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Astrocytes are morphologically complex cells that serve essential roles. They are widely implicated in central nervous system (CNS) disorders, with changes in astrocyte morphology and gene expression accompanying disease. In the Sapap3 knockout (KO) mouse model of compulsive and anxiety-related behaviors related to obsessive-compulsive disorder (OCD), striatal astrocytes display reduced morphology and altered actin cytoskeleton and Gi-G-protein-coupled receptor (Gi-GPCR) signaling proteins. Here, we show that normalizing striatal astrocyte morphology, actin cytoskeleton, and essential homeostatic support functions by targeting the astrocyte Gi-GPCR pathway using chemogenetics corrected phenotypes in Sapap3 KO mice, including anxiety-related and compulsive behaviors. Our data portend an astrocytic pharmacological strategy for rescuing phenotypes in brain disorders that include compromised astrocyte morphology and tissue support., Competing Interests: Declaration of interests UCLA filed a US provisional patent (no. 63/658,760) based on this work. B.S.K. is on the editorial advisory board of Neuron., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Objective: To assess the absence of diabetic foot examination in Brazil and how ethnicity affected this outcome., Design: This is an analysis of a nationwide survey held in Brazil in 2019. Participants with diabetes and that were 15 years of age or older were eligible for inclusion in the analysis. Adjusted Poisson regression with robust variance was used to calculate prevalence ratios (PRs) and 95% confidence intervals (95% CIs) of never having had the foot examined, with separate models according to ethnicity. Stata 14.2 was used for all calculations., Results: We included 6216 individuals with diabetes; 52.1% (95%CI: 50.0%-54.2%) reported never having had their feet examined by a health care professional and 61% self-declared as Black (Black and Brown [Brazilian mixed race]). A higher frequency of negligence was observed among Black individuals (55.3%; 52.5%-58.1%) than among White individuals (48.2%; 45.0%-51.5%). Negligence was higher between 15- to 39-year-old participants (PR = 1.34, 1.14-1.57), lower educational level (PR = 1.37, 1.13-1.65), higher alcohol consumption (PR = 1.18, 1.06-1.31), fair health status (PR = 1.11, 1.01-1.21), and diabetes diagnosis of up to 10 years (PR = 1.42, 1.28-1.57). Among Blacks, tobacco use and other factors increased the frequency of the outcome, whereas participation in the Brazilian Unified Health System primary care program was a protection factor (P<.05)., Conclusion: Black Brazilians with diabetes had higher negligence of foot examination by health care professionals. Strengthening primary care would help mitigate systemic racism in Brazil., Competing Interests: Conflict of Interest: No conflict of interest reported by authors.

Jaboticaba ( Myrciaria jaboticaba ) is a recognizable and unique crop from Brazil. The fruit's byproducts are currently being studied, given their bioactive composition and promising anti-cancer potential. It is not evident, however, if different harvesting seasons can modify the chemical profile and antioxidant capacity of jaboticaba fruit fractions. Furthermore, as there is limited data for jaboticaba's anti-proliferative effects, additional assessments are required to improve the robustness of these findings. Therefore, this study aimed to determine the composition of the peel of jaboticaba collected in two periods (May-off-season, sample 1-and August-October-peak season, sample 2) and test the peel's richest anthocyanin sample against colorectal cancer (CRC) cell lines. To accomplish this, proximate, spectrophotometric, and chromatographic analyses were performed in two freeze-dried samples; and anti-proliferative and/or colony-forming assays were carried out in Caco-2, HT29, and HT29-MTX cells. As a result, sample 2 showed the highest levels of polyphenols overall, including flavonoids and anthocyanins. This sample displayed significative higher contents of cyanidin-3- O -glucoside (48%) and delphinidin-3- O -glucoside (105%), in addition to a superior antioxidant capacity (23% higher). Sample 1 showed higher amounts of total protein, gallic acid (20% higher), and specific carotenoids. An aqueous extract from sample 2 was tested against CRC, showing anti-proliferative effects for Caco-2 cells at 1 and 2 mg/mL concentrations, with IC50 values of 1.2-1.3 mg/mL. Additionally, the extract was able to inhibit cell colony formation when tested at both low and high concentrations. In conclusion, jaboticaba collected in the main season stands out regarding its polyphenol composition and holds potential against cancer cell growth.

Background: Pulmonary hypertension (PH) accompanying COPD (PH-COPD) is associated with worse outcomes than COPD alone. There are currently no approved therapies to treat PH-COPD. The PERFECT study (ClinicalTrials.gov: NCT03496623) evaluated the safety and efficacy of inhaled treprostinil (iTRE) in this patient population., Methods: Patients with PH-COPD (mean pulmonary arterial pressure ≥30 mmHg and pulmonary vascular resistance ≥4 WU) were enrolled in a multicentre, randomised (1:1), double-blind, placebo-controlled, 12-week, crossover study. A contingent parallel design was also prespecified and implemented, based on a blinded interim analysis of missing data. Patients received treatment with iTRE up to 12 breaths (72 µg) 4 times daily or placebo. The primary efficacy end-point was change in peak 6-min walk distance (6MWD) at week 12., Results: In total, 76 patients were randomised, 64 in the original crossover design and 12 in the contingent parallel design; 66 patients received iTRE and 58 received placebo. The study was terminated early at the recommendation of the data and safety monitoring committee based on the totality of evidence that iTRE increased the risk of serious adverse events and suggestive evidence of an increased risk of mortality. The change in 6MWD was numerically worse with iTRE exposure than with placebo exposure., Conclusions: The risk-benefit observations associated with iTRE in patients with PH-COPD did not support continuation of the PERFECT study. The results of this study do not support iTRE as a viable treatment option in patients with PH-COPD., Competing Interests: Conflict of interest: S.D. Nathan is a paid consultant for United Therapeutics. R.G. Argula reports advisory board consulting fees from United Therapeutics, Liquidia Inc., Merck Pharmaceuticals, Janssen and Accordant Health (CVS), and lecture honoraria from United Therapeutics, outside the submitted work. M.G. Trivieri reports advisory board participation with Janssen, outside the submitted work. B. Medarov reports lecture honoraria from Jensen Pharmaceuticals, outside the submitted work. A. Raina reports lecture honoraria from Merck and United Therapeutics, outside the submitted work. M.G. Risbano reports grants from Shadyside Foundation, royalties from Springer (Pulmonary Hypertension: Controversial and Emerging Topics), and advisory board participation with Gilead and Liquidia, outside the submitted work. T. Thenappan reports grants from United Therapeutics, Aerovate, Merck and Aria CV, and consulting fees from Merck and United Therapeutics, outside the submitted work. J.S. Soto reports grants, lecture honoraria, travel support and advisory board participation with GlaxoSmithKline and Genentech Pharmaceuticals, outside the submitted work. H. Bell, V. Lacasse, P. Sista, M. Di Marino, A. Smart, B. Hawkes and E. Nelson are employees of United Therapeutics, the PERFECT study sponsor. T. Bull reports grants from Bayer, Merck, Insmed and Aerovate, lecture honoraria from Merck, payment for expert testimony from Lung Biotechnology, travel support from Lung, advisory board participation with Keros, consultancy for United Therapeutics and a leadership role as PHA SLC chair, outside the submitted work. V. Tapson is a paid consultant for United Therapeutics. A. Waxman reports grants from Aria CV PI, AI Therapeutics and Acceleron/Merck, consultancy for United Therapeutics, and data and safety monitoring board participation with Insmed, outside the submitted work. The remaining authors have no potential conflicts of interest to disclose., (Copyright ©The authors 2024.)

Roads are one of the most widespread structures that drive habitat loss and fragmentation. But they also restrict animal movement and drive landscape-level impacts on biodiversity. The South Pacific of Costa Rica is known for its high levels of biodiversity, but little has been done to reduce road impacts upon wildlife communities. To understand these impacts and advise on possible mitigation action, we used three key data approaches: 1. Camera traps, to survey wildlife activity along two major road sections that dissect the region's protected areas and biological corridors. Seventy-eight camera traps were deployed in secondary forest patches at different distances (between 200 m and 1 km) from the roads for six months and covariates were collected to explain the patterns found. 2. Citizen science data extracted from iNaturalist to identify roadkill "hotspots" along the roads. And 3. Circuitscape analysis, to assess how landscape structure could influence animal movement. Camera traps recorded 30 terrestrial species. Ocelots and agoutis displayed a negative effect of distance from protected area, while the Apex predators displayed a positive effect toward higher forest cover and vegetation density. Circuitscape analysis showed high connectivity throughout most of the area. Only a few locations showed higher flow (bottle neck locations), which coincided with roadkill "hotspots" identified through citizen science direct observations (70 observations of 21 species). Amalgamating data from the different analyses allow us to identify four key wildlife crossing locations (one of less priority) along the Inter-American Highway. We strongly recommend the placement of under/overpasses in these locations, with the aim to ensure wildlife safe movement and connectivity of wildlife populations in the region. Culvert modifications in the area could also be considered to incorporate wildlife underpasses at a reduced cost., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Osa Conservation reports financial support and equipment, drugs, or supplies were provided by Disney Worldwide Conservation Fund. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The central nervous system (CNS) comprises diverse and morphologically complex cells. To understand the molecular basis of their physiology, it is crucial to assess proteins expressed within intact cells. Commonly used methods utilize cell dissociation and sorting to isolate specific cell types such as neurons and astrocytes, the major CNS cells. Proteins purified from isolated cells are identified by mass spectrometry-based proteomics. However, dissociation and cell-sorting methods lead to near total loss of cellular morphology, thereby losing proteins from key relevant subcompartments such as processes, end feet, dendrites and axons. Here we provide a systematic protocol for cell- and subcompartment-specific labeling and identification of proteins found within intact astrocytes and neurons in vivo. This protocol utilizes the proximity-dependent biotinylation system BioID2, selectively expressed in either astrocytes or neurons, to label proximal proteins in a cell-specific manner. BioID2 is targeted genetically to assess the subproteomes of subcellular compartments such as the plasma membrane and sites of cell-cell contacts. We describe in detail the expression methods (variable timing), stereotaxic surgeries for expression (1-2 d and then 3 weeks), in vivo protein labeling (7 d), protein isolation (2-3 d), protein identification methods (2-3 d) and data analysis (1 week). The protocol can be applied to any area of the CNS in mouse models of physiological processes and for disease-related research., (© 2023. Springer Nature Limited.)

Astrocytes are heterogeneous glial cells of the central nervous system 1-3 . However, the physiological relevance of astrocyte diversity for neural circuits and behaviour remains unclear. Here we show that a specific population of astrocytes in the central striatum expresses μ-crystallin (encoded by Crym in mice and CRYM in humans) that is associated with several human diseases, including neuropsychiatric disorders 4-7 . In adult mice, reducing the levels of μ-crystallin in striatal astrocytes through CRISPR-Cas9-mediated knockout of Crym resulted in perseverative behaviours, increased fast synaptic excitation in medium spiny neurons and dysfunctional excitatory-inhibitory synaptic balance. Increased perseveration stemmed from the loss of astrocyte-gated control of neurotransmitter release from presynaptic terminals of orbitofrontal cortex-striatum projections. We found that perseveration could be remedied using presynaptic inhibitory chemogenetics 8 , and that this treatment also corrected the synaptic deficits. Together, our findings reveal converging molecular, synaptic, circuit and behavioural mechanisms by which a molecularly defined and allocated population of striatal astrocytes gates perseveration phenotypes that accompany neuropsychiatric disorders 9-12 . Our data show that Crym-positive striatal astrocytes have key biological functions within the central nervous system, and uncover astrocyte-neuron interaction mechanisms that could be targeted in treatments for perseveration., (© 2024. The Author(s).)

Microorganisms do not live as dispersed single cells but rather they form aggregates with extracellular polymeric substances at interfaces. Biofilms are considered efficient life forms because they shield bacteria from biocides and collect dilute nutrients. This is a big concern in industry since the microorganisms can colonize a wide range of surfaces, accelerating material deterioration, colonizing medical devices, contaminating ultrapure drinking water, increasing energy costs and creating focus of infection. Conventional biocides that target a specific component of the bacteria are not effective in the presence of biofilms. Efficient biofilm inhibitors are based on a multitarget approach interacting with the bacteria and the biofilm matrix. Their rationale design requires a thorough understanding of inhibitory mechanisms that are still largely lacking today. Herein we uncover via molecular modelling the inhibition mechanism of cetrimonium 4-OH cinnamate (CTA-4OHcinn). Simulations show that CTA-4OH micelles can disrupt symmetric and asymmetric bilayers, representative of inner and outer bacterial membranes, following three stages: adsorption, assimilation, and defect formation. The main driving force for micellar attack is electrostatic interactions. In addition to disrupting the bilayers, the micelles work as carriers facilitating the trapping of 4OH cinnamate anions within the bilayer upper leaflet and overcoming electrostatic repulsion. The micelles also interact with extracellular DNA (e-DNA), which is one of the main components of biofilms. It is observed that CTA-4OHcinn forms spherical micelles on the DNA backbone; which hinders their ability to pack. This is demonstrated by modelling the DNA along the hbb histone-like protein, showing that in the presence of CTA-4OHcinn, DNA does not pack properly around hbb. The abilities of CTA-4OHcinn to cause cell death through membrane disruption and to disperse a mature, multi-species biofilm are also confirmed experimentally., (© 2023. The Author(s).)

Calorie restriction can enhance the regenerative capacity of the injured intestinal epithelium. Among other metabolic changes, calorie restriction can activate the autophagy pathway. Although independent studies have attributed the regenerative benefit of calorie restriction to downregulation of mTORC1, it is not known whether autophagy itself is required for the regenerative benefit of calorie restriction. We used mouse and organoid models with autophagy gene deletion to evaluate the contribution of autophagy to intestinal epithelial regeneration following calorie restriction. In the absence of injury, mice with intestinal epithelial-specific deletion of autophagy gene Atg7 ( Atg7 ΔIEC ) exhibit weight loss and histological changes similar to wild-type mice following calorie restriction. Conversely, calorie-restricted Atg7 ΔIEC mice displayed a significant reduction in regenerative crypt foci after irradiation compared with calorie-restricted wild-type mice. Targeted analyses of tissue metabolites in calorie-restricted mice revealed an association between calorie restriction and reduced glycocholic acid (GCA) in wild-type mice but not in Atg7 ΔIEC mice. To evaluate whether GCA can directly modulate epithelial stem cell self-renewal, we performed enteroid formation assays with or without GCA. Wild-type enteroids exhibited reduced enteroid formation efficiency in response to GCA treatment, suggesting that reduced availability of GCA during calorie restriction may be one mechanism by which calorie restriction favors epithelial regeneration in a manner dependent upon epithelial autophagy. Taken together, our data support the premise that intestinal epithelial Atg7 is required for the regenerative benefit of calorie restriction, due in part to its role in modulating luminal GCA with direct effects on epithelial stem cell self-renewal. NEW & NOTEWORTHY Calorie restriction is associated with enhanced intestinal regeneration after irradiation, but the requirement of autophagy for this process is not known. Our data support the premise that intestinal epithelial autophagy is required for the regenerative benefit of calorie restriction. We also report that luminal levels of primary bile acid glycocholic acid are modulated by epithelial cell autophagy during calorie restriction with direct effects on epithelial stem cell function.

Astrocytes and neurons extensively interact in the brain. Identifying astrocyte and neuron proteomes is essential for elucidating the protein networks that dictate their respective contributions to physiology and disease. Here we used cell-specific and subcompartment-specific proximity-dependent biotinylation 1 to study the proteomes of striatal astrocytes and neurons in vivo. We evaluated cytosolic and plasma membrane compartments for astrocytes and neurons to discover how these cells differ at the protein level in their signalling machinery. We also assessed subcellular compartments of astrocytes, including end feet and fine processes, to reveal their subproteomes and the molecular basis of essential astrocyte signalling and homeostatic functions. Notably, SAPAP3 (encoded by Dlgap3), which is associated with obsessive-compulsive disorder (OCD) and repetitive behaviours 2-8 , was detected at high levels in striatal astrocytes and was enriched within specific astrocyte subcompartments where it regulated actin cytoskeleton organization. Furthermore, genetic rescue experiments combined with behavioural analyses and molecular assessments in a mouse model of OCD 4 lacking SAPAP3 revealed distinct contributions of astrocytic and neuronal SAPAP3 to repetitive and anxiety-related OCD-like phenotypes. Our data define how astrocytes and neurons differ at the protein level and in their major signalling pathways. Moreover, they reveal how astrocyte subproteomes vary between physiological subcompartments and how both astrocyte and neuronal SAPAP3 mechanisms contribute to OCD phenotypes in mice. Our data indicate that therapeutic strategies that target both astrocytes and neurons may be useful to explore in OCD and potentially other brain disorders., (© 2023. The Author(s).)

Astrocytes, the most abundant glial cells in the central nervous system, play vital roles in maintaining neuronal function. A new study using focused ion-beam scanning electron microscopy reveals the architecture of astrocytes at the nanoscale and provides new insights on how astrocytes perform their diverse activities., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Huntington's disease (HD) is caused by expanded CAG repeats in the huntingtin gene (HTT) resulting in expression of mutant HTT proteins (mHTT) with extended polyglutamine tracts, including in striatal neurons and astrocytes. It is unknown whether pathophysiology in vivo can be attenuated by lowering mHTT in either cell type throughout the brain, and the relative contributions of neurons and astrocytes to HD remain undefined. We use zinc finger protein (ZFP) transcriptional repressors to cell-selectively lower mHTT in vivo. Astrocytes display loss of essential functions such as cholesterol metabolism that are partly driven by greater neuronal dysfunctions, which encompass neuromodulation, synaptic, and intracellular signaling pathways. Using transcriptomics, proteomics, electrophysiology, and behavior, we dissect neuronal and astrocytic contributions to HD pathophysiology. Remarkably, brain-wide delivery of neuronal ZFPs results in strong mHTT lowering, rescue of HD-associated behavioral and molecular phenotypes, and significant extension of lifespan, findings that support translational development., Competing Interests: Declaration of interests B.S.K. is on the editorial advisory board of Neuron., (Copyright © 2022 The Author(s). Published by Elsevier Inc. All rights reserved.)

Astrocytes, a type of glia, are abundant and morphologically complex cells. Here, we report astrocyte molecular profiles, diversity, and morphology across the mouse central nervous system (CNS). We identified shared and region-specific astrocytic genes and functions and explored the cellular origins of their regional diversity. We identified gene networks correlated with astrocyte morphology, several of which unexpectedly contained Alzheimer's disease (AD) risk genes. CRISPR/Cas9-mediated reduction of candidate genes reduced astrocyte morphological complexity and resulted in cognitive deficits. The same genes were down-regulated in human AD, in an AD mouse model that displayed reduced astrocyte morphology, and in other human brain disorders. We thus provide comprehensive molecular data on astrocyte diversity and mechanisms across the CNS and on the molecular basis of astrocyte morphology in health and disease.

Understanding how human modification of the landscape shapes vertebrate community composition is vital to understanding the current status and future trajectory of wildlife. Using a participatory approach, we deployed the largest camera-trap network in Mesoamerica to date to investigate how anthropogenic disturbance shapes the occupancy and co-occurrence of terrestrial vertebrate species in a tropical biodiversity hotspot: the Osa Peninsula, Costa Rica. We estimated species richness in different categories of land protection with rarefaction analysis and estimated the expected occupancy with a joint species distribution model that included covariates for anthropogenic disturbance, land protection, habitat quality, and habitat availability. Areas with the most stringent land-use protections (e.g., Corcovado National Park, 24 species [95% CI 23-25]) harbored significantly more species than unprotected areas (20 species [19.7-20.3]), mainly due to a reduced presence of large-bodied species of conservation concern in unprotected areas (e.g., jaguar Panthera onca and white-lipped peccary Tayassu pecari). Small-bodied generalist species, such as opossums (Didelphidae) and armadillos (Dasypus novemcinctus), in contrast, were more common at disturbed sites, resulting in a significant difference in vertebrate community composition between sites with low and high disturbance. Co-occurrence of species was also mainly associated with response to disturbance. Similar responses to disturbance create two groups of species, those whose site-level occupancy usually increased as anthropogenic disturbance increased and those whose estimated occupancy decreased. The absence of large-bodied species entails an important loss of ecological function in disturbed areas and can hinder forest development and maintenance. Efforts to protect and restore forested landscapes are likely having a positive effect on the abundance of some threatened species. These efforts, however, must be sustained and expanded to increase connectivity and ensure the long-term viability of the wildlife community., (© 2021 Society for Conservation Biology.)

Helminth parasites are part of almost every ecosystem, with more than 300 000 species worldwide. Helminth infection dynamics are expected to be altered by climate change, but predicting future changes is difficult owing to lacking thermal sensitivity data for greater than 99.9% of helminth species. Here, we compiled the largest dataset to date on helminth temperature sensitivities and used the Metabolic Theory of Ecology to estimate activation energies (AEs) for parasite developmental rates. The median AE for 129 thermal performance curves was 0.67, similar to non-parasitic animals. Although exceptions existed, related species tended to have similar thermal sensitivities, suggesting some helminth taxa are inherently more affected by rising temperatures than others. Developmental rates were more temperature-sensitive for species from colder habitats than those from warmer habitats, and more temperature sensitive for species in terrestrial than aquatic habitats. AEs did not depend on whether helminth life stages were free-living or within hosts, whether the species infected plants or animals, or whether the species had an endotherm host in its life cycle. The phylogenetic conservatism of AE may facilitate predicting how temperature change affects the development of helminth species for which empirical data are lacking or difficult to obtain.

The aim of the study was to molecularly identify virulence and macrolide resistance genes in clinical isolates of Streptococcus agalactiae (GBS), recovered in 2019 from vaginal discharge (n=9) and urine (n=22), from two health facilities in Lima. Identification and antimicrobial susceptibility were determined by the Vitek® 2 automated system, identification was confirmed phenotypically; macrolide resistance was determined by the D-test method. Identification of virulence genes (lmb, bca and rib) and macrolide resistance genes (ermB, ermTR and mefA) was carried out by polymerase chain reaction (PCR). The predominant macrolide resistance phenotype and genotype were cMLSb (12/31) and ermB (11/31); the most frequent virulence gene was lmb (23/31). All were sensitive to penicillin, ampicillin and vancomycin. These findings show the need to implement molecular epidemiology studies that allow adequate knowledge and follow-up of GBS in Peru.

Visual deficits are among the most prevalent symptoms in patients with multiple sclerosis (MS). To understand deficits in the visual pathway during MS and potential treatment effects, we used experimental autoimmune encephalomyelitis (EAE), the most commonly used animal model of MS. The afferent visual pathway was assessed in vivo using optical coherence tomography (OCT), electroretinography (ERG), and visually evoked cortical potentials (VEPs). Inflammation, demyelination, and neurodegeneration were examined by immunohistochemistry ex vivo. In addition, an immunomodulatory, remyelinating agent, the estrogen receptor β ligand chloroindazole (IndCl), was tested for its therapeutic potential in the visual pathway. EAE produced functional deficits in visual system electrophysiology, including suppression of ERG and VEP waveform amplitudes and increased signal latencies. Therapeutic IndCl rescued overall visual system latency by VEP but had little impact on amplitude or ERG findings relative to vehicle. Faster VEP conduction in IndCl-treated mice was associated with enhanced myelin basic protein signal in all visual system structures examined. IndCl preserved retinal ganglion cells (RGCs) and oligodendrocyte density in the prechiasmatic white matter, but similar retinal nerve fiber layer thinning by OCT was noted in vehicle and IndCl-treated mice. Although IndCl differentially attenuated leukocyte and astrocyte staining signal throughout the structures analyzed, axolemmal varicosities were observed in all visual fiber tracts of mice with EAE irrespective of treatment, suggesting impaired axonal energy homeostasis. These data support incomplete functional recovery of VEP amplitude with IndCl, as fiber tracts displayed persistent axon pathology despite remyelination-induced decreases in latencies, evidenced by reduced optic nerve g-ratio in IndCl-treated mice. Although additional studies are required, these findings demonstrate the dynamics of visual pathway dysfunction and disability during EAE, along with the importance of early treatment to mitigate EAE-induced axon damage., (© 2020 The Authors. Brain Pathology published by John Wiley & Sons Ltd on behalf of International Society of Neuropathology.)

Astrocytes tile the central nervous system and are widely implicated in brain diseases, but the molecular mechanisms by which astrocytes contribute to brain disorders remain incompletely explored. By performing astrocyte gene expression analyses following 14 experimental perturbations of relevance to the striatum, we discovered that striatal astrocytes mount context-specific molecular responses at the level of genes, pathways, and upstream regulators. Through data mining, we also identified astrocyte pathways in Huntington's disease (HD) that were reciprocally altered with respect to the activation of striatal astrocyte G protein-coupled receptor (GPCR) signaling. Furthermore, selective striatal astrocyte stimulation of the G i -GPCR pathway in vivo corrected several HD-associated astrocytic, synaptic, and behavioral phenotypes, with accompanying improvement of HD-associated astrocyte signaling pathways, including those related to synaptogenesis and neuroimmune functions. Overall, our data show that astrocytes are malleable, using context-specific responses that can be dissected molecularly and used for phenotypic benefit in brain disorders., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Background: Dravet syndrome (DS) is a developmental and epileptic encephalopathy with onset in the first year of life. At onset, the child displays normal development, but during the second year of life, stagnation/slowing of neurodevelopment is seen. In addition to difficulties with intellectual development, many children display behavioral problems including autistic features, and difficulties with attention and hyperactivity., Aim: The aim of the present study was to systematically review studies that have focused on the prevalence of cognitive/developmental quotients (DQs) consistent with intellectual disability (ID), deficits in adaptive behavior, autism spectrum disorder (ASD), attention-deficit hyperactivity disorder (ADHD), and behavioral difficulties. A secondary aim was to consider possible factors associated with intellectual and behavioral outcomes in individuals with DS., Method: A systematic review using PubMed and Scopus following the Preferred Reporting Items for Systematic Review (PRISMA) guidelines was conducted on the 24th of September 2019. Study quality was rated by two researchers using the National Institutes of Health (NIH) Quality assessment tools., Result: Twenty-nine studies met inclusion criteria. The pooled prevalence of ID was 86% (range across studies: 50%-100%). The pooled prevalence for ASD was 31%. The mean level of adaptive behavior was more than 2 standard deviations (SDs) below average. The prevalence of behavioral difficulties on standardized instruments ranged between 37% and 100%. The only factor consistently associated with lower cognitive scores was age with older children having significantly lower cognitive scores than younger children. For behavioral difficulties, the most consistent association was with low health-related quality of life (HRQoL) with better HRQoL associated with fewer behavioral difficulties. Study quality was almost universally poor or fair - 15/29 studies were rated 'poor', 13/29 studies were 'fair', and 1 was 'good'., Discussion: The prevalence of cognitive scores consistent with ID is very high in DS. Many patients also have significant deficits in adaptive behavior highlighting that the majority of patients with DS will meet criteria for ID. The prevalence of ASD would also appear to be higher than the general population but studies show a wide range. Behavioral difficulties are common, but most studies have not used instruments adapted to patients with low intelligence quotient/DQ (IQ/DQ). No study used standardized instruments to assess ADHD. Few studies have used comprehensive statistical methods to evaluate possible factors associated with worse cognitive and behavioral outcome., Conclusion: It should be routine to screen for cognitive and behavioral difficulties for all patients with DS. There is a need for more robust studies regarding intellectual and behavioral disorders in patients with DS. These should be large population-based or multinational studies that employ standardized instruments., Competing Interests: Declaration of competing interest None of the authors has any conflict of interest to disclose., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Quercetin can affect some pathological manifestations in obesity. The mechanism underlying the presumed therapeutic effect of quercetin is probably related to the influence on the central processes regulating energy homeostasis. Thus, the purpose of this study was to examine the effect of quercetin on the neuromotor and behavioral functions in Zucker (Z) and Wistar (W) rats with genetically and/or diet-induced obesity. Rats of both strains received balanced or high fat and fructose diet (HFCD) in a 62-day experiment or the same diets supplemented with quercetin at the dose of 50 mg/kg body weight per day. The neuromotor function and behavioral responses were examined using the grip strength test, open field test, elevated plus maze test and conditioned passive avoidance response (CPAR) test. The quercetin potentiated a decrease in anxiety in W rats consumed HFCD and this effect was absent in Z rats with a defect in the leptin receptor gene. In contrast, quercetin increased locomotor activity and impaired short-term memory in the CPAR test only in Z rats with the absence of normal leptin reception. Against the background of the identified changes quercetin exerted significant effects on the lipid and nitrogen metabolism indices such as HDL cholesterol, AsAT/AlAT activities ratio, urea level as well as body and fat mass that were different in Z and W rats. The data obtained show that the effects of quercetin on behavior vary significantly between two strains of rat and consequently are mediated by processes of leptin reception., (Copyright © 2019 Elsevier B.V. All rights reserved.)

We compared anxiety, neuromotor, and cognitive functions in mutant rats with different allelic variants of dopamine transporter DAT knockout receiving balanced or excess in fat and fructose diet. The experiments were performed in DAT-/- homozygotes, DAT+/- heterozygotes, and DAT+/+ wild type rats. The genotype of DAT-KO rats was confirmed by restriction analysis of DAT gene compared to behavioral responses in the open field test (OF). Animals in the first groups of each strain were fed a balanced AIN93M diet; and those in the second groups with a high-fat/high-fructose diet. Neuromotor function was studied as grip strength, and behavioral responses were assessed in the elevated plus maze and conditioned passive avoidance response tests. The mass of the internal organs and white and brown fat, as well as selected lipid and nitrogen metabolism parameters in blood plasma were determined at the end of the experiment. DAT-/- had the highest specific grip strength, and showed an increase in initial exploratory activity in comparison with DAT+/- and DAT +/+. The exploratory activity was significantly reduced in the second test compared to the first one in DAT-/- and DAT+/- of first but not second group. Anxiety decreased with age in the second groups of DAT+/- and DAT+/+ (but not in DAT-/-) and was higher in DAT+/+ than in DAT+/- and DAT-/-. Excess fat and fructose resulted in the deterioration of short-term memory in DAT+/+. Lipidomic indices of blood plasma were less responsive to diet in DAT-/- and DAT-/+ in comparison to DAT+/+. The increased AsAT/AlAT activity ratio in DAT-/- compared with those in DAT+/+ suggests the activation of catabolism activity in the mutants. The consumption of excess fat and fructose significantly modified the effects produced by DAT gene allelic variants presumably due to the influence on the processes of dopamine metabolism., (© 2019 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of The Physiological Society and the American Physiological Society.)

The datasets and code presented in this article are related to the research article entitled "Comprehensiveness of conservation of useful wild plants: an operational indicator for biodiversity and sustainable development targets" 1 . The indicator methodology includes five main steps, each requiring and producing data, which are fully described and available here. These data include: species taxonomy, uses, and general geographic information (dataset 1); species occurrence data (dataset 2); global administrative areas data (dataset 3); eco-geographic predictors used in species distribution modeling (dataset 4); a world map raster file (dataset 5); species spatial distribution modeling outputs (dataset 6); ecoregion spatial data used in conservation analyses (dataset 7); protected area spatial data used in conservation analyses (dataset 8); and countries, sub-regions, and regions classifications data (dataset 9). These data are available at http://dx.doi.org/10.17632/2jxj4k32m2.1. In combination with the openly accessible methodology code (https://github.com/CIAT-DAPA/UsefulPlants-Indicator), these data facilitate indicator assessments and serve as a baseline against which future calculations of the indicator can be measured. The data can also contribute to other species distribution modeling, ecological research, and conservation analysis purposes.

During coal surface mining, several activities such as drilling, blasting, loading, and transport produce large quantities of particulate matter (PM) that is directly emitted into the atmosphere. Occupational exposure to this PM has been associated with an increase of DNA damage, but there is a scarcity of data examining the impact of these industrial operations in cytogenetic endpoints frequency and cancer risk of potentially exposed surrounding populations. In this study, we used a Geographic Information Systems (GIS) approach and Inverse Distance Weighting (IDW) methods to perform a spatial and statistical analysis to explore whether exposure to PM 2.5 and PM 10 pollution, and additional factors, including the enrichment of the PM with inorganic elements, contribute to cytogenetic damage in residents living in proximity to an open-pit coal mining area. Results showed a spatial relationship between exposure to elevated concentrations of PM 2.5, PM 10 and micronuclei frequency in binucleated (MNBN) and mononucleated (MNMONO) cells. Active pits, disposal, and storage areas could be identified as the possible emission sources of combustion elements. Mining activities were also correlated with increased concentrations of highly enriched elements like S, Cu and Cr in the atmosphere, corroborating its role in the inorganic elements pollution around coal mines. Elements enriched in the PM 2.5 fraction contributed to increasing of MNBN but seems to be more related to increased MNMONO frequencies and DNA damage accumulated in vivo. The combined use of GIS and IDW methods could represent an important tool for monitoring potential cancer risk associated to dynamically distributed variables like the PM., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Biochemical, vitamin, trace element and immunological changes were searched for the combined nutritional deficiency of vitamins B1, B2, B6 on in vivo models in rats and mice. Female rats of Wistar (W) strain and hybrids of the 1st generation of Dark Aguti and Wistar (DA x W) strains, female mice of BALB/c strain and DBCB tetrahybrids were used in experiment. Animals received for 35 days a balanced diet (control) according to AIN-93 or a similar diet with the exception of vitamins B1, B2, B6 (experimental groups). The content of vitamins B1, B2 in liver, riboflavin blood plasma level and urinary excretion of thiamine, riboflavin and 4-pyridoxic acid were determined, as well as in rats: blood and liver content of α-tocopherol and retinol, blood biochemical indices of lipid and nitrogen metabolism, activity of cytochrome P isoforms-450 (CYP) in liver; in mice: the circulating levels of pro- and anti-inflammatory cytokines of blood plasma, in animals of both species - the content of essential and toxic elements in the kidneys. DAxW rats compared to W and DBCB mice compared to BALB/c were more sensitive to the development of B-vitamin deficiency judging by the B-vitamin status indicators. In the rats of the experimental groups, there were signs of a deterioration in blood and liver levels of vitamin E, multidirectional shifts in vitamin A sufficiency, increased activity of the CYP3A isoform (6β-TG), a decrease in triglycerides, total protein and albumin fraction levels with an increase in urea level. Manifestation degree of these effects depended on the choice of the animal's line. In mice, the B-vitamin deficiency was characterized by an increase in the levels of proinflammatory cytokines TNF-α, IL-10, IL-Ιβ, IL-6 and a decrease in IFN-γ and IL-17A. The content of magnesium, copper, zinc, chromium and silver was lowered, of cesium - was increased in the kidneys of the rats of the experimental groups. In mice, B-vitamin deficiency resulted in diminishment of magnesium, copper, zinc, chromium, selenium, cadmium and lead content, excess accumulation of cobalt and cesium. Some of these biomarkers are supposed to be used in pre-clinical evaluation of the effectiveness of new vitamin complexes, specialized foods and dietary supplements, as well as studies of interactions of various vitamins., Competing Interests: The authors declare no overt and potential conflict of interest related to the publication of this article., (Copyright© GEOTAR-Media Publishing Group.)

Introduction: We review outcomes after management of meatal balanitis xerotica obliterans (BXO). The primary outcome was recurrent meatal BXO., Methods: A database comprising mostly hypospadias patients was queried for meatal BXO. The disease was confirmed histologically in all cases. Management included topical steroids and/or immunosuppressants, and/or surgical excision of BXO with two-stage oral mucosa graft circumferential replacement urethroplasty., Results: A total of 12 patients had meatal BXO (8 boys and 4 adults). Of these, 10 had hypospadias, two presenting without prior surgery and eight returning 5-30 years after one or multiple (n = 2) repairs. Another two boys did not have hypospadias: one developing BXO 10 years after newborn circumcision and the other having persistent meatal BXO following therapeutic circumcision. Topical and intraluminal steroids (1% betamethasone or clobetasol) and tacrolimus were used for ≥12 weeks each as primary therapy or for meatal recurrence in a total of six cases. Complete response with resolution of white discoloration and relief of stranguria only occurred in two of the three receiving clobetasol, with follow up ≤12 weeks. BXO excision and urethroplasty was done in 11 patients, 10 using oral mucosa grafts; one with a focal lesion and a negative frozen section had reoperative TIP. Of the 10 undergoing excision with two-stage replacement urethroplasty, six remain disease free at a mean follow-up of 23 months (8-48 months), and four had recurrent stranguria and visible meatal BXO at a median of 26 months (22-105 months). Three of the four with recurrences had additional treatment and one was lost to follow-up. All initially had topical steroids, and two also used tacrolimus, without clinical resolution. These three then underwent a second BXO excision and two-stage oral graft replacement urethroplasty. In two recurrences, BXO was found invading from the meatus proximally within oral mucosa (Figure). Of these three with secondary urethroplasties, two are free of disease at 6 and 18 months, and the third had another meatal recurrence 6 months after the second stage., Discussion: We found topical steroids and immunosuppressants to have limited efficacy, with two clinical complete responses achieved only with clobetasol in patients with short follow-up. Forty percent of patients recurred at 2-9 years after visually complete BXO excision and two-stage oral mucosa graft replacement urethroplasty, and in two cases disease invaded into oral mucosa, the first well-documented cases of this occurrence., (Copyright © 2016 Journal of Pediatric Urology Company. Published by Elsevier Ltd. All rights reserved.)

Background: There is a need for improved clinical identification of hospitalized patients at risk of aspiration. We evaluated our novel phonetic test in a broad spectrum of patients at risk of aspiration in the ICU or intermediate care unit., Methods: We prospectively enrolled 60 hospitalized patients with aspiration risk, between December 2009 and September 2011, who subsequently underwent audio-recorded three-component phonetic bedside evaluation. The recordings were scored by two blinded speech-language pathologists. The institutional dysphagia admission screening test was performed by a bedside nurse. The primary outcomes, dysphagia and aspiration, were assessed by a videofluoroscopic swallowing study, fiber-optic endoscopic evaluation of swallowing, or both. We assessed the short- and long-term clinical outcomes (length of stay, subsequent aspiration pneumonia and respiratory failure, survival) and how these were associated with the phonetic and swallow assessments., Results: Statistically significant linear associations with dysphagia were noted for all three individual phonetic components. Also, there were statistically significant linear associations with aspiration for diadochokinesis (P = .050) and consensus auditory-perceptual evaluation of voice (P = .025). Diadochokinesis alone predicted dysphagia (area under the curve [AUC], 0.74; P = .001) and aspiration (AUC, 0.67; P = .012). Its predictive ability improved when combined with normalized dysphagia admission screening test results (AUC, 0.79; P = .001). The short- and long-term clinical outcomes were adversely affected by the worse phonetic/swallowing scores, although they were not statistically different., Conclusions: Abnormal phonation among ICU and intermediate care unit patients is associated with dysphagia and aspiration. Future investigative efforts should uncover the most effective combination of evaluations for accurate bedside detection of dysphagia and aspiration risk in a broad spectrum of patients., (Copyright © 2016 American College of Chest Physicians. Published by Elsevier Inc. All rights reserved.)

Anomalous origin of the left coronary artery from the pulmonary artery (ALCAPA) is a rare congenital condition. It responds well to early diagnosis and treatment, but otherwise the prognosis is poor. We present our case series of 12 patients (mean age, 2 ± 2.58 yr; age range, 2 mo-8 yr), emphasizing the diagnostic process and discussing our surgical results. The diagnosis of ALCAPA should be suspected in infants who have dilated cardiomyopathy with electrocardiographic changes that suggest ischemia, and in older children who have isolated mitral regurgitation. When clinical suspicion is high, the results of 2-dimensional echocardiography combined with color-flow Doppler studies in expert hands can establish the diagnosis, thus avoiding angiography in critically ill infants. The treatment of choice in our patients was transfer and reimplantation of the left coronary artery onto the ascending aorta. There were 2 deaths: both were infants in extremis who underwent emergency surgery. An older child with severe ventricular dysfunction was given mechanical ventricular assistance and then heart transplantation. As of this report, all 10 survivors remained well and asymptomatic.

AMP kinase (AMPK) plays an important role in the regulation of energy metabolism in cardiac cells. Furthermore, activation of AMPK protects the heart from myocardial infarction and heart failure. The present study examines whether or not AMPK affects the peroxisome proliferator-activated receptor-α (PPARα)/mitochondria pathway in response to acute oxidative stress in cultured cardiomyocytes. Cultured H9c2 rat embryonic cardioblasts were exposed to H2O2-induced acute oxidative stress in the presence or absence of metformin, compound C (AMPK inhibitor), GW6471 (PPARα inhibitor), or A-769662 (AMPK activator). Results showed that AMPK activation by metformin reverted oxidative stress-induced inactivation of AMPK and prevented oxidative stress-induced cell death. In addition, metformin attenuated reactive oxygen species generation and depolarization of the inner mitochondrial membrane. The antioxidative effects of metformin were associated with the prevention of mitochondrial DNA damage in cardiomyocytes. Coimmunoprecipitation studies revealed that metformin abolished oxidative stress-induced physical interactions between PPARα and cyclophilin D (CypD), and the abolishment of these interactions was associated with inhibition of permeability transition pore formation. The beneficial effects of metformin were not due to acetylation or phosphorylation of PPARα in response to oxidative stress. In conclusion, this study demonstrates that the protective effects of metformin-induced AMPK activation against oxidative stress converge on mitochondria and are mediated, at least in part, through the dissociation of PPARα-CypD interactions, independent of phosphorylation and acetylation of PPARα and CypD., (Copyright © 2015 the American Physiological Society.)