (of true adaptations versus simple inheritance) in pinnipeds (the two dominant groups, phocids and otariids, with varying diving capacities) and in human lineages exposed for varying generational periods to hypobaric hypoxia. Basic principles of the evolution of complex physiological systems first emerged from an analysis of the diving response. We then analyzed human responses to hypobaric hypoxia in three different lineages: lowlanders, Andean natives (Quechuas) and Himalayan natives (Sherpas). As in the pinniped example, we found 'conservative' and 'adaptable' physiological characters involved in human responses to hypoxia. Conservative characters are clearly dominant and are too numerous to outline in detail; three examples are haemoglobin oxygen-affinities, the organization of muscle into different fibre types and the brain's almost exclusive preference for glucose as a fuel. Most notably, we also found evidence for 'adaptable' characters at all levels of organization examined. At the whole-body level in Quechuas and Sherpas, we found (i) that maximum aerobic and anaerobic exercise capacities were down-regulated, (ii) that the acute effect of hypoxia (making up the energy deficit due to oxygen lack; i.e. the Pasteur effect) expected from lowlanders was blunted, and (iii) that acclimation effects were also attenuated. The biochemical behaviour of skeletal muscles was consistent with lowered reliance on glycolytic contributions to energy supply, thus improving the yield of ATP per mole of carbon fuel utilized. Heart adaptations also seemed to rely upon stoichiometric efficiency adjustments, improving the yield of ATP per mole of oxygen consumed (by using glucose in preference to fatty acids). Most of the biochemical and physiological adaptations we noted (both as acute and as acclimation responses) were similar in Sherpas and Quechuas. These two lineages have not shared a common ancestor for approximately one-third of the history of our species, so it is possible that their similar physiological traits arose independently as hypoxia defence adaptations in two different times and places in our history. As in the evolution of exquisite capacities for management of oxygen down to vanishingly low levels in diving animals, the evolution of human hypoxia-tolerance can be described in terms of how two (conservative versus adaptable) categories of physiological characters are assembled in different human lineages and how the assembly changes through generational time. More recent evidence indicating that our species evolved under 'colder, drier and higher' conditions suggests that these adaptations may represent the 'ancestral' physiological condition for humans.


City tumour board Karachi: an innovative step in multidisciplinary consensus meeting and its two years audit.
Management of cancer patients is a team work which usually comprises of surgeons, oncologists, radiologists, pathologists, psychiatrist, nutritionist and a nurse. Any patient who is suffering from any tumour needs a multimodality meeting as cancer treatment is not a single persons job. Most of the time, it is difficult to get the whole team together for a plan discussion due to their busy schedule. This problem was overcome by starting a tumour board meeting early morning of Sunday in Karachi which was named "City Tumour Board (CTB) Karachi". Its first meeting was held on Sunday March 28, 2010 and since then it takes place regularly fortnightly. Till March 2012, 44 sessions were conducted and total 264 cases were discussed. Here we present an audit of these two years. On average, in 60% of cases, tumour was up (36%) or down staged (12%) while in 52% of cases the stage remained unchanged. In 70% of cases (inclusive of above 60%), initial treatment plan was changed after discussion in the tumour board. This data signifies the importance of tumour board especially in a Pakistani setup where patient and even referring persons are not well aware of this disease and its outcome. It is advisable that every case should be discussed in tumour board before embarking on any treatment so that the best treatment plan can be given. It is also important that all relevant specialists should be present in the tumour board when planning for any treatment.


Reproductive strategies and isolation-by-demography in a marine clonal plant along an eutrophication gradient.
Genetic diversity in clonal organisms includes two distinct components, (i) the diversity of genotypes or clones (i.e. genotypic richness) in a population and (ii) that of the alleles (i.e. allelic and gene diversity within populations, and differentiation between populations). We investigated how population differentiation and genotypic components are associated across a gradient of eutrophication in a clonal marine plant. To that end, we combined direct measurements of sexual allocation (i.e. flower and seed counts) and genotypic analyses, which are used as an estimator of effective sexual reproduction across multiple generations. Genetic differentiation across sites was also modelled according to a hypothesis here defined as isolation-by-demography, in which we use population-specific factors, genotypic richness and eutrophication that are hypothesized to affect the source-sink dynamics and thus influence the genetic differentiation between a pair of populations. Eutrophic populations exhibited lower genotypic richness, in agreement with lower direct measurements of sexual allocation and contemporaneous gene flow. Genetic differentiation, while not explained by distance, was best predicted by genotypic richness and habitat quality. A multiple regression model using these two predictors was considered the best model (R(2) = 0.43). In this study, the relationship between environment and effective sexual-asexual balance is not simply (linearly) predicted by direct measurements of sexual allocation. Our results indicate that population-specific factors and the isolation-by-demography model should be used more often to understand genetic differentiation.


Copper Tolerance in Australian Populations of Xanthomonas campestris pv. vesicatoria Contributes to Poor Field Control of Bacterial Spot of Pepper.
Many Australian pepper producers have reported poor control of bacterial spot of pepper (caused by Xanthomonas campestris pv. vesicatoria) despite using copper bactericides at recommended rates. This prompted us to investigate whether copper-tolerant strains of the pathogen are present in Australia and whether they are a contributing factor to poor control. We screened strains of X. campestris pv. vesicatoria in the laboratory, in broths containing different concentrations of CuSO4, and established the maximum copper concentration at which each strain could grow. About one in four strains (75 total) collected between 1999 and 2000 tolerated ≥1.0 mM CuSO4 and could be considered copper tolerant. Only one strain (of 12) collected before 1987 could tolerate 1.0 mM, and comparison of the two populations by fitting regression models revealed that the mortality of strains in the two populations differed significantly across all concentrations of copper. It was necessary to apply higher rates of copper to strains collected between 1999 and 2000 to achieve mortality levels equivalent to those in the older population. This is strong evidence that the prevalence of copper-tolerant strains has increased in Australian populations. We also assessed whether copper-tolerant strains may increase substantially in a field population of X. campestris pv. vesicatoria when copper is repeatedly sprayed. In the field, the proportion of copper-tolerant strains increased after 12 weekly sprays of copper, and they were very prevalent in the population after 21 sprays. This is the first report of copper tolerance in Australian populations of X. campestris pv. vesicatoria.


Characterization of ligand and substrate specificity for the calcium-dependent and calcium-independent protein kinase C isozymes.
Analysis of [3H]phorbol-12,13-dibutyrate (PDBu) binding was performed with protein kinase C (PKC)-alpha, -beta 1, -gamma, -delta, -epsilon, -eta, and -zeta produced in Sf9 insect cells using the baculovirus expression system. With the exception of PKC-zeta, all of the PKC isozymes bound [3H]PDBu with high affinity (Kd < 1 nM), either in the presence or in the absence of calcium. Scatchard analysis using 100% phosphatidylserine vesicles revealed slightly lower affinity for the calcium-independent isozymes (PKC-delta, -epsilon, and -eta) than for the calcium-dependent isozymes (PKC-alpha, -beta, and -gamma). Competition for [3H]PDBu binding by different classes of PKC activators showed that 12-deoxyphorbol esters, mezerein, and octahydromezerein likewise possessed lower affinity for the calcium-independent isozymes. The mezerein analog thymeleatoxin was the most marked example, being almost 20-fold less potent for binding to PKC-epsilon and -eta than to PKC-beta 1. In contrast, the indole alkaloids (-)-indolactam V and (-)-octylindolactam V and the postulated endogenous activator 1,2-diacylglycerol bound with similar affinities to all of the PKC isoforms, suggesting that different residues/configurations in the binding sites of the different PKC isozymes might be involved in interaction with the pharmacophore of the activators. The seven PKC isozymes also showed clearly different substrate specificities with exogenous peptide and protein substrates. The heterogeneous behavior of the different members of the PKC family with ligands and substrates may contribute to the heterogeneity of PKC-mediated pathways at the cellular level.


Thrombophilia and pregnancy.
Pregnancy is hypercoagulable state. The field of thrombophilia; the tendency to thrombosis, has been developed rapidly and has been linked to many aspects of pregnancy. It is recently that severe pregnancy complications such as severe preeclampsia intrauterine growth retardation abruptio placentae and stillbirth has been shown to be associated with thrombophilia. Recurrent miscarriage and has also been associated with thrombophilia. Finally, thromboembolism in pregnancy as in the non-pregnant state is linked to thrombophilia. In this review all aspects of thrombophilia in pregnancy are discussed, and also all prophylactic and therapeutic implications.


Recent reports of Wi-Fi and mobile phone-induced radiation on oxidative stress and reproductive signaling pathways in females and males.
Environmental exposure to electromagnetic radiation (EMR) has been increasing with the increasing demand for communication devices. The aim of the study was to discuss the mechanisms and risk factors of EMR changes on reproductive functions and membrane oxidative biology in females and males. It was reported that even chronic exposure to EMR did not increase the risk of reproductive functions such as increased levels of neoantigens abort. However, the results of some studies indicate that EMR induced endometriosis and inflammation and decreased the number of follicles in the ovarium or uterus of rats. In studies with male rats, exposure caused degeneration in the seminiferous tubules, reduction in the number of Leydig cells and testosterone production as well as increases in luteinizing hormone levels and apoptotic cells. In some cases of male and female infertility, increased levels of oxidative stress and lipid peroxidation and decreased values of antioxidants such as melatonin, vitamin E and glutathione peroxidase were reported in animals exposed to EMR. In conclusion, the results of current studies indicate that oxidative stress from exposure to Wi-Fi and mobile phone-induced EMR is a significant mechanism affecting female and male reproductive systems. However, there is no evidence to this date to support an increased risk of female and male infertility related to EMR exposure.


Poly(phosphoester) ionomers as tissue-engineering scaffolds.
Regenerative medicine requires scaffolds of divergent physicochemical properties for different tissue-engineering applications. To this end, a series of biodegradable poly(phosphoester) ionomers of the general composition [p(BHET-EOP-HOP/TC)] was synthesized, with BHET(bis-hydroxyl ethylene phosphate):EOP(ethylene phosphate):HOP(free phosphate) ratios of 60:20:20, 70:10:20, and 75:5:20, respectively. The 60/20/20 ionomer possessed the best tensile properties, exhibiting an average tensile modulus of 68 MPa and strain at break of 31%. Calcium treatment of the ionomer films led to significantly higher hardness and elastic moduli as measured by indentation. Calcium binding was evident from the increase in glass transition and melting temperatures, and a shift in the P-->O absorption in the FTIR spectrum. Stable N-hydroxysuccinimide ester (NHS) of the ionomers could be synthesized to facilitate derivatization, as demonstrated by conjugation of GRGDS in this study. The polymers conjugated with NHS were hydrolyzed in a biphasic mode, with a fast initial phase occurring in the first few hours, followed by a slower phase in the next few days. These ionomers represent a novel class of biomaterials with readily controllable physical and chemical attributes for tissue engineering.


Pudendal motoneurons of the rat located in separated spinal nuclei possess nicotinic acetylcholine receptors having distinct pharmacological profiles.
Pudendal motoneurons are located in the ventral horn of the caudal lumbar spinal cord and innervate striated pelvic muscles implicated in sexual and eliminative functions. In rats they are distributed in the dorsomedial (DM) and dorsolateral (DL) nucleus. In male rats, dorsomedial motoneurons innervate the bulbocavernosus, the levator ani and the external anal sphincter, whereas dorsolateral motoneurons control the ischiocavernosus and external urethral sphincter. Using spinal cord slices of young male rats and whole-cell patch-clamp recordings, we investigated the sensitivity of pudendal motoneurons to nicotinic cholinergic agonists. Motoneurons were identified following 1,1'-dilinoleyl-3,3,3',3'-tetramethylindocarbocyanine, 4-chlorobenzenesulphonate retrograde labelling. In the presence of atropine, both dorsomedial and dorsolateral motoneurons responded to acetylcholine (ACh) by generating a rapidly activating inward current. By using selective nicotinic antagonists and a nicotinic positive allosteric modulator, we found that nicotinic ACh receptors present in dorsomedial and dorsolateral motoneurons display distinct pharmacological profiles. Whereas the former are of the heteromeric type, the latter are predominantly of the alpha7-containing type. These data were confirmed by light microscopic autoradiography. In young rats, a ligand for heteromeric nicotinic receptors labelled all laminae of the central grey matter, whereas in the ventral part of the central grey, a ligand selective for alpha7-containing nicotinic receptors labelled the DL but not the DM. Dorsolateral and dorsomedial motoneurons innervate two distinct groups of pelvic muscles. A difference in their nicotinic pharmacology may be clinically relevant, as it might allow a selective pharmacological intervention in view of influencing the activity of one or the other set of muscles.


Measurements of Long-range Electronic Correlations During Femtosecond Diffraction Experiments Performed on Nanocrystals of Buckminsterfullerene.
The precise details of the interaction of intense X-ray pulses with matter are a topic of intense interest to researchers attempting to interpret the results of femtosecond X-ray free electron laser (XFEL) experiments. An increasing number of experimental observations have shown that although nuclear motion can be negligible, given a short enough incident pulse duration, electronic motion cannot be ignored. The current and widely accepted models assume that although electrons undergo dynamics driven by interaction with the pulse, their motion could largely be considered 'random'. This would then allow the supposedly incoherent contribution from the electronic motion to be treated as a continuous background signal and thus ignored. The original aim of our experiment was to precisely measure the change in intensity of individual Bragg peaks, due to X-ray induced electronic damage in a model system, crystalline C60. Contrary to this expectation, we observed that at the highest X-ray intensities, the electron dynamics in C60 were in fact highly correlated, and over sufficiently long distances that the positions of the Bragg reflections are significantly altered. This paper describes in detail the methods and protocols used for these experiments, which were conducted both at the Linac Coherent Light Source (LCLS) and the Australian Synchrotron (AS) as well as the crystallographic approaches used to analyse the data.


An oral formulation of angiotensin-(1-7) reverses corpus cavernosum damages induced by hypercholesterolemia.
The renin angiotensin system plays a crucial role in erectile function. It has been shown that elevated angiotensin-II levels contribute to the development of erectile dysfunction (ED). Oppositely, angiotensin-(1-7) (Ang-[1-7]) mediates penile erection by activation of receptor Mas. Recently, we have developed a formulation based on Ang-(1-7) inclusion in cyclodextrin (CyD) [Ang-(1-7)-CyD], which allows for the oral administration of Ang-(1-7). In the present study, we evaluated the effects of chronic treatment with Ang-(1-7)-CyD on penile fibrosis, oxidative stress, and endothelial function in hypercholesterolemic mice. Apolipoprotein(Apo)E-/- mice fed a Western-type diet for 11 weeks received Ang-(1-7)-CyD or vehicle during the final 3 weeks. Collagen content and reactive oxygen species (ROS) production within the corpus cavernosum were evaluated by Sirius red and dihydroethidium staining, respectively. Protein expression of neuronal nitric oxide synthase (nNOS) and endothelial nitric oxide synthase (eNOS), nicotinamide adenine dinucleotide phosphate (NADPH) subunits (p67-phox and p22-phox), and AT1 and Mas receptors in the penis was assessed by Western blotting. Nitric oxide (NO) production was measured by Griess assay in the mice serum. Cavernosal strips were mounted in an isometric organ bath to evaluate the endothelial function. The effect of Ang-(1-7)-CyD treatment on penile fibrosis, oxidative stress, and endothelial function in hypercholesterolemia-induced ED. Ang-(1-7)-CyD treatment reduced collagen content in the corpus cavernosum of ApoE-/- mice. This effect was associated with an attenuation of ROS production and a diminished expression of NADPH. Furthermore, Ang-(1-7)-CyD treatment augmented the expression of nNOS and eNOS in the penis and elevated vascular NO production. Importantly, these effects were accompanied by an improvement in cavernosal endothelial function. Long-term treatment with Ang-(1-7)-CyD reduces penile fibrosis associated with attenuation of oxidative stress. Additionally, cavernosal endothelial function in hypercholesterolemic mice was markedly improved. These results suggest that Ang-(1-7)-CyD might have significant therapeutic benefits for the treatment of erectile dysfunction.


Circulating 25-hydroxyvitamin D3 in pregnancy and infant neuropsychological development.
To investigate whether circulating 25-hydroxyvitamin D(3) [25(OH)D(3)] concentration in pregnancy is associated with neuropsychological development in infants. The Spanish population-based cohort study INfancia y Medio Ambiente Project recruited pregnant women during the first trimester of pregnancy between November 2003 and February 2008. Completed data on 1820 mother-infant pairs were used. Maternal plasma 25(OH)D(3) concentration was measured by high-performance liquid chromatography in pregnancy (mean 13.5 ± 2.1 weeks of gestation). Offspring mental and psychomotor scores were assessed by trained psychologists at age 14 months (range, 11-23) by using the Bayley Scales of Infant Development. β-Coefficients with 95% confidence intervals (CIs) of mental and psychomotor scores associated with continuous or categorical concentrations of maternal plasma 25(OH)D(3) were calculated by using linear regression analysis. The median plasma value of 25(OH)D(3) in pregnancy was 29.6 ng/mL (interquartile range, 21.8-37.3). A positive linear relationship was found between circulating concentrations of maternal 25(OH)D(3) concentrations in pregnancy and mental and psychomotor scores in the offspring. After adjustment for potential confounders, infants of mothers with 25(OH)D(3) concentrations in pregnancy >30 ng/mL showed higher mental score (β = 2.60; 95% CI 0.63-4.56) and higher psychomotor score (β = 2.32; 95% CI 0.36-4.28) in comparison with those of mothers with 25(OH)D(3) concentrations <20 ng/mL. Higher circulating concentration of maternal 25(OH)D(3) in pregnancy was associated with improved mental and psychomotor development in infants.


Senescent cells evade immune clearance via HLA-E-mediated NK and CD8+ T cell inhibition.
Senescent cells accumulate in human tissues during ageing and contribute to age-related pathologies. The mechanisms responsible for their accumulation are unclear. Here we show that senescent dermal fibroblasts express the non-classical MHC molecule HLA-E, which interacts with the inhibitory receptor NKG2A expressed by NK and highly differentiated CD8+ T cells to inhibit immune responses against senescent cells. HLA-E expression is induced by senescence-associated secretary phenotype-related pro-inflammatory cytokines, and is regulated by p38 MAP kinase signalling in vitro. Consistently, HLA-E expression is increased on senescent cells in human skin sections from old individuals, when compared with those from young, and in human melanocytic nevi relative to normal skin. Lastly, blocking the interaction between HLA-E and NKG2A boosts immune responses against senescent cells in vitro. We thus propose that increased HLA-E expression contributes to persistence of senescent cells in tissues, thereby suggesting a new strategy for eliminating senescent cells during ageing.


Involvement of epithelial to mesenchymal transition in the development of pancreatic ductal adenocarcinoma.
Pancreatic ductal adenocarcinoma (PDAC) is an intractable disease as a result of its rapid dissemination and resistance to conventional chemotherapy and radiotherapy. Surgical resection is the only curative therapy, but most of the tumors are unresectable at the time of diagnosis. The molecular mechanisms underlying the biological aggressiveness of this tumor type remain to be clarified. Epithelial to mesenchymal transition (EMT) is a developmental process that leads the phenotype shift from an epithelial morphology to a motile, fibroblast-like morphology. Recent studies showed that EMT is involved in the invasion and metastasis of many types of carcinomas including PDAC. In addition, PDAC cells with the EMT phenotype also exhibit chemoresistance and the cancer stem cell property. Various factors such as cytokines, growth factors, or transcriptional factors were found to promote the EMT program in PDAC cells. In this review, we summarize the current knowledge about the EMT in PDAC cells, focusing on the involvement of this process and its regulatory molecules including microRNA during the development of PDAC cells.


Use and perceived benefits of mobile devices by physicians in preventing adverse drug events in the nursing home.
Although mobile devices equipped with drug reference software may help prevent adverse drug events (ADEs) in the nursing home (NH) by providing medication information at the point of care, little is known about their use and perceived benefits. The goal of this study was to conduct a survey of a nationally representative sample of NH physicians to quantify the use and perceived benefits of mobile devices in preventing ADEs in the NH setting. We surveyed physicians who attended the 2010 American Medical Directors Association Annual Symposium about their use of mobile devices, and beliefs about the effectiveness of drug reference software in preventing ADEs. The overall net valid response rate was 70% (558/800) with 42% (236/558) using mobile devices to assist with prescribing in the NH. Physicians with 15 or fewer years of clinical experience were 67% more likely to be mobile device users, compared with those with more than 15 years of clinical experience (odds ratio = 1.68; 95% confidence interval = 1.17-2.41; P = .005). For those who used a mobile device to assist with prescribing, almost all (98%) reported performing an average of 1 or more drug look-ups per day, performed an average of 1 to 2 lookups per day for potential drug-drug interactions (DDIs), and most (88%) believed that drug reference software had helped to prevent at least 1 potential ADE in the preceding 4-week period. The proportion of NH physicians who use mobile devices with drug reference software, although significant, is lower than in other clinical environments. Our results suggest that NH physicians who use mobile devices equipped with drug reference software believe they are helpful for reducing ADEs. Further research is needed to better characterize the facilitators and barriers to adoption of the technology in the NH and its precise impact on NH ADEs.


pVHL: a multipurpose adaptor protein.
Mutations in the von Hippel-Lindau tumor suppressor gene VHL occur in various inherited and sporadically occurring tumors. The protein encoded by VHL--pVHL--bears no known enzymatic activities but interacts with numerous protein partners. With the identification of distinct pVHL-containing multiprotein complexes, a refined portrait of pVHL tumor suppressor function has arisen. In general, pVHL acts as a multipurpose adaptor protein that controls a diverse array of gene expression programs, as well as extracellular matrix assembly and microtubule-based processes, by linking various target proteins to appropriate enzymatic activities. These findings provide an evermore complex but coherent view of how pVHL functions molecularly and of the consequences of dysregulation of these diverse molecular activities on tumor formation.


Thin magnetically soft wires for magnetic microsensors.
Recent advances in technology involving magnetic materials require development of novel advanced magnetic materials with improved magnetic and magneto-transport properties and with reduced dimensionality. Therefore magnetic materials with outstanding magnetic characteristics and reduced dimensionality have recently gained much attention. Among these magnetic materials a family of thin wires with reduced geometrical dimensions (of order of 1-30 μm in diameter) have gained importance within the last few years. These thin wires combine excellent soft magnetic properties (with coercivities up to 4 A/m) with attractive magneto-transport properties (Giant Magneto-impedance effect, GMI, Giant Magneto-resistance effect, GMR) and an unusual re-magnetization process in positive magnetostriction compositions exhibiting quite fast domain wall propagation. In this paper we overview the magnetic and magneto-transport properties of these microwires that make them suitable for microsensor applications.


Fish oil supplementation during chemotherapy increases posterior time to tumor progression in colorectal cancer.
The authors evaluated clinical outcomes during and after chemotherapy in colorectal cancer patients supplemented with fish oil during the first 9 wk of treatment. Thirty individuals never submitted to chemotherapy were randomized into supplemented group (SG), which received 2 g/day of fish oil (0.6 g/day of EPA and DHA) for 9 wk or control group (CG), which received neither fish oil nor placebo. Outcomes assessed were number of chemotherapy cycles administered; days undergoing chemotherapy; number of delays and interruptions in the administration of chemotherapy; number of hospitalizations during chemotherapy; tumor progression; values of carcinoembryonic antigen (CEA); days until events (death and progression); and 3 yr survival. Time to tumor progression was significantly longer in SG [S593 days (±211.5)] vs. CG [330 days (± 135.1); P = 0.04], other outcomes did not differ between groups. Subjects with advanced cancer who received fish oil presented longer time to tumor progression and lower CEA values after chemotherapy; however these differences were not statistically significant. Supplementation with 2 g/day of fish oil for the first 9 wk of chemotherapy may contribute to delay in tumor progression in colorectal patients, possibly by enhancing the antineoplastic action of the chemotherapeutic drug.


Arginine and the Hofmeister Series: the role of ion-ion interactions in protein aggregation suppression.
L-Arginine hydrochloride is a very important aggregation suppressor for which there has been much attention given regarding elucidating its mechanism of action. Little consideration, however, has been given toward other salt forms besides chloride, even though the counterion likely imparts a large influence per the Hofmeister Series. Here, we report an in depth analysis of the role the counterion plays in the aggregation suppression behavior of arginine. Consistent with the empirical Hofmeister series, we found that the aggregation suppression ability of other arginine salt forms on a model protein (α-chymotrypsinogen) follows the order: H(2)PO(4)(-) > SO(4)(2-) > citrate(2-) > acetate(-) ≈ F(-) ≈ Cl(-) > Br(-) > I(-) ≈ SCN(-). Mechanistically, preferential interaction and osmotic virial coefficient measurements, in addition to molecular dynamics simulations, indicate that attractive ion-ion interactions, particularly attractive interactions between arginine molecules, play a dominate role in the observed behavior. Furthermore, it appears that dihydrogen phosphate, sulfate, and citrate have strong attractive interactions with the guanidinium group of arginine, which seems to contribute to the superior aggregation suppression ability of those salt forms by bridging together multiple arginine molecules into clusters. These results not only further our understanding of how arginine influences protein stability, they also help to elucidate the mechanism behind the Hofmeister Series. This should help to improve biopharmaceutical stabilization through the use of other arginine salts and possibly, the development of novel excipients.


Technical note: Genomic evaluation for crossbred performance in a single-step approach with metafounders.
A single-step genomic BLUP method (ssGBLUP) has been successfully developed and applied for purebred and crossbred performance in pigs. However, it requires phasing the genotypes and inferring the breed origin of alleles in crossbred animals, which is somewhat inconvenient. Recently, a new concept of metafounders that considers the relationship within and across base populations was developed. With this concept of metafounders, regular methods to build and invert the pedigree relationships matrix can be used with only minor modifications and, moreover, genomic relationships and pedigree-based relationships are automatically compatible in the ssGBLUP. In this study, data for the total number of piglets born in Danish Landrace, Yorkshire, and 2-way crossbred pigs and models for purebred and crossbred performance were revisited by use of ssGBLUP with 2 metafounders. Genetic variances and genetic correlations between purebred and crossbred performances were first reestimated. Then, model-based reliabilities of purebred boars for their crossbred performance and predictive abilities for crossbred animals were compared in different scenarios. Results in this study were compared to those in a previous study with identical data but with models that required known breed origin of crossbred genotypes. Results show that relationships for base individuals within Landrace and within Yorkshire are similar and that the ancestor populations for Landrace and Yorkshire are related. In terms of model-based reliabilities and predictive abilities, ssGBLUP with metafounders performs at least as well as the single-step method requiring phasing at a lower complexity.


Differential effects of cortisol and 11-deoxycorticosterone on ion transport protein mRNA levels in gills of two euryhaline teleosts, Mozambique tilapia (Oreochromis mossambicus) and striped bass (Morone saxatilis).
The role of cortisol as the only corticosteroid in fish osmoregulation has recently been challenged with the discovery of a mineralocorticoid-like hormone, 11-deoxycorticosterone (DOC), and necessitates new studies of the endocrinology of osmoregulation in fish. Using an in vitro gill explant incubation approach, DOC-mediated regulation of selected osmoregulatory target genes in the gill was investigated and compared with that of cortisol in two euryhaline teleosts, Mozambique tilapia (Oreochromis mossambicus) and striped bass (Morone saxatilis). The effects were tested in gills from both fresh water (FW)- and seawater (SW)-acclimated fish. Both cortisol and DOC caused an up-regulation of the Na(+),K(+)-ATPase α1 subunit in SW-acclimated tilapia but had no effect in FW-acclimated fish. Cortisol conferred an increase in Na(+),K(+),2Cl(-) cotransporter (NKCC) isoform 1a transcript levels in FW- and SW-acclimated tilapia, whereas DOC had a stimulatory effect only in SW-acclimated fish. Cortisol had no effect on NKCC isoform 1b mRNA levels at both salinities, while DOC stimulated this isoform in SW-acclimated fish. In striped bass, cortisol conferred an up-regulation of Na(+),K(+)-ATPase α1 and NKCC transcript levels in FW- and SW-acclimated fish, whereas DOC resulted in down-regulation of these transcripts in FW-acclimated fish. It was also found that both corticosteroids may rapidly (30 min) alter the mitogen-activated protein kinase signalling pathway in gill, inducing phosphorylation of extracellular signal-regulated kinase 1 (ERK1) and ERK2 in a salinity-dependent manner. The study shows a disparate organisation of corticosteroid signalling mechanisms involved in ion regulation in the two species and adds new evidence to a role of DOC as a mineralocorticoid hormone in teleosts.


Self-harm and life problems: findings from the Multicentre Study of Self-harm in England.
Self-harm is a major clinical problem and is strongly linked to suicide. It is important to understand the problems faced by those who self-harm to design effective clinical services and suicide prevention strategies. We investigated the life problems experienced by patients presenting to general hospitals for self-harm. Data for 2000-2010 from the Multicentre Study of Self-harm in England were used to investigate life problems associated with self-harm and their relationship to patient and clinical characteristics, including age, gender, repeat self-harm and employment status. Of 24,598 patients (36,431 assessed episodes), 57% were female and with a mean age of 33.1 years (SD 14.0 years), 92.6% were identified as having at least one contributing life problem. The most frequently reported problems at first episode of self-harm within the study period were relationship difficulties (especially with partners). Mental health issues and problems with alcohol were also very common (especially in those aged 35-54 years, and those who repeated self-harm). Those who repeated self-harm were more likely to report problems with housing, mental health and dealing with the consequences of abuse. Self-harm usually occurs in the context of multiple life problems. Clinical services for self-harm patients should have access to appropriate care for provision of help for relationship difficulties and problems concerning alcohol and mental health issues. Individualised clinical support (e.g. psychological therapy, interventions for alcohol problems and relationship counselling) for self-harm patients facing these life problems may play a crucial role in suicide prevention.


Skilled care at birth among rural women in Nepal: practice and challenges.
In Nepal, most births take place at home, and many, particularly in rural areas, are not attended by a skilled birth attendant. The main objectives of the study were to assess the use of skilled delivery care and barriers to access such care in a rural community and to assess health problems during delivery and seeking care. This cross-sectional study was carried out in two Village Development Committees in Nepal in 2006. In total, 150 women who had a live birth in the 24 months preceding the survey were interviewed using a structured questionnaire. The sample population included married women aged 15-49 years. Forty-six (31%) women delivered their babies at hospital, and 104 (69%) delivered at home. The cost of delivery at hospital was significantly (p < 0.001) higher than that of a delivery at home. Results of univariate analysis showed that women from Brahmin-Chhetri ethnicity, women with higher education or who were more skilled, whose husbands had higher education and more skilled jobs, had first or second childbirth, and having adverse previous obstetric history were associated with institutional delivery while women with higher education and having an adverse history of pregnancy outcome predicted the uptake of skilled delivery care in Nepal. The main perceived problems to access skilled delivery care were: distance to hospital, lack of transportation, lack of awareness on delivery care, and cost. The main reasons for seeking intrapartum care were long labour, retained placenta, and excessive bleeding. Only a quarter of women sought care immediately after problems occurred. The main reasons seeking care late were: the woman or her family not perceiving that there was a serious problem, distance to health facility, and lack of transport. The use of skilled birth attendants at delivery among rural women in Nepal is very poor. Home delivery by unskilled birth attendants is still a common practice among them. Many associated factors relating to the use of skilled delivery care that were identified included age, education and occupation of women, and education and occupation of husbands. Therefore, the availability of skilled delivery care services at the community, initiation of a primary health centre with skilled staff for delivery, and increasing awareness among women to seek skilled delivery care are the best solution.


The role of health at birth and parental investment in early child development: evidence from the French ELFE cohort.
This paper combines a theoretical and an empirical approach to address how health at birth affects child development. Using a simple theoretical model in which parents invest in their children, we identify the mechanisms through which better health at birth can improve child development. We also emphasise how parental socioeconomic status can shape the effects of health at birth. We perform an empirical analysis on a French cohort of children born in 2011, using a unique dataset ELFE. We identify the effect of birth weight and gestational age on child development at 1 year. The results indicate that only gestational age positively affects early development. We find no empirical evidence for the existence of a severity effect, according to which the adverse effects of poor health at birth are higher for children in low-income families or with poorly educated mothers.


Augmented glucose production is not contingent on high catecholamines in fetal sheep with IUGR.
Fetuses with intrauterine growth restriction (IUGR) have high concentrations of catecholamines, which lowers the insulin secretion and glucose uptake. Here, we studied the effect of hypercatecholaminemia on glucose metabolism in sheep fetuses with placental insufficiency-induced IUGR. Norepinephrine concentrations are elevated throughout late gestation in IUGR fetuses but not in IUGR fetuses with a bilateral adrenal demedullation (IAD) at 0.65 of gestation. Euglycemic (EC) and hyperinsulinemic-euglycemic (HEC) clamps were performed in control, intact-IUGR, and IAD fetuses at 0.87 of gestation. Compared to controls, basal oxygen, glucose, and insulin concentrations were lower in IUGR groups. Norepinephrine concentrations were five-fold higher in IUGR fetuses than in IAD fetuses. During the EC, rates of glucose entry (GER, umbilical + exogenous), glucose utilization (GUR), and glucose oxidation (GOR) were greater in IUGR groups than in controls. In IUGR and IAD fetuses with euglycemia and euinsulinemia, glucose production rates (GPR) remained elevated. During the HEC, GER and GOR were not different among groups. In IUGR and IAD fetuses, GURs were 40% greater than in controls, which paralleled the sustained GPR despite hyperinsulinemia. Glucose-stimulated insulin concentrations were augmented in IAD fetuses compared to IUGR fetuses. Fetal weights were not different between IUGR groups but were less than controls. Regardless of norepinephrine concentrations, IUGR fetuses not only develop greater peripheral insulin sensitivity for glucose utilization but also develop hepatic insulin resistance because GPR was maintained and unaffected by euglycemia or hyperinsulinemia. These findings show that adaptation in glucose metabolism of IUGR fetuses are independent of catecholamines, which implicate that hypoxemia and hypoglycemia cause the metabolic responses.


Frequency-specific coupling between trial-to-trial fluctuations of neural responses and response-time variability.
We assessed intra-individual variability of response times (RT) and single-trial P3 amplitudes following targets in healthy adults during a Flanker/NO-GO task. RT variability and variability of the neural responses coupled at the faster frequencies examined (0.07-0.17 Hz) at Pz, the target-P3 maxima, despite non-significant associations for overall variability (standard deviation, SD). Frequency-specific patterns of variability in the single-trial P3 may help to understand the neurophysiology of RT variability and its explanatory models of attention allocation deficits beyond intra-individual variability summary indices such as SD.


Ionic organic cage-encapsulating phase-transferable metal clusters.
Exploration of metal clusters (MCs) adaptive to both aqueous and oil phases without disturbing their size is promising for a broad scope of applications. The state-of-the-art approach via ligand-binding may perturb MCs' size due to varied metal-ligand binding strength when shuttling between solvents of different polarity. Herein, we applied physical confinement of a series of small noble MCs (<1 nm) inside ionic organic cages (I-Cages), which by means of anion exchange enables reversible transfer of MCs between aqueous and hydrophobic solutions without varying their ultrasmall size. Moreover, the MCs@I-Cage hybrid serves as a recyclable, reaction-switchable catalyst featuring high activity in liquid-phase NH3BH3 (AB) hydrolysis reaction with a turnover frequency (TOF) of 115 min-1.


Pericardial Tamponade Caused by Perforation of Marshall Vein During Left Jugular Central Venous Catheterization.
BACKGROUND The persistence of a vein of Marshall (VoM) from the left subclavian vein to the coronary sinus is a rare cardiac anomaly known as a persistent left superior vena cava (PLVC). This anatomical variant is usually asymptomatic but can lead to serious complications during catheterization via the left subclavian or internal jugular vein, as described here. In our case, the patent vein of Marshall directly connected the coronary sinus with the left subclavian vein discovered in a cardiac arrest patient because of pericardial effusion during the insertion of a central venous catheter (CVC). CASE REPORT A 62-year-old patient required a central line insertion through a left internal jugular vein. The patient immediately went into cardiac arrest after CVC insertion with a pericardial effusion. The patient was successfully resuscitated with the drainage of a pericardial effusion. A chest X-ray revealed that the central venous catheter (CVC) was located along the left border of the mediastinum rather than the right border. It was evident that the central venous catheter was inadvertently placed into the pericardial space, resulting in tamponade. This complication occurred through inadvertent access of a small persistent vein of Marshall. CONCLUSIONS This case illustrates the importance of knowledge of anatomical variants of the persistent vein of Marshall to prevent or correctly interpret and manage procedural complications.


Sugary drink consumption and risk of cancer: results from NutriNet-Santé prospective cohort.
To assess the associations between the consumption of sugary drinks (such as sugar sweetened beverages and 100% fruit juices), artificially sweetened beverages, and the risk of cancer. Population based prospective cohort study. Overall, 101 257 participants aged 18 and over (mean age 42.2, SD 14.4; median follow-up time 5.1 years) from the French NutriNet-Santé cohort (2009-2017) were included. Consumptions of sugary drinks and artificially sweetened beverages were assessed by using repeated 24 hour dietary records, which were designed to register participants' usual consumption for 3300 different food and beverage items. Prospective associations between beverage consumption and the risk of overall, breast, prostate, and colorectal cancer were assessed by multi-adjusted Fine and Gray hazard models, accounting for competing risks. Subdistribution hazard ratios were computed. The consumption of sugary drinks was significantly associated with the risk of overall cancer (n=2193 cases, subdistribution hazard ratio for a 100mL/d increase 1.18, 95% confidence interval 1.10 to 1.27, P<0.0001) and breast cancer (693, 1.22, 1.07 to 1.39, P=0.004). The consumption of artificially sweetened beverages was not associated with the risk of cancer. In specific subanalyses, the consumption of 100% fruit juice was significantly associated with the risk of overall cancer (2193, 1.12, 1.03 to 1.23, P=0.007). In this large prospective study, the consumption of sugary drinks was positively associated with the risk of overall cancer and breast cancer. 100% fruit juices were also positively associated with the risk of overall cancer. These results need replication in other large scale prospective studies. They suggest that sugary drinks, which are widely consumed in Western countries, might represent a modifiable risk factor for cancer prevention. ClinicalTrials.gov NCT03335644.


Developing a falls prevention program for community-dwelling stroke survivors in Singapore: client and caregiver perspectives.
Drawing on the perspectives of stroke survivors, family members and domestic helpers, this study explores participants' experiences of self-perceived fall risk factors after stroke, common fall prevention strategies used, and challenges to community participation after a fall. Semi-structured interviews were conducted in Singapore with community-dwelling stroke survivors with a previous fall (n = 9), family caregivers (n = 4), and domestic helpers (n = 4) who have cared for a stroke survivor with a previous fall. Purposive sampling was used for recruitment; all interviews were audio-recorded with permission and transcribed. Thematic analysis was conducted using NVivo (v10) software. All participants shared their self-perceived intrinsic and extrinsic fall risk factors and main challenges after a fall. For stroke participants and family caregivers, motivational factors in developing safety strategies after a previous fall(s) include social connectedness, independent living and community participation. For family caregivers and domestic helpers, the stroke survivor's safety is their top priority, however this can also lead to over-protective behavior outside of the rehabilitation process. Reducing the risk of falls in community-dwelling stroke survivors seems to be more important than promoting community participation among caregivers. The study findings highlight that a structured and client-centered fall prevention program targeting stroke survivors and caregivers is needed in Singapore. Implications for rehabilitation Falls after stroke can lead to functional decline in gait and mobility and restricted self-care activities. Community-dwelling stroke survivors develop adaptive safety strategies after a fall and want to be socially connected. However, caregivers see the safety of the stroke survivors as their top priority and demonstrate over-protective behaviors. Fall prevention programs for community-dwelling stroke survivors should target both stroke survivors and their caregivers. A structured and client-centered fall prevention program targeting at multiple risk factors post-stroke is needed for community-living stroke survivors.


ARTD1-induced poly-ADP-ribose formation enhances PPARγ ligand binding and co-factor exchange.
PPARγ-dependent gene expression during adipogenesis is facilitated by ADP-ribosyltransferase D-type 1 (ARTD1; PARP1)-catalyzed poly-ADP-ribose (PAR) formation. Adipogenesis is accompanied by a dynamic modulation of the chromatin landscape at PPARγ target genes by ligand-dependent co-factor exchange. However, how endogenous PPARγ ligands, which have a low affinity for the receptor and are present at low levels in the cell, can induce sufficient co-factor exchange is unknown. Moreover, the significance of PAR formation in PPARγ-regulated adipose tissue function is also unknown. Here, we show that inhibition of PAR formation in mice on a high-fat diet reduces weight gain and cell size of adipocytes, as well as PPARγ target gene expression in white adipose tissue. Mechanistically, topoisomerase II activity induces ARTD1 recruitment to PPARγ target genes, and ARTD1 automodification enhances ligand binding to PPARγ, thus promoting sufficient transcriptional co-factor exchange in adipocytes. Thus, ARTD1-mediated PAR formation during adipogenesis is necessary to adequately convey the low signal of endogenous PPARγ ligand to effective gene expression. These results uncover a new regulatory mechanism of ARTD1-induced ADP-ribosylation and highlight its importance for nuclear factor-regulated gene expression.


Genome-wide surveillance of mismatched alleles for graft-versus-host disease in stem cell transplantation.
Acute graft-versus-host disease (aGVHD) represents one of the major complications in allogeneic stem cell transplantation and is primarily caused by genetic disparity between the donor and recipient. In HLA-matched transplants, the disparity is thought to be determined by loci encoding minor histocompatibility antigens (minor H antigens), which are presented by specific HLA molecules. We performed a genome-wide association study (GWAS) to identify minor H antigen loci associated with aGVHD. A total of 500 568 single nucleotide polymorphisms (SNPs) were genotyped for donors and recipients from 1589 unrelated bone marrow transplants matched for HLA-A, -B, -C, -DRB1, and -DQB1, followed by the imputation of unobserved SNPs. We interrogated SNPs whose disparity between the donor and recipient was significantly associated with aGVHD development. Without assuming HLA unrestriction, we successfully captured a known association between HLA-DPB1 disparity (P = 4.50 × 10(-9)) and grade II-IV aGVHD development, providing proof of concept for the GWAS design aimed at discovering genetic disparity associated with aGVHD. In HLA-restricted analyses, whereby association tests were confined to major subgroups sharing common HLA alleles to identify putative minor H antigen loci, we identified 3 novel loci significantly associated with grade III-IV aGVHD. Among these, rs17473423 (P = 1.20 × 10(-11)) at 12p12.1 within the KRAS locus showed the most significant association in the subgroup, sharing HLA-DQB1*06:01. Our result suggested that a GWAS can be successfully applied to identify allele mismatch associated with aGVHD development, contributing to the understanding of the genetic basis of aGVHD.


The effect of divided attention on novices and experts in laparoscopic task performance.
Attention is important for the skilful execution of surgery. The surgeon's attention during surgery is divided between surgery and outside distractions. The effect of this divided attention has not been well studied previously. We aimed to compare the effect of dividing attention of novices and experts on a laparoscopic task performance. Following ethical approval, 25 novices and 9 expert surgeons performed a standardised peg transfer task in a laboratory setup under three randomly assigned conditions: silent as control condition and two standardised auditory distracting tasks requiring response (easy and difficult) as study conditions. Human reliability assessment was used for surgical task analysis. Primary outcome measures were correct auditory responses, task time, number of surgical errors and instrument movements. Secondary outcome measures included error rate, error probability and hand specific differences. Non-parametric statistics were used for data analysis. 21109 movements and 9036 total errors were analysed. Novices had increased mean task completion time (seconds) (171 ± 44SD vs. 149 ± 34, p < 0.05), number of total movements (227 ± 27 vs. 213 ± 26, p < 0.05) and number of errors (127 ± 51 vs. 96 ± 28, p < 0.05) during difficult study conditions compared to control. The correct responses to auditory stimuli were less frequent in experts (68 %) compared to novices (80 %). There was a positive correlation between error rate and error probability in novices (r (2) = 0.533, p < 0.05) but not in experts (r (2) = 0.346, p > 0.05). Divided attention conditions in theatre environment require careful consideration during surgical training as the junior surgeons are less able to focus their attention during these conditions.


Mini-implant supported canine retraction with micro-osteoperforation: A split-mouth randomized clinical trial.
To investigate, using a split-mouth randomized clinical design, the effect of micro-osteoperforation (MOP) on mini-implant supported canine retraction using fixed appliances. Thirty subjects (seven males and 23 females) with a mean age of 22.2 (3.72) years were randomized into three canine retraction groups: Group 1 (MOP 4-weekly maxilla/8-weekly mandible; n = 10); Group 2 (MOP 8-weekly maxilla/12-weekly mandible; n = 10) and Group 3 (MOP 12-weekly maxilla/4-weekly mandible; n = 10) measured at 4-week intervals over 16 weeks. Subjects also completed pain (5-point Likert scale) and pain impact (Visual Analogue Scale) questionnaires. The primary outcome was the amount of canine retraction over 16 weeks at MOP (experimental) and non-MOP (control) sites. Mean overall canine retraction was 4.16 (1.62) mm with MOP and 3.06 (1.64) mm without. After adjusting for differences between jaws, all MOP groups exhibited significantly higher canine distalization than the control group: 0.89 mm more (95% confidence interval [CI] = 0.19 to 1.59 mm; P = .01) in the MOP-4 group, 1.08 mm more (95% CI = 0.49 to 1.68 mm; P = .001) in the MOP-8 group and 1.33 mm more (95% CI = 0.55 to 2.10 mm; P = .002) in the MOP-12 group. All subjects reported pain associated with MOP with 60% classifying it as moderate and 15% severe. The main impact of this reported pain was related to chewing and speech. MOP can increase overall mini-implant supported canine retraction over a 16-week period of observation but this difference is unlikely to be clinically significant.


Oxidative stress regulates expression of VEGFR1 in myeloid cells: link to tumor-induced immune suppression in renal cell carcinoma.
Metastatic renal cell carcinoma (RCC) associates with overproduction of vascular endothelial growth factor (VEGF) due to the mutation/inactivation of the von Hippel-Lindau (VHL) tumor suppressor gene. Herein we demonstrate that implantation of human RCC tumor cells into athymic nude mice promotes the appearance of VEGF receptor 1 (VEGFR1)/CD11b double-positive myeloid cells in peripheral blood. Avastin-mediated VEGF neutralization was capable of significantly reducing the numbers of circulating VEGFR1+ myeloid cells. Conversely, up-regulation of VEGFR1 by myeloid cells could also be achieved in vitro by coculturing bone marrow cells with RCC-conditioned medium or by short-term exposure of naive myeloid cells to oxidative stress. Treatment of myeloid cells with H2O2, lipid peroxidation product 4-hydroxy-2(E)-nonenal, or an inhibitor of thioredoxin reductase all resulted in increased expression of VEGFR1. Furthermore, after exposure to oxidative stress, myeloid cells acquire immunosuppressive features and become capable of inhibiting T cell proliferation. Data suggest that tumor-induced oxidative stress may promote both VEGFR1 up-regulation and immunosuppressive function in bone marrow-derived myeloid cells. Analysis of tumor tissue and peripheral blood from patients with metastatic RCC revealed that VEGFR1+ cells can be also found in cancer patients. Restoration of immunocompetence in metastatic RCC patients by pharmacological elimination of VEGFR1+ cells may have a significant impact on the therapeutic efficacy of cancer vaccines or other immune-based therapies.


Advancing neodymium single-band nanothermometry.
Near-infrared (NIR) emitting contrast agents with integrated optical temperature sensing are highly desirable for a variety of biomedical applications, particularly when subcutaneous target visualization and measurement of its thermodynamic properties are required. To that end, the possibility of using Nd3+ doped LiLuF4 rare-earth nanoparticles (RENPs) as NIR photoluminescent nanothermometers is explored. These RENPs are relatively small, have narrow size distribution, and can easily be core/shell engineered - all combined, these features meet the requirements of biologically relevant and multifunctional nanoprobes. The LiLuF4 host allows to observe the fine Stark structure of the 4F3/2→4I9/2, 4I11/2, and 4I13/2 optical transitions, each of which can then be used for single-band NIR nanothermometry. The thermometric parameter defined for the most intense Nd3+ emission around 1050 nm, shows high temperature sensitivity (∼0.49% °C-1), and low temperature uncertainty (0.3 °C) as compared to the thermometric parameters defined for the 880 and 1320 nm Nd3+ emissions. Additionally, transient temperature measurements through tissue show that these RENPs can be used to assess fast temperature changes at a tissue depth of 3 mm, while slower temperature changes can be measured at even greater depths. Nd3+ doped LiLuF4 RENPs represent a significant improvement for Nd3+ based single-band photoluminescence nanothermometry, with the possibility of its integration within more sophisticated multifunctional theranostic nanostructures.


Evaluation of occupational therapy interventions for elderly patients in Swedish acute care: a pilot study.
The aim was to evaluate whether occupational therapy interventions in acute care could improve the elderly patient's perception of ability to manage at home after discharge. A pilot study was performed, including 22 patients in the experimental group and 19 in the control group. Occupational therapy interventions were conducted in the experimental group concerning personal care, information, prescription of assistive devices, planning of discharge, and reporting to primary care or community care. The control group was given no occupational therapy interventions. Structured interviews were performed on discharge and at a follow-up in about 14 weeks after discharge. The two groups were comparable concerning gender, age, days of care, and diagnoses. Patients in the experimental group scored lower on mental health and were more anxious on discharge. However, there was no difference between the groups in managing at home after discharge. Patients in the control group had greater need of further contacts with healthcare after discharge. Due to the small sample interpretations must be made with caution. The findings indicate that occupational therapy interventions in acute care might have a positive effect from the perspective of the elderly patient. These results need to be confirmed in a larger study.


Structural determination of biomolecular interfaces by nuclear magnetic resonance of proteins with reduced proton density.
Protein interactions are important for understanding many molecular mechanisms underlying cellular processes. So far, interfaces between interacting proteins have been characterized by NMR spectroscopy mostly by using chemical shift perturbations and cross-saturation via intermolecular cross-relaxation. Although powerful, these techniques cannot provide unambiguous estimates of intermolecular distances between interacting proteins. Here, we present an alternative approach, called REDSPRINT (REDduced/Standard PRoton density INTerface identification), to map protein interfaces with greater accuracy by using multiple NMR probes. Our approach is based on monitoring the cross-relaxation from a source protein (or from an arbitrary ligand that need not be a protein) with high proton density to a target protein (or other biomolecule) with low proton density by using isotope-filtered nuclear Overhauser spectroscopy (NOESY). This methodology uses different isotropic labeling for the source and target proteins to identify the source-target interface and also determine the proton density of the source protein at the interface for protein-protein or protein-ligand docking. Simulation indicates significant gains in sensitivity because of the resultant relaxation properties, and the utility of this technique, including a method for direct determination of the protein interface, is demonstrated for two different protein-protein complexes.


Three-dimensional (3D)-printed devices composed of hydrophilic cap and hydrophobic body for improving buoyancy and gastric retention of domperidone tablets.
The aim of this study was to produce capsule-shaped floating devices (CFD) using a fused deposition modeling (FDM) three-dimensional (3D) printer, for controlling the release and gastric retention of domperidone (DOM) tablets (Motilium-MⓇ). In order to enhance the buoyancy of the devices, a hollow cap with different wall thicknesses (1.2-1.5 mm) was printed with a hydrophilic (polyvinyl alcohol, PVA) filament. The body of the device was made from a hydrophobic (polylactic acid, PLA) filament. Bodies with aperture sizes (1-2 mm) were produced to investigate how this would affect drug release. Morphology, weight variation, ex vivo and in vivo floating time and drug release characteristics were examined. The results revealed that increasing the cap thickness of the devices (1.2 to 1.3 mm) increased the total floating time (TFT). The maximum TFT (10 h) with floating lag time (FLT) < 5 s was observed from Motilium-MⓇ incorporated CFD3-5 (cap with 1.3-mm wall thickness). Decreasing the size of the holes on the devices led to the sustained release of DOM. The CFD5 (cap with 1.3-mm cap thickness and 1.5-mm hole width) delivered approximately 98% release in 10 h, and the release kinetics fit well with the zero-order kinetics (R2 > 0.95). In vivo floating studies in rabbits showed that the floating time of CFD5 was more than 10 h. These results demonstrated that the CFD was successfully designed to provide gastro-retentive drug delivery with the capacity to float and provide sustained drug release.


Biomechanics of ankle instability. Part 1: Reaction time to simulated ankle sprain.
To test the hypothesis that ankles with functional instability will demonstrate slower muscular reaction times than their contralateral stable ankle (SA) and stable healthy controls to a simulated nonpathological ankle sprain mechanism. Nineteen male volunteers with a history of unilateral ankle sprain and functional ankle instability (FAI) and 19 healthy male controls performed reaction time tests on a purpose-built platform that simulated a nonpathological combined inversion/plantarflexion ankle sprain mechanism. Participants provided informed written consent. Reaction time and muscle activity magnitude data were reported for the FAI group's unstable (UA) and stable ankles (SA) and the control group's dominant (DA) and nondominant ankles (NDA) to unilateral simulated ankle sprain (USAS). The reaction times of the peroneus longus (PL), peroneus brevis (PB), and tibialis anterior (TA) in the UA were significantly slower (P < 0.025) than the SA and control group's DA in the limb experiencing USAS. The reaction times of the support limb PL, TA, and extensor digitorum longus (EDL) muscles of the UA were slower than the DA (P < 0.025). The magnitude of EMG response was not different between the SA and UA (P > 0.025). Results demonstrate a deficit (slower reaction time) in ankles with FAI when acting in support and when exposed to a simulated sprain compared to stable healthy controls. As a result of slower reaction times, acting to support the UA may put the contralateral SA at an increased risk of ankle sprain. This suggests that rehabilitation of a lateral ankle sprain should include strengthening the evertors (peroneals and EDL) at the subtalar joint and the dorsiflexors (TA and EDL) at the talocrural joint.


Variation in suspected cancer referral pathways in primary care: comparative analysis across the International Benchmarking Cancer Partnership.
International variations in cancer outcomes persist and may be influenced by differences in the accessibility and organisation of cancer patient pathways. More evidence is needed to understand to what extent variations in the structure of primary care referral pathways for cancer investigation contribute to differences in the timeliness of diagnoses and cancer outcomes in different countries. To explore the variation in primary care referral pathways for the management of suspected cancer across different countries. Descriptive comparative analysis using mixed methods across the International Cancer Benchmarking Partnership (ICBP) countries. Schematics of primary care referral pathways were developed across 10 ICBP jurisdictions. The schematics were initially developed using the Aarhus statement (a resource providing greater insight and precision into early cancer diagnosis research) and were further supplemented with expert insights through consulting leading experts in primary care and cancer, existing ICBP data, a focused review of existing evidence on the management of suspected cancer, published primary care cancer guidelines, and evaluations of referral tools and initiatives in primary care. Referral pathway schematics for 10 ICBP jurisdictions were presented alongside a descriptive comparison of the organisation of primary care management of suspected cancer. Several key areas of variation across countries were identified: inflexibility of referral pathways, lack of a managed route for non-specific symptoms, primary care practitioner decision-making autonomy, direct access to investigations, and use of emergency routes. Analysing the differences in referral processes can prompt further research to better understand the impact of variation on the timeliness of diagnoses and cancer outcomes. Studying these schematics in local contexts may help to identify opportunities to improve care and facilitate discussions on what may constitute best referral practice.


A randomized clinical trial of endometrial perfusion with granulocyte colony-stimulating factor in in vitro fertilization cycles: impact on endometrial thickness and clinical pregnancy rates.
To investigate whether granulocyte colony-stimulating factor (G-CSG) affects endometrial thickness, implantation rates, and clinical pregnancy rates in routine, unselected IVF cycles. Registered, individually randomized, two-group, parallel double-blinded placebo-controlled clinical trial. Academically affiliated private clinical and research center. 141 consecutive, unselected, consenting women with no history of renal disease, sickle cell disease, or malignancy who were undergoing IVF. Sealed, numbered, opaque envelopes assigned 73 patients to receive G-CSF (Filgrastim, Amgen, 300 μg/1.0 mL) and 68 to receive placebo (saline). Endometrial thickness, clinical pregnancy, and embryo implantation rates. The mean age for the whole study group was 39.59 ± 5.56 years (G-CSF: 39.79 ± 5.13 years; placebo: 39.38 ± 6.03 years). Endometrial thickness statistically significantly increased over the 5-day observation period for the whole group by approximately 1.36 mm. The increase in the G-CSF group was not statistically significantly different from the control group. Statistical models looking at treatment effects on clinical pregnancy and implantation rates demonstrated no effect of G-CSF treatment. There were no adverse events for either treatment group. In normal IVF patients, G-CSF does not affect endometrial thickness, implantation rates, or clinical pregnancy rates. Because these results were obtained in an older patient population, they may not necessarily apply to younger women. NCT01202656.


Studying Catabolism of Protein ADP-Ribosylation.
Protein ADP-ribosylation is a conserved posttranslational modification that regulates many major cellular functions, such as DNA repair, transcription, translation, signal transduction, stress response, cell division, aging, and cell death. Protein ADP-ribosyl transferases catalyze the transfer of an ADP-ribose (ADPr) group from the β-nicotinamide adenine dinucleotide (β-NAD+) cofactor onto a specific target protein with the subsequent release of nicotinamide. ADP-ribosylation leads to changes in protein structure, function, stability, and localization, thus defining the appropriate cellular response. Signaling processes that are mediated by modifications need to be finely tuned and eventually silenced and one of the ways to achieve this is through the action of enzymes that remove (reverse) protein ADP-ribosylation in a timely fashion such as PARG, TARG1, MACROD1, and MACROD2. Here, we describe several basic methods used to study the enzymatic activity of de-ADP-ribosylating enzymes.


Brain correlates of aesthetic judgment of beauty.
Functional MRI was used to investigate the neural correlates of aesthetic judgments of beauty of geometrical shapes. Participants performed evaluative aesthetic judgments (beautiful or not?) and descriptive symmetry judgments (symmetric or not?) on the same stimulus material. Symmetry was employed because aesthetic judgments are known to be often guided by criteria of symmetry. Novel, abstract graphic patterns were presented to minimize influences of attitudes or memory-related processes and to test effects of stimulus symmetry and complexity. Behavioral results confirmed the influence of stimulus symmetry and complexity on aesthetic judgments. Direct contrasts showed specific activations for aesthetic judgments in the frontomedian cortex (BA 9/10), bilateral prefrontal BA 45/47, and posterior cingulate, left temporal pole, and the temporoparietal junction. In contrast, symmetry judgments elicited specific activations in parietal and premotor areas subserving spatial processing. Interestingly, beautiful judgments enhanced BOLD signals not only in the frontomedian cortex, but also in the left intraparietal sulcus of the symmetry network. Moreover, stimulus complexity caused differential effects for each of the two judgment types. Findings indicate aesthetic judgments of beauty to rely on a network partially overlapping with that underlying evaluative judgments on social and moral cues and substantiate the significance of symmetry and complexity for our judgment of beauty.


Factors associated with human papillomavirus (HPV) test acceptability in primary screening for cervical cancer: A mixed methods research synthesis.
Primary screening for cervical cancer is transitioning from the longstanding Pap smear towards implementation of an HPV-DNA test, which is more sensitive than Pap cytology in detecting high-risk lesions and offers greater protection against invasive cervical carcinomas. Based on these results, many countries are recommending and implementing HPV testing-based screening programs. Understanding what factors (e.g., knowledge, attitudes) will impact on HPV test acceptability by women is crucial for ensuring adequate public health practices to optimize cervical screening uptake. We used mixed methods research synthesis to provide a categorization of the relevant factors related to HPV primary screening for cervical cancer and describe their influence on women's acceptability of HPV testing. We searched Medline, Embase, PsycINFO, CINAHL, Global Health and Web of Science for journal articles between January 1, 1980 and October 31, 2017 and retained 22 empirical articles. Our results show that while most factors associated with HPV test acceptability are included in the Health Belief Model and/or Theory of Planned Behavior (e.g., attitudes, knowledge), other important factors are not encompassed by these theoretical frameworks (e.g., health behaviors, negative emotional reactions related to HPV testing). The direction of influence of psychosocial factors on HPV test acceptability was synthesized based on 14 quantitative studies as: facilitators (e.g., high perceived HPV test benefits), barriers (e.g., negative attitudes towards increased screening intervals), contradictory evidence (e.g., sexual history) and no impact (e.g., high perceived severity of HPV infection). Further population-based studies are needed to confirm the impact of these factors on HPV-based screening acceptability.


Pharmacoepigenetics of the role of DNA methylation in μ-opioid receptor expression in different human brain regions.
Exposure to opioids has been associated with epigenetic effects. Studies in rodents suggested a role of varying degrees of DNA methylation in the differential regulation of μ-opioid receptor expression across the brain. In a translational investigation, using tissue acquired postmortem from 21 brain regions of former opiate addicts, representing a human cohort with chronic opioid exposure, μ-opioid receptor expression was analyzed at the level of DNA methylation, mRNA and protein. While high or low μ-opioid receptor expression significantly correlated with local OPRM1 mRNA levels, there was no corresponding association with OPRM1 methylation status. Additional experiments in human cell lines showed that changes in DNA methylation associated with changes in μ-opioid expression were an order of magnitude greater than differences in brain. Hence, different degrees of DNA methylation associated with chronic opioid exposure are unlikely to exert a major role in the region-specificity of μ-opioid receptor expression in the human brain.


Cerebral venous sinus thrombosis: a report of two cases.
Cerebral venous sinus thrombosis is an uncommon yet potentially life threatening condition which is often under-diagnosed or misdiagnosed because of the variability of the clinical signs and symptoms. The imaging findings are often subtle thus the need for a high index of suspicion. The superior sagittal and transverse sinuses are the most frequently involved. The correct diagnosis of cerebral venous thrombosis relies on neuroimaging studies. The two cases reported highlight the fact that the radiologist may be the first clinician to suspect and diagnose this condition. This ultimately affects prognosis as early diagnosis and treatment reduces morbidity and mortality.


Distribution of substance P and calcitonin gene-related peptide in the spinal cord of Cavalier King Charles Spaniels affected by symptomatic syringomyelia.
The causes of clinical signs associated with syringomyelia in the Cavalier King Charles Spaniel (CKCS) are incompletely understood. In this study we compared expression of two pain-related neuropeptides: substance P (SP) and calcitonin gene related peptide (CGRP), in the spinal cord dorsal horn of normal dogs with that in CKCS with and without clinical signs of syringomyelia. There was a decrease in expression of both peptides in CKCS with 'symptomatic' syringomyelia that was also associated with significant asymmetry in SP-I and similar, though non-significant, asymmetry in CGRP-I compared with other groups. The asymmetric distribution of these pain-related peptides may be a consequence of syrinx-associated damage to grey matter but may also play a role in generation of pain.


Morphological relationship between GnIH and GnRH neurons in the brain of the neotropical cichlid fish Cichlasoma dimerus.
Reproduction is regulated by the hypothalamic-pituitary-gonadal axis. The first neuropeptide identified that regulates this function was the decapeptide gonadotropin-releasing hormone (GnRH). Nowadays, in gnatostomates, a number of GnRH variants have been identified and classified into three different types: GnRH1, GnRH2, and GnRH3. Almost 30 years later, a new peptide that inhibits gonadotropin synthesis and secretion was discovered and thus named as gonadotropin-inhibitory hormone (GnIH). In avians and mammals, the interaction and regulation between GnRH and GnIH neurons has been widely studied; however, in other vertebrate groups there is little information about the relationship between these neurons. In previous works, three GnRH variants and a GnIH propeptide were characterized in Cichlasoma dimerus, and it was demonstrated that GnIH inhibited gonadotropins release in this species. Because no innervation was detected at the pituitary level, we speculate that GnIH would inhibit gonadotropins via GnRH. Thus, the aim of the present study was to evaluate the anatomical relationship between neurons expressing GnIH and the three GnRH variants by double labelling confocal immunofluorescence in adults of C. dimerus. Our results showed no apparent contacts between GnIH and GnRH1, fiber to fiber interactions between GnIH and GnRH2, and co-localization of GnIH and GnRH3 variant in neurons of the nucleus olfacto-retinalis. In conclusion, whether GnIH regulates the expression or secretion of GnRH1 in this species, an indirect modulation seems more plausible. Moreover, the present results suggest an interaction between GnIH and GnRH2 systems. Finally, new clues were provided to investigate the role of nucleus olfacto-retinalis cells and putative GnIH and GnRH3 interactions in the modulation of the reproductive network in teleost fish.


Placebo-controlled trials in schizophrenia: are they ethical? Are they necessary?
The current controversy as to the proper role of the placebo control in the evaluation of new treatments for schizophrenia requires an analysis that is sensitive to both ethical and scientific issues. Clinical equipoise, widely regarded as the moral foundation of the randomized controlled trial (RCT), requires the use of best available treatment as the control in RCT. Scientific criticisms of the use of an active control are examined and none present an insuperable barrier to the use of an active control. Indeed, scrutiny of the most recent argument for the use of placebo controls, 'assay sensitivity', suggests that the use of placebo may be the cause of the problem pointed to. Scientific, regulatory, ethical and legal advantages of the use of an active control are described. While the use of a placebo control may be acceptable in carefully defined circumstances, in most cases the use of an active control in schizophrenia research is ethically and scientifically preferable.


Conceptualizing and quantifying body condition using structural equation modelling: A user guide.
Body condition is an important concept in behaviour, evolution and conservation, commonly used as a proxy of an individual's performance, for example in the assessment of environmental impacts. Although body condition potentially encompasses a wide range of health state dimensions (nutritional, immune or hormonal status), in practice most studies operationalize body condition using a single (univariate) measure, such as fat storage. One reason for excluding additional axes of variation may be that multivariate descriptors of body condition impose statistical and analytical challenges. Structural equation modelling (SEM) is used in many fields to study questions relating multidimensional concepts, and we here explain how SEM is a useful analytical tool to describe the multivariate nature of body condition. In this 'Research Methods Guide' paper, we show how SEM can be used to resolve different challenges in analysing the multivariate nature of body condition, such as (a) variable reduction and conceptualization, (b) specifying the relationship of condition to performance metrics, (c) comparing competing causal hypothesis and (d) including many pathways in a single model to avoid stepwise modelling approaches. We illustrated the use of SEM on a real-world case study and provided R-code of worked examples as a learning tool. We compared the predictive power of SEM with conventional statistical approaches that integrate multiple variables into one condition variable: multiple regression and principal component analyses. We show that model performance on our dataset is higher when using SEM and led to more accurate and precise estimates compared to conventional approaches. We encourage researchers to consider SEM as a flexible framework to describe the multivariate nature of body condition and thus understand how it affects biological processes, thereby improving the value of body condition proxies for predicting organismal performance. Finally, we highlight that it can be useful for other multidimensional ecological concepts as well, such as immunocompetence, oxidative stress and environmental conditions.


A Computational Study of RNA Tetraloop Thermodynamics, Including Misfolded States.
An important characteristic of RNA folding is the adoption of alternative configurations of similar stability, often referred to as misfolded configurations. These configurations are considered to compete with correctly folded configurations, although their rigorous thermodynamic and structural characterization remains elusive. Tetraloop motifs found in large ribozymes are ideal systems for an atomistically detailed computational quantification of folding free energy landscapes and the structural characterization of their constituent free energy basins, including nonnative states. In this work, we studied a group of closely related 10-mer tetraloops using a combined parallel tempering and metadynamics technique that allows a reliable sampling of the free energy landscapes, requiring only knowledge that the stem folds into a canonical A-RNA configuration. We isolated and analyzed unfolded, folded, and misfolded populations that correspond to different free energy basins. We identified a distinct misfolded state that has a stability very close to that of the correctly folded state. This misfolded state contains a predominant population that shares the same structural features across all tetraloops studied here and lacks the noncanonical A-G base pair in its loop portion. Further analysis performed with biased trajectories showed that although this competitive misfolded state is not an essential intermediate, it is visited in most of the transitions from unfolded to correctly folded states. Moreover, the tetraloops can transition from this misfolded state to the correctly folded state without requiring extensive unfolding.


Genetic variation in human NPY expression affects stress response and emotion.
Understanding inter-individual differences in stress response requires the explanation of genetic influences at multiple phenotypic levels, including complex behaviours and the metabolic responses of brain regions to emotional stimuli. Neuropeptide Y (NPY) is anxiolytic and its release is induced by stress. NPY is abundantly expressed in regions of the limbic system that are implicated in arousal and in the assignment of emotional valences to stimuli and memories. Here we show that haplotype-driven NPY expression predicts brain responses to emotional and stress challenges and also inversely correlates with trait anxiety. NPY haplotypes predicted levels of NPY messenger RNA in post-mortem brain and lymphoblasts, and levels of plasma NPY. Lower haplotype-driven NPY expression predicted higher emotion-induced activation of the amygdala, as well as diminished resiliency as assessed by pain/stress-induced activations of endogenous opioid neurotransmission in various brain regions. A single nucleotide polymorphism (SNP rs16147) located in the promoter region alters NPY expression in vitro and seems to account for more than half of the variation in expression in vivo. These convergent findings are consistent with the function of NPY as an anxiolytic peptide and help to explain inter-individual variation in resiliency to stress, a risk factor for many diseases.


Body composition of female road and track endurance cyclists: Normative values and typical changes.
The aims of this study were to describe normative values and seasonal variation of body composition in female cyclists comparing female road and track endurance cyclists, and to validate the use of anthropometry to monitor lean mass changes. Anthropometric profiles (seven site skinfolds) were measured over 16 years from 126 female cyclists. Lean mass index (LMI) was calculated as body weight × skinfolds(-x). The exponent (x) was calculated as the slope of the natural logarithm of body weight and skinfolds. Percentage changes in LMI were compared to lean mass changes measured using dual-energy X-ray absorptiometry (DXA) in a subset of 25 road cyclists. Compared to sub-elite and elite cyclists, world class cyclists were (mean [95% CI]) 1.18 kg [0.46, 1.90] and 0.60 kg [0.05, 1.15] lighter and had skinfolds that were 7.4 mm [3.8, 11.0] and 4.6 mm [1.8, 7.4] lower, respectively. Body weight (0.41 kg [0.04, 0.77]) and skinfolds (4.0 mm [2.1, 6.0]) were higher in the off-season compared to the early-season. World class female road cyclists had lower body weight (6.04 kg [2.73, 9.35]) and skinfolds (11.5 mm [1.1, 21.9]) than track endurance cyclists. LMI (mean exponent 0.15 [0.13, 0.18]) explained 87% of the variance in DXA lean mass. In conclusion, higher performing female cyclists were lighter and leaner than their less successful peers, road cyclists were lighter and leaner than track endurance cyclists, and weight and skinfolds were lowest early in the season. LMI appears to be a reasonably valid tool for monitoring lean mass changes.


Hypothalamic-pituitary dysfunction following traumatic brain injury affects functional improvement during acute inpatient rehabilitation.
To evaluate the occurrence of hypothalamic-pituitary dysfunction following a traumatic brain injury (TBI) and to determine its effect on functional improvement in acute inpatient rehabilitation. A retrospective chart review identified male patients with a primary diagnosis of TBI with or without a skull fracture, an onset date within 6 months prior to admission, and were 16 years of age or older. The percentage of individuals in this population with abnormal hormone levels was determined on the basis of the established normal reference range for each hormone assay. The functional independence measure, which assesses functional outcomes in acute inpatient rehabilitation, was used to examine the relationship between hormone levels and functional improvement. Hypothalamic-pituitary dysfunction was identified in nearly 70% of men following TBI. Hypogonadism, or low testosterone levels, was observed in 66% of the patients, followed by low levels of free T4 in 46% and low levels of insulin growth factor-1 in 26% of patients. Hypopituitarism associated with impaired functional recovery. Specifically, the functional independence measure change per day was significantly lower in patients with low levels of testosterone and insulin growth factor-1. These findings suggest the importance of testosterone and insulin growth factor-1 activity in the early stages of physical and cognitive rehabilitation.