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Anticoagulant rodenticides (ARs) have a long history of successful use in controlling vertebrate pest and invasive species. Despite regulatory efforts to mitigate risk, non-target wildlife may be unintentionally exposed to ARs through various trophic pathways, and depending on dose, exposure can result in adverse effects and mortality. Second-generation ARs (SGARs) are mixtures of cis- and trans-diastereoisomers (each including two stereoisomers) that exhibit similar in vitro inhibitory potency for vitamin K epoxide reductase in rodent microsomal assay systems. Some diastereoisomers and hence some individual stereoisomers are preferentially metabolized in vivo, resulting in residue patterns in exposed target rodents that differ from the bait formulations. Use of less persistent but equally potent SGAR stereoisomers in baits results in lower tissue residues in target rodents, which in turn constitutes lower risk when consumed by non-target wildlife. The toxicity of two brodifacoum formulations with stereoisomers having markedly different elimination half-lives in rats (Formulation A containing the two least persistent stereoisomers, and Formulation B containing the most persistent stereoisomer) were tested in a 7-day dietary feeding trial with American kestrels. Based on previous kestrel studies using commercially available brodifacoum, Formulations A and B were each provided at three dietary concentrations (0.05, 0.1 and 0.5 µg/g diet, 4 kestrels/dose level) predicted to cause a range of toxicity. Compared to unexposed controls, all kestrels that ingested 0.5 µg/g diet of the longer-lived Formulation B exhibited extreme coagulopathy. In contrast, the 0.5 µg/g diet of the shorter-lived Formulation A yielded only a modest lengthening of clotting time in just 1 of the 4 exposed kestrels. These findings support the notion that SGAR baits enriched with less persistent stereoisomers may pose lower hazard and ultimately risk to non-target wildlife.

Abstract Semen quality and fertility has declined over the last 50 years, corresponding to ever-increasing environmental stressors. However, the cellular mechanisms involved and their impact on sperm functions remain unknown. In a repeated sampling human cohort study, we identify a significant effect of prior perceived stress to increase sperm motility 2-3 months following stress, timing that expands upon our previous studies revealing significant stress-associated changes in sperm RNA important for fertility. We mechanistically examine this post-stress timing in mice using an in vitro stress model in the epididymal epithelial cells responsible for sperm maturation and find 7282 differentially H3K27me3 bound DNA regions involving genes critical for mitochondrial and metabolic pathways. Further, prior stress exposure significantly changes the composition and size of epithelial cell-secreted extracellular vesicles that when incubated with mouse sperm, increase mitochondrial respiration and sperm motility, adding to our prior work showing impacts on embryo development. Together, these studies identify a time-dependent, translational signaling pathway that communicates stress experience to sperm, ultimately affecting reproductive functions.

Abstract Background & objectives Tumor grade determines prognosis in urothelial carcinoma. The classification of low and high grade is based on nuclear morphological features that include nuclear size, hyperchromasia and pleomorphism. These features are subjectively assessed by the pathologists and are not numerically measured, which leads to high rates of interobserver variability. The purpose of this study is to assess the value of a computer-based image analysis tool for identifying predictors of tumor grade in bladder cancer. Methods Four hundred images of urothelial tumors were graded by five pathologists and two expert genitourinary pathologists using a scale of 1 (lowest grade) to 5 (highest grade). A computer algorithm was used to automatically segment the nuclei and to provide morphometric parameters for each nucleus, which were used to establish the grading algorithm. Grading algorithm was compared to pathologists’ agreement. Results Comparison of the grading scores of the five pathologists with the expert genitourinary pathologists score showed agreement rates between 88.5% and 97.5%.The agreement rate between the two expert genitourinary pathologists was 99.5%. The quantified algorithm based conventional parameters that determine the grade (nuclear size, pleomorphism and hyperchromasia) showed > 85% agreement with the expert genitourinary pathologists. Surprisingly, the parameter that was most associated with tumor grade was the 10th percentile of the nuclear area, and high grade was associated with lower 10th percentile nuclei, caused by the presence of more inflammatory cells in the high-grade tumors. Conclusion Quantitative nuclear features could be applied to determine urothelial carcinoma grade and explore new biologically explainable parameters with better correlation to grade than those currently used.

A cesárea é uma intervenção que salva vidas, mas seu uso sem indicação obstétrica está relacionado a complicações a curto e longo prazo. O Brasil é conhecido internacionalmente por suas altas taxas desse procedimento, ainda mais elevadas no setor privado. Para reverter esse problema, a Agência Nacional de Saúde Suplementar lançou o Programa Parto Adequado, e este estudo tem como objetivo analisar a sua efetividade. Neste estudo retrospectivo, incluímos a totalidade de nascimentos em maternidades privadas ocorridos entre 2014 e 2019 para comparar a evolução das taxas de cesárea de hospitais participantes e não participantes do projeto, a partir das bases de dados públicas do Sistema de Informações sobre Nascidos Vivos (SINASC). Foram analisados 277.747 nascimentos, sendo observada uma redução na taxa de cesárea nos dois grupos, mais acentuada entre os hospitais participantes. Não foi observada redução da chance de cesárea antes do lançamento do programa (2014), mas notou-se uma tendência constante de redução após 2014, até se tornar significativa em 2018. Esse resultado ocorreu de forma independente das variáveis demográficas, maternas e dos grupos de Robson. A taxa de cesárea dos hospitais participantes do programa foi de 83,8% para 72,3% (intervalo de 95% de confiança - IC95%: 71,7-72,9). Apesar da redução, permanece bem acima das taxas esperadas de acordo com a ferramenta c-model, que seria de 45,2% (IC95%: 33,9-56,5) para essa população. Os resultados deste estudo mostraram que uma política pública bem conduzida com o envolvimento de instituições privadas pode mudar o cenário da atenção ao parto e ao nascimento, promovendo a redução das altas taxas de cesárea.

Abstract The genetic material encoded on X and Y chromosomes provides the foundation by which biological sex differences are established. Epigenetic regulators expressed on these sex chromosomes, including Kdm6a (Utx), Kdm5c, and Ddx3x have far-reaching impacts on transcriptional control of phenotypic sex differences. Although the functionality of UTY (Kdm6c, the Y-linked homologue of UTX), has been supported by more recent studies, its role in developmental sex differences is not understood. Here we test the hypothesis that UTY is an important transcriptional regulator during development that could contribute to sex-specific phenotypes and disease risks across the lifespan. We generated a random insertion Uty transgenic mouse (Uty-Tg) to overexpress Uty. By comparing transcriptomic profiles in developmental tissues, placenta and hypothalamus, we assessed potential UTY functional activity, comparing Uty-expressing female mice (XX + Uty) with wild-type male (XY) and female (XX) mice. To determine if Uty expression altered physiological or behavioral outcomes, adult mice were phenotypically examined. Uty expression masculinized female gene expression patterns in both the placenta and hypothalamus. Gene ontology (GO) and gene set enrichment analysis (GSEA) consistently identified pathways including immune and synaptic signaling as biological processes associated with UTY. Interestingly, adult females expressing Uty gained less weight and had a greater glucose tolerance compared to wild-type male and female mice when provided a high-fat diet. Utilizing a Uty-overexpressing transgenic mouse, our results provide novel evidence as to a functional transcriptional role for UTY in developing tissues, and a foundation to build on its prospective capacity to influence sex-specific developmental and health outcomes.

Abstract Background Community members in Quibdó (Choco, Colombia) are highly vulnerable to psychosocial problems associated with the internal armed conflict, poverty, and insufficient public services, and exacerbated by the COVID-19 pandemic. A pilot study was conducted with conflict-affected adults in Quibdó to assess feasibility and outcomes of a community-based psychosocial support group intervention using three different intervention modalities: in-person, remote (conducted online), and hybrid (half of sessions in-person, half-remote). This group model integrated problem-solving and culturally based expressive activities and was facilitated by local community members with supervision by mental health professionals. Methods This study utilized a mixed-explanatory sequential design (a quantitative phase deriving in a qualitative phase) with 39 participants and 8 staff members. Participants completed quantitative interviews before and after an eight-week group intervention. A subset of 17 participants also completed in-depth qualitative interviews and a focus group discussion was conducted with staff at post-intervention. Results From pre- to post-intervention, participants in all modalities demonstrated improved wellbeing and reduced symptoms of generalized distress, anxiety, depression, and posttraumatic stress. Use of coping skills varied across modalities, with remote groups associated with a decrease in some forms of coping, including use of social support. In qualitative interviews and the focus group discussion, participants and staff described logistical challenges and successes, as well as facilitators of change such as problem resolution, emotional regulation and social support with variations across modalities, such that remote groups provided fewer opportunities for social support and cohesion. Conclusions Results offer preliminary evidence that this model can address psychosocial difficulties across the three modalities, while also identifying potential risks and challenges, therefore providing useful guidance for service delivery in conflict-affected settings during the COVID-19 pandemic and other challenging contexts. Implications of this study for subsequent implementation of a Randomized Control Trial (RCT) are discussed.

BackgroundSpiroplasma citri comprises a bacterial complex that cause diseases in citrus, horseradish, carrot, sesame, and also infects a wide array of ornamental and weed species. S. citri is transmitted in a persistent propagative manner by the beet leafhopper, Neoaliturus tenellus in North America and Circulifer haematoceps in the Mediterranean region. Leafhopper transmission and the pathogen's wide host range serve as drivers of genetic diversity. This diversity was examined in silico by comparing the genome sequences of seven S. citri strains from the United States (BR12, CC-2, C5, C189, LB 319, BLH-13, and BLH-MB) collected from different hosts and times with other publicly available spiroplasmas.ResultsPhylogenetic analysis using 16S rRNA sequences from 39 spiroplasmas obtained from NCBI database showed that S. citri strains, along with S. kunkelii and S. phoeniceum, two other plant pathogenic spiroplasmas, formed a monophyletic group. To refine genetic relationships among S. citri strains, phylogenetic analyses with 863 core orthologous sequences were performed. Strains that clustered together were: CC-2 and C5; C189 and R8-A2; BR12, BLH-MB, BLH-13 and LB 319. Strain GII3-3X remained in a separate branch. Sequence rearrangements were observed among S. citri strains, predominantly in the center of the chromosome. One to nine plasmids were identified in the seven S. citri strains analyzed in this study. Plasmids were most abundant in strains isolated from the beet leafhopper, followed by strains from carrot, Chinese cabbage, horseradish, and citrus, respectively. All these S. citri strains contained one plasmid with high similarity to plasmid pSci6 from S. citri strain GII3-3X which is known to confer insect transmissibility. Additionally, 17 to 25 prophage-like elements were identified in these genomes, which may promote rearrangements and contribute to repetitive regions.ConclusionsThe genome of seven S. citri strains were found to contain a single circularized chromosome, ranging from 1.58 Mbp to 1.74 Mbp and 1597-2232 protein-coding genes. These strains possessed a plasmid similar to pSci6 from the GII3-3X strain associated with leafhopper transmission. Prophage sequences found in the S. citri genomes may contribute to the extension of its host range. These findings increase our understanding of S. citri genetic diversity.

Abstract Background Trials of tyrosine kinase inhibitors (TKI) have not demonstrated dramatic benefits in advanced colorectal cancer (CRC), and this may be a function of poor patient selection. TKI‐induced hypertension is reportedly a surrogate marker for treatment benefit for some tumor types. Our objective was to determine whether hypertension was associated with benefit in the context of CRC treatment, and also to gain insight on the pathogenesis of TKI‐induced hypertension by monitoring associated changes in the circulating metabolome. Patients and Methods Clinical data were acquired from clinical trial patients with metastatic CRC randomized to cetuximab ± the TKI brivanib (N = 750). Outcomes were evaluated as a function of treatment‐induced hypertension. For metabolomic studies, plasma samples were taken at baseline, as well as at 1, 4, and 12 weeks after treatment initiation. Samples were submitted to gas chromatography–mass spectrometry to identify treatment‐related metabolomic changes associated with TKI‐induced hypertension, compared to pre‐treatment baseline. A model based on changes in metabolite concentrations was generated using orthogonal partial least squares discriminant analysis (OPLS‐DA). Results In the brivanib treated group, 95 patients had treatment‐related hypertension within 12 weeks of initiating treatment. TKI‐induced hypertension was not associated with a significantly higher response rate, nor was it associated with improved progression‐free or overall survival. In metabolomic studies, 386 metabolites were identified. There were 29 metabolites that changed with treatment and distinguished patients with and without TKI‐induced hypertension. The OPLS‐DA model for brivanib‐induced hypertension was significant and robust (R2Y score = 0.89, Q2Y score = 0.70, CV‐ANOVA = 2.01 e‐7). Notable metabolomic features previously reported in pre‐eclampsia and associated with vasoconstriction were found. Conclusion TKI‐induced hypertension was not associated with clinical benefit in metastatic CRC. We have identified changes in the metabolome that are associated with the development of worsening brivanib‐induced hypertension that may be useful in future efforts of characterizing this toxicity.

Abstract Background Increasing cesarean section (CS) rates are a global concern because they are related to higher maternal and neonatal complication rates and do not provide positive childbirth experiences. In 2019, Brazil ranked second globally, given its overall CS rate of 57%. According to the World Health Organization (WHO), populational CS rates of 10–15% are associated with decreased maternal, neonatal, and infant mortality rates. This study aimed to investigate whether multidisciplinary care following evidence-based protocols associated with a high motivation of both women and professionals for a vaginal birth leads to less overuse of CS in a Brazilian private practice (PP). Methods This cross-sectional study evaluated CS rates by Robson group for women who sought vaginal birth in a private practice in Brazil comparing with Swedish data. Collaborative care of midwives and obstetricians who adopted evidence-based guidelines was offered. CS rates, overall and by Robson group, contribution of each Robson group to the overall CS rate, clinical and nonclinical interventions, vaginal birth, pre-labor CS, and intrapartum CS proportions were estimated. The expected CS rate was calculated using the World Health Organization C-model tool. The analysis used Microsoft Excel and R Studio (version 1.2.1335. 2009–2019). Results The PP overall CS rate was 15.1% (95%CI, 13.4–17.1%) versus the 19.8% (95%CI, 14.8–24.7%) rate expected by the WHO C-model tool. The population included 43.7% women in Robson Group 1 (nulliparous, single, cephalic, at term, spontaneous labor), 11.4% in Group 2 (nulliparous, single, cephalic, at term, induced labor or CS before labor), and 14.9% in Group 5 (multiparous women with previous CS), the greatest contributors to higher CS rates (75.4% of them). The Swedish overall CS rate was 17.9% (95%CI, 17.6–18.1%) in a population of 27% women in Robson Group 1, 10.7% in Group 2, and 9.2% in Group 5. Conclusions Multidisciplinary care following evidence-based protocols, associated with high motivation of both women and professionals for vaginal birth, may lead to a significant and safe reduction of CS rates even in contexts such as Brazil, with high medicalization of obstetric care and excess CS.

Abstract Background Community-based psychosocial support (CB-PSS) interventions utilizing task sharing and varied (in-person, remote) modalities are essential strategies to meet mental health needs, including during the COVID-19 pandemic. However, knowledge gaps remain regarding feasibility and effectiveness. Methods This study assesses feasibility, acceptability and preliminary effectiveness of a CB-PSS intervention for conflict-affected adults in Colombia through parallel randomized controlled trials, one delivered in-person (n = 165) and the other remotely (n = 103), implemented during the COVID-19 pandemic and national protests. Interventions were facilitated by nonspecialist community members and consisted of eight problem-solving and expressive group sessions. Findings Attendance was moderate and fidelity was high in both modalities. Participants in both modalities reported high levels of satisfaction, with in-person participants reporting increased comfort expressing emotions and more positive experiences with research protocols. Symptoms of depression, anxiety and posttraumatic stress disorder improved among in-person participants, but there were no significant changes for remote participants in comparison to waitlist controls. Implications This CB-PSS intervention appears feasible and acceptable in both in-person and remote modalities and associated with reduction in some forms of distress when conducted in-person but not when conducted remotely. Methodological limitations and potential explanations and areas for future research are discussed, drawing from related studies.

Abstract Four solution-processable, linear conjugated polymers of intrinsic porosity are synthesised and tested for gas phase carbon dioxide photoreduction. The polymers’ photoreduction efficiency is investigated as a function of their porosity, optical properties, energy levels and photoluminescence. All polymers successfully form carbon monoxide as the main product, without the addition of metal co-catalysts. The best performing single component polymer yields a rate of 66 μmol h−1 m−2, which we attribute to the polymer exhibiting macroporosity and the longest exciton lifetimes. The addition of copper iodide, as a source of a copper co-catalyst in the polymers shows an increase in rate, with the best performing polymer achieving a rate of 175 μmol h−1 m−2. The polymers are active for over 100 h under operating conditions. This work shows the potential of processable polymers of intrinsic porosity for use in the gas phase photoreduction of carbon dioxide towards solar fuels.

Dame Cicely Saunders’ conceptualization of ‘total pain’, or ‘total suffering’, is one of her most significant and lasting contributions to the field of palliative care. It was Saunders’ unique combination of knowledge and experiences as a trained social worker, nurse and physician that influenced her understanding of suffering specific to a life-limiting illness as being multi-dimensional: that suffering may be simultaneously physical, psychological, emotional, social, spiritual and/or existential in nature. ‘Total pain’ remains a highly relevant and significant concept within palliative care and Saunders’ lasting contributions are to be revered. This paper invites us to reconsider one particular aspect of Saunders’ conceptualization: that patients’ ‘mental reactions’ to their anticipated dying/death is a key contributor to their ‘total pain’. Drawing upon Saunders’ works from the late 1950s to the early 2000s, this paper details the socio-historical manifestation of this aspect of ‘total pain’ within Saunders’ writings, including influences from her Christian religion and Viktor Frankl, and its enduring impact on palliative care philosophy, practice, and discourse. Then, drawing upon patient stories rooted in my own clinical experiences over a 10 year period as a hospice social worker, I suggest that this particular feature of Saunders’ ‘total pain’ may, unintentionally, work to pathologize both the patient for whom suffering persists and remains unsolvable, and the palliative care clinician who may struggle to relieve it — and why it therefore stands to be revisited. It is my sincere hope and intention that ongoing reverence for Saunders’ significant contributions can sit alongside respectful reconsideration.

Wild-grown mushroom species chanterelle (Cantharellus cibarius) and porcini mushroom (Boletus edulis), available at Austrian local markets, were evaluated regarding microbial and sensory quality, content of biogenic amines, and proximate chemical composition. Chemical composition varied widely with species, but showed no correlation with sensory quality and microbial load. Chanterelles were characterized by high levels of aerobic mesophilic counts, regardless of sensory quality. In both mushroom species, poor sensory quality was associated with increased counts of Enterobacteriaceae. In porcini mushrooms, the level of ß-phenylethylamine, cadaverine, histamine, and tyramine correlated to the count and type of microorganism. The level of biogenic amines does not pose a risk to healthy individuals. Almost half of the samples purchased were sensorically deficient, indicating handling under poor hygienic conditions or being offered for sale for too long. To obtain wild mushrooms of excellent quality, all relevant stakeholders at harvest and retail level should be trained in hygienic handling of mushrooms.

Abstract Homeostatic regulation of the maternal milieu during pregnancy is critical for maternal and fetal health. The placenta facilitates critical communication between maternal and fetal compartments, in part, through the production of extracellular vesicles (EVs). EVs enable tissue synchrony via cell–cell and long-distance communication and are at their highest circulating concentration during pregnancy. While much work has been done investigating how physiological challenges in pregnancy affect the fetus, the role of placental communication in maternal health has not been well examined. We previously identified placental O-glycosyl transferase (OGT), a glucose-sensing enzyme, as a target of maternal stress where OGT levels and activity affected the O-glycosylation of proteins critical for EV cargo loading and secretion. Here, we hypothesized that placental OGT plays an essential role in maternal homeostatic regulation during pregnancy via its regulation of maternal circulating EV concentrations. Our studies found that changes to key metabolic factors over the circadian cycle, including glucocorticoids, insulin, and glucose, were significantly associated with changes in circulating EV concentration. Targeting placental OGT in mice, we found a novel significant positive relationship between placental OGT and maternal circulating EV concentration that was associated with improving maternal glucose tolerance during pregnancy. Finally, an intravenous elevation in EVs, matching the concentration of EVs during pregnancy, shifted non-pregnant female glucose sensitivity, blunted glucose variance, and improved synchrony of glucose uptake. These data suggest an important and novel role for circulating EVs as homeostatic regulators important in maternal health during pregnancy.

Abstract Extracellular vesicles (EVs) are a unique mode of intercellular communication capable of specificity in transmitting signals and cargo to coordinate local and distant cellular functions. A key example of this is the essential role that EVs secreted by epithelial cells lining the lumen of the male reproductive tract play in post-spermatogenic sperm maturation. We recently showed in a preclinical mouse model that this fundamental process had a causal role in somatic-to-germline transmission of biological information regarding prior stress experience capable of altering the rate of fetal development. However, critical mechanistic questions remain unanswered as to the processes by which signaling occurs between EVs and sperm, and whether EVs or their cargo are delivered at conception and are detectable in the early embryo. Unfortunately, notable methodological limitations shared across EV biology, particularly in the isolation and labeling of EVs, complicate efforts to answer these important questions as well as questions on EV targeting specificity and mechanisms. In our current studies, we developed a novel approach to track EVs using a conditional transgenic construct designed to label EVs via conditional Cre-induced hemagglutinin (HA) tagging of the EV endogenous tetraspanin, CD63. In our exhaustive validation steps, this internal small molecular weight tag did not affect EV secretion or functionality, a common problem found in the previous design of EV tags using larger molecular weight proteins, including fluorescent proteins. Utilizing a stably transfected immortalized epididymal epithelial cell line, we first validated key parameters of the conditional HA-tagged protein packaged into secreted EVs. Importantly, we systematically confirmed that expression of the CD63-HA had no impact on the production, size distribution, or surface charge of secreted EVs, nor did it alter the tetraspanin or miRNA composition of these EVs. We also utilized the CD63-HA EVs to verify physical interactions with sperm. Finally, using in vitro fertilization we produced some of the first images confirming sperm delivered EV cargo at conception and still detectable in the early-stage embryo. As such, this construct serves as a methodological advance and as a valuable tool, with applications in the study of EV function across biomedical research areas.

Pathogenic microorganisms surviving in dry products have regularly led to recalls and foodborne disease outbreaks. Therefore, the microbiological quality of 61 dried mushrooms samples purchased online and in supermarkets were analyzed. Counts of aerobic mesophiles (AMCs), Enterobacteriaceae (EB), yeasts and molds, presumptive Bacillus cereus (pBC), the presence of Salmonella spp., and L. monocytogenes were investigated. Isolates of pBC were screened for their partial panC gene sequences and their toxin genes’ profiles. The microbiological quality of the dried mushrooms investigated in this study was generally found to be acceptable. Average AMCs, EB, yeasts, and molds were 3.9 log, 1.1 log, 1.6 log, and 1.5 log cfu/g, respectively. All mushroom samples tested negative for Salmonella spp. and L. monocytogenes. Presumptive BC were detected in 59.0% of the samples, but the contamination level was low (1.0 to 3.4 log cfu/g). None of the isolates were positive for the ces gene. Incomplete labeling was found in 45.9% of the samples, mainly in the form of missing heating instructions (31.1%) and/or country of origin (16.3%). Contamination by pathogens can occur in dried mushrooms. Adequate information on home cooking practices is essential to reduce the risk of foodborne illness to the consumer and to provide a safe food product.

Bacterial meningitis is a major cause of morbidity and mortality, especially among infants and the elderly. Here, we study mice to assess the response of each of the major meningeal cell types to early postnatal E. coli infection using single nucleus RNA sequencing (snRNAseq), immunostaining, and genetic and pharamacologic perturbations of immune cells and immune signaling. Flatmounts of the dissected leptomeninges and dura were used to facilitiate high-quality confocal imaging and quantification of cell abundances and morphologies. Upon infection, the major meningeal cell types – including endothelial cells (ECs), macrophages, and fibroblasts – exhibit distinctive changes in their transcriptomes. Additionally, ECs in the leptomeninges redistribute CLDN5 and PECAM1, and leptomeningeal capillaries exhibit foci with reduced blood-brain barrier integrity. The vascular response to infection appears to be largely driven by TLR4 signaling, as determined by the nearly identical responses induced by infection and LPS administration and by the blunted response to infection in Tlr4-/- mice. Interestingly, knocking out Ccr2, encoding a major chemoattractant for monocytes, or acute depletion of leptomeningeal macrophages, following intracebroventricular injection of liposomal clodronate, had little or no effect on the response of leptomeningeal ECs to E. coli infection. Taken together, these data imply that EC responses to infection are largely driven by the intrinsic EC response to LPS.