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Abstract Pollinator-driven evolution of floral traits is thought to be a major driver of angiosperm speciation and diversification. Ophrys orchids mimic female insects to lure male pollinators into pseudocopulation. This strategy, called sexual deception, is species-specific, thereby providing strong premating reproductive isolation. Identifying the genomic architecture underlying pollinator adaptation and speciation may shed light on the mechanisms of angiosperm diversification. Here, we report the 5.2 Gb chromosome-scale genome sequence of Ophrys sphegodes. We find evidence for transposable element expansion that preceded the radiation of the O. sphegodes group, and for gene duplication having contributed to the evolution of chemical mimicry. We report a highly differentiated genomic candidate region for pollinator-mediated evolution on chromosome 2. The Ophrys genome will prove useful for investigations into the repeated evolution of sexual deception, pollinator adaptation and the genomic architectures that facilitate evolutionary radiations.

Summary: Despite extensive global research into genetic predisposition for severe COVID-19, knowledge on the role of rare host genetic variants and their relation to other risk factors remains limited. Here, 52 genes with prior etiological evidence were sequenced in 1,772 severe COVID-19 cases and 5,347 population-based controls from Spain/Italy. Rare deleterious TLR7 variants were present in 2.4% of young (

Abstract BackgroundChemsex prevalence is still not well known, and both the lack of homogeneity and cultural component of chemsex practices are usually overlooked. ObjectiveThis study aims to estimate the proportion of sexual minority men (SMM) engaging in chemsex sessions, while understanding the cultural dimension of chemsex, and to analyze distinct session typologies with potential risk differences and the sociodemographic factors associated with engaging in them. MethodsA total of 5711 SMM residing throughout Spain participated in an anonymous web-based survey that assessed chemsex session engagement and characteristics, drug use, and sociodemographic variables. We measured the association of sociodemographic factors with engaging in chemsex sessions by calculating adjusted prevalence ratios, using multivariate Poisson regression analysis. Chemsex typologies were analyzed using latent class analysis, and sociodemographic factors were associated with the different risk classes. ResultsOur results determined that 21.1% (1205/5711; 95% CI 20.0%‐22.1%) of SMM engaged in chemsex sessions during their lifetime. Participating in sessions was significantly associated with being a migrant, not having a comfortable financial situation, openly living their sexuality, residing in bigger municipalities, older age, using steroids, and living with HIV (adjusted prevalence ratio: range 1.17-2.01; all P ConclusionsChemsex is relatively common among SMM in Spain, but it is important to differentiate typologies of sessions with varying risks, and the proportion of SMM engaging in high-risk sessions is low. Chemsex is highly associated with sociodemographic factors. Chemsex should be prioritized in public health programs, which should consider the different forms of sessions with their varying risks and prevalence, while also considering the cultural dimension inherent to chemsex.

Reflexionar sobre la materialidad, las texturas, la experiencia táctil, los colores, el peso de la materia y su poética, se ha vuelto imprescindible para un considerable número de artistas. Explorar procesos y materiales de manera no convencional es esencial para generar resultados innovadores. También lo es relacionarse con otras disciplinas y culturas, pues nos brinda la oportunidad de conocer nuevos materiales y nos ofrece infinitas posibilidades para crear. Nos enfrentamos a obras abiertas a una modificación constante, donde la experimentación y el proceso a lo largo de toda la elaboración, adquieren más importancia que la pieza acabada. Palpar la materia, interactuar con las herramientas, jugar, hacer, reflexionar, observar, deshacer y rehacer, constituyen una manera de abordar la práctica creativa desde la acción. Al mismo tiempo, el arte puede convertirse en el contexto idóneo para repensar las formas de producir, donde la elección de materiales respetuosos con el medio ambiente sea algo fundamental en un mundo preocupado por la sostenibilidad. Una estrategia de producción que establece un diálogo entre la práctica y el pensamiento, entre el procedimiento y el concepto, donde es esencial comprender y explorar los materiales, permitiéndoles que nos cautiven y estimulen nuestra imaginación

Abstract Lipid nanoparticles have demonstrated utility in hepatic delivery of a range of therapeutic modalities and typically deliver their cargo via low-density lipoprotein receptor-mediated endocytosis. For patients lacking sufficient low-density lipoprotein receptor activity, such as those with homozygous familial hypercholesterolemia, an alternate strategy is needed. Here we show the use of structure-guided rational design in a series of mouse and non-human primate studies to optimize a GalNAc-Lipid nanoparticle that allows for low-density lipoprotein receptor independent delivery. In low-density lipoprotein receptor-deficient non-human primates administered a CRISPR base editing therapy targeting the ANGPTL3 gene, the introduction of an optimized GalNAc-based asialoglycoprotein receptor ligand to the nanoparticle surface increased liver editing from 5% to 61% with minimal editing in nontargeted tissues. Similar editing was noted in wild-type monkeys, with durable blood ANGPTL3 protein reduction up to 89% six months post dosing. These results suggest that GalNAc-Lipid nanoparticles may effectively deliver to both patients with intact low-density lipoprotein receptor activity as well as those afflicted by homozygous familial hypercholesterolemia.

ABSTRACT Purpose: To evaluate the effects of the experimental subcutaneous Walker-256 tumor and L-glutamine supplementation, an antioxidant, on the glomerular morphology of rats. Methods: Twenty Wistar rats were distributed into four groups (n = 5): control (C); control treated with 2% L-glutamine (CG); rats with Walker-256 tumor (WT); and rats with Walker-256 tumor treated with 2% L-glutamine (WTG). Renal histological samples were submitted to periodic acid-Schiff and Masson’s Trichrome staining to analyze glomerular density, morphometry of glomerular components and glomerulosclerosis; and to immunohistochemistry for fibroblast growth factor-2 (FGF-2). Results: WT showed 50% reduction in body mass gain and cachexia index > 10%, while WTG demonstrated reduction in cachexia (p < 0.05). WT revealed reduction of glomerular density, increase in the glomerular tuft area, mesangial area, matrix in the glomerular tuft, decrease in the urinary space and synechia, and consequently higher glomerulosclerosis (p < 0.05). L-glutamine supplementation in the WTG improved glomerular density, and reduced glomerular tuft area, urinary space, mesangial area, and glomerulosclerosis compared to WT(p < 0.05). WT showed higher collagen area and FGF-2 expression compared to C (p < 0.05). WTG presented lower collagen fibers and FGF-2 expression compared to WT (p < 0.05). Conclusions: L-glutamine supplementation reduced cachexia and was beneficial for glomerular morphology of the rats, as well as it reduced kidney damage and improved the remaining glomeruli morphology.

Abstract Background There has been an increase in the integration of qualitative studies in randomised controlled trials. The purpose of this article is to reflect on our experience of carrying out a rapid qualitative study during a feasibility trial of goal-directed fluid therapy (GDFT) in patients with acute pancreatitis, including our sharing of emerging findings and the use of these findings by the trial team. Methods The study was designed as a rapid feedback evaluation and combined interviews with staff and patients who took part in the trial. Findings The rapid qualitative study pointed to common problems in trial recruitment among multiple sites, where lack of engagement of clinical teams across sites might impact negatively on patient recruitment. The article describes how the use of rapid feedback loops can be used as the trial is ongoing to inform changes in implementation. It also covers the potential challenges of working rapidly and collaborative with the trial team. Conclusions Rapid feedback evaluations can be used to generate findings across all stages of trial design and delivery. Additional research is required to explore the implementation of this research design in other settings and trial designs.

ObjectiveThe management of cardiotoxicity concerning the use of oral antineoplastic agents (OAAs) is a challenge for healthcare professionals. Our objective was to create a comprehensive medication management guide with dose adjustment recommendations on OAAs concerning cardiotoxic and lipid metabolic adverse events (AEs) to assist healthcare professionals when prescribing OAAs.Materials and methodsA review of the available information on all dose adjustments necessary to safely prescribe and dispense OAAs concerning cardiotoxicity was conducted. In January 2023, we identified all OAAs authorized by the European Medicines Agency (EMA). For each drug, the latest summary of product characteristics (SPC) approved by the EMA and the tertiary data source Lexicomp® were reviewed. Cardiotoxic AEs were recorded, namely, QT interval prolongation, decrease in left ventricular ejection fraction (LVEF), imbalances in blood pressure (hypertension and hypotension), alterations in heart rate (tachycardia and bradycardia), and thrombosis. Any available dose adjustment recommendations in case of an occurrence of these adverse events were collected.ResultsIn all, 93 different OAAs had been approved by the EMA and were reviewed. Among them, 51.6% have recognized cardiotoxic AEs and 10.8% can cause alterations in lipid metabolism. A total of 27 (29.0%) OAAs had specific recommendations regarding QT prolongation; 88.9% were listed in the SPC and 59.3% in Lexicomp®. Eight OAAs (9.68%) have reported a decrease in LVEF, and four of these drugs, namely, encorafenib, lorlatinib, ripretinib, and sunitinib, have specific management recommendations. Almost half (49.5%) of currently approved OAAs can potentially alter blood pressure; 34 (36.6%) of them have been reported to cause hypertension and 12 (12.9%) are related to hypotension. Tachycardia and/or bradycardia are associated with 22.6% and 8.6% of the evaluated drugs, respectively. Regarding thrombosis, 30 (32.3%) of the drugs analyzed included the appearance of a thrombus as a possible AE.ConclusionsMore than half of the OAAs can produce cardiotoxic effects, with the most frequent being blood pressure alteration and QT interval prolongation with a non-depreciable incidence of LV dysfunction or thrombosis. Before starting the treatment, it is necessary to stratify baseline cardiovascular risk, plan a surveillance schedule, and consider referral to cardio-oncology units.

Abstract The use of remote measurement technologies (RMTs) across mobile health (mHealth) studies is becoming popular, given their potential for providing rich data on symptom change and indicators of future state in recurrent conditions such as major depressive disorder (MDD). Understanding recruitment into RMT research is fundamental for improving historically small sample sizes, reducing loss of statistical power, and ultimately producing results worthy of clinical implementation. There is a need for the standardisation of best practices for successful recruitment into RMT research. The current paper reviews lessons learned from recruitment into the Remote Assessment of Disease and Relapse- Major Depressive Disorder (RADAR-MDD) study, a large-scale, multi-site prospective cohort study using RMT to explore the clinical course of people with depression across the UK, the Netherlands, and Spain. More specifically, the paper reflects on key experiences from the UK site and consolidates these into four key recruitment strategies, alongside a review of barriers to recruitment. Finally, the strategies and barriers outlined are combined into a model of lessons learned. This work provides a foundation for future RMT study design, recruitment and evaluation.

In Rwanda, cattle and milk hold a cultural and historical significance, providing an opportunity for pro-dairy governmental policies aimed to alleviate food insecurity, malnutrition, and improve livelihoods. The government of Rwanda has identified strategies to grow the dairy sector through strategic investment to achieve these goals. It is estimated two-thirds of lactating cows in Rwanda have clinical or subclinical mastitis, which reduces milk production and increases the risk of milk as a source for zoonotic disease if the milk is consumed undercooked or unpasteurized. This case study outlines the implementation of a One Health framework that integrates education, research, and outreach in Rwanda to improve food safety and food security, for the social, economic, and health benefit of Rwandans and their livestock. Twenty-five Rwandan Extension Specialists participated in the Dairy Dynamic Management education, research, and outreach program. Once trained, the extension specialists supported 30 small holder dairy farmers in performing proper husbandry and animal health practices for mastitis control and reduction of bacterial counts in the udder. Over the 16-week program, 30 small holder dairy farmers and 100 dairy cows were surveyed weekly for animal husbandry, animal health, and mastitis indicators. Outcomes were evaluated by monitoring animal health, foodborne pathogens in milk, and compliance to animal husbandry protocols. Quarter milk samples were collected weekly and evaluated for the presence of bacteria that are common causes of mastitis. We found a statistically significant reduction of mean total bacterial counts and prevalence of bacterial species in quarters over the 16-week training (P ≤ .01). Smallholders were monitored through observing farmers performing hygienic milking protocols. Farmers conducted the protocol correctly greater than 90% of the time by the end of the 16-week program for 5 of 7 steps for proper hygienic milking procedures, indicating farmers were eager to learn and adopt the procedures. However, follow-up and retraining with Extension Specialists is vital to continued success. We demonstrate that an integrative One Health education, research, and outreach program can be successful in improving animal health, food safety, and food security and this framework can be applied to other agricultural sectors and geographic regions.

The aim of this study was to determine whether the soil faunal-microbial interaction complexity (SFMIC) is a significant factor influencing the soil microbial communities and the willow growth in the context of PAH contamination. The SFMIC treatment had eight levels: just the microbial community, or the microbial community with nematodes, springtails, earthworms and all the possible combinations. SFMIC affected the height and biomass of willows after eight weeks or growth. SFMIC affected the structure and the composition of the bacterial, archaeal and fungal communities, with significant effects of SFMIC on the relative abundance of fungal genera such as Sphaerosporella, a known willow symbiont during phytoremediation, and bacterial phyla such as Actinobacteriota, containing many polycyclic aromatic hydrocarbons (PAH) degraders. These SFMIC effects on microbial communities were not clearly reflected in the community structure and abundance of PAH degraders, even though some degraders related to Actinobacteriota and the diversity of Gram-negative degraders were affected by the SFMIC treatments. Over 95% of PAH was degraded in all pots at the end of the experiment. Overall, our results suggest that, under our experimental conditions, SFMIC changes willow phytoremediation outcomes.

El propósito fue comprender en qué medida los Proyecto de Inversión Pública realizados se relacionan en el desarrollo de las actividades turísticas en la Provincia de Huánuco - 2015. El método fue cuantitativa, aplicada, deductiva, de diseño no experimental- corte transversal, la muestra se conformó por 167 PIP de la zona estudio. Se tuvo en cuenta los siguientes instrumentos: las fichas de registros, referencia electrónica y la observación. La validación estadística se dio mediante el alfa de Cronbach, con 96 % de confiabilidad. Del total de PIP realizados, según el grado de evaluación de influencia del desarrollo del turismo, se presenta un grado bajo y medio. Del total de proyectos, según las evaluaciones del grado de influencia en el desarrollo del turismo, se obtuvo un grado medio y alto. Del total de proyectos ejecutados por los gobiernos locales, según la evaluación del grado de influencia en el desarrollo del turismo, tienen un grado medio y alto. Se concluye que los PIP influyen en el crecimiento de las actividades turísticas.

IntroductionAstrocytic Aquaporin 4 (AQP4) and Transient receptor potential vanilloid 4 (TRPV4) channels form a functional complex that likely influences cell volume regulation, the development of brain edema, and the severity of the ischemic injury. However, it remains to be fully elucidated whether blocking these channels can serve as a therapeutic approach to alleviate the consequences of having a stroke.Methods and resultsIn this study, we used in vivo magnetic resonance imaging (MRI) to quantify the extent of brain lesions one day (D1) and seven days (D7) after permanent middle cerebral artery occlusion (pMCAO) in AQP4 or TRPV4 knockouts and mice with simultaneous deletion of both channels. Our results showed that deletion of AQP4 or TRPV4 channels alone leads to a significant worsening of ischemic brain injury at both time points, whereas their simultaneous deletion results in a smaller brain lesion at D1 but equal tissue damage at D7 when compared with controls. Immunohistochemical analysis 7 days after pMCAO confirmed the MRI data, as the brain lesion was significantly greater in AQP4 or TRPV4 knockouts than in controls and double knockouts. For a closer inspection of the TRPV4 and AQP4 channel complex in the development of brain edema, we applied a real-time iontophoretic method in situ to determine ECS diffusion parameters, namely volume fraction (α) and tortuosity (λ). Changes in these parameters reflect alterations in cell volume, and tissue structure during exposure of acute brain slices to models of ischemic conditions in situ, such as oxygen-glucose deprivation (OGD), hypoosmotic stress, or hyperkalemia. The decrease in α was comparable in double knockouts and controls when exposed to hypoosmotic stress or hyperkalemia. However, during OGD, there was no decrease in α in the double knockouts as observed in the controls, which suggests less swelling of the cellular components of the brain.ConclusionAlthough simultaneous deletion of AQP4 and TRPV4 did not improve the overall outcome of ischemic brain injury, our data indicate that the interplay between AQP4 and TRPV4 channels plays a critical role during neuronal and non-neuronal swelling in the acute phase of ischemic injury.

Cells convert electrical signals into chemical outputs to facilitate the active transport of information across larger distances. This electrical-to-chemical conversion requires a tightly regulated expression of ion channels. Alterations of ion channel expression provide landmarks of numerous pathological diseases, such as cardiac arrhythmia, epilepsy, or cancer. Although the activity of ion channels can be locally regulated by external light or chemical stimulus, it remains challenging to coordinate the expression of ion channels on extended spatial–temporal scales. Here, we engineered yeast Saccharomyces cerevisiae to read and convert chemical concentrations into a dynamic potassium channel expression. A synthetic dual-feedback circuit controls the expression of engineered potassium channels through phytohormones auxin and salicylate to produce a macroscopically coordinated pulses of the plasma membrane potential. Our study provides a compact experimental model to control electrical activity through gene expression in eukaryotic cell populations setting grounds for various cellular engineering, synthetic biology, and potential therapeutic applications.

Background and aim: Genome wide association studies have scaled up both in terms of sample size and range of complex disorders investigated, but these have explained relatively little phenotypic variance. Of the several reasons, phenotypic heterogeneity seems to be a likely contributor for missing out genetic associations of large effects. Ayurveda, the traditional Indian system of medicine is one such tool which adopts a holistic deep phenotyping approach and classifies individuals based on their body constitution/prakriti. We hypothesized that Ayurveda based phenotypic stratification of healthy and diseased individuals will allow us to achieve much desired homogeneous cohorts which would facilitate detection of genetic association of large effects. In this proof of concept study, we performed a genome wide association testing of clinically diagnosed rheumatoid arthritis patients and healthy controls, who were re-phenotyped into Vata, Pitta and Kapha predominant prakriti sub-groups. Experimental procedure: Genotypes of rheumatoid arthritis cases (Vata = 49; Pitta = 117; Kapha = 78) and controls (Vata = 33; Pitta = 175; Kapha = 85) were retrieved from the total genotype data, used in a recent genome-wide association study performed in our laboratory. A total of 528461 SNPs were included after quality control. Prakriti-wise genome-wide association analysis was employed. Results and conclusion: This study identified (i) prakriti-specific novel disease risk genes of high effect sizes; (ii) putative candidates of novel therapeutic potential; and (iii) a good correlation between genetic findings and clinical knowledge in Ayurveda. Adopting Ayurveda based deep phenotyping may facilitate explaining hitherto undiscovered heritability in complex traits and may propel much needed progress in personalized medicine.

Abstract Metastasis is the process of cancer cell dissemination from primary tumors to different organs being the bone the preferred site for metastatic homing of prostate cancer (PCa) cells. Prostate tumorigenesis is a multi-stage process that ultimately tends to advance to become metastatic PCa. Once PCa patients develop skeletal metastases, they eventually succumb to the disease. Therefore, it is imperative to identify essential molecular drivers of this process to develop new therapeutic alternatives for the treatment of this devastating disease. Here, we have identified MAP4K4 as a relevant gene for metastasis in PCa. Our work shows that genetic deletion of MAP4K4 or pharmacological inhibition of its encoded kinase, HGK, inhibits metastatic PCa cells migration and clonogenic properties. Hence, MAP4K4 might promote metastasis and tumor growth. Mechanistically, our results indicate that HGK depleted cells exhibit profound differences in F-actin organization, increasing cell spreading and focal adhesion stability. Additionally, HGK depleted cells fails to respond to TNF-α stimulation and chemoattractant action. Moreover, here we show that HGK upregulation in PCa samples from TCGA and other databases correlates with a poor prognosis of the disease. Hence, we suggest that it could be used as prognostic biomarker to predict the appearance of an aggressive phenotype of PCa tumors and ultimately, the appearance of metastasis. In summary, our results highlight an essential role for HGK in the dissemination of PCa cells and its potential use as prognostic biomarker.

Summary: Repeat elements can be dysregulated at a genome-wide scale in human diseases. For example, in Ewing sarcoma, hundreds of inert GGAA repeats can be converted into active enhancers when bound by EWS-FLI1. Here we show that fusions between EWS and GGAA-repeat-targeted engineered zinc finger arrays (ZFAs) can function at least as efficiently as EWS-FLI1 for converting hundreds of GGAA repeats into active enhancers in a Ewing sarcoma precursor cell model. Furthermore, a fusion of a KRAB domain to a ZFA can silence GGAA microsatellite enhancers genome wide in Ewing sarcoma cells, thereby reducing expression of EWS-FLI1-activated genes. Remarkably, this KRAB-ZFA fusion showed selective toxicity against Ewing sarcoma cells compared with non-Ewing cancer cells, consistent with its Ewing sarcoma-specific impact on the transcriptome. These findings demonstrate the value of ZFAs for functional annotation of repeats and illustrate how aberrant microsatellite activities might be regulated for potential therapeutic applications.

The correct interpretation of relative abundance indices provided by different sampling methods is essential to correctly estimate population size. Although multiple indices and models have been proposed, their ability to estimate relative abundances and their performance in models explaining abundance trends remains unclear. We used the red fox (Vulpes vulpes) as a model species to compare the relationship and derived models of relative abundance between three indices of relative abundance: RAI (number of captures/total occasions); NI (number of photo-identified individuals) obtained by camera-trapping, and NSE (number of segments with scats) obtained by the scat census sampling method. In addition, we modelled the relationship between a set of habitat predictors and fox relative abundance for each of the three estimated relative abundance indices. We compared the relative abundance models explained for each index against N-mixture models that estimate abundance controlled for variation in detection. Results showed a positive correlation between the RAI and NI indices, while both indices showed a negative relationship with the NSE index. Relative abundance models and N-Mixture models showed a different selection of predictors to explain abundance trends. NSE and RAI indices selected predictors that could explain variability in fox detection rather than fox abundance. In contrast, the NI index and N-Mixture models selected the same predictors to explain fox abundance. Our results suggest the use of the NI index for abundance models without the need to control for variation in detection. Relative abundance indices based on scats and captures per occasion are suboptimal indices for species abundance studies due to possible bias caused by animal behaviour. If count-based methods on captures per occasion (RAI) are selected, we suggest using session-based data processing to incorporate detectability variation in N-mixture models.

Most individuals affected with DYT1 dystonia have a heterozygous 3-bp deletion in the TOR1A gene (c.907_909delGAG). The mutation appears to act through a dominant-negative mechanism compromising normal torsinA function, and it is proposed that reducing mutant torsinA may normalize torsinA activity. In this study, we used an engineered Cas9 variant from Streptococcus pyogenes (SpCas9-VRQR) to target the mutation in the TOR1A gene in order to disrupt mutant torsinA in DYT1 patient fibroblasts. Selective targeting of the DYT1 allele was highly efficient with most common non-homologous end joining (NHEJ) edits, leading to a predicted premature stop codon with loss of the torsinA C terminus (delta 302–332 aa). Structural analysis predicted a functionally inactive status of this truncated torsinA due to the loss of residues associated with ATPase activity and binding to LULL1. Immunoblotting showed a reduction of the torsinA protein level in Cas9-edited DYT1 fibroblasts, and a functional assay using HSV infection indicated a phenotypic recovery toward that observed in control fibroblasts. These findings suggest that the selective disruption of the mutant TOR1A allele using CRISPR-Cas9 inactivates mutant torsinA, allowing the remaining wild-type torsinA to exert normal function.

Psychopathy is a personality disorder characterized by a mix of traits belonging to four facets: affective (e.g., callous/lack of empathy), interpersonal (e.g., grandiosity), behavioral instability (e.g., impulsivity, poor behavioral controls), and social deviance (e.g., juvenile delinquency, criminal versatility). Several scholars have argued that early childhood maltreatment impacts the development of psychopathy, although views regarding its role in the four facets differ. We conducted a meta-analysis including 47 studies comprising a total of 389 effect sizes and 12,737 participants, to investigate the association between psychopathy and four types of child maltreatment: physical abuse, emotional abuse, neglect, and sexual abuse. We found support for a moderate link between overall psychopathy and childhood physical abuse, emotional abuse, and neglect, as well as overall childhood maltreatment. The link between psychopathy and childhood sexual abuse was small, but still statistically significant. These associations were stronger for the behavioral and antisocial facets than for the affective and interpersonal facets of psychopathy, but nearly all associations were statistically significant. Our findings are consistent with recently developed theories on the role of complex trauma in the development of severe personality disorders. Trauma-focused preventive and therapeutic interventions can provide further tests of the trauma-psychopathy hypothesis.

ABSTRACT BACKGROUND: Serotonin (5-HT) is present in the epithelial enterochromaffin cells (EC), mast cells of the lamina propria and enteric neurons. The 5-HT is involved in regulating motility, secretion, gut sensation, immune system and inflammation. OBJECTIVE: Evaluate the effects of diabetes and quercetin supplementation on serotoninergic cells and its cell loss by apoptosis in jejunal mucosa of streptozotocin-induced diabetic rats (STZ-rats). METHODS: Twenty-four male Wistar rats were divided into four groups: normoglycemic (C), normoglycemic supplemented with 40 mg/day quercetin (Q), diabetic (D) and diabetic supplemented with 40 mg/day quercetin (DQ). After 120 days, the jejunum was collected and fixated in Zamboni’s solution for 18 h. After obtaining cryosections, immunohistochemistry was performed to label 5-HT and caspase-3. Quantification of 5-HT and caspase-3 immunoreactive (IR) cells in the lamina propria, villi and crypts were performed. RESULTS: The diabetic condition displayed an increase of the number of 5-HT-IR cells in villi and crypts, while decreased number of these cells was observed in lamina propria in the jejunum of STZ-rats. In the diabetic animals, an increased density of apoptotic cells in epithelial villi and crypts of the jejunum was observed, whereas a decreased number of caspase-3-IR cells was observed in lamina propria. Possibly, quercetin supplementation slightly suppressed the apoptosis phenomena in the epithelial villi and crypts of the STZ-rats, however the opposite effect was observed on the 5-HT-IR cells of the lamina propria. Quercetin supplementation on healthy animals promoted few changes of serotoninergic function and apoptotic stimuli. CONCLUSION: These results suggest that quercetin supplementation mostly improved the serotonergic function affected by diabetes maybe due to antioxidant and anti-inflammatory properties of quercetin.

Abstract Background Recent innovations in single-cell Assay for Transposase Accessible Chromatin using sequencing (scATAC-seq) enable profiling of the epigenetic landscape of thousands of individual cells. scATAC-seq data analysis presents unique methodological challenges. scATAC-seq experiments sample DNA, which, due to low copy numbers (diploid in humans), lead to inherent data sparsity (1–10% of peaks detected per cell) compared to transcriptomic (scRNA-seq) data (10–45% of expressed genes detected per cell). Such challenges in data generation emphasize the need for informative features to assess cell heterogeneity at the chromatin level. Results We present a benchmarking framework that is applied to 10 computational methods for scATAC-seq on 13 synthetic and real datasets from different assays, profiling cell types from diverse tissues and organisms. Methods for processing and featurizing scATAC-seq data were compared by their ability to discriminate cell types when combined with common unsupervised clustering approaches. We rank evaluated methods and discuss computational challenges associated with scATAC-seq analysis including inherently sparse data, determination of features, peak calling, the effects of sequencing coverage and noise, and clustering performance. Running times and memory requirements are also discussed. Conclusions This reference summary of scATAC-seq methods offers recommendations for best practices with consideration for both the non-expert user and the methods developer. Despite variation across methods and datasets, SnapATAC, Cusanovich2018, and cisTopic outperform other methods in separating cell populations of different coverages and noise levels in both synthetic and real datasets. Notably, SnapATAC is the only method able to analyze a large dataset (> 80,000 cells).

In-depth characterization of adeno-associated virus (AAV)-mediated CRISPR delivery is still lacking. Here, the authors show high levels of integration into Cas9-induced double-strand breaks (DSBs) in therapeutically relevant genes in vivo.

Sexually transmitted infections (STIs) are a major public health challenge. Although theoretically informed public health campaigns are more effective for changing behaviour, there is little evidence of their use when campaigns are commissioned to the commercial sector. This study describes the implementation of the COM-B model to a sexual health campaign that brought together expertise from academics, sexual healthcare, and marketing and creative professionals. Insights were gathered following a review of the relevant academic literature. Barriers and facilitators to condom use and STI testing were explored with the use of the COM-B model and the Behaviour Change Wheel in a workshop attended by academics, behavioural scientists, healthcare experts and creative designers. Feedback on the creative execution of the campaign was obtained from healthcare experts and via surveys. Barriers to psychological capability, automatic and reflective motivation, and social opportunity were identified as targets for the campaign, and creative solutions to these barriers were collaboratively devised. The final sexual health campaign was rated positively in its ability to change attitudes and intentions regarding the use of condoms and STI testing. This study describes the implementation of the COM-B model of behaviour change to a public sexual health campaign that brought together academics, public and commercial sector expertise. The barriers and facilitators identified in this collaborative process represent potential targets for future public health communication campaigns.

Timely identification of COVID-19 patients at high risk of mortality can significantly improve patient management and resource allocation within hospitals. This study seeks to develop and validate a data-driven personalized mortality risk calculator for hospitalized COVID-19 patients. De-identified data was obtained for 3,927 COVID-19 positive patients from six independent centers, comprising 33 different hospitals. Demographic, clinical, and laboratory variables were collected at hospital admission. The COVID-19 Mortality Risk (CMR) tool was developed using the XGBoost algorithm to predict mortality. Its discrimination performance was subsequently evaluated on three validation cohorts. The derivation cohort of 3,062 patients has an observed mortality rate of 26.84%. Increased age, decreased oxygen saturation (≤ 93%), elevated levels of C-reactive protein (≥ 130 mg/L), blood urea nitrogen (≥ 18 mg/dL), and blood creatinine (≥ 1.2 mg/dL) were identified as primary risk factors, validating clinical findings. The model obtains out-of-sample AUCs of 0.90 (95% CI, 0.87-0.94) on the derivation cohort. In the validation cohorts, the model obtains AUCs of 0.92 (95% CI, 0.88-0.95) on Seville patients, 0.87 (95% CI, 0.84-0.91) on Hellenic COVID-19 Study Group patients, and 0.81 (95% CI, 0.76-0.85) on Hartford Hospital patients. The CMR tool is available as an online application at covidanalytics.io/mortality_calculator and is currently in clinical use. The CMR model leverages machine learning to generate accurate mortality predictions using commonly available clinical features. This is the first risk score trained and validated on a cohort of COVID-19 patients from Europe and the United States.

The genotype of a single SNP, rs12913832, is the primary predictor of blue and brown eye colours. The genotypes rs12913832:AA and rs12913832:GA are most often observed in individuals with brown eye colours, whereas rs12913832:GG is most often observed in individuals with blue eye colours. However, approximately 3% of Europeans with the rs12913832:GG genotype have brown eye colours. The purpose of the study presented here was to identify variants that explain brown eye colour formation in individuals with the rs12913832:GG genotype. Genes and regulatory regions surrounding SLC24A4, TYRP1, SLC24A5, IRF4, TYR, and SLC45A2, as well as the upstream region of OCA2 within the HERC2 gene were sequenced in a study comprising 40 individuals with the rs12913832:GG genotype. Of these, 24 individuals were considered to have blue eye colours and 16 individuals were considered to have brown eye colours. We identified 211 variants within the SLC24A4, TYRP1, IRF4, and TYR target regions associated with eye colour. Based on in silico analyses of predicted variant effects we recognized four variants, TYRP1 rs35866166:C, TYRP1 rs62538956:C, SLC24A4 rs1289469:C, and TYR rs1126809:G, to be the most promising candidates for explanation of brown eye colour in individuals with the rs12913832:GG genotype. Of the 16 individuals with brown eye colours, 14 individuals had four alleles, whereas the alleles were rare in the blue eyed individuals. rs35866166, rs62538956, and rs1289469 were for the first time found to be associated with pigmentary traits, whilst rs1126809 was previously found to be associated with pigmentary variation. To improve prediction of eye colours we suggest that future eye colour prediction models should include rs35866166, rs62538956, rs1289469, and rs1126809.

Globally, our society faces an enormous challenge to feed, house, and provide a healthy life for the growing human population while preserving the environment and natural resources for the benefit of future generations. In order to meet these challenges, sustainable food production and environmental stewardship is paramount and will require a One Health approach. One Health is the concept that the health of humans, animals, and the environment are inextricably linked. This approach can be applied to food safety, sustainable food production, and environmental stewardship by bringing together interdisciplinary teams to create a One Health network to address these challenges. In order to achieve food security for the global population, preserve natural resources, and improve health through safeguarding food safety, there is a need for increased awareness among academics, producers, consumers, and government agencies in the following topics: (1) One Health Initiatives for Sustainable Food Systems, Food Safety and Food Security, (2) Brief History of Food Safety in the United States, (3) Food Safety in the Twenty First Century: The Need for a New Perspective, (4) Food Security in the Twenty First Century: Disasters and Transboundary Disease. These topics cover the need for incorporating One Health education into curriculum for scientific, engineering, and humanities programs to build capacity in One Health competencies with the goal of established networks that will work toward improving public health, food safety, and sustainable agriculture by establishing new perspectives on interactions among plants, animals, and humans and recognizing the threat of disasters and transboundary diseases to food security.

Purpose – The purpose of this paper is to highlight the importance of the humanitarian supply chain (HSC) as the backbone of the humanitarian operations. It further proposes feasible ways to overcome some of the main supply chain challenges identified by practitioners to achieve cost efficient and effective operations. Design/methodology/approach – The challenges that the HSC faces and proposed changes to overcome them are gathered from interviews with nearly 40 practitioners. Findings – Five critical issues that affect the future of HSCs are identified along with recommendations to address them. Social implications – It supports the fulfillment of the agenda for humanity’s five core responsibilities: global leadership to prevent and end conflict, uphold the norms that safeguard humanity, leave no one behind, change people’s lives – from delivering aid to ending need, and invest in humanity. Originality/value – The original report was presented at the first World Humanitarian Summit in Istanbul in May 2016. The reader can find it via the following link www.logcluster.org/sites/default/files/whs_humanitarian_supply_chain_paper_final_24_may.pdf

Purpose – Following a large-scale disaster, medical assistance is a critical component of the emergency response. The paper aims to discuss this issue. Design/methodology/approach – Academic and practitioner literature is used to develop a framework studying the effectiveness of the humanitarian medical supply chain (HMSC). The framework is validated by using the findings of interviews conducted with experts and the case study of a serious humanitarian medical crisis (Ebola outbreak in 2014). Findings – The factors affecting the effectiveness of the HMSC are identified. Research limitations/implications – To get an expert opinion on the major logistical challenges of the medical assistance in emergencies only 11 interviews with practitioners were conducted. Originality/value – While the existing academic literature discusses the distribution of various supplies needed by the affected population, limited research focuses specifically on studying the HMSC aspect of the response. This paper closes this gap by describing the HMSC in the case of disaster response, and identifying the factors affecting its effectiveness, especially focusing on the factors that are unique to the medical aspect of the humanitarian supply chain.

Loss of muscle tone is a distinguishing feature of paradoxical or REM sleep (PS) and is disrupted in REM sleep behavior disorder. Here the authors report that GABA/glycine inhibitory neurons in the ventromedial medulla are essential for producing PS muscle atonia without affecting PS quantity.

Abstract Ingested fluoride (F) is absorbed mainly in the small intestine, which is controlled by the Enteric Nervous System (ENS). Although important intestinal symptomatology has been described after excessive F exposure, there have been no studies reporting the effects of F on the ENS. In this study, the effects of chronic F exposure were evaluated on the duodenums of rats through proteomic and morphological analyses. Concentrations of 0, 10, or 50 ppm of F were applied to the drinking water for 30 days. Immunofluorescence techniques were performed in the myenteric plexus of the duodenum to detect HuC/D, neuronal nitric oxide (nNOS), vasoactive intestinal peptide (VIP), calcitonin gene related peptide (CGRP), and substance P (SP). The 50 ppm F group presented a significant decrease in the density of nNOS-IR neurons. Significant morphological alterations were also observed in HUC/D-IR and nNOS-IR neurons; VIP-IR, CGRP-IR, and SP-IR varicosities for both groups (10 and 50 ppm F). Proteomic analysis of the duodenum demonstrated alterations in the expression of several proteins, especially those related to important biological processes, such as protein polymerization, which helps to explain the downregulation of many proteins upon exposure to 50 ppm of F.

A doença de Parkinson é uma doença neurodegenerativa caracterizada pela depleção da dopamina, morte dos neurônios dopaminérgicos da substância negra da via nigroestriatal e presença de inclusão neurofibrilares nos neurônios ativos. A substância negra esta interconectada com todas as regiões do cérebro, estas recebem inervações glutamatérgicas e enviam projeções diretas e indiretas, modulando a atividade dos neurônios dopaminérgicos, assim, a perda de dopamina no estriado causa alterações químicas monoaminérgicas levando ao aumento e diminuição da atividade neuronal nas áreas motoras do córtex cerebral. Este estudo tem por objetivo compreender a relação dos neurônios dopaminérgicos com os sintomas clínicos na doença de Parkinson, investigar a disfunção fisiológica e apontar as alterações morfológicas do sistema nervoso central. Foi realizada uma pesquisa descritiva do tipo revisão bibliográfica, através de documentos disponíveis nas bases de dados: LILACS, SCIELO, PUBMED e Google Acadêmico, utilizando-se os seguintes descritores: doença de Parkinson, dopamina, gânglios da base e neurônios dopaminérgicos. Foram selecionados documentos que incluíssem: dissertações, teses, artigos e livros que abordassem o tema, entre os anos de 2008 a 2018, disponíveis na integra nos idiomas português e inglês. Revisamos como a dopamina se adéqua no SNC e os prejuízos causados devido sua disfunção na doença de Parkinson. De forma geral, este estudo mostra a importância do equilíbrio fisiológico entre as vias neuronais, estabelecendo a relação das estruturas cerebrais com os neurotransmissores responsáveis pelo planejamento e execução do movimento juntamente com as alterações físicas que ela provoca.

As the global population approaches 9.7 billion inhabitants by the year 2050, humanity faces enormous challenges to feed, house, and provide basic living requirements for the growing population while preserving the health of wildlife and the ecosystem. Dairy source foods play an important part in providing nutrient and energy dense sources of calories and establishing Bifidobacterium as a keystone species in the gut for positive health outcomes in infants and children. In developed countries, dairy products have a high food safety record when pasteurized and properly processed. However, when milk is consumed unpasteurized, as often occurs in developing countries where regulation and oversight of the dairy industry is lacking, dairy can serve as a vector for zoonotic transmission of disease and can contain adulterants such as antibiotic residues. Here we provide an overview for the importance of dairy source foods for nutrition and with a One Health perspective and discuss the historical events that have resulted in a high standard of dairy food safety in the United States. This review article covers the Origins of One Health, the role of milk in transmission of disease, management practices and regulations to ensure safe dairy products reach consumers, current challenges facing the dairy industry and impacts on public health, and how these standards can be employed in low and middle income countries to improve public health, nutrition and economic benefits to farmers. Keywords: Dairy, Food safety, Dairy production medicine, One health, Public health

PurposeEpigenetic regulating proteins like histone methyltransferases produce variations in several functions, some of them associated with the generation of oncogenic processes. Mutations of genes involved in these functions have been recently associated with cancer, and strategies to modulate their activity are currently in clinical development.MethodsBy using data extracted from the METABRIC study, we searched for mutated genes linked with detrimental outcome in invasive breast carcinoma (n = 772). Then, we used downstream signatures for each mutated gene to associate that signature with clinical prognosis using the online tool "Genotype-2-Outcome" (http://www.g-2-o.com). Next, we performed functional annotation analyses to classify genes by functions, and focused on those associated with the epigenetic machinery.ResultsWe identified KMT2D, SETD1A and SETD2, included in the lysine methyltransferase activity function, as linked with poor prognosis in invasive breast cancer. KMT2D which codes for a histone methyltransferase that acts as a transcriptional regulator was mutated in 6% of triple negative breast tumors and found to be linked to poor survival. Genes regulated by KMT2D included RAC3, KRT23, or KRT14, among others, which are involved in cell communication and signal transduction. Finally, low expression of KMT2D at the transcriptomic level, which mirror what happens when KMT2D is mutated and functionally inactive, confirmed its prognostic value.ConclusionIn the present work, we describe epigenetic modulating genes which are found to be mutated in breast cancer. We identify the histone methyltransferase KMT2D, which is mutated in 6% of triple negative tumors and linked with poor survival.