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Successful conservation of rare, threatened, or endangered (RTE) species is dependent upon rapid and accurate assessment of their distribution and abundance. However, assessments are challenging as RTE species typically exist as numerically small populations in often fragmented habitats and can possess complex natural histories. Environmental DNA (eDNA) analysis may provide a rapid, cost‐effective means of assessing RTE species presence/absence in viable habitat patches. We evaluated the efficacy of eDNA surveillance for the Louisiana Pinesnake (Pituophis ruthveni), an elusive, semi‐fossorial, nonvenomous colubroid snake endemic to Louisiana and Texas, USA, that has dramatically declined in both distribution and abundance. We developed two quantitative polymerase chain reaction (qPCR) assays that target the mitochondrial cytochrome c oxidase subunit I (COI) and mitochondrially encoded ATP synthase membrane subunit 6 (ATP6) genes. We validated each assay in silico, in vitro, and in situ, and investigated the influence of eDNA extraction method and genetic marker on assay performance. Both assays were highly sensitive and successfully detected the Louisiana Pinesnake under artificial and field conditions, including bedding samples collected from captive snake enclosures (100%), soil samples from Louisiana Pinesnake release sites (100%), and soil samples from sites where Louisiana Pinesnakes were documented via radio telemetry (45%). Although differences between genetic markers were negligible, assay performance was strongly influenced by eDNA extraction method. Informed by our results, we discuss methodological and environmental factors influencing Louisiana Pinesnake eDNA detection and quantification, broader implications for management and conservation of the Louisiana Pinesnake and other terrestrial reptiles and provide recommendations for future research. We suggest that eDNA surveys can more effectively assess Louisiana Pinesnake occupancy than conventional sampling, highlighting the need for comprehensive eDNA monitoring initiatives to better identify suitable habitat that will promote persistence of this imperiled species going forward. Environmental DNA effectively detected the federally threatened Louisiana Pinesnake. We provide an optimized approach for a comprehensive eDNA survey for the species. We also contextualize these results in the broader constellation of snake eDNA studies.

Environmental DNA (eDNA) analysis is an effective and non‐invasive technique for surveying and monitoring rare, threatened, or endangered (RTE) species. Compared to conventional capture‐based sampling, eDNA analysis may offer a more cost‐effective approach for surveying RTE species, yet few studies have compared their cost‐efficiency—a critical consideration for conservation planning. We compared the costs, effort, and relative performance of aquatic eDNA sampling and conventional trapping for detecting the Alligator Snapping Turtle, Macrochelys temminckii Troost, 1835, in southwest Louisiana, United States. Environmental DNA was sampled quarterly over 1 year (2018–2019) at 19 streams, including three streams where M. temminckii presence had been previously confirmed via conventional trapping efforts (2012–2013). Water samples from each stream were analyzed using quantitative polymerase chain reaction (qPCR) to assess M. temminckii eDNA presence/absence. Time and costs (i.e., labor, travel, wages, and supplies) per detection via eDNA analysis and trapping were calculated and compared. Environmental DNA analysis documented the presence of M. temminckii DNA at two of the three streams where individuals had previously been trapped and yielded detections (qPCR amplifications) at 16 additional streams not previously sampled, expanding M. temminckii's documented distribution at our study sites by 84%. Environmental DNA analysis returned a detection rate (per site) 5.55 times higher than conventional trapping and was 18.7% less expensive. Our results provide evidence that strategically deployed eDNA surveys may be an effective and cost‐efficient approach for detecting freshwater RTE species. With eDNA analysis, additional resources can be invested toward expanding survey coverage and increasing sampling frequency, allowing managers to more effectively target subsequent intensive monitoring efforts. Successful integration of a new method into a standard management regime requires understanding both its relative efficacy and cost‐effectiveness when compared against conventional sampling methods. We successfully detected M. temminckii in Louisiana streams using both eDNA surveys and conventional trapping with eDNA surveys yielding 5.55 times higher detection rates, 40% fewer labor hours per detection, and 84% more streams surveyed per year than conventional trapping. Our results suggest that eDNA presents a promising and cost‐efficient technique for M. temminckii surveys.

Protein kinase D (PKD) enzymes regulate cofilin-driven actin reorganization and directed cell migration through both p21-activated kinase 4 (PAK4) and the phosphatase slingshot 1L (SSH1L). The relative contributions of different endogenous PKD isoforms to both signaling pathways have not been elucidated, sufficiently. We here analyzed two cell lines (HeLa and MDA-MB-468) that express the subtypes protein kinase D2 (PKD2) and protein kinase D3 (PKD3). We show that under normal growth conditions both isoforms can form a complex, in which PKD3 is basally-active and PKD2 is inactive. Basal activity of PKD3 mediates PAK4 activity and downstream signaling, but does not significantly inhibit SSH1L. This signaling constellation was required for facilitating directed cell migration. Activation of PKD2 and further increase of PKD3 activity leads to additional phosphorylation and inhibition of endogenous SSH1L. Net effect is a dramatic increase in phospho-cofilin and a decrease in cell migration, since now both PAK4 and SSH1L are regulated by the active PKD2/PKD3 complex. Our data suggest that PKD complexes provide an interface for both cofilin regulatory pathways. Dependent on the activity of involved PKD enzymes signaling can be balanced to guarantee a functional cofilin activity cycle and increase cell migration, or imbalanced to decrease cell migration. Our data also provide an explanation of how PKD isoforms mediate different effects on directed cell migration.

ABSTRACT Aim Freshwater mussels are considered among the most at‐risk taxa in the world. As such, comprehensive monitoring assessments of what abiotic and biotic factors influence mussel occupancy will be vital for guiding effective conservation. Here, we analysed vertebrate and mussel environmental DNA (eDNA) metabarcoding data to explore the influence of biotic (i.e., host fish diversity, predator presence, and community composition) and abiotic (i.e., drainage size, forest cover, and stream order) factors on freshwater mussel populations. Location This study utilised water samples and tactile survey data collected from streams throughout Fort Johnson, Louisiana. Methods We first evaluated the effectiveness of eDNA metabarcoding for characterising freshwater communities based on previous conventional tactile surveys. Next, we used eDNA metabarcoding analysis for freshwater mussels and vertebrate species alongside GIS‐derived satellite remote sensing data to assess how various biotic and abiotic variables impact freshwater mussel eDNA occupancy. Results Our eDNA metabarcoding survey largely agreed with both historical and contemporary surveys on Fort Johnson, while uniquely detecting Louisiana pigtoe (Pleurobema riddellii), a proposed threatened species under the US Endangered Species Act. We also found that eDNA detections and occupancy had strong seasonal variation, with increased read abundance and diversity in the spring. Vertebrate, fish, and predator diversity (as a function of habitat quality) were strongly predictive of mussel occupancy, supporting the concept of land managers focusing on the entire ecosystem for mussel conservation. Lastly, we found that percent forest cover and drainage basin size influenced mussel eDNA occupancy, informing habitat associations for mussel species of interest (i.e., the mussels occupied larger drainage sizes and perennial streams). Conclusions Our results demonstrate that combining eDNA metabarcoding of target and non‐target species with occupancy modelling can provide insights into the ecology of freshwater mussels and is a useful tool to improve their conservation and management.

Pituophis ruthveni (Louisiana Pinesnake) is considered to be one of the rarest snake species in North America and, accordingly, was federally listed as threatened under the Endangered Species Act in 2018. Although much previous work has demonstrated the species' tight association with its primary prey, Geomys breviceps (Baird's Pocket Gopher), relatively little work has been done on habitat selection, hampering management and conservation efforts. Here we use a variety of technologies, including traditional radio-telemetry, automated radio-telemetry, and subsurface temperature loggers, to monitor Louisiana Pinesnake behavior and habitat use for one of the few remaining populations of the species. We find support for previous studies indicating that Louisiana Pinesnakes exhibit bimodal seasonal activity patterns and preferences for areas with Baird's Pocket Gophers. However, we also document extensive individual variation in behavior and habitat use with some individuals exhibiting small home ranges (8.36 ha) in relatively open habitats and others with much larger home ranges (166.83 ha) in relatively closed habitats. These results suggest that Louisiana Pinesnake habitat selection, although largely restricted to areas with pocket gophers, can be extremely variable in other habitat features. Based on subsurface temperatures, our results suggest that snakes are most often found relatively close to the surface (88% of observations within 15 cm of surface) when utilizing burrows. Although we document relatively high annual survival (56%) compared to previous studies, we also detected Ophidiomyces ophiodiicola, the fungal causative agent for snake fungal disease, in 3 of our 7 individuals included in the study, highlighting the continued threats facing the species.

The seventh edition of the most authoritative and comprehensive book published on lung function, now completely revised and restructured Lung function assessment is the central pillar of respiratory diagnosis. Most hospitals have lung function laboratories where patients are tested with a variety of physiological methods. The tests and techniques used are specialized and utilize the expertise of respiratory physicians, physiologists, and technicians. This new edition of the classic text on lung function is a theoretical textbook and practical manual in one that gives a comprehensive account of lung function and its assessment in healthy persons and those with all types of respiratory disorder, against a background of respiratory, exercise, and environmental physiology. It incorporates the technical and methodological recommendations for lung function testing of the American Thoracic Society and European Respiratory Society. Cotes' Lung Function, 7th Edition is filled with chapters covering respiratory surveys, respiratory muscles, neonatal assessment, exercise, sleep, high altitude, hyperbaria, the effects of cold and heat, respirable dusts, fumes and vapors, anesthesia, surgery, and respiratory rehabilitation. It also offers a compendium of lung function in selected individual diseases and is filled with more diagrams and illustrative cases than previous editions. * The only text to cover lung function assessment from first principles including methodology, reference values, and interpretation * Completely re-written in a contemporary style—includes user-friendly equations and more diagrams * Covers the latest advances in the treatment of lung function, including a stronger clinical and practical bias and more on new techniques and equipment * Keeps mathematical treatments to a minimum Cotes' Lung Function is an ideal guide for respiratory physicians and surgeons, staff of lung function laboratories, and others who have a professional interest in the function of the lungs at rest or on exercise and how it may be assessed. Physiologists, anthropologists, pediatricians, anesthetists, occupational physicians, explorers, epidemiologists, and respiratory nurses should also find the book useful.

Excessive heat exposure reduces intestinal integrity and post-absorptive energetics that can inhibit wellbeing and be fatal. Therefore, our objectives were to examine how acute heat stress (HS) alters intestinal integrity and metabolism in growing pigs. Animals were exposed to either thermal neutral (TN, 21°C; 35-50% humidity; n=8) or HS conditions (35°C; 24-43% humidity; n=8) for 24 h. Compared to TN, rectal temperatures in HS pigs increased by 1.6°C and respiration rates by 2-fold (P<0.05). As expected, HS decreased feed intake by 53% (P<0.05) and body weight (P<0.05) compared to TN pigs. Ileum heat shock protein 70 expression increased (P<0.05), while intestinal integrity was compromised in the HS pigs (ileum and colon TER decreased; P<0.05). Furthermore, HS increased serum endotoxin concentrations (P=0.05). Intestinal permeability was accompanied by an increase in protein expression of myosin light chain kinase (P<0.05) and casein kinase II-α (P=0.06). Protein expression of tight junction (TJ) proteins in the ileum revealed claudin 3 and occludin expression to be increased overall due to HS (P<0.05), while there were no differences in claudin 1 expression. Intestinal glucose transport and blood glucose were elevated due to HS (P<0.05). This was supported by increased ileum Na(+)/K(+) ATPase activity in HS pigs. SGLT-1 protein expression was unaltered; however, HS increased ileal GLUT-2 protein expression (P=0.06). Altogether, these data indicate that HS reduce intestinal integrity and increase intestinal stress and glucose transport.

1. Climate change is reported to have caused widespread changes to species' populations and ecological communities. Warming has been associated with population declines in long-distance migrants and habitat specialists, and increases in southerly distributed species. However, the specific climatic drivers behind these changes remain undescribed. 2. We analysed annual fluctuations in the abundance of 59 breeding bird species in England over 45 years to test the effect of monthly temperature and precipitation means upon population trends. 3. Strong positive correlations between population growth and both winter and breeding season temperature were identified for resident and short-distance migrants. Lagged correlations between population growth and summer temperature and precipitation identified for the first time a widespread negative impact of hot, dry summer weather. Resident populations appeared to increase following wet autumns. Populations of long-distance migrants were negatively affected by May temperature, consistent with a potential negative effect of phenological mismatch upon breeding success. There was evidence for some nonlinear relationships between monthly weather variables and population growth. 4. Habitat specialists and cold-associated species showed consistently more negative effects of higher temperatures than habitat generalists and southerly distributed species associated with warm temperatures. Results suggest that previously reported changes in community composition represent the accumulated effects of spring and summer warming. 5. Long-term population trends were more significantly correlated with species' sensitivity to temperature than precipitation, suggesting that warming has had a greater impact on population trends than changes in precipitation. Months where there had been the greatest warming were the most influential drivers of long-term change. There was also evidence that species with the greatest sensitivity to extremes of precipitation have tended to decline. 6. Our results provide novel insights about the impact of climate change on bird communities. Significant lagged effects highlight the potential for altered species' interactions to drive observed climate change impacts, although some community changes may have been driven by more immediate responses to warming. In England, resident and short-distance migrant populations have increased in response to climate change, but potentially at the expense of long-distance migrants, habitat specialists and cold-associated species.

Precise neurostimulation can revolutionize therapies for neurological disorders. Electrode-based stimulation devices face challenges in achieving precise and consistent targeting due to the immune response and the limited penetration of electrical fields. Ultrasound can aid in energy propagation, but transcranial ultrasound stimulation in the deep brain has limited spatial resolution caused by bone and tissue scattering. Here, we report an implantable piezoelectric ultrasound stimulator (ImPULS) that generates an ultrasonic focal pressure of 100 kPa to modulate the activity of neurons. ImPULS is a fully-encapsulated, flexible piezoelectric micromachined ultrasound transducer that incorporates a biocompatible piezoceramic, potassium sodium niobate [(K,Na)NbO 3 ]. The absence of electrochemically active elements poses a new strategy for achieving long-term stability. We demonstrated that ImPULS can i) excite neurons in a mouse hippocampal slice ex vivo, ii) activate cells in the hippocampus of an anesthetized mouse to induce expression of activity-dependent gene c-Fos, and iii) stimulate dopaminergic neurons in the substantia nigra pars compacta to elicit time-locked modulation of nigrostriatal dopamine release. This work introduces a non-genetic ultrasound platform for spatially-localized neural stimulation and exploration of basic functions in the deep brain. Ultrasound neuromodulation overcomes limitations of electrode-based stimulation through improved targeting and long-term stability for treating neurological disorders. Here, authors present a hair-thin, implantable piezoelectric stimulator that selectively modulates neurons in the deep brain.

Loneliness is a relatively common problem in young people (14–24 years) and predicts the onset of depression and anxiety. Interventions to reduce loneliness thus have significant potential as active ingredients in strategies to prevent or alleviate anxiety and depression among young people. Previous reviews have focused on quantitative evidence and have not examined potential mechanisms that could be targets for intervention strategies. To build on this work, in this review we aimed to combine qualitative and quantitative evidence with stakeholder views to identify interventions that appear worth testing for their potential effectiveness in reducing loneliness, anxiety and depression in young people aged 14–24 years, and provide insights into the potential mechanisms of action. We conducted a Critical Interpretative Synthesis, a systematic review method that iteratively synthesises qualitative and quantitative evidence and is explicitly focused on building theory through a critical approach to the evidence that questions underlying assumptions. Literature searches were performed using nine databases, and eight additional databases were searched for theses and grey literature. Charity and policy websites were searched for content relevant to interventions for youth loneliness. We incorporated elements of Rapid Realistic Review approaches by consulting with young people and academic experts to feed into search strategies and the resulting conceptual framework, in which we aimed to set out which interventions appear potentially promising in terms of theoretical and empirical underpinnings and which fit with stakeholder views. We reviewed effectiveness data and quality ratings for the included randomised controlled trials only. Through synthesising 27 studies (total participants n = 105,649; range 1–102,072 in different studies) and grey literature, and iteratively consulting with stakeholders, a conceptual framework was developed. A range of ‘Intrapersonal’ (e.g. therapy that changes thinking and behaviour), ‘Interpersonal’ (e.g. improving social skills), and ‘Social’ Strategies (e.g. enhancing social support, and providing opportunities for social contact) seem worth testing further for their potential to help young people address loneliness, thereby preventing or alleviating depression and/or anxiety. Such strategies should be co-designed with young people and personalised to fit individual needs. Plausible mechanisms of action are facilitating sustained social support, providing opportunities for young people to socialise with peers who share similar experiences, and changing thinking and behaviour, for instance through building positive attitudes to themselves and others. The most convincing evidence of effectiveness was found in support of Intrapersonal Strategies: two randomised controlled studies quality-rated as ‘good’ found decreases in loneliness associated with different forms of therapy (Cognitive Behavioural Therapy or peer network counselling), although power calculations were not reported, and effect sizes were small or missing. Strategies to address loneliness and prevent or alleviate anxiety and depression need to be co-designed and personalised. Promising elements to incorporate into these strategies are social support, including from peers with similar experiences, and psychological therapy.