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Abstract The study of microbial communities of the plant phyllosphere in remote locations using DNA-based approaches is limited by the challenges associated with their preservation in the field and during transportation. Freezing is a common DNA preservation strategy, but it may be unsuitable for leaf samples, or inaccessible in some locations. Other methods such as desiccation, ethanol or commercial preservatives are potential alternative DNA preservation methods for ambient temperature storage. In this study, we assessed the efficacy of desiccation (with silica gel packs), and of three preservation solutions (95% ethanol, RNAlater, LifeGuard) for the preservation of epiphytic phyllosphere communities of Populus tremuloides and Picea glauca at ambient indoor temperature (21 °C) for up to three weeks. We assessed effects on DNA concentration and quality and used metabarcoding to detect changes in bacterial and fungal communities between treatments over time. A secondary study was conducted on leaves of Populus grandidentata to further test the ability of the desiccation treatment to resolve differences between sampling sites. Silica gel packs were identified as effective ambient temperature preservative of phyllosphere bacterial and fungal communities. There were some changes in the communities compared to immediate extraction due to this treatment, but these changes did not affect the ability to distinguish tree species and sampling locations. Overall, our study supports the use of silica gel pack short term preservation at ambient temperature for phyllosphere samples intended for DNA-based microbial community analyses.

This article describes the evaluation of a pilot Montessori-based home visiting program called "Mi Pequeño Mundo" (MPM) aimed at promoting optimal child development in Spanish-speaking Latine children from birth to three years old in Hennepin County, Minnesota. Phase I included World Cafés and interviews to gather Centro Tyrone Guzman (CTG) stakeholder feedback on program design. Thematic analysis informed the development of the program and implementation approach. Primary input included topics for curriculum development. Phase II included training three bilingual/bicultural community members for home visiting (Conectores), recruiting families, data collection, and family visits. Feasibility and acceptability measures plus pre--post assessment of knowledge, self-efficacy, mindfulness parenting, child assessment, and process data were collected. Data were analyzed using descriptive statistics. Qualitative summaries highlighted themes from open-ended questions. Phase II included training of three Conectores who worked with 15 families for up to 20 weekly remote visits each (due to COVID-19 protocols). Participating families were 100% satisfied with the program and said they would recommend it to a friend. Program outputs/goals were met upon the completion of the pilot (Phase II) indicating the program was feasible and acceptable. The full implementation phase of MPM was modified based on the first two phases and programming was scaled to reach the broader Spanish-speaking Latine community living in Minnesota.

Thermophily is a trait scattered across the fungal tree of life, with its highest prevalence within three fungal families (Chaetomiaceae, Thermoascaceae, and Trichocomaceae), as well as some members of the phylum Mucoromycota. We examined 37 thermophilic and thermotolerant species and 42 mesophilic species for this study and identified thermophily as the ancestral state of all three prominent families of thermophilic fungi. Thermophilic fungal genomes were found to encode various thermostable enzymes, including carbohydrate-active enzymes such as endoxylanases, which are useful for many industrial applications. At the same time, the overall gene counts, especially in gene families responsible for microbial defense such as secondary metabolism, are reduced in thermophiles compared to mesophiles. We also found a reduction in the core genome size of thermophiles in both the Chaetomiaceae family and the Eurotiomycetes class. The Gene Ontology terms lost in thermophilic fungi include primary metabolism, transporters, UV response, and O-methyltransferases. Comparative genomics analysis also revealed higher GC content in the third base of codons (GC3) and a lower effective number of codons in fungal thermophiles than in both thermotolerant and mesophilic fungi. Furthermore, using the Support Vector Machine classifier, we identified several Pfam domains capable of discriminating between genomes of thermophiles and mesophiles with 94% accuracy. Using AlphaFold2 to predict protein structures of endoxylanases (GH10), we built a similarity network based on the structures. We found that the number of disulfide bonds appears important for protein structure, and the network clusters based on protein structures correlate with the optimal activity temperature. Thus, comparative genomics offers new insights into the biology, adaptation, and evolutionary history of thermophilic fungi while providing a parts list for bioengineering applications.

Abstract Thermophily is a trait scattered across the fungal tree of life, with its highest prevalence within three fungal families (Chaetomiaceae, Thermoascaceae, and Trichocomaceae), as well as some members of the phylum Mucoromycota. We examined 37 thermophilic and thermotolerant species and 42 mesophilic species for this study and identified thermophily as the ancestral state of all three prominent families of thermophilic fungi. Thermophilic fungal genomes were found to encode various thermostable enzymes, including carbohydrate-active enzymes such as endoxylanases, which are useful for many industrial applications. At the same time, the overall gene counts, especially in gene families responsible for microbial defense such as secondary metabolism, are reduced in thermophiles compared to mesophiles. We also found a reduction in the core genome size of thermophiles in both the Chaetomiaceae family and the Eurotiomycetes class. The Gene Ontology terms lost in thermophilic fungi include primary metabolism, transporters, UV response, and O-methyltransferases. Comparative genomics analysis also revealed higher GC content in the third base of codons (GC3) and a lower effective number of codons in fungal thermophiles than in both thermotolerant and mesophilic fungi. Furthermore, using the Support Vector Machine classifier, we identified several Pfam domains capable of discriminating between genomes of thermophiles and mesophiles with 94% accuracy. Using AlphaFold2 to predict protein structures of endoxylanases (GH10), we built a similarity network based on the structures. We found that the number of disulfide bonds appears important for protein structure, and the network clusters based on protein structures correlate with the optimal activity temperature. Thus, comparative genomics offers new insights into the biology, adaptation, and evolutionary history of thermophilic fungi while providing a parts list for bioengineering applications.

Despite concerted efforts, LGBTQ + inclusivity is not fostered across all sporting contexts. Interpersonal relationships with teammates and coaches are particularly significant when it comes to cultivating positive and inclusive LGBTQ + experiences within sport. Using online qualitative data collected from 741 LGBTQ + adults, this study examined how relationships with coaches, teammates and sport-related staff impacted sport experiences and participation. Participant responses were independently coded using reflexive thematic analysis. Data were used to develop a new visual model to understand different types of relationships LGBTQ + individuals experience in sport and how this impacted their sport experiences. Exclusive sport relationships were characterised by experiences of discrimination and were found to greatly influence how participants viewed sport and physical activity contexts as adults. Inert or neutral sport relationships were maintained through protective self-distancing techniques. Inclusive relationships with coaches and teammates were primarily based on acceptance and associated with viewing physical activity positively. Identity-concealment strategies and athletic ability were identified as factors that impacted LGBTQ + adults' relationships within sport. Our overall findings demonstrate how discrimination in the form of overt or subtle sexual and gender prejudice can thwart feelings of relatedness among LGBTQ + athletes.

La jeunesse engagée est consciente des enjeux qui affectent nos sociétés et cherche à trouver des solutions et contribuer à développer un modèle économique plus juste et solidaire. On assiste à une montée de l’engouement pour des modèles alternatifs d’entreprises, comme les entreprises d’économie sociale. Cependant, on constate que le passage de l’idée à la concrétisation d’un projet est un écueil difficile à contourner pour de nombreux porteurs de projets. Le programme SISMIC, porté par les pôles régionaux d’économie sociale, offre un soutien aux jeunes dans leurs démarches de démarrage d’entreprises d’économie sociale. Bien que l’appui offert par les pôles soit des plus pertinents au regard des nombreux projets accompagnés, on observe que la proportion de constitutions d’entreprises collectives issues des incubateurs SISMIC demeure faible. En se basant sur sa perspective régionale, le Pôle des entreprises d’économie sociale de la région de la Capitale-Nationale (Pôle CN) expose dans cet article les raisons de la difficile concrétisation des projets collectifs portés par les jeunes.

Objective: The early consumption of calorie-rich diet disrupts circadian rhythms and has adverse effects on memory, yet the effects of time-restricted feeding (TRF) and the underlying molecular mechanisms are unknown. Here, we set out to identify the behavioral and molecular circadian rhythms disruptions generated by juvenile obesogenic diet consumption and their restoration by TRF in male mice. Methods: Metabolic rhythms were measured by indirect calorimetry and memory performances by behavioral tasks. Hippocampal translatome (pS6_TRAP), enrichment and co-regulated gene network analyses were conducted to identify the molecular pathways involved in memory impairments and their restoration by TRF. Differential exon usage analyses, mass spectrometry and pharmacological intervention were used to confirm thyroid hormone signaling involvement. Results: We show that four weeks of TRF restore the rhythmicity of metabolic parameters and prevents memory impairments in mice fed a high fat-high sucrose (HFS) diet since weaning, independently of body fat levels. Hippocampal translatome and differential exon usage analyses indicate that impaired memory of mice under ad libitum HFS diet is accompanied by reduced thyroid hormone signaling and altered expression of astrocytic genes regulating glutamate neurotransmission. TRF restored the diurnal expression variation of part of these genes and intra-hippocampal infusion of T3, the active form of thyroid hormone, rescues memory performances and astrocytic gene expression of ad libitum HFS diet-fed mice. Conclusions: Thus, thyroid hormones contribute to the TRF positive effects on both metabolism and memory in mice fed an obesogenic diet, highlighting this nutritional approach as a powerful tool in addressing obesity brain comorbidities and paving the way for further mechanistic studies on hippocampal thyroid signaling.

The univoltine leaf beetle Psylliodes attenuata (Koch, 1803) is a pest of Cannabis and Humulus (Cannabaceae) and native to the Palaearctic Region, known from eastern Asia to western Europe.First North American records are presented for P. attenuata from Canada: Ontario and Québec. Adult beetle feeding damage to hops Humulus lupulus L. (Cannabacaea) plants is recorded from Québec. Diagnostic information is presented to distinguish P. attenuata from other North American Chrysomelidae and a preliminary assessment of its potential to spread in North America is presented. While our climate analysis is limited by a lack of data, it appears P. attenuata is physiologically capable of persisting throughout the range of Humulus in North America.The United States of America and Canada are now known to be home to 71 or more species of adventive Chrysomelidae.

Summary: Tissue clearing enables deep imaging of long biological structures using light microscopy approaches. Protocols such as iDISCO are not currently available for human skin. Here, we present Skin-iDISCO+, a tissue-clearing and labeling protocol for morphometric analysis of human cutaneous vasculature. We describe steps for processing and pre-treating tissue, immunostaining, clearing, and imaging using confocal and light-sheet fluorescence microscopy. We then detail procedures for the reconstruction of networks and the extraction of morphometric parameters of the skin vasculature.For complete details on the use and execution of this protocol, please refer to Renier et al.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

IntroductionEncouraging the activation of brain repair mechanisms and fostering spontaneous functional recovery in stroke patients hold great promise for alleviating the global burden of this condition and allowing an extended therapeutic time window. Cell-based regenerative therapy (with mesenchymal stem/stromal cells, such as adipose-derived stem cells [ADSCs]) is particularly attractive considering its excellent safety profile, low immunogenicity after allogeneic application, and well-established functional benefits on stroke recovery in animal models. This study aims to assess the efficacy and safety effects of intravenous (IV) infusion of freshly cultured allogeneic ADSCs on recovery after ischemic stroke.Population and methodsRESSTORE is a multicentric, randomized 1:1 controlled double-blind clinical trial. Eighty patients will be enrolled in nine French stroke centers. The main inclusion criteria are ≥18 years of age, acute hemispheric ischemic stroke, and a National Institutes of Health Stroke Scale (NIHSS) score of ≥7, including a motor subscore of ≥3. According to the previous dose-escalation safety trial data, the maximum tolerated dose of 3 million ADSCs/kg was selected. IV infusion was performed within 10 days following stroke onset, with a follow-up over 2 years.OutcomesThe primary endpoint is the motor NIHSS subscore, computed as the sum of the upper limb, lower limb, and hand scores, measured 6 months after stroke onset to assess motor recovery. The secondary outcomes are the occurrence of death/serious adverse events, clinical scores (the detailed NIHSS scores, Montreal Cognitive Assessment scores, modified Rankin Scale scores, Aphasia Handicap Scores, Depression Intensity Scale Circles scores, Fatigue Scale scores, etc.), immune monitoring (for the first 30 patients), and multimodal biomarkers derived from diffusion and functional magnetic resonance imaging.DiscussionThis study may provide some evidence for the effects of freshly cultured allogenic ADSCs IV infusion in subacute stroke that may help design a larger international randomized controlled trial.

Aims Navigating a Serious Incident (SI) investigation and participating in a Coroner's Court proceedings can pose challenges for psychiatry trainees. The Higher Training curriculum emphasizes active participation in activities that enhance patient safety and care quality. This project aims to enhance patient safety and trainee confidence by improving training on SI investigations and Coroners Court proceedings. Methods Using Quality Improvement (QI) methodology, in the first cycle an initial survey was distributed to all psychiatry trainees and middle grade doctors working in Kent and Medway (n = 67) to establish baseline knowledge and confidence levels in areas related to risk assessment & management, SI investigations and Coroner's Inquests. In response to the identified need for training, we organized the Initial Training Event with support from Deputy Chief Medical Officer for Quality and Safety, Patient Safety Team and Medical Education Department. The half-day, in-person event was opened to all doctors and featured 5 sessions: Serious Incident Investigation Process, Thematic Review of Suicides, Systems Engineering and Human Factors in Patient Safety, Learning from Mortality and Structured Judgement Review along with ‘Being Involved in Investigation – An Investigator's Guide’. Data from a survey of attendees (n = 47) informed the development of a tailored training session for psychiatry Core and Higher Trainees. Results The initial survey received 32 responses (response rate: 47.76%). 71.88% of respondents had little to no understanding of SI investigation processes. Remarkably, 87.5% expressed strong interest in receiving training on conducting SI investigations. 90.62% were extremely or very interested in receiving training on participating in a Coroner's Inquest. 47 doctors attended the Initial Training Event. 30 responded to the feedback questionnaire (47.76%). All doctors found the training useful, with over 90% rating it ‘very’ or ‘extremely’ useful. 97% felt that the training would improve their clinical practice in terms of patients’ safety. After the training, 60% understood the process of conducting an SI investigation a moderate amount; 33.33% understood the process a lot or to a great extent. Nevertheless 92.86% felt a need for additional training in SI investigations. 63.33% suggested making training available yearly, and 36.67% favoured making it mandatory training. Conclusion This project identified a significant need for training in SI investigations and Coroner's Court proceedings among psychiatric trainees. An Initial Training Event developed from the first QI cycle survey data received positive feedback. The next phase involves developing a tailored training program that addresses identified knowledge gaps. Further considerations include making this training a regular event.

Aims Antipsychotic medications are commonly used in the management of psychiatric disorders, including in older adults. However, the use of these medications in older adults can be associated with a higher risk of adverse effects such as cardiovascular event and extrapyramidal symptoms. This retrospective audit aimed to evaluate adherence to antipsychotic prescribing and monitoring guidelines in a Psychiatric Unit for Older Adult Females in Kent and Medway NHS and Social Care Partnership Trust (KMPT). Methods The audit criteria encompassed various aspects of documentation and medication management, including diagnosis documentation, indication, age, comorbidities, consent, baseline assessments, monitoring, review, and follow-up care. Data from two months’ records were analysed leading to an action plan with slight amendments to the user-friendly template for ward round and a physical health monitoring poster for junior doctors and ward staff. These initiatives aim to improve patient care, streamline documentation, while accommodating the rotation of junior doctors. A re-audit is planned post implementation. This audit’s limitations included the study’s single-site nature, potential sample size constraints and reliance on accurate documentation. Results The audit achieved 100% compliance in documenting patient age and MHA status, meeting legal requirements. Weight, BMI, and baseline blood pressure exhibited full compliance. Baseline ECGs had an 86.66% compliance rate, while QTc interval documentation reached 100%. Antipsychotic indication and weekly reviews were documented at 100%, with an 83.33% rate for rationale documentation for medication changes. Comorbidities were fully documented, but extrapyramidal symptom and sedation monitoring showed a 46.66% compliance rate. Baseline blood tests, including glucose, bA1c, lipid profile, electrolytes, renal and liver function, thyroid function, and prolactin levels, generally had high compliance, but lipid profile and liver function achieved 73.33%. Repeat blood tests varied, with electrolytes and renal function at 100%, while thyroid function and prolactin levels scored lower at 26.66% and 46.66%. Continued monitoring of weight, BMI, and blood pressure remained fully compliant. Compliance for repeating ECGs within recommended timeframes reached 53.33%, and recommendations to GPs for yearly ECGs and blood monitoring achieved 50%. Conclusion In summary, the audit identified areas of commendable high and medium compliance with antipsychotic prescribing guidelines in a Psychiatric Unit for Older Adult Females in KMPT. An action plan has been formulated to not only enhance patient care but also to refine the documentation process positively further, fostering continued progress in the provision of high-quality care.

Drawing on a review of international literature published between 2005 and 2021, the present article discusses interventions tailored for Indigenous people in vocational education and training. A critical analysis of this literature was carried out to gain a deeper understanding of the scope of these interventions in bolstering success among Indigenous people in vocational education. Three axes of analysis were used to examine 24 academic papers: the interventions featured in these papers, the conception of social justice in education (i.e. equality of place, equality of opportunity, equity and inclusion, and decolonisation) underpinning these interventions, and their effects on success. The findings paint a picture of the relevance and scope of initiatives that can foster Indigenous student success in vocational education and at the same time reveal the limitations of these studies. They also show which conception of social justice in education appears the most promising. The article concludes with a brief discussion of a few fundamental elements that need to be kept in mind when developing initiatives for Indigenous people in vocational education.