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Introdução: O constante surgimento e a disseminação de fenótipos bacterianos de resistência a múltiplas drogas têm se tornado problemas de saúde pública em todo o mundo. Objetivo: Investigar a ocorrência de fenótipos críticos de resistência aos antimicrobianos em bactérias isoladas de culturas de vigilância. Método: Os dados microbiológicos foram coletados a partir de culturas de vigilância realizadas entre janeiro e dezembro de 2015 em um serviço terciário de saúde. Foram analisadas 1.590 culturas de vigilância utilizando o sistema VITEK 2 para identificação fenotípica e suscetibilidade aos antimicrobianos dos isolados. A detecção de β-lactamases de espectro estendido (ESBL) foi realizada por método de disco aproximação e a resistência aos carbapenêmicos e Staphylococcus aureus resistente à meticilina (MRSA) por métodos de disco difusão. A detecção de bactérias portadoras dos genes blaKPC, blaNDM e blaOXA-48 foi realizada através da reação em cadeia da polimerase in house. Resultados: Foram identificadas 201 (12,64%) fenótipos de ESBL, 173 (11,87%) de resistência aos carbapenêmicos e 3 (0,18%) de MRSA. Foram detectadas 34 cepas portadoras de genes codificadores de carbapenemases, onde 23 (67,64%) carreavam o gene blaKPC, oito (23,53%) o blaNDM e três (8,82%) ambos os genes. Conclusões: É um desafio controlar a disseminação de fenótipos críticos de resistência bacterianos. O predomínio de ESBL no referente hospital, assim como a ocorrência de genes codificadores de carbapenemases até então não relatados, ratificam a importância da vigilância epidemiológica e do incentivo a pesquisas subsequentes.

Bacterial resistance is an emerging public health issue that is disseminated worldwide. Silver nanocomposite can be an alternative strategy to avoid Gram-positive and Gram-negative bacteria growth, including multidrug-resistant strains. In the present study a silver nanocomposite was synthesized, using a new green chemistry process, by the addition of silver nitrate (1.10−3 mol·L−1) into a fermentative medium of Xanthomonas spp. to produce a xanthan gum polymer. Transmission electron microscopy (TEM) was used to evaluate the shape and size of the silver nanoparticles obtained. The silver ions in the nanocomposite were quantified by flame atomic absorption spectrometry (FAAS). The antibacterial activity of the nanomaterial against Escherichia coli (ATCC 22652), Enterococcus faecalis (ATCC 29282), Pseudomonas aeruginosa (ATCC 27853) and Staphylococcus aureus (ATCC 25923) was carried out using 500 mg of silver nanocomposite. Pseudomonas aeruginosa and Acinetobacter baumannii multidrug-resistant strains, isolated from hospitalized patients were also included in the study. The biosynthesized silver nanocomposite showed spherical nanoparticles with sizes smaller than 10 nm; 1 g of nanocomposite contained 49.24 µg of silver. Multidrug-resistant strains of Pseudomonas aeruginosa and Acinetobacter baumannii, and the other Gram-positive and Gram-negative bacteria tested, were sensitive to the silver nanocomposite (10–12.9 mm of inhibition zone). The biosynthesized silver nanocomposite seems to be a promising antibacterial agent for different applications, namely biomedical devices or topical wound coatings.

Background: As a group, autistic children with high support needs (with adaptive functioning in the range of an intellectual disability) are at risk of significant literacy difficulties. We investigated the parent-reported home literacy environment of this group of children., Method: Sixty-two parents of autistic children (4.5 to 18.25 years) attending an autism-specific school completed a home literacy survey reporting on their child's: (1) alphabet knowledge, (2) interest in reading, (3) activities/interactions around books, (4) reading ability, and (5) writing ability., Results: We found significant positive correlations between parent-reported child interest in reading and literacy-related interactions and skills, but not with child age. Children using spoken words to communicate obtained significantly greater scores on four home-literacy subscales, but not on reading interest., Conclusions: A better understanding of the home literacy activities of autistic children with high-support needs is needed to inform educational practices aimed at promoting literacy development in this vulnerable population., (© 2024 The Author(s). Journal of Applied Research in Intellectual Disabilities published by John Wiley & Sons Ltd.)

Background and Aim: Although it is well established that hormones like glucagon stimulates gluconeogenesis via the PKA-mediated phosphorylation of CREB and dephosphorylation of the cAMP-regulated CREB coactivators CRTC2, the role of neural signals in the regulation of gluconeogenesis remains uncertain., Methods and Results: Here, we characterize the noradrenergic bundle architecture in mouse liver; we show that the sympathoexcitation induced by acute cold exposure promotes hyperglycemia and upregulation of gluconeogenesis via triggering of the CREB/CRTC2 pathway. Following its induction by dephosphorylation, CRTC2 translocates to the nucleus and drives the transcription of key gluconeogenic genes. Rodents submitted to different models of sympathectomy or knockout of CRTC2 do not activate gluconeogenesis in response to cold. Norepinephrine directly acts in hepatocytes mainly through a Ca 2+ -dependent pathway that stimulates CREB/CRTC2, leading to activation of the gluconeogenic program., Conclusion: Our data demonstrate the importance of the CREB/CRTC2 pathway in mediating effects of hepatic sympathetic inputs on glucose homeostasis, providing new insights into the role of norepinephrine in health and disease., Competing Interests: Declaration of competing interest The authors declare no competing interests., (Crown Copyright © 2024. Published by Elsevier Inc. All rights reserved.)

Introduction: The modified Brain Injury Guidelines (mBIG) support a subset of low-risk patients to be managed without repeat head computed tomography (RHCT), neurosurgical consult (NSC), or hospital transfer/admission. This pilot aimed to assess mBIG implementation at a single facility to inform future systemwide implementation., Methods: Single cohort pilot trial at a level I trauma center, December 2021-August 2022. Adult patients included if tICH meeting BIG 1 or 2 criteria. BIG 3 patients excluded., Results: No patients required neurosurgical intervention. 72 RHCT and 83 NSC were prevented. 21 isolated BIG 1 were safely discharged home from the ED. No hospital readmissions for tICH. Protocol adherence rate was 92%., Conclusion: Implementation of the mBIG at a single trauma center is feasible and optimizes resource utilization. This pilot study will inform an implementation trial of the mBIG across a 24-hospital integrated health system., Competing Interests: Declaration of competing interest None of the authors have anything to disclose., (Copyright © 2023 Elsevier Inc. All rights reserved.)

At the present time, no viable treatment exists for cognitive and olfactory deficits in Down syndrome (DS). We show in a DS model (Ts65Dn mice) that these progressive nonreproductive neurological symptoms closely parallel a postpubertal decrease in hypothalamic as well as extrahypothalamic expression of a master molecule that controls reproduction-gonadotropin-releasing hormone (GnRH)-and appear related to an imbalance in a microRNA-gene network known to regulate GnRH neuron maturation together with altered hippocampal synaptic transmission. Epigenetic, cellular, chemogenetic, and pharmacological interventions that restore physiological GnRH levels abolish olfactory and cognitive defects in Ts65Dn mice, whereas pulsatile GnRH therapy improves cognition and brain connectivity in adult DS patients. GnRH thus plays a crucial role in olfaction and cognition, and pulsatile GnRH therapy holds promise to improve cognitive deficits in DS.

There is evidence that early variations in the development of an approximate number system (ANS) and symbolic number understanding are both influences on the later development of formal arithmetic skills. We report a large-scale (N = 552) longitudinal study of the predictors of arithmetic spanning a critical developmental period (the first 3 years of formal education). Variations in early knowledge of symbolic representations of number and the ordinal associations between them are direct predictors of later arithmetic skills. The development of number ordering ability is in turn predicted by earlier variations in arithmetic, the ANS (numerosity judgments), and rapid automatized naming (RAN). These findings have important implications for theories of numerical and arithmetical development and potentially for the teaching of these skills., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The production effect-whereby reading words aloud improves memory for those words relative to reading them silently-was investigated in two experiments with 7- to 10-year-old children residing in Brisbane, Australia. Experiment 1 (n = 41) involved familiar printed words, with words read aloud or silently appearing either in mixed- or blocked-list formats in a within-subject design. Recognition for words read aloud was better than for those read silently, an effect consistent across both list formats. These results were confirmed in Experiment 2 (n = 40) using longer lists of printed novel nonwords. Final analyses indicated that the production effect was comparable for words and nonwords. Findings are discussed in relation to the distinctiveness account and the use of production as a mnemonic in children., (© 2019 Society for Research in Child Development.)

Congenital hypogonadotropic hypogonadism (CHH) is a rare genetic disorder characterized by infertility and the absence of puberty. Defects in GnRH neuron migration or altered GnRH secretion and/or action lead to a severe gonadotropin-releasing hormone (GnRH) deficiency. Given the close developmental association of GnRH neurons with the olfactory primary axons, CHH is often associated with anosmia or hyposmia, in which case it is defined as Kallmann syndrome (KS). The genetics of CHH are heterogeneous, and >40 genes are involved either alone or in combination. Several CHH-related genes controlling GnRH ontogeny encode proteins containing fibronectin-3 (FN3) domains, which are important for brain and neural development. Therefore, we hypothesized that defects in other FN3-superfamily genes would underlie CHH. Next-generation sequencing was performed for 240 CHH unrelated probands and filtered for rare, protein-truncating variants (PTVs) in FN3-superfamily genes. Compared to gnomAD controls the CHH cohort was statistically enriched for PTVs in neuron-derived neurotrophic factor (NDNF) (p = 1.40 × 10 -6 ). Three heterozygous PTVs (p.Lys62 ∗ , p.Tyr128Thrfs ∗ 55, and p.Trp469 ∗ , all absent from the gnomAD database) and an additional heterozygous missense mutation (p.Thr201Ser) were found in four KS probands. Notably, NDNF is expressed along the GnRH neuron migratory route in both mouse embryos and human fetuses and enhances GnRH neuron migration. Further, knock down of the zebrafish ortholog of NDNF resulted in altered GnRH migration. Finally, mice lacking Ndnf showed delayed GnRH neuron migration and altered olfactory axonal projections to the olfactory bulb; both results are consistent with a role of NDNF in GnRH neuron development. Altogether, our results highlight NDNF as a gene involved in the GnRH neuron migration implicated in KS., (Copyright © 2019. Published by Elsevier Inc.)

Increased body weight is a major factor that interferes with smoking cessation. Nicotine, the main bioactive compound in tobacco, has been demonstrated to have an impact on energy balance, since it affects both feeding and energy expenditure at the central level. Among the central actions of nicotine on body weight, much attention has been focused on its effect on brown adipose tissue (BAT) thermogenesis, though its effect on browning of white adipose tissue (WAT) is unclear. Here, we show that nicotine induces the browning of WAT through a central mechanism and that this effect is dependent on the κ opioid receptor (KOR), specifically in the lateral hypothalamic area (LHA). Consistent with these findings, smokers show higher levels of uncoupling protein 1 (UCP1) expression in WAT, which correlates with smoking status. These data demonstrate that central nicotine-induced modulation of WAT browning may be a target against human obesity.

Numerosity discrimination tasks (judging which of two random dotarrays contains the larger number) have been widely used as a measure of the efficiency of an approximate number system (ANS) and are a correlate of early arithmetic skills. Recently, it has been suggested that the relationship between numerosity discrimination and arithmeticis explained by inhibition rather than the ANS. We assessed this hypothesis in astudy of 496 children (mean age = 81.23 months) using numerosity discrimination tasks that manipulated the congruency between surface area and numerosity. Numerosity discrimination for incongruent arrays (which are postulated to require inhibition due to a conflict between judgments based on surface area rather than numerosity) was more difficult than that for congruent arrays. However, all numerosity discrimination tasks showed substantial correlations with each other and correlated with arithmetic. A latent variable path model showed that a general numerosity judgment factorcorrelated witharithmetic even after controlling for a measure of response inhibition. In contrast, numerosity discrimination for incongruent arrays showed no unique relationship with arithmetic ability. Our results do not support the view that the relationship between numerosity discrimination and arithmetic is largely attributable to inhibition; rather, theyare consistent withthe view that numerosity discrimination tasks tap the operation of an ANS., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Congenital hypogonadotropic hypogonadism (CHH) is a condition characterized by absent puberty and infertility due to gonadotropin releasing hormone (GnRH) deficiency, which is often associated with anosmia (Kallmann syndrome, KS). We identified loss-of-function heterozygous mutations in anti-Müllerian hormone ( AMH ) and its receptor, AMHR2 , in 3% of CHH probands using whole-exome sequencing. We showed that during embryonic development, AMH is expressed in migratory GnRH neurons in both mouse and human fetuses and unconvered a novel function of AMH as a pro-motility factor for GnRH neurons. Pathohistological analysis of Amhr2 -deficient mice showed abnormal development of the peripheral olfactory system and defective embryonic migration of the neuroendocrine GnRH cells to the basal forebrain, which results in reduced fertility in adults. Our findings highlight a novel role for AMH in the development and function of GnRH neurons and indicate that AMH signaling insufficiency contributes to the pathogenesis of CHH in humans., Competing Interests: SM, GP, AM, NM, ST, MI, CA, IC, JA, DC, CX, RQ, GS, PP, LA, LM, JM, VP, NP, PG No competing interests declared, (© 2019, Malone et al.)

According to the Triple Code Model, early arithmetic development depends on learning the mappings between non-verbal representations of magnitude (quantity) and symbolic verbal (number words) and visual (Arabic numerals) representations of number. We examined this hypothesis in a sample of 166 4- to 7-year old children. Children completed 4 paired-associate learning tasks and a broad range of measures assessing early numerical (symbolic and non-symbolic magnitude comparison, digit writing, arithmetic) and reading skills (letter-sound knowledge, phoneme awareness, rapid automatized naming, word reading). A path model showed that paired-associate learning tasks involving mapping magnitudes onto verbal or visual stimuli predicted arithmetic performance over and above other well-established predictors. This relationship was specific to arithmetic and, consistent with the Triple Code Model, highlights that mapping between non-symbolic magnitude representations and the corresponding symbolic forms (verbal and visual) is important to the development of arithmetic skills., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Participants consisted of 496 children (mean age = 6 years; 9 months) recruited from 11 schools in Brisbane, Australia. Children were assessed on the addition and subtraction subtests of the Test of Basic Arithmetic and Number Skills (TOBANS) , an adapted version of the Head-Toes-Knees-Shoulders task to measure inhibition, numerosity discrimination using eight subtests varying ratio (2:3 or 5:6) and congruency, and non-verbal cognitive ability using an adapted version of Raven's Coloured Progressive Matrices . Information on children's demographics (gender, English as an additional language, and learning difficulty status) is also provided. All assessments were administered during the second year of formal schooling (i.e. Grade 1). Findings regarding the impact of inhibition on the relation between numerosity discrimination and arithmetic are reported elsewhere [1].

Background: Weak or inconsistent hand preference has been postulated to be a risk factor for developmental language delay. Following on from our Registered Stage 1 report this study assessed the extent to which variations in language skills are associated with the strength of hand preference. Methods: Data are drawn from a large sample ( N = 569) of 6- to 7-year-old children unselected for ability, assessed at two time points, 6 months apart. Hand preference was assessed using the Quantitative Hand Preference (QHP) task and five uni-manual motor tasks. Language skills (expressive and receptive vocabulary, receptive grammar, and morphological awareness) were assessed with standardized measures. Results: We found QHP scores did not distinguish children with weaker language skills from those with stronger language skills and the correlation between QHP scores and language ability was negligible in this study. Hand preference on the QHP task was significantly stronger among right-handed than left-handed children and left-handed children were typically inconsistent in the hand used across different tasks.  Conclusions: The findings presented here fail to provide any support for the theory that weak cerebral lateralisation (as assessed here by the QHP task) places children at risk of language difficulties . Stage 1 report:  https://doi.org/10.12688/wellcomeopenres.15077.1., Competing Interests: No competing interests were disclosed.

Background: Mitochondrial disease is a family of genetic disorders characterized by defects in the generation and regulation of energy. Epilepsy is a common symptom of mitochondrial disease, and in the vast majority of cases, refractory to commonly used antiepileptic drugs. Ferroptosis is a recently-described form of iron- and lipid-dependent regulated cell death associated with glutathione depletion and production of lipid peroxides by lipoxygenase enzymes. Activation of the ferroptosis pathway has been implicated in a growing number of disorders, including epilepsy. Given that ferroptosis is regulated by balancing the activities of glutathione peroxidase-4 (GPX4) and 15-lipoxygenase (15-LO), targeting these enzymes may provide a rational therapeutic strategy to modulate seizure. The clinical-stage therapeutic vatiquinone (EPI-743, α-tocotrienol quinone) was reported to reduce seizure frequency and associated morbidity in children with the mitochondrial disorder pontocerebellar hypoplasia type 6. We sought to elucidate the molecular mechanism of EPI-743 and explore the potential of targeting 15-LO to treat additional mitochondrial disease-associated epilepsies., Methods: Primary fibroblasts and B-lymphocytes derived from patients with mitochondrial disease-associated epilepsy were cultured under standardized conditions. Ferroptosis was induced by treatment with the irreversible GPX4 inhibitor RSL3 or a combination of pharmacological glutathione depletion and excess iron. EPI-743 was co-administered and endpoints, including cell viability and 15-LO-dependent lipid oxidation, were measured., Results: EPI-743 potently prevented ferroptosis in patient cells representing five distinct pediatric disease syndromes with associated epilepsy. Cytoprotection was preceded by a dose-dependent decrease in general lipid oxidation and the specific 15-LO product 15-hydroxyeicosatetraenoic acid (15-HETE)., Conclusions: These findings support the continued clinical evaluation of EPI-743 as a therapeutic agent for PCH6 and other mitochondrial diseases with associated epilepsy., Competing Interests: The authors have read the journal’s policy and the authors of this manuscript have the following competing interests: The following authors declare financial competing interests as paid current or former employees and/or equity holders of BioElectron Technology Corporation, Inc.: AA, JJB, AH, KGH, CRH, AHKK, VK, MBK, YK, JCL, CIM, SAM, JJM, WDS, SS, JKT, LWang. EPI-743 is a commercial product in development by BioElectron Technology Corporation, Inc., from which employees may benefit. The following authors declare patent applications (pending or actual) belonging to BioElectron Technology Corporation, from which they may benefit: JJB, AH, CRH, AHKK, JCL, SAM, WDS, JKT. The following authors declare grant support from BioElectron Technology Corporation pertaining to the work under consideration forpublication: GME, CD-V, DM. WG declares a cooperative research agreement between NHGRI and BioElectron pertaining to the scope of the work under consideration, as well as to activities outside the submitted work. RPS is a site principal investigator for two studies sponsored by BioElectron Technology Corporation. GME is a site principal investigator for an emergency protocol sponsored by BioElectron Technology Corporation. Outside the scope of the submitted work, EAA declares he is a Co-Founder of Personalis Inc., and DeepCell, Inc. CD-V declares grant support from Actelion and Sanofi Genzyme, as well as personal fees from Actelion, CureVac, Moderna Therapeutics, Logic- Biotherapeutics, Nutricia, Orphan Europe, Medifood, Promethera, Sanofi Genzyme, and SOBI. GME declares grant support and serving as site principal investigator for a clinical trial sponsored by Stealth Therapeutics. DM declares personal fees from SOBI. Thefollowing authors declared no competing interests: EB, RC, KAC, SCJ, MTW, LWolfe. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Weak or inconsistent hand preference may be a risk factor for developmental language delay.  This study will test the extent to which variations in language skills are associated with the strength of hand preference. Data are drawn from a large sample (n = 569) of 6- to 7-year-old children unselected for ability, assessed at two time points, 6 months apart. Hand preference is assessed using the Quantitative Hand Preference task (QHP) and five uni-manual motor tasks. Language skills (expressive and receptive vocabulary, receptive grammar, and morphological awareness) are assessed with standardized measures. If weak cerebral lateralisation (as assessed by the QHP task) is a risk factor for language difficulties, it should be possible to detect such effects in the large representative sample of children examined here., Competing Interests: No competing interests were disclosed.

Since its initial discovery in the 1940s, research into the physiological actions of anti-Müllerian hormone (AMH), from its eponymous role in male developmental biology to its routine clinical use in female reproductive health, has undergone a paradigm shifting change. With several exciting studies recently reporting hitherto unforeseen AMH actions at all levels in the hypogonadal-pituitary-gonadal axis, the importance of this hormone for both hypothalamic and pituitary reproductive control is finding increasing support and significance. In this review, we will briefly summarize what is known about the traditional roles and biology of AMH and how this could be integrated with new findings of AMH actions at the level of the hypothalamic-pituitary axis. We also synthesize the important findings from these new studies and discuss their potential impact and significance to our understanding of one of the most common reproductive disorders currently affecting women, polycystic ovary syndrome., (© 2019 The Author(s) Published by S. Karger AG, Basel.)

Ferroptosis is a form of programmed cell death associated with inflammation, neurodegeneration, and ischemia. Vitamin E (alpha-tocopherol) has been reported to prevent ferroptosis, but the mechanism by which this occurs is controversial. To elucidate the biochemical mechanism of vitamin E activity, we systematically investigated the effects of its major vitamers and metabolites on lipid oxidation and ferroptosis in a striatal cell model. We found that a specific endogenous metabolite of vitamin E, alpha-tocopherol hydroquinone, was a dramatically more potent inhibitor of ferroptosis than its parent compound, and inhibits 15-lipoxygenase via reduction of the enzyme's non-heme iron from its active Fe3+ state to an inactive Fe2+ state. Furthermore, a non-metabolizable isosteric analog of vitamin E which retains antioxidant activity neither inhibited 15-lipoxygenase nor prevented ferroptosis. These results call into question the prevailing model that vitamin E acts predominantly as a non-specific lipophilic antioxidant. We propose that, similar to the other lipophilic vitamins A, D and K, vitamin E is instead a pro-vitamin, with its quinone/hydroquinone metabolites responsible for its anti-ferroptotic cytoprotective activity., Competing Interests: The authors have read the journal’s policy and the authors of this manuscript have the following competing interests: All authors are employees of, and/or hold equity in BioElectron Technology Corporation, Inc. This does not alter the authors’ adherence to PLOS ONE policies on sharing data and materials.

Polycystic ovary syndrome (PCOS) is the main cause of female infertility worldwide and corresponds with a high degree of comorbidities and economic burden. How PCOS is passed on from one generation to the next is not clear, but it may be a developmental condition. Most women with PCOS exhibit higher levels of circulating luteinizing hormone, suggestive of heightened gonadotropin-releasing hormone (GnRH) release, and anti-Müllerian hormone (AMH) as compared to healthy women. Excess AMH in utero may affect the development of the female fetus. However, as AMH levels drop during pregnancy in women with normal fertility, it was unclear whether their levels were also elevated in pregnant women with PCOS. Here we measured AMH in a cohort of pregnant women with PCOS and control pregnant women and found that AMH is significantly more elevated in the former group versus the latter. To determine whether the elevation of AMH during pregnancy in women with PCOS is a bystander effect or a driver of the condition in the offspring, we modeled our clinical findings by treating pregnant mice with AMH and followed the neuroendocrine phenotype of their female progeny postnatally. This treatment resulted in maternal neuroendocrine-driven testosterone excess and diminished placental metabolism of testosterone to estradiol, resulting in a masculinization of the exposed female fetus and a PCOS-like reproductive and neuroendocrine phenotype in adulthood. We found that the affected females had persistently hyperactivated GnRH neurons and that GnRH antagonist treatment in the adult female offspring restored their neuroendocrine phenotype to a normal state. These findings highlight a critical role for excess prenatal AMH exposure and subsequent aberrant GnRH receptor signaling in the neuroendocrine dysfunctions of PCOS, while offering a new potential therapeutic avenue to treat the condition during adulthood.

Neurons expressing nitric oxide (NO) synthase (nNOS) and thus capable of synthesizing NO play major roles in many aspects of brain function. While the heterogeneity of nNOS-expressing neurons has been studied in various brain regions, their phenotype in the hypothalamus remains largely unknown. Here we examined the distribution of cells expressing nNOS in the postnatal and adult female mouse hypothalamus using immunohistochemistry. In both adults and neonates, nNOS was largely restricted to regions of the hypothalamus involved in the control of bodily functions, such as energy balance and reproduction. Labeled cells were found in the paraventricular, ventromedial, and dorsomedial nuclei as well as in the lateral area of the hypothalamus. Intriguingly, nNOS was seen only after the second week of life in the arcuate nucleus of the hypothalamus (ARH). The most dense and heavily labeled population of cells was found in the organum vasculosum laminae terminalis (OV) and the median preoptic nucleus (MEPO), where most of the somata of the neuroendocrine neurons releasing GnRH and controlling reproduction are located. A great proportion of nNOS-immunoreactive neurons in the OV/MEPO and ARH were seen to express estrogen receptor (ER) α. Notably, almost all ERα-immunoreactive cells of the OV/MEPO also expressed nNOS. Moreover, the use of EYFP Vglut2 , EYFP Vgat , and GFP Gad67 transgenic mouse lines revealed that, like GnRH neurons, most hypothalamic nNOS neurons have a glutamatergic phenotype, except for nNOS neurons of the ARH, which are GABAergic. Altogether, these observations are consistent with the proposed role of nNOS neurons in physiological processes., (© 2017 Wiley Periodicals, Inc.)

Generating a precise cellular and molecular cartography of the human embryo is essential to our understanding of the mechanisms of organogenesis in normal and pathological conditions. Here, we have combined whole-mount immunostaining, 3DISCO clearing, and light-sheet imaging to start building a 3D cellular map of the human development during the first trimester of gestation. We provide high-resolution 3D images of the developing peripheral nervous, muscular, vascular, cardiopulmonary, and urogenital systems. We found that the adult-like pattern of skin innervation is established before the end of the first trimester, showing important intra- and inter-individual variations in nerve branches. We also present evidence for a differential vascularization of the male and female genital tracts concomitant with sex determination. This work paves the way for a cellular and molecular reference atlas of human cells, which will be of paramount importance to understanding human development in health and disease. PAPERCLIP., (Copyright © 2017 Elsevier Inc. All rights reserved.)

We live in a digital society that provides a range of opportunities for virtual interaction. Consequently, emojis have become popular for clarifying online communication. This presents an exciting opportunity for psychologists, as these prolific online behaviours can be used to help reveal something unique about contemporary human behaviour., (Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.)

Following on from ideas developed by Gerstmann, a body of work has suggested that impairments in finger gnosis may be causally related to children's difficulties in learning arithmetic. We report a study with a large sample of typically developing children (N=197) in which we assessed finger gnosis and arithmetic along with a range of other relevant cognitive predictors of arithmetic skills (vocabulary, counting, and symbolic and nonsymbolic magnitude judgments). Contrary to some earlier claims, we found no meaningful association between finger gnosis and arithmetic skills. Counting and symbolic magnitude comparison were, however, powerful predictors of arithmetic skills, replicating a number of earlier findings. Our findings seriously question theories that posit either a simple association or a causal connection between finger gnosis and the development of arithmetic skills., (Crown Copyright © 2016. Published by Elsevier Inc. All rights reserved.)

Fertility in mammals is controlled by hypothalamic neurons that secrete gonadotropin-releasing hormone (GnRH). These neurons differentiate in the olfactory placodes during embryogenesis and migrate from the nose to the hypothalamus before birth. Information regarding this process in humans is sparse. Here, we adapted new tissue-clearing and whole-mount immunohistochemical techniques to entire human embryos/fetuses to meticulously study this system during the first trimester of gestation in the largest series of human fetuses examined to date. Combining these cutting-edge techniques with conventional immunohistochemistry, we provide the first chronological and quantitative analysis of GnRH neuron origins, differentiation and migration, as well as a 3D atlas of their distribution in the fetal brain. We reveal not only that the number of GnRH-immunoreactive neurons in humans is significantly higher than previously thought, but that GnRH cells migrate into several extrahypothalamic brain regions in addition to the hypothalamus. Their presence in these areas raises the possibility that GnRH has non-reproductive roles, creating new avenues for research on GnRH functions in cognitive, behavioral and physiological processes., (© 2016. Published by The Company of Biologists Ltd.)

Adults reason by exclusivity to identify the meanings of novel words. However, it is debated whether, like children, they extend this strategy to disambiguate other referential expressions (e.g., facts about objects). To further inform this debate, this study tested 41 adults on four conditions of a disambiguation task: label/label, fact/fact, label/fact, and fact/label (Scofield & Behrend, ). Participants also provided a verbal explanation for their referent selections to tease apart the underlying processes. Results indicated that adults successfully discerned the target object in the label/label and label/fact condition, yet not the remaining two conditions. Verbal reports indicated that the strategy utilized to disambiguate differed depending upon communicative context. These findings confirm that the tendency to reason by exclusivity becomes restricted to word-learning situations with growing linguistic and communicative experience., (Copyright © 2015 Cognitive Science Society, Inc.)

Reproductive competence in mammals depends on the projection of gonadotropin-releasing hormone (GnRH) neurons to the hypothalamic median eminence (ME) and the timely release of GnRH into the hypothalamic-pituitary-gonadal axis. In adult rodents, GnRH neurons and the specialized glial cells named tanycytes periodically undergo cytoskeletal plasticity. However, the mechanisms that regulate this plasticity are still largely unknown. We demonstrate that Semaphorin7A, expressed by tanycytes, plays a dual role, inducing the retraction of GnRH terminals and promoting their ensheathment by tanycytic end feet via the receptors PlexinC1 and Itgb1, respectively. Moreover, Semaphorin7A expression is regulated during the oestrous cycle by the fluctuating levels of gonadal steroids. Genetic invalidation of Semaphorin7A receptors in mice induces neuronal and glial rearrangements in the ME and abolishes normal oestrous cyclicity and fertility. These results show a role for Semaphorin7A signalling in mediating periodic neuroglial remodelling in the adult ME during the ovarian cycle.

Background: Antiretroviral therapy has become a central component of combination in HIV prevention efforts. Defining the individual exposure of commercially available antiretroviral therapy in genital secretions and vulnerable mucosal tissues is paramount to designing future prevention interventions., Methods: A pharmacokinetic (PK) study was performed in 12 HIV-negative men receiving 600 mg of darunavir, 100 mg of ritonavir, and 200 mg of etravirine orally, twice daily for 8 days. Seven blood plasma (BP) samples were collected over 12 hours on day 1 (PK1) and days 7 and 8 (PK2). One rectal tissue (RT) sample from each subject was collected during PK1 and PK2. During PK1, 2 seminal plasma (SP) samples were collected from each subject. During PK2, 6 SP samples were collected from each subject over 2 days., Results: Antiretrovirals were detected in SP and RT within 1 hour after a single dose. Over PK1 and PK2, SP exposures were lower than BP by 80%-92% (DRV), 89-95% (RTV), and 83-88% (ETR). However, protein binding in SP (14% for darunavir, 70% for ritonavir, and 97% for etravirine) was lower than in BP. Rectal tissue exposures were higher than BP by 39- to 155-fold for darunavir, 12- to 61-fold for ritonavir, and 20- to 40-fold for etravirine., Conclusions: Lower SP protein binding resulted in higher pharmacologically active darunavir and etravirine concentrations compared with BP. High RT concentrations may also be favorable for suppressing viral replication in the gastrointestinal mucosa. The high protein-unbound exposures in SP and total exposures in RT support further investigations of darunavir plus ritonavir and etravirine in secondary prevention.

Background: Antiretroviral pharmacology in seminal plasma (SP) and rectal tissue (RT) may provide insight into antiretroviral resistance and the prevention of sexual transmission of human immunodeficiency virus (HIV). Saliva may be of utility for noninvasively measuring adherence., Methods: A pharmacokinetic study was performed in 12 HIV-negative men receiving maraviroc 300 mg twice daily for 8 days. Seven time-matched pairs of blood plasma (BP) and saliva samples were collected over 12 h on day 1 (PK1) and days 7 and 8 (PK2). One RT sample from each subject was collected during PK1 and PK2. Two SP samples were collected from each subject during PK1, and 6 SP samples were collected from each subject during PK2., Results: SP AUCs were ∼50% lower than BP. However, protein binding in SP ranged from 4% to 25%, resulting in protein-free concentrations >2-fold higher than BP. RT AUCs were 7.5- to 26-fold higher than BP. Maraviroc saliva AUCs were ∼70% lower than BP, but saliva concentrations correlated with BP (r(2) = 0.58)., Conclusions: More pharmacologically available maraviroc was found in SP than BP. High RT concentrations are promising for preventing rectal HIV acquisition. Saliva correlation with BP suggests that this may be useful for monitoring adherence., Clinical Trials Registration: NCT00775294.

Many approaches to word learning argue for the importance of joint attention and other social-pragmatic abilities. This study explored word learning in children with autism (CWA), by examining it in ostensive and non-ostensive contexts, tested through both comprehension and elicited production. Novel nouns were taught to 17 CWA and 13 children with moderate learning difficulties (MLD) using an adapted version of Tomasello and Barton's (Developmental Psychology, 30: 639-650, 1994) search paradigm. In elicited production for words learnt within an ostensive context, CWA performed at a significantly higher level than MLD children. This is contrary to prior findings and suggests that word learning abilities in CWA have been underestimated.

Lamotrigine concentrations were measured simultaneously (as far as was feasible) in stimulated and unstimulated saliva samples, and in plasma, from seven adult volunteers over a 32 h period following a single 50 mg dose of the drug, and in 20 children and adolescents during the course of routine antiepileptic therapy. In individuals there was a close correlation between the measurements at least 2 h after ingestion of the drug. Concentrations in stimulated and unstimulated saliva were similar; the stimulation produced little change in the saliva secretion rate. The saliva-to-plasma concentration ratio increased linearly by 0.78% for each 1 mg/L plasma lamotrigine concentration, with a mean value of 48.8% at a plasma lamotrigine concentration of 10 mg/L. With appropriate precautions as to the timing of saliva collections, and a single plasma lamotrigine concentration measurement to calibrate the salivary values in the individual, salivary lamotrigine concentration measurement appears to be a practicable approach to therapeutic drug monitoring. This has significant implications for the elucidation of the pharmacokinetics of lamotrigine in the paediatric population.

Ferritins are iron-storage proteins capable of holding up to 4500 Fe(3+) ions within a single water-soluble protein shell made from 24 polypeptide chains. The Glu128Arg/Glu135Arg mutants of Escherichia coli and Rhodobacter capsulatus bacterioferritins are unable to associate into 24-meric structures, with dimers of polypeptide chains being their stable forms. The aerobic addition to these of up to 8-10 or 14-20 Fe(2+) ions per dimer, respectively, results in the oxidation of the added Fe(2+) to Fe(3+). Gel permeation chromatography and sedimentation equilibrium studies confirm that the Fe(3+) ions are associated with the polypeptide dimer, and the lack of intense EPR signals from magnetically isolated Fe(3+) ions confirms the formation of one or more antiferromagnetically coupled clusters of Fe(3+) ions. The effect of Fe(3+) chelators on iron-loaded subunit dimers is to remove the Fe(3+) from the protein, but to do so slowly, consistent with it not being merely adventitiously associated with protein. These data provide experimental support for the presence of nucleation centers for the mineral cores in bacterioferritins and indicate that these proteins are not simply acting as vessels in which hydrolysis of Fe(3+) occurs independent from the protein surface. From analyses of X-ray structures and amino acid sequence comparisons, possible nucleation sites are identified.

A senior registrar 'shadowed' a district general manager and a unit general manager for one week each as a management training exercise. The three principal participants describe the exercise from their different perspectives. Consequent issues and the value of such 'shadowing' as a mode of training are discussed.

The effects of 1 hour of mild and moderate reductions in coronary blood flow on myocardial high-energy phosphate levels were evaluated. Thirty anesthetized pigs were instrumented with left anterior descending arterial and venous catheters, crystals for instantaneous wall thickness, and a fluid-filled occluder. Measurement of myocardial blood flow was performed with microspheres, and a series of myocardial biopsies also was performed. In 10 pigs, overall coronary blood flow was lowered by 22%, with a fall in subendocardial-to-subepicardial flow ratio from 1.11 to 0.54 and in wall thickening from 33% to 15%. Subendocardial flow fell 48%. Coronary blood flow and thickening were constant during 1 hour of ischemia. Phosphocreatine (mumol/g wet wt) in the subendocardial third of the ischemic zone fell from 7.6 to 3.8 at 5 minutes of ischemia (p less than 0.005 versus control) and returned to normal (7.9) at 60 minutes (p = NS), despite ongoing ischemia. Subendocardial ATP (mumol/g wet wt) fell slowly from 4.3 and leveled off at 2.1 at 60 minutes of ischemia (p less than 0.001 versus control). Similar regeneration of phosphocreatine was found in seven additional pigs, with a 43% transmural reduction in coronary blood flow and a 66% reduction in subendocardial flow. No significant changes in ATP and phosphocreatine were noted in two different control groups (n = 13 pigs). The regeneration of phosphocreatine despite ongoing ischemia and low ATP levels was not related to changes in myocardial oxygen demand or consumption, or in regional function during the period of ischemia. This may reflect 1) a successful downregulation of the energy needs of the ischemic myocardium to maintain cell viability, or 2) a metabolic abnormality in the ability of the cells to produce ATP primarily or by use of phosphocreatine.

To explore the relation between myocardial and vascular injury in the generation of the no-reflow phenomenon, the pressure-flow relation during maximal vasodilation after coronary artery reperfusion was studied in the open-chest porcine model. During both endogenous and maximal vasodilation with intracoronary adenosine, pressure-flow (P/Q) plots were constructed before and after 20-minute (n = 9) or 40-minute (n = 17) circumflex artery occlusions. Decreases in circumflex vascular bed conductance were represented by downward shifts in P/Q plot regression lines. No significant change occurred in P/Q line slope or pressure at zero flow 30 minutes after release of the 20-minute occlusion, and no infarction was found. After release of the 40-minute occlusion, a small but insignificant decrease in P/Q line slope occurred during endogenous vasodilation. However, during maximal vasodilation, a significant (p less than 0.01) decrease in P/Q line slope was present during reperfusion compared with preocclusion corresponding to a decrease in vasodilatory reserve (P/Q line slope = 1.52 +/- 0.14 ml/min/mm Hg preocclusion vs. 1.03 +/- 0.13 at 15 minutes reperfusion). Pretreatment with aspirin did not prevent this decrease in vascular conductance during maximal vasodilation. Total circumflex, as well as subendocardial, midmyocardial, and subepicardial blood flows, was measured with radioactive microspheres. There was a good correlation between the extent of infarction measured by triphenyltetrazolium chloride staining and the decrease in vascular conductance during maximal vasodilation for all three myocardial layers as well as for the total circumflex vascular bed. Hence, the degree of no-reflow correlates closely with the extent of infarction during maximal vasodilation (but not during endogenous vasodilation) and is not altered by aspirin therapy.

Pretransfusion blood samples were routinely tested for ABO group, Rh type, and the presence of unexpected red blood cell antibodies. Patients who had unexpected red blood cell antibodies received transfusions of blood that was crossmatched using an immediate spin test, a 37 degrees C incubation step, and an indirect antiglobulin test. Patients who did not have unexpected red blood cell antibodies received transfusions with blood that was crossmatched by an immediate spin crossmatch only. Because the average immediate spin crossmatch required only 3.25 minutes to be performed, crossmatches were not done for patients without unexpected antibodies until blood was actually requested to be issued for transfusion. During the first 8 1/2 months this protocol was used, 27,742 crossmatches were performed and 46,959 unnecessary crossmatches were avoided, thus reducing direct costs by at least +49,300. This protocol also allowed for optimal blood inventory control and minimized the outdating of units of blood to only 0.19%.