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Background It remains largely unknown where and how infections with soil-transmitted helminths (STHs; Ascaris, Trichuris, Necator and Ancylostoma) occur. We therefore aimed to identify possible sources of infection by assessing the environmental contamination in an STH-endemic area. Methods We first performed a series of laboratory experiments designed to optimize a soil straining-flotation method to detect and quantify Ascaris and Trichuris eggs in soil, and to validate the diagnostic performance of the optimized method when followed by microscopy and qPCR. In a second phase, we applied this method to assess the level of STH contamination in 399 environmental samples collected from 10 school compounds, 50 households and 9 open markets in Jimma Town (Ethiopia). Subsequently, we explored associations between the environmental contamination and both the corresponding STH epidemiology at the level of the schools and the household characteristics. Finally, we assessed the knowledge, attitude and practice (KAP) towards STHs in school children. Principal findings Our soil straining-flotation method has an analytical sensitivity of 50 eggs per 100 grams of soil and egg recovery rate of 36.0% (Ascaris) and 8.0% (Trichuris). The analysis of field samples with both microscopy and qPCR revealed the presence of 8 different helminth species of medical importance, including but not limited to the human STHs. There was a significant association between the environmental contamination and prevalence of any STH infections at the school level only. The KAP indicated a lack of knowledge and awareness of STHs. Conclusions/Significance Our optimized straining-flotation method has a moderate diagnostic performance and revealed that life stages of helminths are ubiquitous in the environment, which might be due to the poor sanitary facilities at both the schools and the households, and a poor level of KAP towards STHs. Further research is required to gain more insights into the contribution of these life stages to transmission.

Changes in motor activity are core symptoms of mood episodes in bipolar disorder. The manic state is characterized by increased variance, augmented complexity and irregular circadian rhythmicity when compared to healthy controls. No previous studies have compared mania to euthymia intra-individually in motor activity. The aim of this study was to characterize differences in motor activity when comparing manic patients to their euthymic selves. Motor activity was collected from 16 bipolar inpatients in mania and remission. 24-h recordings and 2-h time series in the morning and evening were analyzed for mean activity, variability and complexity. Lastly, the recordings were analyzed with the similarity graph algorithm and graph theory concepts such as edges, bridges, connected components and cliques. The similarity graph measures fluctuations in activity reasonably comparable to both variability and complexity measures. However, direct comparisons are difficult as most graph measures reveal variability in constricted time windows. Compared to sample entropy, the similarity graph is less sensitive to outliers. The little-understood estimate Bridges is possibly revealing underlying dynamics in the time series. When compared to euthymia, over the duration of approximately one circadian cycle, the manic state presented reduced variability, displayed by decreased standard deviation (p= 0.013) and augmented complexity shown by increased sample entropy (p= 0.025). During mania there were also fewer edges (p= 0.039) and more bridges (p= 0.026). Similar significant changes in variability and complexity were observed in the 2-h morning and evening sequences, mainly in the estimates of the similarity graph algorithm. Finally, augmented complexity was present in morning samples during mania, displayed by increased sample entropy (p= 0.015). In conclusion, the motor activity of mania is characterized by altered complexity and variability when compared within-subject to euthymia.

Background Metformin is prescribed to women with polycystic ovary syndrome (PCOS) to prevent pregnancy complications. Children exposed to metformin vs. placebo in utero, have increased head circumference at birth and are more overweight and obese at 8 years of age. Also, maternal PCOS-status seems to alter the long-term cardio-metabolic health of offspring. We hypothesized that the long-term effects of metformin-exposure and/or maternal PCOS may be mediated by circulatory adaptations during fetal life. Material and methods This is a sub-study of a larger double-blinded, placebo-controlled trial, where women with PCOS were randomized to metformin (2g/day) or placebo in pregnancy, a total of 487 women. A sub-group of participants (N = 58) took part in this sub-study and had an extended ultrasound examination at gestational week 32, including blood flow velocity and diameter measurements of the umbilical vein (UV), the ductus venosus (DV) and the portal vein (PV). Blood flow volume was calculated and adjusted for estimated fetal weight (EFW) (normalized flow). Metformin exposed fetuses were compared to placebo exposed fetuses. Fetuses of mothers with PCOS (metformin [n = 30] and placebo [n = 28]) were compared to a low-risk reference population (N = 160) by z-score statistics. Results There was no difference in fetal liver flow between metformin vs. placebo-exposed fetuses. Fetuses of mothers with PCOS had higher EFW (0.63 [95% CI 0.44–0.83] p Conclusion Metformin during pregnancy did not affect fetal liver blood-flow. In our population, maternal PCOS-status was associated with reduced total venous liver blood-flow, which may explain altered growth and metabolism later in life.

Background The burden of non-obstructive coronary artery disease (CAD) in the society is high, and there is currently limited evidence-based recommendation for risk stratification and treatment. Previous studies have demonstrated an association between increasing extent of non-obstructive CAD and cardiovascular events. Whether hypertension, a modifiable cardiovascular risk factor, is associated with extensive non-obstructive CAD in patients with symptomatic chronic coronary syndrome (CCS) remains unclear. Methods We included 1138 patients (mean age 62±11 years, 48% women) with symptomatic CCS and non-obstructive CAD (1–49% lumen diameter reduction) by coronary computed tomography angiography (CCTA) from the Norwegian Registry for Invasive Cardiology (NORIC). The extent of non-obstructive CAD was assessed as coronary artery segment involvement score (SIS), and extensive non-obstructive CAD was adjudicated when SIS >4. Hypertension was defined as known hypertension or use of antihypertensive medication. Results Hypertension was found in 45% of patients. Hypertensive patients were older, with a higher SIS, calcium score, and prevalence of comorbidities and statin therapy compared to the normotensive (all p Conclusion In symptomatic CCS, hypertension was associated with extensive non-obstructive CAD by CCTA. Whether hypertension may be a new treatment target in symptomatic non-obstructive CAD needs to be explored in future studies. Clinical trial registration ClinicalTrials.gov: Identifier NCT 04009421.

Background In the context of the COVID-19 pandemic, many individuals have been found to endorse conspiracy beliefs. Socio-demographic variables, personality functioning, anxiety, and loneliness could be risk factors for this endorsement. Methods In a representative sample of the German population (N = 2,503) measures of conspiracy mentality, conspiracy-related beliefs toward COVID-19, personality functioning (OPD-SQS), anxiety (HADS), and loneliness (UCLA) were assessed. Pearson product-moment correlations and multiple linear regression analyses were conducted. Results Conspiracy mentality and conspiracy-related beliefs toward COVID-19 were strongly correlated. Regression analyses found younger age, male gender, lower education, and lower income to be associated with conspiracy mentality. The subscales relationship model and self-perception of the OPD-SQS were positively related to conspiracy mentality whereas interpersonal contact was negatively associated. Higher levels of anxiety were statistically predictive for conspiracy mentality. Conclusion Our findings indicate a contribution of personality functioning to the understanding of conspiracy mentality and thus to the advancement of interventions during the pandemic.

The capability to establish accurate predictions is an integral part of learning. Whether predictions about different dimensions of a stimulus interact with each other, and whether such an interaction affects learning, has remained elusive. We conducted a statistical learning study with EEG (electroencephalography), where a stream of consecutive sound triplets was presented with deviants that were either: (a) statistical, depending on the triplet ending probability, (b) physical, due to a change in sound location or (c) double deviants, i.e. a combination of the two. We manipulated the predictability of stimulus-onset by using random stimulus-onset asynchronies. Temporal unpredictability due to random onsets reduced the neurophysiological responses to statistical and location deviants, as indexed by the statistical mismatch negativity (sMMN) and the location MMN. Our results demonstrate that the predictability of one stimulus attribute influences the processing of prediction error signals of other stimulus attributes, and thus also learning of those attributes.

Mammalian adenoviruses (AdVs) comprise more than ~350 types including over 100 human (HAdVs) and just three mouse AdVs (MAdVs). While most HAdVs initiate infection by high affinity/avidity binding of their fiber knob (FK) protein to either coxsackievirus AdV receptor (CAR), CD46 or desmoglein (DSG)-2, MAdV-1 (M1) infection requires arginine-glycine-aspartate (RGD) binding integrins. To identify the receptors mediating MAdV infection we generated five novel reporter viruses for MAdV-1/-2/-3 (M1, M2, M3) transducing permissive murine (m) CMT-93 cells, but not B16 mouse melanoma cells expressing mCAR, human (h) CD46 or hDSG-2. Recombinant M1 or M3 FKs cross-blocked M1 and M3 but not M2 infections. Profiling of murine and human cells expressing RGD-binding integrins suggested that αvβ6 and αvβ8 heterodimers are associated with M1 and M3 infections. Ectopic expression of mβ6 in B16 cells strongly enhanced M1 and M3 binding, infection, and progeny production comparable with mαvβ6-positive CMT-93 cells, whereas mβ8 expressing cells were more permissive to M1 than M3. Anti-integrin antibodies potently blocked M1 and M3 binding and infection of CMT-93 cells and hαvβ8-positive M000216 cells. Soluble integrin αvβ6, and synthetic peptides containing the RGDLXXL sequence derived from FK-M1, FK-M3 and foot and mouth disease virus coat protein strongly interfered with M1/M3 infections, in agreement with high affinity interactions of FK-M1/FK-M3 with αvβ6/αvβ8, determined by surface plasmon resonance measurements. Molecular docking simulations of ternary complexes revealed a bent conformation of RGDLXXL-containing FK-M3 peptides on the subunit interface of αvβ6/β8, where the distal leucine residue dips into a hydrophobic pocket of β6/8, the arginine residue ionically engages αv aspartate215, and the aspartate residue coordinates a divalent cation in αvβ6/β8. Together, the RGDLXXL-bearing FKs are part of an essential mechanism for M1/M3 infection engaging murine and human αvβ6/8 integrins. These integrins are highly conserved in other mammals, and may favour cross-species virus transmission., PLoS Pathogens, 17 (12), ISSN:1553-7374, ISSN:1553-7366

Source monitoring refers to the ability to identify the origin of a memory, for example, whether you remember saying something or thinking about it, and confusions of these sources have been associated with the experience of auditory verbal hallucinations (AVHs). Both AVHs and source confusions are reported to originate from dysfunctional brain activations in the prefrontal cortex (PFC) and the superior temporal gyrus (STG); specifically, it is assumed that a hypoactive PFC and a hyperactive STG gives rise to AVHs and source confusions. We set out to test this assumption by trying to mimic this hypertemporal/hypofrontal model in healthy individuals with transcranial direct current stimulation (tDCS): the inhibitory cathode was placed over the left PFC and the excitatory anode over the left dorsolateral STG. Participants completed a reality monitoring task (distinguishing between external and internal memory sources) and an internal source monitoring task (distinguishing between two or more internal memory sources) in two separate experiments (offline vs. online tDCS). In the offline experiment (n = 34), both source monitoring tasks were completed after tDCS stimulation, and in the online experiment (n = 27) source monitoring tasks were completed while simultaneously being stimulated with tDCS. We found that internal source monitoring abilities were significantly enhanced during active online tDCS, while reality monitoring abilities were unaffected by stimulation in both experiments. We speculate, based on combining the present findings with previous studies, that there might be different brain areas involved in reality and internal source monitoring. While internal source monitoring seems to involve speech production areas, specifically Broca’s area, as suggested in the present study, reality monitoring seems to rely more on the STG and DLPFC, as shown in other studies of the field.

BackgroundPlatelet indices (PIs) are platelet parameters that are correlated with platelet activity. Despite being widely available, inexpensive, and feasible; their use in clinical settings is limited. Recently, we developed a new score (EgyDAS), which relies on PIs and assesses disease activity in rheumatoid arthritis (RA).ObjectivesThis study explored the practicability and validity of EgyDAS in RA with ankle involvement, considering that ankle is neglected in the commonly used DAS28 score.MethodsThis comparative case-control study included 2-groups of RA patients, group1 (control): without and group 2: with ankle involvement.ResultsAnkle involvement in RA showed no gender or age differences, however, it was associated with higher platelet count, erythrocyte sedimentation rate (ESR), C-reactive protein (CRP), platelet distribution width (PDW), visual analogue scale (VAS), tender joint count (TJC), and lower hemoglobin (Hb) and mean platelet volume (MPV). DAS28 categorized a higher proportion of patients to have high disease activity compared with EgyDAS; moreover, it did not detect those in remission in group 2 patients. Highly significant differences in the 2-scores were observed between the two groups. Further analyses revealed superiority of EgyDAS in assessing disease activity in group 2 patients. Finally, both scores were found correlated together in the study groups.ConclusionsOver or underestimation of RA disease activity could occur when using DAS28. PIs were found correlated with ankle involvement in RA. PIs and EgyDAS are the best tools to assess disease activity in RA patients with ankle involvement. However, the study recommended the use of both scores together.

Sperm morphology is generally uniform within a species due to selective pressures that act to achieve better fertilization outcomes under postcopulatory competitive circumstances. Therefore, polyandry that intensifies post-mating sperm competition should constrain intraspecific sperm polymorphism. Contrary to this paradigm, we previously found that a polyandrous squid, Heterololigo bleekeri, produces dimorphic eusperm (flagellum length dimorphism; FLD), which is closely associated with alternative reproductive tactics (ARTs); large males (consorts) transfer their spermatophores inside the female’s mantle cavity, while small males (sneakers) do so outside the mantle. Thus, FLD was considered as the consequence of different insemination strategies that arise from different modes of sperm competition, sperm storage and the fertilization environment. However, in other squid species showing ARTs, the choice of mating behaviour is rather conditional (i.e., switching mating tactic between consorts and sneakers), which poses the question of whether sperm FLD could have evolved. Here, we investigated five species in the family Loliginidae that exhibit ARTs and found that all species showed sneaker-biased FLD. However, in a species with conditional ARTs, we found FLD rather ambiguous and the testicular somatic index to be nearly continuous among individuals at transitional state, suggesting that plasticity in mating behaviour compromises the disruptive selection on a sperm morphological trait.

Background The provision of post-discharge malaria chemoprevention (PMC) in children recently admitted with severe anemia reduces the risk of death and re-admissions in malaria endemic countries. The main objective of this trial was to identify the most effective method of delivering dihydroartemesinin-piperaquine to children recovering from severe anemia. Methods This was a 5-arm, cluster-randomized trial among under-5 children hospitalized with severe anemia at Zomba Central Hospital in Southern Malawi. Children were randomized to receive three day treatment doses of dihydroartemesinin-piperaquine monthly either; 1) in the community without a short text reminder; 2) in the community with a short message reminder; 3) in the community with a community health worker reminder; 4) at the facility without a short text reminder; or 5) at the facility with a short message reminder. The primary outcome measure was adherence to all treatment doses of dihydroartemesinin-piperaquine and this was assessed by pill-counts done by field workers during home visits. Poisson regression was utilized for analysis. Results Between March 2016 and October 2018, 1460 clusters were randomized. A total of 667 children were screened and 375 from 329 clusters were eligible and enrolled from the hospital. Adherence was higher in all three community-based compared to the two facility-based delivery (156/221 [70·6%] vs. 78/150 [52·0%], IRR = 1·24,95%CI 1·06–1·44, p = 0·006). This was observed in both the SMS group (IRR = 1·41,1·21–1·64, p Interpretation Community-based delivery of dihydroartemesinin-piperaquine for post-discharge malaria chemoprevention in children recovering from severe anemia resulted in higher adherence compared to facility-based methods. Trial registration NCT02721420; ClinicalTrials.gov.

The cilium, the sensing centre for the cell, displays an extensive repertoire of receptors for various cell signalling processes. The dynamic nature of ciliary signalling indicates that the ciliary entry of receptors and associated proteins must be regulated and conditional. To understand this process, we studied the ciliary localisation of the odour-receptor coreceptor (Orco), a seven-pass transmembrane protein essential for insect olfaction. Little is known about when and how Orco gets into the cilia. Here, using Drosophila melanogaster, we show that the bulk of Orco selectively enters the cilia on adult olfactory sensory neurons in two discrete, one-hour intervals after eclosion. A conditional loss of heterotrimeric kinesin-2 during this period reduces the electrophysiological response to odours and affects olfactory behaviour. We further show that Orco binds to the C-terminal tail fragments of the heterotrimeric kinesin-2 motor, which is required to transfer Orco from the ciliary base to the outer segment and maintain within an approximately four-micron stretch at the distal portion of the ciliary outer-segment. The Orco transport was not affected by the loss of critical intraflagellar transport components, IFT172/Oseg2 and IFT88/NompB, respectively, during the adult stage. These results highlight a novel developmental regulation of seven-pass transmembrane receptor transport into the cilia and indicate that ciliary signalling is both developmentally and temporally regulated., PLoS Genetics, 17 (8), ISSN:1553-7390, ISSN:1553-7404

Sarcopenia is intricately related to aging associated diseases, such as neuropsychiatric disorders, oral status, and chronic diseases. Dementia and depression are interconnected and also related to sarcopenia. The preliminary shift from robust to sarcopenia (i.e., pre-sarcopenia) is an important albeit underdiscussed stage and is the focus of this study. Identifying factors associated with pre-sarcopenia may lead to sarcopenia prevention. To separately examine the effects of dementia and depression on pre-sarcopenia/sarcopenia, we conducted multiple analyses. This cross-sectional study used health checkup data from a rural Japanese island. The participants were aged 60 years and above, and the data included muscle mass, gait speed, handgrip strength, oral status (teeth and denture), chronic diseases (e.g., hypertension), dementia (cognitive assessment for dementia, iPad Version), and depression (self-rating depression scale). A total of 753 older adult participants were divided into the sarcopenia (n = 30), pre-sarcopenia (n = 125), and robust (n = 598) groups. An ordered logit regression analysis indicated that age and depression were positively correlated with sarcopenia, while hypertension was negatively associated with it. A multiple logistic regression analysis between the robust and pre-sarcopenia groups showed significant associations between the same three variables. Depression was associated with pre-sarcopenia, but not dementia. There was also a significant association between hypertension and pre-sarcopenia. Further research is needed to reveal whether the management of these factors can prevent sarcopenia.

Background It was shown that single repetition, contraction-phase specific and total time-under-tension (TUT) can be extracted reliably and validly from smartphone accelerometer-derived data of resistance exercise machines using user-determined resistance exercise velocities at 60% one repetition maximum (1-RM). However, it remained unclear how robust the extraction of these mechano-biological descriptors is over a wide range of movement velocities (slowversus fast-movement velocity) and intensities (30% 1-RM versus 80% 1-RM) that reflect the interindividual variability during resistance exercise. Objective In this work, we examined whether the manipulation of velocity or intensity would disrupt an algorithmic extraction of single repetitions, contraction-phase specific and total TUT. Methods Twenty-seven participants performed four sets of three repetitions of their 30% and 80% 1- RM with velocities of 1 s, 2 s, 6 s and 8 s per repetition, respectively. An algorithm extracted the number of repetitions, single repetition, contraction-phase specific and total TUT. All exercises were video-recorded. The video recordings served as the gold standard to which algorithmically-derived TUT was compared. The agreement between the methods was examined using Limits of Agreement (LoA). The Pearson correlation coefficients were used to calculate the association, and the intraclass correlation coefficient (ICC 2.1) examined the interrater reliability. Results The calculated error rate for the algorithmic detection of the number of single repetitions derived from two smartphones accelerometers was 1.9%. The comparison between algorithmically- derived, contraction-phase specific TUT against video, revealed a high degree of correlation (r > 0.94) for both exercise machines. The agreement between the two methods was high on both exercise machines, intensities and velocities and was as follows: LoA ranged from -0.21 to 0.22 seconds for single repetition TUT (2.57% of mean TUT), from -0.24 to 0.22 seconds for concentric contraction TUT (6.25% of mean TUT), from -0.22 to 0.24 seconds for eccentric contraction TUT (5.52% of mean TUT) and from -1.97 to 1.00 seconds for total TUT (5.13% of mean TUT). Interrater reliability for single repetition, contraction- phase specific TUT was high (ICC > 0.99). Conclusion Neither intensity nor velocity disrupts the proposed algorithmic data extraction approach. Therefore, smartphone accelerometers can be used to extract scientific mechano-biological descriptors of dynamic resistance exercise with intensities ranging from 30% to 80% of the 1-RM with velocities ranging from 1 s to 8 s per repetition, respectively, thus making this simple method a reliable tool for resistance exercise mechano-biological descriptors extraction., PLoS ONE, 16 (7), ISSN:1932-6203

A new styracosternan ornithopod genus and species is described based on the right dentary of a single specimen from the Mirambell Formation (Early Cretaceous, early Barremian) at the locality of Portell, (Castellón, Spain).Portellsaurus sosbaynatigen. et sp. nov. is diagnosed by two autapomorphic features as well as a unique combination of characters. The autapomorphies include: the absence of a bulge along the ventral margin directly ventral to the base of the coronoid process and the presence of a deep oval cavity on the medial surface of the mandibular adductor fossa below the eleventh-twelfth tooth position. Phylogenetic analyses reveal that the new Iberian form is more closely related to the African taxonOuranosaurus nigeriensisthan to its synchronic Iberian taxaMagnamanus soriaensisandIguanodon galvensis. In addition,Portellsaurus sosbaynatiis less related to other Iberian taxa such asIguanodon bernissartensisandProa valdearinnoensisthan to the other Early Cretaceous Iberian styracosternansMantellisaurus atherfieldensisandMorelladon beltrani. A new phylogenetic hypothesis is proposed that resolvesIguanodon(I.bernissartensis,I.galvensis) with the ValanginianBarilium dawsoniinto a monophyletic clade (Iguanodontoidea). The recognition ofPortellsaurus sosbaynatigen. et sp. nov. as the first styracosternan dinosaur species identified from the Margas de Mirambell Formation (early Barremian–early late Barremian) in the Morella sub-basin (Maestrat Basin, eastern Spain) indicates that the Iberian Peninsula was home to a highly diverse assemblage of medium-to-large bodied styracosternan hadrosauriforms during the Early Cretaceous.

AimThe lockdown of sports infrastructure due to the COVID-19 pandemic has substantially shifted people's physical activity towards public green spaces. With Germany's lockdown as one of the more severe governmentally imposed epidemic-control-decisions, we tested to what extent the frequency of outdoor cycling activities changed from March to June 2020.MethodsUser behaviour and frequency in 15 urban and 7 rural German public green spaces was quantified using cycling data from the fitness application Strava. Changes in cycling activities were analysed with four different generalised linear models, correcting for factors like weather conditions and temporal changes in the user base of the fitness application.ResultsWe found a clear increase in outdoor cycling sport activities in urban public green spaces in response to epidemic-control decisions (e.g. increase by 81% in April relative to the expected value (95% CI [48%, 110%])). In contrast, biking in rural areas showed no significant change with epidemic-control-decisions in place.ConclusionFitness App data, e.g. from Strava, can be used to monitor visitor behaviour and frequency. The increase in outdoor cycling activities during epidemic control decisions likely reflects a shift of sport activities from indoor and team sports to outdoor and individual sports. This highlights the importance of accessible green space for maintaining physical fitness and health. Beyond this shift, it is likely that outdoor activities may be of particularly importance for stress relief in times of crisis such as the current COVID-19 pandemic.

PDkit is an open source software toolkit supporting the collaborative development of novel methods of digital assessment for Parkinson’s Disease, using symptom measurements captured continuously by wearables (passive monitoring) or by high-use-frequency smartphone apps (active monitoring). The goal of the toolkit is to help address the current lack of algorithmic and model transparency in this area by facilitating open sharing of standardised methods that allow the comparison of results across multiple centres and hardware variations. PDkit adopts the information-processing pipeline abstraction incorporating stages for data ingestion, quality of information augmentation, feature extraction, biomarker estimation and finally, scoring using standard clinical scales. Additionally, a dataflow programming framework is provided to support high performance computations. The practical use of PDkit is demonstrated in the context of the CUSSP clinical trial in the UK. The toolkit is implemented in the python programming language, the de facto standard for modern data science applications, and is widely available under the MIT license., Author summary Parkinson’s Disease is the fastest growing neurological condition affecting millions of people across the world. People with Parkinson’s suffer from a variety of symptoms that result in diminished ability to move, eat, remember or sleep. Research in new treatments are limited because the clinical tools used to assess its symptoms are subjective, require considerable time to perform and specialised skills and can only detect coarse-grain changes. To address this situation, clinicians are turning to smartphone apps and wearables to create new ways to assess symptoms that are more sensitive to change and can be applied frequently at home by patients and their carers. In this paper, we discuss PDkit, an open source toolkit that we developed to help address this current lack of algorithmic and model transparency. Adopting PDkit facilitates the open sharing of standardised methods and can accelerate the development of new methods and system to assess Parkinson’s and enables research groups to innovate. The toolkit provides funcionality that support data ingestion, quality of information augmentation, feature extraction, biomarker estimation and finally, scoring using standard clinical scales. The practical use of PDkit is demonstrated via its use by the CUSSP clinical trial conducted in the UK.

Seabirds are one of the most at-risk groups, with many species in decline. In Scandinavia, seabirds are at a heightened risk of extinction due to accelerated global warming. Norway is home to significant portion of the European Atlantic Puffin (Fratercula arctica) populations, but Norwegian populations have declined significantly during the last decades. In this paper we use biometric data from modern and archaeologicalF.arcticaspecimens to investigate patterns in body size variation over time of this iconic species. We aimed to set out a baseline for our archaeological comparison by firstly investigating whether modern subspecies ofF.arcticaare reflected in the osteological characters and are enough to distinguish subspecies from the bones alone. We then investigated if archaeological remains ofF.arcticadiffer in size from the modern subspecies. Our results show that the subspeciesFratercula arctica naumanniwas distinctly larger than the other subspecies. However,Fratercula arctica arcticaandFratercula arctica grabaewere difficult to separate based on size. This generally supports ornithological observations. Post-MedievalF.arcticabones from Måsøy were similar to modernF.a.arcticapopulations. The mid-Holocene remains from Dollsteinhola overlaps with the modern size ranges ofF.a.arcticaandF.a.grabaebut are generally shorter and more robust. Dollsteinhola is located close to the borders of the modern breeding ranges of bothF.a.arcticaandF.a.grabae. We consider it therefore likely that given the mid-Holocene climatic oscillations, breeding ranges of the two subspecies shifted north or south accordingly. However, this does not explain the different proportions of the Dollsteinhola specimens. Our data provide the first evidence for shifting distributions in ancient Atlantic Puffins and represent the first osteological analysis ofFratercula arcticasubspecies.

Background Vancomycin-resistant enterococci (VRE) represent several types of transferable vancomycin resistance gene clusters. The vanD type, associated with moderate to high level vancomycin resistance, has only sporadically been described in clinical isolates. The aim of this study was to perform a genetic characterization of the first VanD-type VRE strains detected in Norway. Methods The VanD-type VRE-strains (n = 6) from two patient cases were examined by antimicrobial susceptibility testing and whole genome sequencing (WGS) to uncover Van-phenotype, strain phylogeny, the vanD gene clusters, and their genetic surroundings. The putative transferability of vanD was examined by circularization PCR and filter mating. Results The VanD-type Enterococcus faecium (n = 4) and Enterococcus casseliflavus (n = 2) strains recovered from two cases (A and B), expressed moderate to high level vancomycin resistance (MIC 64—>256 mg/L) and various levels of teicoplanin susceptibility (MIC 2—>256 mg/L). WGS analyses revealed phylogenetically different E. faecium strains (A1, A2, and A3 of case A and B1 from case B) as well as vanD gene clusters located on different novel genomic islands (GIs). The E. casseliflavus strains (B2 and B3 of case B) were not clonally related, but harbored nearly identical novel GIs. The vanD cluster of case B strains represents a novel vanD-subtype. All the vanD-GIs were integrated at the same chromosomal site and contained genes consistent with a Clostridiales origin. Circular forms of the vanD-GIs were detected in all strains except B1. Transfer of vanD to an E. faecium recipient was unsuccessful. Conclusions We describe the first VanD-type E. casseliflavus strains, a novel vanD-subtype, and three novel vanD-GIs with a genetic content consistent with a Clostridiales order origin. Despite temporal occurrence, case A and B E. faecium strains were phylogenetically diverse and harbored different vanD subtypes and vanD-GIs.

Copyright: © 2021 Lampert et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited., Introduction: There has been growing interest in community gardens as an effective and affordable health promotion strategy. However, most available evidence is derived from qualitative studies, whereas quantitative research on this subject is limited. Objectives: To synthetize the literature about physical and mental health outcomes associated with community gardening. Two main questions were addressed: a) is there evidence, from quantitative studies, that community gardening is associated to physical and mental health and well-being of non-institutionalized individuals? b) Does community gardening provokes any discomfort in terms of physical health, i.e., bodily pain, to their beneficiaries? Methods: A systematic review of the literature was carried out following PRISMA guidelines by searching relevant electronic databases (PubMed, Scopus, and Web of Science). Empirical, quantitative studies published in English with no restrictions concerning the date of publication were considered eligible. The quality of the evidence was appraised using the tool developed by the National Heart, Lung, and Blood Institute of the National Institutes of Health for Observational Cohort and Cross-Sectional Studies. Results: Overall, 8 studies were considered eligible, of which seven studies were rated as having good methodological quality (one scored as fair). Community gardeners had significantly better health outcomes than their neighbours not engaged in gardening activities in terms of life satisfaction, happiness, general health, mental health, and social cohesion. Conclusion: Community gardens are associated to health gains for their users, irrespective of age, being an affordable and efficient way of promoting physical and mental health and well-being. To encourage the design, maintenance, and prospective evaluation of supportive urban environments promoting healthy and, at the same time, sustainable lifestyles, is essential to achieve public health gains and environmental sustainability., This literature review was funded by the Fundação para a Ciência e a Tecnologia and by the Empresa Municipal de Ambiente de Cascais through a doctoral grant to the first author of this paper (PDE/BDE/122672/2016). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Aerobic fitness (AF) and lipoprotein subclasses associate to each other and to cardiovascular health. Adiposity and physical activity (PA) influence the association pattern of AF to lipoproteins almost inversely making it difficult to assess their independent and joint influence on the association pattern. This study, including 841 children (50% boys) 10.2 ± 0.3 years old with BMI 18.0 ± 3.0 kg/m2 from rural Western Norway, aimed at examining the association pattern of AF to the lipoprotein subclasses and to estimate the independent and joint influence of PA and adiposity on this pattern. We used multivariate analysis to determine the association pattern of a profile of 26 lipoprotein features to AF with and without adjustment for three measures of adiposity and a high-resolution PA descriptor of 23 intensity intervals derived from accelerometry. For data not adjusted for adiposity or PA, we observed a cardioprotective lipoprotein pattern associating to AF. This pattern withstood adjustment for PA, but the strength of association to AF was reduced by 58%, while adjustment for adiposity weakened the association of AF to the lipoproteins by 85% and with strongest changes in the associations to a cardioprotective high-density lipoprotein subclass pattern. When adjusted for both adiposity and PA, the cardioprotective lipoprotein pattern still associated to AF, but the strength of association was reduced by 90%. Our results imply that the (negative) influence of adiposity on the cardioprotective association pattern of lipoproteins to AF is considerably stronger than the (positive) contribution of PA to this pattern. However, our analysis shows that PA contributes also indirectly through a strong inverse association to adiposity. The trial was registered 7 May, 2014 in clinicaltrials.gov with trial reg. no.: NCT02132494 and the URL is https://clinicaltrials.gov/ct2/results?term=NCT02132494&cntry=NO.

PLOS ONE 16(6), 0251894 (2021). doi:10.1371/journal.pone.0251894, Published by PLOS, San Francisco, California, US

B-cell lymphoproliferative disorders exhibit a diverse spectrum of diagnostic entities with heterogeneous behaviour. Multiple efforts have focused on the determination of the genomic drivers of B-cell lymphoma subtypes. In the meantime, the aggregation of diverse tumors in pan-cancer genomic studies has become a useful tool to detect new driver genes, while enabling the comparison of mutational patterns across tumors. Here we present an integrated analysis of 354 B-cell lymphoid disorders. 112 recurrently mutated genes were discovered, of whichKMT2D,CREBBP,IGLL5andBCL2were the most frequent, and 31 genes were putative new drivers. Mutations inCREBBP,TNFRSF14andKMT2Dpredominated in follicular lymphoma, whereas those inBTG2,HTA-AandPIM1were more frequent in diffuse large B-cell lymphoma. Additionally, we discovered 31 significantly mutated protein networks, reinforcing the role of genes such asCREBBP,EEF1A1,STAT6,GNA13andTP53, but also pointing towards a myriad of infrequent players in lymphomagenesis. Finally, we report aberrant expression of oncogenes and tumor suppressors associated with novel noncoding mutations (DTX1andS1PR2), and new recurrent copy number aberrations affecting immune check-point regulators (CD83,PVR) and B-cell specific genes (TNFRSF13C). Our analysis expands the number of mutational drivers of B-cell lymphoid neoplasms, and identifies several differential somatic events between disease subtypes.

In response to cold or diet, fatty acids are dissipated into heat through uncoupling protein 1 (UCP1) in brown adipose tissue (BAT). This process is termed non-shivering thermogenesis, which is important for body temperature maintenance and contributes to obesity pathogenesis. Thermogenic enhancement has been considered a promising anti-obesity strategy. Ghrelin and its receptor Growth Hormone Secretagogue Receptor (GHS-R) have critical roles in energy intake, nutrient sensing, and lipid metabolism. We previously reported that global Ghsr-knockout mice have increased energy expenditure due to enhanced thermogenesis. To determine the site of action for GHS-R mediated thermogenesis, we generated brown adipocyte-specific Ghsr knockout mice (UCP1-CreER/Ghsrf/f) and assessed thermogenic responses under regular diet (RD) fed homeostatic metabolic state or high-fat diet (HFD) fed metabolically-impaired obese state, under normal or cold housing environment. Under a RD-feeding, UCP1-CreER/Ghsrf/f mice showed increased body fat and a slightly elevated core body temperature under cold but not under normal temperature. Consistently, the expression of thermogenic genes in BAT of RD-fed UCP1-CreER/Ghsrf/f mice was increased in reposes to cold. Under HFD feeding, HFD-fed UCP1-CreER/Ghsrf/f mice showed no difference in body fat or body temperature under either normal or cold exposure. Interestingly, the expression of thermogenic genes in BAT of HFD-fed UCP1-CreER/Ghsrf/f mice was upregulated under normal temperature but downregulated under cold exposure. Overall, our data show that GHS-R has cell-autonomous effect in brown adipocytes, and GHS-R regulates BAT thermogenic activity in a temperature- and metabolic state-dependent manner. The thermogenic effect of GHS-R in BAT is more pronounced in cold environment and differentially variable based on metabolic state; under cold exposure, GHS-R inhibition in BAT activates thermogenesis under homeostatic state but suppresses thermogenesis under obese state. Our finding collectively suggests that GHS-R in BAT, acting as a “metabolic thermostat”, differentially regulates thermogenesis in response to different metabolic and thermal stimuli., PLoS ONE, 16 (4), ISSN:1932-6203

PLOS ONE 16(1), e0246182 (2021). doi:10.1371/journal.pone.0246182, Published by PLOS, San Francisco, California, US

Patellofemoral pain (PFP) is one of the most prevalent injuries in runners. Unfortunately, a substantial part of injured athletes do not recover fully from PFP in the long-term. Although previous studies have shown positive effects of gait retraining in this condition, retraining protocols often lack clinical applicability because they are time-consuming, costly for patients and require a treadmill. The primary objective of this study will be to compare the effects of two different two-week partially supervised gait retraining programs, with a control intervention; on pain, function and lower limb kinematics of runners with PFP. It will be a single-blind randomized clinical trial with six-month follow-up. The study will be composed of three groups: a group focusing on impact (group A), a group focusing on cadence (group B), and a control group that will not perform any intervention (group C). The primary outcome measure will be pain assessed using the Visual Analog Pain scale during running. Secondary outcomes will include pain during daily activities (usual), symptoms assessed using the Patellofemoral Disorders Scale and lower limb running kinematics in the frontal (contralateral pelvic drop; hip adduction) and sagittal planes (foot inclination; tibia inclination; ankle dorsiflexion; knee flexion) assessed using the MyoResearch 3.14—MyoVideo (Noraxon U.S.A. Inc.). The study outcomes will be evaluated before (t0), immediately after (t2), and six months (t24) after starting the protocol. Our hypothesis is that both partially supervised gait retraining programs will be more effective in reducing pain, improving symptoms, and modifying lower limb kinematics during running compared with the control group, and that the positive effects from these programs will persist for six months. Also, we believe that one gait retraining group will not be superior to the other. Results from this study will help improve care in runners with PFP, while maximizing clinical applicability as well as time and cost-effectiveness.

Este artículo se encuentra disponible en la siguiente URL: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0250686 The aim of the present clinical trial is to evaluate the efficacy of kinesio taping on patients with chronic low back pain, when the exploration identifies skin/fascia mobilization as a factor that could modify the treatment effect. This study is a randomized controlled trial with intention-totreat analysis. Sixty-two participants with chronic low back pain were therefore recruited from a tertiary referral hospital. Targeted kinesio taping, according to skin/fascia mobility exploration, was applied in the experimental group (17 female/13 male; 49.47 ± 11.15 years) once a week for four sessions. The control group (17 female/14 male; 48.87 ± 9.09 years) underwent a placebo taping application. At post-treatment time there was a statistically significant reduction both in disability (Roland-Morris Disability Questionnaire) and pain (Numeric Pain Rating Scale) in the experimental group (disability: −2.88, 95% confidence interval [CI] −4.56 to −1.21, P < .001; pain: −1.58, 95% CI −2.67 to −0.54 P = .001) and the control group (disability: −1.82, 95% CI −3.46 to −0.17 P = .025; pain: −1.30, 95% CI −2.32 to −0.28 P = .008). However, at six months, these changes only remained significant in the experimental group (disability: −2.95, 95% CI −4.72 to −1.18, P < .001; pain: −1.06, 95% CI −2.07 to −0.04, P < .05). As a conclusion, the application of targeted kinesio taping produced a significant reduction in pain and disability, at 4 weeks and at 6 moths follow-up, although there were no differences between groups at any measurement time point.

Experimental Zika virus infection in non-human primates results in acute viral load dynamics that can be well-described by mathematical models. The inoculum dose that would be received in a natural infection setting is likely lower than the experimental infections and how this difference affects the viral dynamics and immune response is unclear. Here we study a dataset of experimental infection of non-human primates with a range of doses of Zika virus. We develop new models of infection incorporating both an innate immune response and viral interference with that response. We find that such a model explains the data better than models with no interaction between virus and the immune response. We also find that larger inoculum doses lead to faster dynamics of infection, but approximately the same total amount of viral production., Author summary The relationship between the infecting dose of a pathogen and the subsequent viral dynamics is unclear in many disease settings, and this relationship has implications for both the timing and the required efficacy of antiviral therapy. Since experimental challenge studies often employ higher doses of virus than would generally be present in natural infection assessment of this relationship is particularly important for translation of findings. In this study we used mathematical modelling of viral load data from a multi-dose study of Zika virus infection in a macaque model to describe the impact of varying the dose of Zika virus on model parameters, and developed a novel mathematical model incorporating viral interference with the innate immune response.

The emergence of new pathogens is a major threat to public and veterinary health. Changes in bacterial habitat such as a switch in host or disease tropism are typically accompanied by genetic diversification. Staphylococcus aureus is a multi-host bacterial species associated with human and livestock infections. A microaerophilic subspecies, Staphylococcus aureus subsp. anaerobius, is responsible for Morel’s disease, a lymphadenitis restricted to sheep and goats. However, the evolutionary history of S. aureus subsp. anaerobius and its relatedness to S. aureus are unknown. Population genomic analyses of clinical S. aureus subsp. anaerobius isolates revealed a highly conserved clone that descended from a S. aureus progenitor about 1000 years ago before differentiating into distinct lineages that contain African and European isolates. S. aureus subsp. anaerobius has undergone limited clonal expansion, with a restricted population size, and an evolutionary rate 10-fold slower than S. aureus. The transition to its current restricted ecological niche involved acquisition of a pathogenicity island encoding a ruminant host-specific effector of abscess formation, large chromosomal re-arrangements, and the accumulation of at least 205 pseudogenes, resulting in a highly fastidious metabolism. Importantly, expansion of ~87 insertion sequences (IS) located largely in intergenic regions provided distinct mechanisms for the control of expression of flanking genes, including a novel mechanism associated with IS-mediated anti-anti-sense decoupling of ancestral gene repression. Our findings reveal the remarkable evolutionary trajectory of a host-restricted bacterial pathogen that resulted from extensive remodelling of the S. aureus genome through an array of diverse mechanisms in parallel., Author summary The emergence of new pathogens is a major threat to public and veterinary health. Some bacteria such as Staphylococcus aureus, have the capacity to infect many different host species including humans and livestock while others such as the closely-related S. aureus subsp. anaerobius, associated with a single type of pathology called Morel’s disease in small ruminants, are highly niche-restricted. However, our understanding of the genetic basis for such differences in bacterial host-tropism is very limited. Here, we discovered that S. aureus subsp. anaerobius evolved from an S. aureus ancestor and underwent an array of extensive changes to its genome that accompanied the transition to its current restricted lifestyle. We observed genome decay involving loss of function of hundreds of genes, large intra-chromosomal rearrangements affecting most of the genome, acquisition of a pathogenicity island, and expansion of large numbers of insertion sequences that are inserted at intergenic sites around the genome. Importantly, we found that IS elements affect the expression of neighbouring genes in different ways including a novel mechanism of IS-enabled disruption of ancestral gene repression. Taken together, we provide a remarkable example of radical genomic changes associated with evolutionary transition from a multi-host to highly restricted host ecology.

Este artículo se encuentra disponible en la página web de la revista en la siguiente URL: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0248499 En este artículo también participan: Isabelle Naemi Kaneko, Laura Adriane de Moraes Pinto, Antônio Alves de Melo Filho, Vany Perpétua Ferraz, Gisele Maria Fagundes y James Pierre Muir. We evaluated the effects of the inclusion of copaı´ba (Copaifera officinalis L.) essential oil at 0.05 and 0.1% as a possible replacement of synthetic additive butylated hydroxytoluene (BHT) in sheep burgers during 14 days of storage in a refrigerated display case (4˚C). During the shelf life days, analyzes of antioxidant activity, lipid oxidation, pH, color, cooking loss, texture and consumer acceptability were carried out on refrigerated burgers. The addition of copaı´ba essential oil showed an antioxidant effect in sheep burgers (P > 0.05), reducing lipid oxidation. Copaiba essential oil added at 0.05% showed the highest antioxidant activity, decreased cooking losses and delaying discoloration (loss of redness) during storage; it further improved, the tenderness of sheep burgers (P < 0.05). The treatments had no effect (P > 0.05) on consumer acceptability. Copaiba essential oil is a promising natural antioxidant to increase the shelf life of meat products, as well as being a viable solution to replace synthetic antioxidant BHT, thereby promoting the fresh-like quality appeal of sheep burgers.

Purpose Organizational context is recognized as important for facilitating evidence-based practice and improving patient outcomes. Organizational context is a complex construct to measure and appropriate instruments that can quantify and measure context are needed. The aim of this study was to translate and cross-culturally adapt the Alberta Context Tool (ACT) to Norwegian, and to test the reliability and structural validity among registered nurses (RNs) and licenced practice nurses (LPNs) working in nursing homes. Methods This study was a validation study utilizing a cross-sectional design. The sample consisted of n = 956 healthcare personnel from 28 nursing homes from a municipality in Norway. In the first stage, the ACT was translated before being administered in 28 nursing homes. In the second stage, internal consistency and structural validity were explored using Cronbach’s alpha and confirmatory factor analysis. Results A rigorous forward-and-back translation process was performed including a team of academics, experts, professional translators and the copyright holders, before an acceptable version of the ACT was piloted and finalized. The Norwegian version of the ACT showed good internal consistency with Chronbachs alpha above .75 for all concepts except for Formal interactions where the alpha was .69. Structural validity was acceptable for both RNs and LPNs with factors loadings more than .4 for most items. Conclusions The Norwegian version of the ACT is a valid measure of organizational context in Norwegian nursing homes among RNs and LPNs.

This study determined the association between respiratory symptoms and death from respiratory causes over a period of 45 years. In four cohorts of random samples of Norwegian populations with 103,881 participants, 43,731 persons had died per 31 December 2016. In total, 5,949 (14%) had died from respiratory diseases; 2,442 (41%) from lung cancer, 1,717 (29%) chronic obstructive pulmonary disease (COPD), 1,348 (23%) pneumonia, 119 (2%) asthma, 147 (2%) interstitial lung disease and 176 (3%) other pulmonary diseases. Compared with persons without respiratory symptoms the multivariable adjusted hazard ratio (HR) for lung cancer deaths increased with score of breathlessness on effort and cough and phlegm, being 2.6 (95% CI 2.1–3.2) for breathlessness score 3 and 2.1 (95% CI 1.7–2.5) for cough and phlegm score 5. The HR of COPD death was 6.4 (95% CI 5.4–7.7) for breathlessness score 3 and 3.0 (2.4–3.6) for cough and phlegm score 5. Attacks of breathlessness and wheeze score 2 had a HR of 1.6 (1.4–1.9) for COPD death. The risk of pneumonia deaths increased also with higher breathlessness on effort score, but not with higher cough and phlegm score, except for score 2 with HR 1.5 (1.2–1.8). In this study with >2.4 million person-years at risk, a positive association was observed between scores of respiratory symptoms and deaths due to COPD and lung cancer. Respiratory symptoms are thus important risk factors, which should be followed thoroughly by health care practitioners for the benefit of public health.

Copyright: © 2021 Machado et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited., Adipose tissue is one of the major reservoirs of Trypanosoma brucei parasites, the causative agent of sleeping sickness, a fatal disease in humans. In mice, the gonadal adipose tissue (AT) typically harbors 2-5 million parasites, while most solid organs show 10 to 100-fold fewer parasites. In this study, we tested whether the AT environment responds immunologically to the presence of the parasite. Transcriptome analysis of T. brucei infected adipose tissue revealed that most upregulated host genes are involved in inflammation and immune cell functions. Histochemistry and flow cytometry confirmed an increasingly higher number of infiltrated macrophages, neutrophils and CD4+ and CD8+ T lymphocytes upon infection. A large proportion of these lymphocytes effectively produce the type 1 effector cytokines, IFN-γ and TNF-α. Additionally, the adipose tissue showed accumulation of antigen-specific IgM and IgG antibodies as infection progressed. Mice lacking T and/or B cells (Rag2-/-, Jht-/-), or the signature cytokine (Ifng-/-) displayed a higher parasite load both in circulation and in the AT, demonstrating the key role of the adaptive immune system in both compartments. Interestingly, infections of C3-/- mice showed that while complement system is dispensable to control parasite load in the blood, it is necessary in the AT and other solid tissues. We conclude that T. brucei infection triggers a broad and robust immune response in the AT, which requires the complement system to locally reduce parasite burden., This work was supported by the European Research Council (FatTryp, ref. 771714) awarded to LMF, by Fundação para a Ciência e Tecnologia (CEECIND/03322/2018) awarded to LMF, (PTDC/MED-IMU/30948/2017 and CEECIND/00697/2018) awarded to KS, (PD/BD/128286/2017) awarded to HM, (SFRH/BPD/89833/2012) awarded to ST, (IMM/BI/83-2017 through PTDC/BIM-MET/4471/2014) awarded to TB-R and by the National Institutes of Health (NIGMS K99GM132557) awarded to FR-F. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Background Electronic Health Record Systems (EHRs) are being rolled out nationally in many low- and middle-income countries (LMICs) yet assessing actual system usage remains a challenge. We employed a nominal group technique (NGT) process to systematically develop high-quality indicators for evaluating actual usage of EHRs in LMICs. Methods An initial set of 14 candidate indicators were developed by the study team adapting the Human Immunodeficiency Virus (HIV) Monitoring, Evaluation, and Reporting indicators format. A multidisciplinary team of 10 experts was convened in a two-day NGT workshop in Kenya to systematically evaluate, rate (using Specific, Measurable, Achievable, Relevant, and Time-Bound (SMART) criteria), prioritize, refine, and identify new indicators. NGT steps included introduction to candidate indicators, silent indicator ranking, round-robin indicator rating, and silent generation of new indicators. 5-point Likert scale was used in rating the candidate indicators against the SMART components. Results Candidate indicators were rated highly on SMART criteria (4.05/5). NGT participants settled on 15 final indicators, categorized as system use (4); data quality (3), system interoperability (3), and reporting (5). Data entry statistics, systems uptime, and EHRs variable concordance indicators were rated highest. Conclusion This study describes a systematic approach to develop and validate quality indicators for determining EHRs use and provides LMICs with a multidimensional tool for assessing success of EHRs implementations.

Despite the increasing personalization of medicine, surprisingly ~37.0°C (98.6°F) continues as the estimate of normal temperature. We investigated between-subject and within-subject thermal variability, whether a significant percentage of individuals have a low mean oral temperature, and whether these differ by sex, age, time of day, ethnicity, body mass index (BMI), or menstrual phase. Oral temperature was measured by Life Brand® Fast-Read Digital Oral Thermometers and sampled 14 times over two weeks, seven morning and seven evening readings. The volunteer sample consisted of 96 adults (42 men, 54 women; 27 couples, 42 singletons), ages 18–67 years. We found sizeable individual differences in body temperature and that the normal temperature of many individuals is considerably lower than 37.0°C (98.6°F). Mean temperatures ranged from 35.2°C (95.4°F) to 37.4°C (99.3°F). The mean temperature across all participants was 36.1°C (97.0°F)—lower than most studies have reported, consistent with recent evidence of temperature declining over decades. 77% had mean temperatures at least 0.55°C (1°F) lower than 37.0°C (98.6°F). Mean temperature did not differ by age, but women had higher temperatures than men, even within a couple with room temperature and warmth of clothing equated. Although oral temperature varied widely across individuals, it showed marked stability within individuals over days. Variability of temperature over days did not differ by sex, but was larger among younger adults. Using 37.0°C (98.6°F) as the assumed normal temperature for everyone can result in healthcare professionals failing to detect a serious fever in individuals with a low normal temperature or obtaining false negatives for those individuals when using temperature to screen for COVID-19, mistaking their elevated temperature as normal. Some have called for lowering the estimate of normal temperature slightly (e.g., 0.2°C [0.36°F]). That still seems an overly high estimate. More important, using any standardized “normal” temperature will lead to errors for many people. Individual differences are simply too great. Personalizing body temperature is needed. Temperature could be measured at yearly doctor visits, as blood pressure is now. That would be simple to implement. Since our results show marked thermal stability within an individual, sampling temperature only once yearly could provide an accurate indication of a person’s normal temperature at that time of day. Such records over time would also provide a more accurate understanding of how temperature changes over the lifespan.

Increasing body of experimental evidence suggests that anticancer and antimicrobial therapies may themselves promote the acquisition of drug resistance by increasing mutability. The successful control of evolving populations requires that such biological costs of control are identified, quantified and included to the evolutionarily informed treatment protocol. Here we identify, characterise and exploit a trade-off between decreasing the target population size and generating a surplus of treatment-induced rescue mutations. We show that the probability of cure is maximized at an intermediate dosage, below the drug concentration yielding maximal population decay, suggesting that treatment outcomes may in some cases be substantially improved by less aggressive treatment strategies. We also provide a general analytical relationship that implicitly links growth rate, pharmacodynamics and dose-dependent mutation rate to an optimal control law. Our results highlight the important, but often neglected, role of fundamental eco-evolutionary costs of control. These costs can often lead to situations, where decreasing the cumulative drug dosage may be preferable even when the objective of the treatment is elimination, and not containment. Taken together, our results thus add to the ongoing criticism of the standard practice of administering aggressive, high-dose therapies and motivate further experimental and clinical investigation of the mutagenicity and other hidden collateral costs of therapies., Author summary Evolution of drug resistance to anticancer and antimicrobial therapies is widespread among cancer and pathogen cell populations. Classical theory posits strictly that genetic and phenotypic variation is generated in evolving populations independently of the selection pressure. However, recent experimental findings among antimicrobial agents, traditional cytotoxic chemotherapies and targeted cancer therapies suggest that treatment not only imposes selection but can also affect the rate of adaptation by increasing mutability. Here we analyse a model with drug-induced increase in mutation rate and explore its consequences for treatment optimisation. We argue that the true biological cost of treatment is not limited to the harmful side-effects, but instead realises even more profoundly by fundamentally changing the underlying eco-evolutionary dynamics within the microenvironment. Using the concept of evolutionary rescue, we formulate the treatment as an optimal control problem and solve the optimal elimination strategy, which minimises the probability of evolutionary rescue. We show that aggressive elimination strategies, which aim at eradication as fast as possible and which represent the current standard of care, can be detrimental even with modest drug-induced increases (fold change ≤10) to the baseline mutation rate.

For functional MRI with a multi-channel receiver RF coil, images are often reconstructed channel by channel, resulting into multiple images per time frame. The final image to analyze usually is the result of the covariance Sum-of-Squares (covSoS) combination across these channels. Although this reconstruction is quasi-optimal in SNR, it is not necessarily the case in terms of temporal SNR (tSNR) of the time series, which is yet a more relevant metric for fMRI data quality. In this work, we investigated tSNR optimality through voxel-wise RF coil combination and its effects on BOLD sensitivity. An analytical solution for an optimal RF coil combination is described, which is somewhat tied to the extended Krueger-Glover model involving both thermal and physiological noise covariance matrices. Compared experimentally to covSOS on four volunteers at 7T, the method yielded great improvement of tSNR but, surprisingly, did not result into higher BOLD sensitivity. Solutions to improve the method such as for example the t-score for the mean recently proposed are also explored, but result into similar observations once the statistics are corrected properly. Overall, the work shows that data-driven RF coil combinations based on tSNR considerations alone should be avoided unless additional and unbiased assumptions can be made.

The aim of the study was to examine the test-retest reliability and agreement across methods for assessing individual force-velocity (FV) profiles of the lower limbs in athletes. Using a multicenter approach, 27 male athletes completed all measurements for the main analysis, with up to 82 male and female athletes on some measurements. The athletes were tested twice before and twice after a 2- to 6-month period of regular training and sport participation. The double testing sessions were separated by ~1 week. Individual FV-profiles were acquired from incremental loading protocols in squat jump (SJ), countermovement jump (CMJ) and leg press. A force plate, linear encoder and a flight time calculation method were used for measuring force and velocity during SJ and CMJ. A linear regression was fitted to the average force and velocity values for each individual test to extrapolate the FV-variables: theoretical maximal force (F0), velocity (V0), power (Pmax), and the slope of the FV-profile (SFV). Despite strong linearity (R2>0.95) for individual FV-profiles, the SFV was unreliable for all measurement methods assessed during vertical jumping (coefficient of variation (CV): 14–30%, interclass correlation coefficient (ICC): 0.36–0.79). Only the leg press exercise, of the four FV-variables, showed acceptable reliability (CV:3.7–8.3%, ICC:0.82–0.98). The agreement across methods for F0 and Pmax ranged from (Pearson r): 0.56–0.95, standard error of estimate (SEE%): 5.8–18.8, and for V0 and SFV r: -0.39–0.78, SEE%: 12.2–37.2. With a typical error of 1.5 cm (5–10% CV) in jump height, SFV and V0 cannot be accurately obtained, regardless of the measurement method, using a loading range corresponding to 40–70% of F0. Efforts should be made to either reduce the variation in jumping performance or to assess loads closer to the FV-intercepts. Coaches and researchers should be aware of the poor reliability of the FV-variables obtained from vertical jumping, and of the differences across measurement methods.

Background Prediction models should be externally validated to assess their performance before implementation. Several prediction models for coronavirus disease-19 (COVID-19) have been published. This observational cohort study aimed to validate published models of severity for hospitalized patients with COVID-19 using clinical and laboratory predictors. Methods Prediction models fitting relevant inclusion criteria were chosen for validation. The outcome was either mortality or a composite outcome of mortality and ICU admission (severe disease). 1295 patients admitted with symptoms of COVID-19 at Kings Cross Hospital (KCH) in London, United Kingdom, and 307 patients at Oslo University Hospital (OUH) in Oslo, Norway were included. The performance of the models was assessed in terms of discrimination and calibration. Results We identified two models for prediction of mortality (referred to as Xie and Zhang1) and two models for prediction of severe disease (Allenbach and Zhang2). The performance of the models was variable. For prediction of mortality Xie had good discrimination at OUH with an area under the receiver-operating characteristic (AUROC) 0.87 [95% confidence interval (CI) 0.79–0.95] and acceptable discrimination at KCH, AUROC 0.79 [0.76–0.82]. In prediction of severe disease, Allenbach had acceptable discrimination (OUH AUROC 0.81 [0.74–0.88] and KCH AUROC 0.72 [0.68–0.75]). The Zhang models had moderate to poor discrimination. Initial calibration was poor for all models but improved with recalibration. Conclusions The performance of the four prediction models was variable. The Xie model had the best discrimination for mortality, while the Allenbach model had acceptable results for prediction of severe disease.

Background Midpelvic vacuum extractions are controversial due to reports of increased risk of maternal and perinatal morbidity and high failure rates. Prospective studies of attempted midpelvic vacuum outcomes are scarce. Our main aims were to assess frequency, failure rates, labor characteristics, maternal and neonatal complications of attempted midpelvic vacuum deliveries, and to compare labor characteristics and complications between successful and failed midpelvic vacuum deliveries. Study design Clinical data were obtained prospectively from all attempted vacuum deliveries (n = 891) over a one-year period with a total of 6903 births (overall cesarean section rate 18.2% (n = 1258). Student’s t-test, Mann-Whitney U-test or Chi-square test for group differences were used as appropriate. Odds ratios and 95% confidence intervals are given as indicated. The uni- and multivariable analysis were conducted both as a complete case analysis and with a multiple imputation approach. A p-value of Results Attempted vacuum extractions from midpelvic station constituted 36.7% (n = 319) of all attempted vacuum extractions (12.9% (n = 891) of all births). Of these 319 midpelvic vacuum extractions, 11.3% (n = 36) failed and final delivery mode was cesarean section in 86.1% (n = 31) and forceps in the remaining 13.9% (n = 5). Successful completion of midpelvic vacuum by 3 pulls or fewer was achieved in 67.1%. There were 3.9% third-degree and no fourth-degree perineal tears. Cup detachments were associated with a significantly increased failure rate (adjusted OR 6.13, 95% CI 2.41–15.56, p< 0.001). Conclusion In our study, attempted midpelvic vacuum deliveries had relatively low failure rate, the majority was successfully completed within three pulls and they proved safe to perform as reflected by a low rate of third-degree perineal tears. We provide data for nuanced counseling of women on vacuum extraction as a second stage delivery option in comparable obstetric management settings with relatively high vacuum delivery rates and low cesarean section rates.

This paper presents regulation of an asynchronous induction motor so as to create a stable vacuum milk pump using Variable Frequency Drive (VFD). Contribution includes providing information about the usage of the VFD, which regulates the activity of an asynchronous induction motor so that the vacuum pump milking machine creates stable vacuum. The paper describes the functional and time dependence of input values and output parameters of frequency converters at changing electric motor speed. For simulation and verification the milking process a mathematical model of the milking machine was created. The simulation was verified in Matlab/Simulink software. The constructed mathematical model showed symmetric regulation. Control model symmetry was verified at the laboratory of milking machine. The possibility to remove the control valve from milking equipment was proven using the measured data. It was found that constant vacuum values can be maintained. A constant vacuum can be maintained by changing vacuum pump speed. This control is of an accepted standard (ISO 5707: 2007). The power saving control values (on the milking equipment) of the VFD were positive throughout the measuring range. The performance of the milking vacuum pump is normally designed from the maximum air consumption of the milking machine at nominal vacuum (50 kPa), and a performance reserve is added to this. This means that the pump is operated between the ranges 7.53 and 15.06 dm3 s−1. By using a vacuum pump controlled by a VFD, power savings can be achieved from 32.50% to 54.02% compared to a control valve.

Background Early diagnosis and treatment are one of the key strategies of tuberculosis control globally, and there are strong efforts in detecting and treating tuberculosis cases in Ethiopia. Smear microscopy examination has been a routine diagnostic test for pulmonary tuberculosis diagnosis in resource-constrained settings for decades. Recently, many countries, including Ethiopia, are scaling up the use of Gene Xpert without the evaluation of the cost and cost-effectiveness implications of this strategy. Therefore, this study evaluated the cost and cost-effectiveness of Gene Xpert (MTB/RIF) and smear microscopy tests to diagnosis tuberculosis patients in Ethiopia. Methods We compared the costs and cost-effectiveness of tuberculosis diagnosis using smear microscopy and Gene Xpert among 1332 patients per intervention in the Arsi zone. We applied combinations of top-down and bottom-up costing approaches. The costs were estimated from the health providers’ perspective within one year (2017–2018). We employed “cases detected” as an effectiveness measure, and the incremental cost-effectiveness ratio was calculated by dividing the changes in cost and change in effectiveness. All costs and incremental cost-effectiveness ratio were reported in 2018 US$. Results The unit cost per test for Gene Xpert was $12.9 whereas it is $3.1 for AFB smear microscopy testing. The cost per TB case detected was $77.9 for Gene Xpert while it was $55.8 for the smear microscopy method. The cartridge kit cost accounted for 42% of the overall Gene Xpert’s costs and the cost of the reagents and consumables accounted for 41.3% ($1.3) of the unit cost for the smear microscopy method. The ICER for the Gene Xpert strategy was $20.0 per tuberculosis case detected. Conclusion Using Gene Xpert as a routine test instead of standard care (smear microscopy) can be potentially cost-effective. In the cost scenario analysis, the price of the cartridge, the number of tests performed per day, and the life span of the capital equipment were the drivers of the unit cost of the Gene Xpert method. Therefore, Gene Xpert can be a part of the routine TB diagnostic testing strategy in Ethiopia.

As genomic architectures become more complex, they begin to accumulate degenerate and redundant elements. However, analyses of the molecular mechanisms underlying these genetic architecture features remain scarce, especially in compact but sufficiently complex genomes. In the present study, we followed a proteomic approach together with a computational network analysis to reveal molecular signatures of protein function degeneracy from a plant virus (as virus-host protein-protein interactions). We employed affinity purification coupled to mass spectrometry to detect several host factors interacting with two proteins of Citrus tristeza virus (p20 and p25) that are known to function as RNA silencing suppressors, using an experimental system of transient expression in a model plant. The study was expanded by considering two different isolates of the virus, and some key interactions were confirmed by bimolecular fluorescence complementation assays. We found that p20 and p25 target a common set of plant proteins including chloroplastic proteins and translation factors. Moreover, we noted that even specific targets of each viral protein overlap in function. Notably, we identified argonaute proteins (key players in RNA silencing) as reliable targets of p20. Furthermore, we found that these viral proteins preferentially do not target hubs in the host protein interactome, but elements that can transfer information by bridging different parts of the interactome. Overall, our results demonstrate that two distinct proteins encoded in the same viral genome that overlap in function also overlap in their interactions with the cell proteome, thereby highlighting an overlooked connection from a degenerate viral system., This work was supported by the grants PROMETEO/2019/012 from the Valencian Regional Government (to SA), as well as grants AGL2010-20221/AGR, BIO2017-83184-R, and PGC2018-101410-B-I00 from the Spanish Ministry of Science, Innovation, and Universities (to SA, JAD, and GR, respectively). The two latter grants were co-financed by the European Regional Development Fund.

Objective This study aims to estimate the incidence of severe binocular vision impairment and blindness (SVI/B) and to identify eye diseases and regional risk factors of persons with SVI/B at ages 50 years and older. Methods We designed an observational cohort study based on longitudinal, multifactorial, and administrative information of a random sample of 250,000 persons at ages 50+. All individuals were included in the process-produced health claims register of the Allgemeine Ortskrankenkasse in 2004, and were followed until 2015. We analyzed ten selected eye diseases and regional characteristics as risk factors for SVI/B using Cox models, adjusting for demographic characteristics and multi-morbidity. Results The age-standardized incidence was 79 new diagnoses of SVI/B per 100,000 person-years (95%-CI: 76-82); 77 for males (72-82) and 81 for females (77-85). By adjusting for multiple factors, the model revealed and confirmed that individuals who were very old (Hazard ratio90+: 6.67; 3.59-12.71), male (1.18; 1.01-1.38), had multi-morbidities (three+ diseases: 3.36; 2.51-4.49), or had diabetes (1.26; 1.07-1.49) had an increased risk of SVI/B. Compared to persons without the particular eye disease (all p Conclusion The study revealed and confirmed some profound risk factors of SVI/B at both the individual and the macro level. The sizes of the effects of the characteristics of the living context were smaller than those of the individual characteristics, especially for some severe eye diseases. While urbanity and access to health services had no effect, regional economic wealth was a risk factor for SVI/B. Future health care measures and advice by physicians should take these dimensions of inequalities in SVI/B into account.

The contemporaneous association between higher socioeconomic position and better health is well established. Life course research has also demonstrated a lasting effect of childhood socioeconomic conditions on adult health and well-being. Yet, little is known about the separate health effects of intergenerational mobility—moving into a different socioeconomic position than one’s parents—among early adults in the United States. Most studies on the health implications of mobility rely on cross-sectional datasets, which makes it impossible to differentiate between health selection and social causation effects. In addition, understanding the effects of social mobility on health at a relatively young age has been hampered by the paucity of health measures that reliably predict disease onset. Analysing 4,713 respondents aged 25 to 32 from the National Longitudinal Study of Adolescent Health’s Waves I and IV, we use diagonal reference models to separately identify the effects of socioeconomic origin and destination, as well as social mobility on allostatic load among individuals in the United States. Using a combined measure of educational and occupational attainment, and accounting for individuals’ initial health, we demonstrate that in addition to health gradient among the socially immobile, individuals’ socioeconomic origin and destination are equally important for multi-system physiological dysregulation. Short-range upward mobility also has a positive and significant association with health. After mitigating health selection concerns in our observational data, this effect is observed only among those reporting poor health before experiencing social mobility. Our findings move towards the reconciliation of two theoretical perspectives, confirming the positive effect of upward mobility as predicted by the “rags to riches” perspective, while not contradicting potential costs associated with more extensive upward mobility experiences as predicted by the dissociative thesis.

The endothelial glycocalyx (EG) is essential for proper function of the endothelium and for vascular integrity, but its role in premature atherogenesis in rheumatoid arthritis (RA) has not been studied yet. EG impairment can play a role in pathogenesis of vascular disease, and one of its characteristics is shedding of syndecan-1 from endothelial cells. Syndecan-1 shedding is mediated by matrix metalloproteinase-9 (MMP-9) and counteracted by tissue inhibitor of metalloproteinases (TIMP)-1. Cardiovascular disease risk in RA is reversible by disease modifying antirheumatic drugs (DMARDs), but the exact modes of action are still unclear. Therefore, we examined effects of DMARDs on syndecan-1, MMP-9 and TIMP-1 in RA patients, and searched for associations between these parameters and inflammatory activity. From the observational PSARA study, we examined 39 patients starting with methotrexate (MTX) monotherapy (in MTX naïve patients, n = 19) or tumor necrosis factor inhibitors (TNFi) in combination with MTX (in MTX non-responders, n = 20) due to active RA. Serum syndecan-1, MMP-9 and TIMP-1 were measured at baseline and after six weeks of treatment. Serum syndecan-1 (p = 0.008) and TIMP-1 (p

Objective Whether use of nonsteroidal anti-inflammatory drugs (NSAIDs) reduce the risk of incident Parkinson’s disease (PD) remains unresolved. Here, we employed the Norwegian Prescription Database to examine whether NSAID use is associated with a lower incidence of PD. Methods We compared the incidence of PD among users of NSAIDs in a population-based retrospective study using the Norwegian Prescription Database from 2004 to 2017. In total 7580 PD patients were identified using dopaminergic therapy over time as proxy for PD diagnosis. Analyses were performed with minimum 90 and 365 defined daily dose (DDD) NSAID exposure, respectively. Time-dependent Cox regression model and a binary logistic regression analysis with a 5-year lag until PD diagnosis were performed for all NSAIDs. Results There was overall no decrease in incidence of PD among NSAID users compared to controls. Using a minimum of 90 or 365 DDD threshold of exposure produced similar results. Analysis of individual NSAIDs did not show difference in PD incidence compared to controls Age-specific incidence rates of PD were comparable to reported age-specific incidence rates in previous studies. Interpretation Our findings provide no evidence that cumulative high exposure to NSAIDs affects the risk of developing PD.

Extensive non-pharmaceutical and physical distancing measures are currently the primary interventions against coronavirus disease 2019 (COVID-19) worldwide. It is therefore urgent to estimate the impact such measures are having. We introduce a Bayesian epidemiological model in which a proportion of individuals are willing and able to participate in distancing, with the timing of distancing measures informed by survey data on attitudes to distancing and COVID-19. We fit our model to reported COVID-19 cases in British Columbia (BC), Canada, and five other jurisdictions, using an observation model that accounts for both underestimation and the delay between symptom onset and reporting. We estimated the impact that physical distancing (social distancing) has had on the contact rate and examined the projected impact of relaxing distancing measures. We found that, as of April 11 2020, distancing had a strong impact in BC, consistent with declines in reported cases and in hospitalization and intensive care unit numbers; individuals practising physical distancing experienced approximately 0.22 (0.11–0.34 90% CI [credible interval]) of their normal contact rate. The threshold above which prevalence was expected to grow was 0.55. We define the “contact ratio” to be the ratio of the estimated contact rate to the threshold rate at which cases are expected to grow; we estimated this contact ratio to be 0.40 (0.19–0.60) in BC. We developed an R package ‘covidseir’ to make our model available, and used it to quantify the impact of distancing in five additional jurisdictions. As of May 7, 2020, we estimated that New Zealand was well below its threshold value (contact ratio of 0.22 [0.11–0.34]), New York (0.60 [0.43–0.74]), Washington (0.84 [0.79–0.90]) and Florida (0.86 [0.76–0.96]) were progressively closer to theirs yet still below, but California (1.15 [1.07–1.23]) was above its threshold overall, with cases still rising. Accordingly, we found that BC, New Zealand, and New York may have had more room to relax distancing measures than the other jurisdictions, though this would need to be done cautiously and with total case volumes in mind. Our projections indicate that intermittent distancing measures—if sufficiently strong and robustly followed—could control COVID-19 transmission. This approach provides a useful tool for jurisdictions to monitor and assess current levels of distancing relative to their threshold, which will continue to be essential through subsequent waves of this pandemic.

***this preprint has since been published: https://journals.plos.org/plosone/article?id=10.1371/journal.pone.0247997 (Open Access)*** During medical pandemics, protective behaviors need to be motivated by effective communication, where finding predictors of fear and perceived health is of critical importance. The varying trajectories of the COVID-19 pandemic in different countries afford the opportunity to assess the unique influence of ‘macro-level’ environmental factors and ‘micro-level’ psychological variables on both fear and perceived health. Here, we investigate predictors of fear and perceived health using machine learning as lockdown restrictions in response to the COVID-19 pandemic were introduced in Austria, Spain, Poland and Czech Republic. Over a seven-week period, 533 participants completed weekly self-report surveys which measured the target variables subjective fear of the virus and perceived health, in addition to potential predictive variables related to psychological factors, social factors, perceived vulnerability to disease (PVD), and economic circumstances. Viral spread, mortality and governmental responses were further included in the analysis as potential environmental predictors. Results revealed that our models could accurately predict fear of the virus (accounting for approximately 23% of the variance) using predictive factors such as worrying about shortages in food supplies, and perceived vulnerability to disease (PVD), where interestingly, environmental factors such as spread of the virus and governmental restrictions did not contribute to this prediction. Furthermore, our results revealed that perceived health could be predicted using PVD, physical exercise, attachment anxiety and age as input features, albeit with smaller effect sizes. Taken together, our results emphasize the importance of ‘micro-level’ psychological factors, as opposed to ‘macro-level’ environmental factors, when predicting fear and perceived health, and offer a starting point for more extensive research on the influences of pathogen threat and governmental restrictions on the psychology of fear and health.

[Abstract] Our current knowledge on the crocodyliform evolution is strongly biased towards the skull morphology, and the postcranial skeleton is usually neglected in many taxonomic descriptions. However, it is logical to expect that it can contribute with its own phylogenetic signal. In this paper, the changes in the tree topology caused by the addition of the postcranial information are analysed for the family Allodaposuchidae, the most representative eusuchians in the latest Cretaceous of Europe. At present, different phylogenetic hypotheses have been proposed for this group without reaching a consensus. The results of this paper evidence a shift in the phylogenetic position when the postcranium is included in the dataset, pointing to a relevant phylogenetic signal in the postcranial elements. Finally, the phylogenetic relationships of allodaposuchids within Eusuchia are reassessed; and the internal relationships within Allodaposuchidae are also reconsidered after an exhaustive revision of the morphological data. New and improved diagnoses for each species are here provided. This work was supported by postdoctoral grants funded by the program “Axudas á etapa postdoutoral da Xunta de Galicia 2017 – Modalidade A” (ED481B 2017/027) (https://www.xunta.gal/) and the program “Ayudas Juan de la Cierva – Formación 2019” (FJC2019-042583-I) (https://www.ciencia.gob.es/). Additional support to visit palaeontological collections was received from the program “Axudas á investigación 2017 e 2018” of the Universidade da Coruña (https://www.udc.es/), and from the SYNTHESYS+ Project (DE-TAF-2604) which is financed by the European Commission via the H2020 Research Infrastructure programme (https://www.synthesys.info/). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript Xunta de Galicia; ED481B 2017/027