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Edith M Heintjes,1 Irene D Bezemer,1 Daniel Prieto-Alhambra,2,3 Elisabeth Smits,1 Helen P Booth,4 Daniel Dedman,4 Ying He,3 Fabian Hoti,5 Minna Vehkala,5 Stefan de Vogel,6 Noah Jamie Robinson,6 Kwame Appenteng,7 Fernie JA Penning-van Beest1 1PHARMO Institute for Drug Outcomes Research, Utrecht, the Netherlands; 2Idiap Jordi Gol Primary Care Research Institute and CIBERFes, Universitat Autonoma de Barcelona and Instituto de Salud Carlos III, Barcelona, Catalonia, Spain; 3Centre for Statistics in Medicine, NDORMS, University of Oxford, Oxford, England, UK; 4Clinical Practice Research Datalink (CPRD), London, UK; 5Statfinn - EPID Research, Espoo, Finland; 6Astellas Pharma Europe B.V., Leiden, Netherlands; 7Astellas Pharma Global Development, Inc., Northbrook, IL, USACorrespondence: Edith M HeintjesPHARMO Institute for Drug Outcomes Research, Van Deventerlaan 30-40, Utrecht, AE 3528, the NetherlandsTel +31 30 7440 800Email pharmo@pharmo.nlBackground: Mirabegron, indicated for the treatment of overactive bladder, is contraindicated in patients with severe uncontrolled hypertension (systolic blood pressure ≥ 180 mm Hg and/or diastolic blood pressure ≥ 110 mm Hg). In September 2015, a Direct Healthcare Professional Communication (DHPC) letter was disseminated as an additional risk minimisation measure.Purpose: To assess the effectiveness of the DHPC in reducing the proportions of patients with severe or non-severe uncontrolled hypertension at mirabegron initiation.Methods: An observational multi-database cohort study was undertaken using routinely collected healthcare data (December 2012–December 2016) from the PHARMO Database Network (Netherlands), SIDIAP database (Spain), CPRD (United Kingdom, UK) and national healthcare registers and electronic medical records from Finland. DHPC effectiveness was evaluated using interrupted time series analyses comparing trends and changes in monthly proportions of severe or non-severe uncontrolled hypertensive mirabegron initiations relative to the timing of the DHPC dissemination.Results: The study population comprised 52,078 patients. Prior to DHPC dissemination, across the four databases, 0.3– 1.3% had severe uncontrolled hypertension. Estimated absolute changes (EAC) in proportions of severe uncontrolled hypertension post-DHPC indicated a tendency towards a lower proportion in the Netherlands (EAC − 0.36%, p=0.053), unchanged proportions in Spain and the UK and a higher proportion in Finland (EAC +0.73%, p=0.016). For non-severe uncontrolled hypertension (13– 16% pre-DHPC), post-DHPC proportions tended to be lower in the Netherlands (EAC − 2.02%, p=0.038) and Spain (EAC − 1.04%, p=0.071), and unchanged in the UK and Finland.Conclusion: Severe uncontrolled hypertension prior to mirabegron initiation was uncommon in these four European countries even before DHPC dissemination. This suggests that other risk minimisation communications (prior to the DHPC dissemination) had worked adequately with respect to minimising mirabegron use among patients with severe uncontrolled hypertension. No strong and consistent evidence of further risk minimisation after the DHPC dissemination was observed in this study.Keywords: drug utilisation study, direct healthcare professional communication, risk minimization, interrupted time series analysis

BackgroundIn Kenya, Community Health Committees (CHC) were established to enhance community participation in health services. Their role is to provide leadership, oversight in delivery of community health services, promote social accountability and mobilize resources for community health. CHCs form social networks with other actors, with whom they exchange health information for decision-making and accountability. This case study aimed to explore the structure of a rural and an urban CHC network and to analyze how health-related information flowed in these networks. Understanding the pathways of information in community settings may provide recommendations for strategies to improve the role and functioning of CHCs.MethodsIn 2017, we conducted 4 focus group discussions with 27 community discussants and 10 semi-structured interviews with health professionals in a rural area and an urban slum. Using social network analysis, we determined the structure of their social networks and how health related information flowed in these networks.ResultsBoth CHCs were composed of respected persons nominated by their communities. Each social network had 12 actors that represented both community and government institutions. CHCs were not central actors in the exchange of health-related information. Health workers, community health volunteers and local Chiefs in the urban slum often passed information between the different groups of actors, while CHCs hardly did this. Therefore, CHCs had little control over the flow of health-related information. Although CHC members were respected persons who served in multiple roles within their communities, this did not enhance their centrality. It emerged that CHCs were often left out in the flow of health-related information and decision-making, which led to demotivation. Community health volunteers were more involved by other actors such as health managers and non-governmental organizations as a conduit for health-related information.ConclusionSocial network analysis demonstrated how CHCs played a peripheral role in the flow of health-related information. Their perception of being left out of the information flow led to demotivation, which hampered their ability to facilitate community participation in community health services; hence challenging effective participation through CHCs.

BackgroundClose-to-community (CTC) providers of health care are a crucial workforce for delivery of high-quality and universal health coverage. There is limited evidence on the effect of training supervisors of this cadre in supportive supervision. Our study aimed to demonstrate the effects of a training intervention on the approach to and frequency of supervision of CTC providers of health care.MethodsWe conducted a context analysis in 2013 in two Kenyan counties to assess factors that influenced delivery of community health services. Supervision was identified a priority factor that needed to be addressed to improve community health services. Supervision was inadequate due to lack of supervisor capacity in supportive approaches and lack of supervision guidelines. We designed a six-day training intervention and trained 48 purposively selected CTC supervisors on the educative, administrative and supportive components of supportive supervision, problem solving and advocacy and provided them with checklists to guide supervision sessions. We administered quantitative questionnaires to supervisors to assess changes in supervision frequency before and after the training and then explored perspectives on the intervention with community health volunteers (CHVs) and their supervisors using qualitative in-depth interviews.ResultsSix months after the intervention, we observed that supervisors had shifted the supervision approach from being controlling and administrative to coaching, mentorship and problem solving. Changes in the frequency of supervision were found in Kitui only, whereby significant decreases in group supervision were met with increases in accompanied home visit supervision. Supervisors and CHVs reported the intervention was helpful and it responded to capacity gaps in supervision of CHVs.ConclusionOur intervention responded to capacity gaps in supervision and contributed to enhanced supervision capacity among supervisors. Supervisors found the curriculum acceptable and useful in improving supervision skills.

Background: Unprotected heterosexual contact in the presence of other sexually transmitted diseases (STD) enhances the probability of HIV transmission. The objective of this study was to estimate the proportion of HIV infections attributable to STD in rural Uganda.Methods: Simulation modelling scenarios of the transmission dynamics of HIV infection and of ulcerative and non-ulcerative STD were employed to address this objective, drawing on data from a specific rural population cohort of 10,000 in south-west Uganda.Results: In simulations of the initial 10-year period of the HIV epidemic (1980-1990), over 90% of HIV infections were attributed to STD. Even given conservative assumptions about the prevalence of STD and about their enhancing effects on HIV transmission, STD played a critical role in the rapid and extensive spread of HIV infection. The role of STD decreased with progression of the HIV epidemic.Conclusions: In developing countries, control of the spread of HIV infection may benefit substantially from successful STD intervention programmes, and particularly in areas where HIV infection is not already well established. [ABSTRACT FROM AUTHOR]

BACKGROUND:The MSM population in Kenya contributes to 15% of HIV incidence. This calls for innovative HIV prevention interventions. Pre-exposure prophylaxis (PrEP) has been efficacious in preventing HIV among MSM in trials. There is limited data on the willingness to take daily oral PrEP in sub-Sahara Africa. PrEP has not been approved for routine use in most countries globally. This study aimed to document the willingness to take PrEP and barriers to uptake and adherence to PrEP in Kenya. The findings will inform the design of a PrEP delivery program as part of the routine HIV combination prevention. METHODS:Eighty MSM were recruited in 2 Counties in December 2013. Quantitative data on sexual behaviour and willingness to take PrEP were collected using semi-structured interviews and analysed using SPSS. Qualitative data on knowledge of PrEP, motivators and barriers to uptake and adherence to PrEP were collected using in-depth interviews and FGDs and analysed using Nvivo. Analysis of data in willingness to take PrEP was conducted on the HIV negative participants (n = 55). RESULTS:83% of MSM were willing to take daily oral HIV PrEP. Willingness to take PrEP was higher among the bi-sexual and younger men. Motivators for taking PrEP were the need to stay HIV negative and to protect their partners. History of poor medication adherence, fear of side effects and HIV stigma were identified as potential barriers to adherence. Participants were willing to buy PrEP at a subsidized price. CONCLUSIONS:There is willingness to take PrEP among MSM in Kenya and there is need to invest in targeted education and messaging on PrEP to enhance adherence, proper use and reduce stigma in the general population and among policy makers.

Promoting human contact with wildlife can be harmful to animal conservation and welfare by exposing animals to unsafe situations and driving demand for wildlife tourism and exotic pets. Conservationists and researchers justifiably use social media to raise awareness, but professionals posting pictures of themselves with animals can have unintended negative consequences for conservation. Though the International Union for Conservation of Nature Best Practice Guidelines for Responsible Images of Non-Human Primates suggests researchers and animal professionals provide context in captions of images of humans interacting with primates, there is little research investigating whether this approach is effective. We investigated whether informative captions affect viewers' desires to have primates as pets and attitudes toward wildlife conservation. Using 4 mock Instagram posts depicting human-gorilla and human-slender loris interactions, we surveyed 2977 respondents to assess the effect of captions on viewers' perceptions of the images and primate conservation. Likert scale response data were analyzed with ordered probit regression models. Captions clearly contextualizing an image as research resulted in a significantly higher agreement that posts depicted wildlife research (gorilla β = 0.28 [SE 0.06], p < 0.001; loris β = 0.18 [0.06], p = 0.002), but such captions resulted in no significant difference in responses regarding viewers' desires to own primates as pets or questions regarding the primates' conservation statuses. Although most participants agreed the primates were endangered, more than 56% and 59%, respectively, stated they would have a gorilla or loris as a pet, that they would make good pets, or both, further supporting the conclusion that captions do not minimize harmful impacts of images of human-primate interactions., (© 2024 The Authors. Conservation Biology published by Wiley Periodicals LLC on behalf of Society for Conservation Biology.)

Life on our planet likely evolved in the ocean, and thus exo-oceans are key habitats to search for extraterrestrial life. We conducted a data-driven bibliographic survey on the astrobiology literature to identify emerging research trends with marine science for future synergies in the exploration for extraterrestrial life in exo-oceans. Based on search queries, we identified 2592 published items since 1963. The current literature falls into three major groups of terms focusing on (1) the search for life on Mars, (2) astrobiology within our Solar System with reference to icy moons and their exo-oceans, and (3) astronomical and biological parameters for planetary habitability. We also identified that the most prominent research keywords form three key-groups focusing on (1) using terrestrial environments as proxies for Martian environments, centred on extremophiles and biosignatures, (2) habitable zones outside of "Goldilocks" orbital ranges, centred on ice planets, and (3) the atmosphere, magnetic field, and geology in relation to planets' habitable conditions, centred on water-based oceans., Competing Interests: The authors declare no conflicts of interest.

Background: Many factors contribute to developing and conducting a successful multi-data source, non-interventional, post-authorization safety study (NI-PASS) for submission to multiple health authorities. Such studies are often large undertakings; evaluating and sharing lessons learned can provide useful insights to others considering similar studies., Objectives: We discuss challenges and key methodological and organizational factors that led to the delivery of a successful post-marketing requirement (PMR)/PASS program investigating the risk of cardiovascular and cancer events among users of mirabegron, an oral medication for the treatment of overactive bladder., Results: We provide context and share learnings, including sections on research program collaboration, scientific transparency, organizational approach, mitigation of uncertainty around potential delays, validity of study outcomes, selection of data sources and optimizing patient numbers, choice of comparator groups and enhancing precision of estimates of associations, potential confounding and generalizability of study findings, and interpretation of results., Conclusions: This large PMR/PASS program was a long-term commitment from all parties and benefited from an effective coordinating center and extensive scientific interactions across research partners, scientific advisory board, study sponsor, and health authorities, and delivered useful learnings related to the design and organization of multi-data source NI-PASS., (© 2024 The Authors. Pharmacoepidemiology and Drug Safety published by John Wiley & Sons Ltd.)

Booralana nickorum sp. nov. is described from the deep-water slope of the Exuma Sound, The Bahamas, from depths of 540 to 560 metres. It is the fourth species to be assigned to the genus and the second species described from the Western North Atlantic. The species can be distinguished from Booralana tricarinata Camp and Heard, 1988 and the other species by the sub-triangular pleotelson and the uropodal exopod of mature males being far longer than endopod, with both rami extending well beyond the posterior margin of the pleotelson. Additionally, pleopods 3 and 4 lack a prominent angle at midpoint of ramus.

Sea turtles are frequently proposed as indicator species for assessing ocean health. To faciliate the use of these species as bioindicators requires the development of tools for rapidly and effectively assessing individual health. Here, we collected 104 blood samples from 69 loggerhead sea turtles, Caretta caretta, undergoing rehabilitation to determine the connection between health status, the activity of B-esterases, and other biochemical parameters. To determine the optimal assay protocol for B-esterases, we measured the activity and kinetics of cholinesterases-(ChEs) and carboxylesterases (CEs) using 3 and 5 commercial substrates, respectively, at different assay conditions. IC50 values for the activity of B-esterases were calculated within a concentration range for model pesticide inhibitors. Turtles' health status was determined via routine veterinary procedures. During rehabilitation (which was associated with improving health status), we observed a decrease in the activity of most enzymes (especially in acetylcholinesterase) alongside an increase in CE when using p-nitrophenyl acetate as a substrate. As such, it is possible that the activity rates of plasmatic B-esterases could serve as an indicator of health status. There is also high potential that B-esterases could be specifically sensitive to marine pollutants although to further validate this would require future studies to specifically correlate B-esterarse activities to pollutant concentrations in blood or excreta., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Ocean conditions near the grounding zones of Antarctica's ice shelves play a key role in controlling the outflow and mass balance of the ice sheet. However, ocean observations in these regions are largely absent. Here, we present a detailed spatial survey collected with an underwater vehicle in a basal crevasse located in the ocean cavity at the Ross Ice Shelf grounding zone. The observations depict fine-scale variability in ocean forcing that drives asymmetric melting along the lower crevasse sidewalls and freezing in the upper reaches of the crevasse. Freshwater release from melting at depth and salt rejection from freezing above drives an overturning circulation. This vertical circulation pattern overlays a dominant throughflow jet, which funnels water parallel to the coastline, orthogonal to the direction of tidal currents. Importantly, these data reveal that basal crevasses influence ocean circulation and mixing at ice shelf grounding zones to an extent previously unknown.

Tissue and blood gas embolism (GE) associated with fisheries bycatch are likely a widespread, yet underestimated, cause of sea turtle mortality. Here, we evaluated risk factors associated with tissue and blood GE in loggerhead turtles caught incidentally by trawl and gillnet fisheries on the Valencian coastline of Spain. Of 413 turtles (303 caught by trawl, 110 by gillnet fisheries), 54% ( n  = 222) exhibited GE. For sea turtles caught in trawls, the probability and severity of GE increased with trawl depth and turtle body mass. In addition, trawl depth and the GE score together explained the probability of mortality (P[mortality]) following recompression therapy. Specifically, a turtle with a GE score of 3 caught in a trawl deployed at 110 m had a P[mortality] of ~50%. For turtles caught in gillnets, no risk variables were significantly correlated with either the P[GE] or GE score. However, gillnet depth or GE score, separately, explained P[mortality], and a turtle caught at 45 m or with a GE score between 3 and 4 had a P[mortality] of 50%. Differences in the fishery characteristics precluded direct comparison of GE risk and mortality between these gear types. Although P[mortality] is expected to be significantly higher in untreated turtles released at sea, our findings can improve estimates of sea turtle mortality associated with trawls and gillnets, and help guide associate conservation efforts., Competing Interests: None of the authors have conflicting interests., (© The Author(s) 2023. Published by Oxford University Press and the Society for Experimental Biology.)

An intense, nonresolving airway inflammatory response leads to destructive lung disease in cystic fibrosis (CF). Dysregulation of macrophage immune function may be a key facet governing the progression of CF lung disease, but the underlying mechanisms are not fully understood. We used 5' end centered transcriptome sequencing to profile P. aeruginosa LPS-activated human CF macrophages, showing that CF and non-CF macrophages deploy substantially distinct transcriptional programs at baseline and following activation. This includes a significantly blunted type I IFN signaling response in activated patient cells relative to healthy controls that was reversible upon in vitro treatment with CFTR modulators in patient cells and by CRISPR-Cas9 gene editing to correct the F508del mutation in patient-derived iPSC macrophages. These findings illustrate a previously unidentified immune defect in human CF macrophages that is CFTR dependent and reversible with CFTR modulators, thus providing new avenues in the search for effective anti-inflammatory interventions in CF.

The acquisition of Co II by the corrin component of vitamin B 12 follows one of two distinct pathways, referred to as early or late Co II insertion. The late insertion pathway exploits a Co II metallochaperone (CobW) from the COG0523 family of G3E GTPases, while the early insertion pathway does not. This provides an opportunity to contrast the thermodynamics of metalation in a metallochaperone-requiring and a metallochaperone-independent pathway. In the metallochaperone-independent route, sirohydrochlorin (SHC) associates with the CbiK chelatase to form Co II -SHC. Co II -buffered enzymatic assays indicate that SHC binding enhances the thermodynamic gradient for Co II transfer from the cytosol to CbiK. In the metallochaperone-dependent pathway, hydrogenobyrinic acid a,c -diamide (HBAD) associates with the CobNST chelatase to form Co II -HBAD. Here, Co II -buffered enzymatic assays indicate that Co II transfer from the cytosol to HBAD-CobNST must somehow traverse a highly unfavorable thermodynamic gradient for Co II binding. Notably, there is a favorable gradient for Co II transfer from the cytosol to the Mg II GTP-CobW metallochaperone, but further transfer of Co II from the GTP-bound metallochaperone to the HBAD-CobNST chelatase complex is thermodynamically unfavorable. However, after nucleotide hydrolysis, Co II transfer from the chaperone to the chelatase complex is calculated to become favorable. These data reveal that the CobW metallochaperone can overcome an unfavorable thermodynamic gradient for Co II transfer from the cytosol to the chelatase by coupling this process to GTP hydrolysis., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

Telomeres are structures that cap the ends of linear chromosomes and play critical roles in maintaining genome integrity and establishing the replicative lifespan of cells. In stem and cancer cells, telomeres are actively elongated by either telomerase or the alternative lengthening of telomeres (ALT) pathway. This pathway is characterized by several hallmark features, including extrachromosomal C-rich circular DNAs that can be probed to assess ALT activity. These so-called C-circles are the product of ALT-associated DNA damage repair processes and simultaneously serve as potential templates for iterative telomere extension. This bifunctional nature makes C-circles highly sensitive and specific markers of ALT. Here, we describe a C-circle assay, adapted from previous reports, that enables the quantitation of C-circle abundance in mammalian cells subjected to a wide range of experimental perturbations. This protocol combines the Quick C-circle Preparation (QCP) method for DNA isolation with fluorometry-based DNA quantification, rolling circle amplification (RCA), and detection of C-circles using quantitative PCR. Moreover, the inclusion of internal standards with well-characterized telomere maintenance mechanisms (TMMs) allows for the reliable benchmarking of cells with unknown TMM status. Overall, our work builds upon existing protocols to create a generalizable workflow for in vitro C-circle quantitation and ascertainment of TMM identity., Competing Interests: Competing interestsThe authors declare that they have no compe ting interests., (Copyright © 2023 The Authors; This is an open access article under the CC BY-NC license (https://creativecommons.org/licenses/by-nc/4.0/).)

The burrowing crustacean decapod Nephrops norvegicus is a significant species in European Atlantic and Mediterranean fisheries. Research over the decades has mainly focused on behavioral and physiological aspects related to the burrowing lifestyle, since animals can only be captured by trawls when engaged in emergence on the seabed. Here, we performed a global bibliographic survey of all the scientific literature retrieved in SCOPUS since 1965, and terminology maps were produced with the VOSviewer software to reveal established and emerging research areas. We produced three term-map plots: term clustering, term citation, and term year. The term clustering network showed three clusters: fishery performance, assessment, and management; biological cycles in growth, reproduction, and behavior; and finally, physiology and ecotoxicology, including food products. The term citation map showed that intense research is developed on ecotoxicology and fishery management. Finally, the term year map showed that the species was first studied in its morphological and physiological aspects and more recently in relation to fishery and as a food resource. Taken together, the results indicate scarce knowledge on how burrowing behavior and its environmental control can alter stock assessment, because of the poor use of current and advanced monitoring technologies.

Metalation, the acquisition of metals by proteins, must avoid mis-metalation with tighter binding metals. This is illustrated by four selected proteins that require different metals: all show similar ranked orders of affinity for bioavailable metals, as described in a universal affinity series (the Irving-Williams series). Crucially, cellular protein metalation occurs in competition with other metal binding sites. The strength of this competition defines the intracellular availability of each metal: its magnitude has been estimated by calibrating a cells' set of DNA-binding, metal-sensing, transcriptional regulators. This has established that metal availabilities (as free energies for forming metal complexes) are maintained to the inverse of the universal series. The tightest binding metals are least available. With these availabilities, correct metalation is achieved., (© 2022 The Authors. FEBS Letters published by John Wiley & Sons Ltd on behalf of Federation of European Biochemical Societies.)

Sea turtles generally lay several clutches of eggs in a single nesting season. While a negative correlation between water temperatures and the time required between constitutive nesting events (termed the internesting interval) has been previously reported in loggerhead Caretta caretta and green turtles Chelonia mydas, it is not understood whether this relationship remains constant across other sea turtle species. Here, we expanded upon these previous studies on loggerhead and green turtles by using larger sample sizes and including data from species with a wider range of body-sizes; specifically: hawksbill Eretmochelys imbricata, leatherback Dermochelys coriacea, and olive ridley turtles Lepidochelys olivacea. In total, we compiled temperature data from biologgers deployed over internesting intervals on 23 loggerhead, 22 green, 7 hawksbill, 26 leatherback and 11 olive ridley turtles from nesting sites in 8 different countries. The relationship between the duration of the internesting interval and water temperatures in green and loggerhead turtles were statistically similar yet it differed between all other turtle species. Specifically, hawksbill turtles had much longer internesting intervals than green or loggerhead turtles even after controlling for temperature. In addition, both olive ridley and leatherback turtles exhibited thermal independence of internesting intervals presumably due to the large body-size of leatherback turtles and the unique capacity of ridley turtles to delay oviposition. The observed interspecific differences in the relationship between the length of the internesting interval and water temperatures indicate the complex and variable responses that each sea turtle species may exhibit due to environmental fluctuations and climate change., Competing Interests: Declaration of competing interest The authors declare no conflicts of interest., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objectives: The COVID-19 pandemic saw major changes to small animal veterinary practice, many of which may have had an impact on stress in cats presented to the clinic. The aim of this study was to examine the nature of feline outpatient visits before and during the pandemic, and examine signs of stress noted in cats before, during and after these visits., Methods: A questionnaire was used to gather data on cat owner experiences of visits to the veterinary clinic. Data were gathered on the owner's most recent experience of a consultation, with consultations occurring in February 2020 or earlier coded as a standard consultation, and consultations occurring in March 2020 or later coded as COVID-19 consultations., Results: A total of 371 responses were received, with 210 coded as standard consultations and 161 coded as COVID-19 consultations. Consultation type varied significantly between standard and COVID-19 consultations ( P <0.001), with emergency consultations more frequent and preventive healthcare consultations less frequent during the COVID-19 pandemic. The area in which the owner and their cat waited also varied significantly between standard and COVID-19 consultations ( P <0.001), with standard consultations more likely to involve time in a waiting room while COVID-19 consultations were often called straight in or waited outside the practice. Most owners notedbehaviours associated with stress in their cats, regardless of consultation type, although trying to hide or escape were noted more frequently for cats seen prior to the COVID-19 pandemic., Conclusions and Relevance: The findings suggest that outpatient visits to the veterinary clinic were stressful for cats both prior to and during the COVID-19 pandemic, but some measures taken during the pandemic, for example less use of waiting rooms, could be used alongside existing cat friendly measures to help to reduce stress in feline patients.

Objectives: The importance of the effects of stress on cats is well recognised, with visits to the veterinary clinic a common cause of stress. The aim of this study was to explore owners' experiences of stress, both for themselves and their cat, during veterinary visits, and to gather data on owners' perception of resources within the clinic to reduce stress in their cats., Methods: A questionnaire aimed at cat owners with recent experience of an outpatient visit to the veterinary clinic was developed and distributed. Questions covered demographics; their most recent veterinary visit; their general experience of visits over the past 3 years; measures taken by the clinic to reduce stress; and awareness of the Cat Friendly Clinic programme. There were various question types, with owners often asked to rate stress from 1 (least stressful) to 10 (most stressful)., Results: A total of 277 respondents gave details about their experience of consultations over the past 3 years. Owners rated the stress of travelling to the clinic, other animals in the waiting room and the consultation itself (all median 6, interquartile range 4-8) as the most stressful elements. Most owners reported gentle methods of removing the cat from their carrier; however, almost a third (n = 81/263; 30.8%) had seen their cat scruffed during a consultation. Cat-only waiting rooms were viewed as the most effective measure to reduce stress in the clinic., Conclusions and Relevance: Veterinary clinics are already taking steps to address stress in cats, and owners have a good awareness of stress in their cats. Future work should focus on trialling specific interventions to determine their effectiveness in reducing stress in feline patients, and measures currently perceived by owners to be highly effective, such as cat-only waiting rooms, should be used where feasible.

The recent commercialization of unoccupied aerial vehicles (UAVs) has facilitated their incorporation into a variety of ecological studies. While UAVs are able to provide accurate visual data of marine species from an aerial perspective, these devices have some limitations that make measuring marine animals below the surface challenging. Many marine organisms are often visible from the air, but are deeper in the water column, and current methods cannot measure animals below the surface. Here, we developed and tested a stereo-video camera (SVC) system that was mounted onto a commercially-available UAV. We used the SVC-UAV to conduct remote body-size measurements for two marine species: the green sea turtle (Chelonia mydas) and the nurse shark (Ginglymostoma cirratum). When comparing SVC measurements to those taken by hand, the SVC-UAV had a mean absolute error (MAE) of 4.44 cm (n = 6; mean percent error (MPE) = 10.6%) for green sea turtles and 7.16 cm absolute error (n = 1; PE = 3.6%) for the nurse shark. Using a linear model, we estimated the slope of the SVC versus hand measurements for green sea turtles to be 1.085 (±0.099 SE), and accounting for the standard error, a measurement bias was not apparent. Using model selection, based on a global model predicting MAE from animal distance to the SVC and body size, the top ranked model was the intercept-only model. This indicates that neither animal distance nor body size strongly influenced measurement error. Incorporating SVC systems into UAVs can allow for relatively accurate measurements of near surface-dwelling marine species. To our knowledge, there is no other stand-alone SVC for UAVs available that offers similar accuracy and utility., Competing Interests: The authors have declared that no competing interests exist.

Our understanding of the importance of microbiomes on large aquatic animals-such as whales, sea turtles and manatees-has advanced considerably in recent years. The latest observations indicate that epibiotic diatom communities constitute diverse, polyphyletic, and compositionally stable assemblages that include both putatively obligate epizoic and generalist species. Here, we outline a successful approach to culture putatively obligate epizoic diatoms without their hosts. That some taxa can be cultured independently from their epizoic habitat raises several questions about the nature of the interaction between these animals and their epibionts. This insight allows us to propose further applications and research avenues in this growing area of study. Analyzing the DNA sequences of these cultured strains, we found that several unique diatom taxa have evolved independently to occupy epibiotic habitats. We created a library of reference sequence data for use in metabarcoding surveys of sea turtle and manatee microbiomes that will further facilitate the use of environmental DNA for studying host specificity in epizoic diatoms and the utility of diatoms as indicators of host ecology and health. We encourage the interdisciplinary community working with marine megafauna to consider including diatom sampling and diatom analysis into their routine practices., (© 2022. The Author(s).)

Three Web-based calculators, and three analogous spreadsheets, have been generated that predict in vivo metal occupancies of proteins based on known metal affinities. The calculations exploit estimates of the availabilities of the labile buffered pools of different metals inside a cell. Here, metal availabilities have been estimated for a strain of Escherichia coli that is commonly used in molecular biology and biochemistry research, e.g. in the production of recombinant proteins. Metal availabilities have been examined for cells grown in Luria-Bertani (LB) medium aerobically, anaerobically, and in response to H2O2 by monitoring the abundance of a selected set of metal-responsive transcripts by quantitative polymerase chain reaction (qPCR). The selected genes are regulated by DNA-binding metal sensors that have been thermodynamically characterized in related bacterial cells enabling gene expression to be read out as a function of intracellular metal availabilities expressed as free energies for forming metal complexes. The calculators compare these values with the free energies for forming complexes with the protein of interest, derived from metal affinities, to estimate how effectively the protein can compete with exchangeable binding sites in the intracellular milieu. The calculators then inter-compete the different metals, limiting total occupancy of the site to a maximum stoichiometry of 1, to output percentage occupancies with each metal. In addition to making these new and conditional calculators available, an original purpose of this article was to provide a tutorial that discusses constraints of this approach and presents ways in which such calculators might be exploited in basic and applied research, and in next-generation manufacturing., (© The Author(s) 2022. Published by Oxford University Press.)

Background: There has been increasing provision of veterinary telemedicine consultations, particularly during the COVID-19 pandemic; however, little evidence currently exists examining these remote consultations. The aim of this cross-sectional study was to explore veterinary and cat owner experiences of telemedicine consultations during the pandemic., Methods: Two questionnaires, one aimed at veterinary professionals and one at cat owners, were launched in September 2020. Questions explored the type of consultation conducted remotely, the perceived advantages and disadvantages of telemedicine, and the role of telemedicine in the future of veterinary practice., Results: Responses were received from 242 veterinary professionals and 98 owners with experience of telemedicine. Monitoring and advice consultations were felt to be most suited to telemedicine. Reduced stress for owners/cats was seen as an advantage of telemedicine, while lack of clinical examination and risk of misdiagnosis were viewed as disadvantages. Most respondents (85.7% [n = 84/98] of owners; 67.4% [n = 163/242] of veterinary professionals) felt practices should continue to offer telemedicine consultations., Conclusion: With increasing pet ownership and practice workload, telemedicine may play a crucial role in the future of veterinary practice. Future work should focus on a strategic approach to feline telemedicine, integrating it alongside face-to-face visits and developing technologies to maximise its advantages., (© 2022 British Veterinary Association.)

In this study, we aimed to investigate the effect of Think Aloud (TA) on performance in trained and untrained participants, using functional Near Infrared Spectroscopy, during incrementally paced cycling. A mixed design was implemented with cycling expertise (10 untrained vs. 9 trained) as the between groups variable and trial stage (5 stages of increasing effort), and condition (silent vs. TA) as within groups independent variables (IVs). Dependent measures were changes in cortical oxygenation (O 2 Hb) in 12 areas of the prefrontal cortex (PFC) and physiological indicators of percentage heart rate maximum (%HRmax), average power output (APO), peak power output (PPO), rate of perceived exertion (RPE) and blood lactate ([La]b) over time. Trained cyclists had higher APO and significantly higher PPO from stages 2-5, in addition to a greater increase in PPO over the duration of the test (range 168W-480 W vs. 133W-313 W). There were significant main effects of stage on %HRmax, Bla and RPE ( p < .001), with effect sizes (ήp 2 ) ranging from .31 to .97. On average, HRmax%, [La]b and RPE were significantly lower after stage 2 onwards within the TA trial than the silent trial, even though similar power outputs were obtained. Thus, the TA trial elicited a better pacing strategy. There was no main effect of group on changes in O 2 Hb, though O 2 Hb did change as a function of stage in four areas of the PFC, and as a function of condition in one area. In this first study to assess the effects of TA on performance during self-paced cycling, TA did not disrupt performance outcomes at low through to high levels of physical exertion for either untrained or trained participants.

During the process of malignant transformation, cells undergo a series of genetic, epigenetic, and phenotypic alterations, including the acquisition and propagation of genomic aberrations that impart survival and proliferative advantages. These changes are mediated in part by the induction of replicative immortality that is accompanied by active telomere elongation. Indeed, telomeres undergo dynamic changes to their lengths and higher-order structures throughout tumor formation and progression, processes overseen in most cancers by telomerase. Telomerase is a multimeric enzyme whose function is exquisitely regulated through diverse transcriptional, post-transcriptional, and post-translational mechanisms to facilitate telomere extension. In turn, telomerase function depends not only on its core components, but also on a suite of binding partners, transcription factors, and intra- and extracellular signaling effectors. Additionally, telomerase exhibits telomere-independent regulation of cancer cell growth by participating directly in cellular metabolism, signal transduction, and the regulation of gene expression in ways that are critical for tumorigenesis. In this review, we summarize the complex mechanisms underlying telomere maintenance, with a particular focus on both the telomeric and extratelomeric functions of telomerase. We also explore the clinical utility of telomeres and telomerase in the diagnosis, prognosis, and development of targeted therapies for primary, metastatic, and recurrent cancers.

Inorganic metals supplement the chemical repertoire of organic molecules, especially proteins. This requires the correct metals to associate with proteins at metalation. Protein mismetalation typically occurs when excesses of unbound metals compete for a binding site ex vivo. However, in biology, excesses of metal-binding sites typically compete for limiting amounts of exchangeable metals. Here, we summarise mechanisms of metal homeostasis that sustain optimal metal availabilities in biology. We describe recent progress to understand metalation by comparing the strength of metal binding to a protein versus the strength of binding to competing sites inside cells., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this article., (Copyright © 2021 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Tumor dormancy is a major contributor to the lethality of metastatic disease, especially for cancer patients who develop metastases years-to-decades after initial diagnosis. Indeed, tumor cells can disseminate during early disease stages and persist in new microenvironments at distal sites for months, years, or even decades before initiating metastatic outgrowth. This delay between primary tumor remission and metastatic relapse is known as "dormancy," during which disseminated tumor cells (DTCs) acquire quiescent states in response to intrinsic (i.e., cellular) and extrinsic (i.e., microenvironmental) signals. Maintaining dormancy-associated phenotypes requires DTCs to activate transcriptional, translational, and post-translational mechanisms that engender cellular plasticity. RNA processing is emerging as an essential facet of cellular plasticity, particularly with respect to the initiation, maintenance, and reversal of dormancy-associated phenotypes. Moreover, dysregulated RNA processing, particularly that associated with alternative RNA splicing and expression of noncoding RNAs (ncRNAs), can occur in DTCs to mediate intrinsic and extrinsic metastatic dormancy. Here we review the pathophysiological impact of alternative RNA splicing and ncRNAs in promoting metastatic dormancy and disease recurrence in human cancers., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Movement is a fundamental characteristic of life, yet some invertebrate taxa, such as barnacles, permanently affix to a substratum as adults. Adult barnacles became 'sessile' over 500 Ma; however, we confirm that the epizoic sea turtle barnacle, Chelonibia testudinaria , has evolved the capacity for self-directed locomotion as adults. We also assess how these movements are affected by water currents and the distance between conspecifics. Finally, we microscopically examine the barnacle cement. Chelonibia testudinaria moved distances up to 78.6 mm yr -1 on loggerhead and green sea turtle hosts. Movements on live hosts and on acrylic panels occasionally involved abrupt course alterations of up to 90°. Our findings showed that barnacles tended to move directly against water flow and independent of nearby conspecifics. This suggests that these movements are not passively driven by external forces and instead are behaviourally directed. In addition, it indicates that these movements function primarily to facilitate feeding, not reproduction. While the mechanism enabling movement remained elusive, we observed that trails of cement bore signs of multi-layered, episodic secretion. We speculate that proximal causes of movement involve one or a combination of rapid shell growth, cement secretion coordinated with basal membrane lifting, and directed contraction of basal perimeter muscles.

Sea turtles, like other air-breathing diving vertebrates, commonly experience significant gas embolism (GE) when incidentally caught at depth in fishing gear and brought to the surface. To better understand why sea turtles develop GE, we built a mathematical model to estimate partial pressures of N 2 (PN 2 ), O 2 (PO 2 ), and CO 2 (PCO 2 ) in the major body-compartments of diving loggerheads ( Caretta caretta ), leatherbacks ( Dermochelys coriacea ), and green turtles ( Chelonia mydas ). This model was adapted from a published model for estimating gas dynamics in marine mammals and penguins. To parameterize the sea turtle model, we used values gleaned from previously published literature and 22 necropsies. Next, we applied this model to data collected from free-roaming individuals of the three study species. Finally, we varied body-condition and cardiac output within the model to see how these factors affected the risk of GE. Our model suggests that cardiac output likely plays a significant role in the modulation of GE, especially in the deeper diving leatherback turtles. This baseline model also indicates that even during routine diving behavior, sea turtles are at high risk of GE. This likely means that turtles have additional behavioral, anatomical, and/or physiologic adaptions that serve to reduce the probability of GE but were not incorporated in this model. Identifying these adaptations and incorporating them into future iterations of this model will further reveal the factors driving GE in sea turtles., Competing Interests: AF was employed without salary by the company Global Diving Research Inc. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Robinson, García-Párraga, Stacy, Costidis, Blanco, Clyde-Brockway, Haas, Harms, Patel, Stacy and Fahlman.)

The maintenance of telomere length supports repetitive cell division and therefore plays a central role in cancer development and progression. Telomeres are extended by either the enzyme telomerase or the alternative lengthening of telomeres (ALT) pathway. Here, we found that the telomere-associated protein SLX4IP dictates telomere proteome composition by recruiting and activating the E3 SUMO ligase PIAS1 to the SLX4 complex. PIAS1 SUMOylated the telomere-binding protein RAP1, which disrupted its interaction with the telomere-binding protein TRF2 and facilitated its nucleocytoplasmic shuttling. In the cytosol, RAP1 bound to IκB kinase (IKK), resulting in activation of the transcription factor NF-κB and its induction of Jagged-1 expression, which promoted Notch signaling and the institution of ALT. This axis could be targeted therapeutically in ALT-driven cancers and in tumor cells that develop resistance to antitelomerase therapies. Our results illuminate the mechanisms underlying SLX4IP-dependent telomere plasticity and demonstrate the role of telomere proteins in directly coordinating intracellular signaling and telomere maintenance dynamics., (Copyright © 2021 The Authors, some rights reserved; exclusive licensee American Association for the Advancement of Science. No claim to original U.S. Government Works.)

Although ultrafast manipulation of magnetism holds great promise for new physical phenomena and applications, targeting specific states is held back by our limited understanding of how magnetic correlations evolve on ultrafast timescales. Using ultrafast resonant inelastic X-ray scattering we demonstrate that femtosecond laser pulses can excite transient magnons at large wavevectors in gapped antiferromagnets and that they persist for several picoseconds, which is opposite to what is observed in nearly gapless magnets. Our work suggests that materials with isotropic magnetic interactions are preferred to achieve rapid manipulation of magnetism., Competing Interests: The authors declare no competing interest.

Rationale and Objectives: Three-dimensional (3D) visualization has been shown to benefit new generations of medical students and physicians-in-training in a variety of contexts. However, there is limited research directly comparing student performance after using 3D tools to those using two-dimensional (2D) screens., Materials and Methods: A CT was performed on a donated cadaver and a 3D CT hologram was created. A total of 30 first-year medical students were randomly assigned into two groups to review head and neck anatomy in a teaching session that incorporated CT. The first group used an augmented reality headset, while the second group used a laptop screen. The students were administered a five-question anatomy test before and after the session. Two-tailed t-tests were used for statistical comparison of pretest and posttest performance within and between groups. A feedback survey was distributed for qualitative data., Results: Pretest vs. posttest comparison of average percentage of questions answered correctly demonstrated both groups showing significant in-group improvement (p < 0.05), from 59% to 95% in the augmented reality group, and from 57% to 80% in the screen group. Between-group analysis indicated that posttest performance was significantly better in the augmented reality group (p = 0.022, effect size = 0.73)., Conclusion: Immersive 3D visualization has the potential to improve short-term anatomic recall in the head and neck compared to traditional 2D screen-based review, as well as engage millennial learners to learn better in anatomy laboratory. Our findings may reflect additional benefit gained from the stereoscopic depth cues present in augmented reality-based visualization., (Copyright © 2020 The Association of University Radiologists. Published by Elsevier Inc. All rights reserved.)

Point 1: Stereo-video camera systems (SVCSs) are a promising tool to remotely measure body size of wild animals without the need for animal handling. Here, we assessed the accuracy of SVCSs for measuring straight carapace length (SCL) of sea turtles. Point 2: To achieve this, we hand captured and measured 63 juvenile, subadult, and adult sea turtles across three species: greens, Chelonia mydas ( n  = 52); loggerheads, Caretta caretta ( n  = 8); and Kemp's ridley, Lepidochelys kempii ( n  = 3) in the waters off Eleuthera, The Bahamas and Crystal River, Florida, USA, between May and November 2019. Upon release, we filmed these individuals with the SVCS. We performed photogrammetric analysis to extract stereo SCL measurements (eSCL), which were then compared to the (manual) capture measurements (mSCL). Point 3: mSCL ranged from 25.9 to 89.2 cm, while eSCL ranged from 24.7 to 91.4 cm. Mean percent bias of eSCL ranged from -0.61% (±0.11 SE ) to -4.46% (±0.31 SE ) across all species and locations. We statistically analyzed potential drivers of measurement error, including distance of the turtle to the SVCS, turtle angle, image quality, turtle size, capture location, and species. Point 4: Using a linear mixed effects model, we found that the distance between the turtle and the SVCS was the primary factor influencing measurement error. Our research suggests that stereo-video technology enables high-quality measurements of sea turtle body size collected in situ without the need for hand-capturing individuals. This study contributes to the growing knowledge base that SVCS are accurate for body size measurements independent of taxonomic clade., Competing Interests: The authors declare no conflict of interest., (© 2021 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Protein metal-occupancy (metalation) in vivo has been elusive. To address this challenge, the available free energies of metals have recently been determined from the responses of metal sensors. Here, we use these free energy values to develop a metalation-calculator which accounts for inter-metal competition and changing metal-availabilities inside cells. We use the calculator to understand the function and mechanism of GTPase CobW, a predicted Co II -chaperone for vitamin B 12 . Upon binding nucleotide (GTP) and Mg II , CobW assembles a high-affinity site that can obtain Co II or Zn II from the intracellular milieu. In idealised cells with sensors at the mid-points of their responses, competition within the cytosol enables Co II to outcompete Zn II for binding CobW. Thus, Co II is the cognate metal. However, after growth in different [Co II ], Co II -occupancy ranges from 10 to 97% which matches CobW-dependent B 12 synthesis. The calculator also reveals that related GTPases with comparable Zn II affinities to CobW, preferentially acquire Zn II due to their relatively weaker Co II affinities. The calculator is made available here for use with other proteins.

Vitamin B 12 , cobalamin, is a cobalt-containing ring-contracted modified tetrapyrrole that represents one of the most complex small molecules made by nature. In prokaryotes it is utilised as a cofactor, coenzyme, light sensor and gene regulator yet has a restricted role in assisting only two enzymes within specific eukaryotes including mammals. This deployment disparity is reflected in another unique attribute of vitamin B 12 in that its biosynthesis is limited to only certain prokaryotes, with synthesisers pivotal in establishing mutualistic microbial communities. The core component of cobalamin is the corrin macrocycle that acts as the main ligand for the cobalt. Within this review we investigate why cobalt is paired specifically with the corrin ring, how cobalt is inserted during the biosynthetic process, how cobalt is made available within the cell and explore the cellular control of cobalt and cobalamin levels. The partitioning of cobalt for cobalamin biosynthesis exemplifies how cells assist metalation., (Copyright © 2020 The Authors. Published by Elsevier B.V. All rights reserved.)