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This is a report on how 1H NMR-based metabonomics was employed to discriminate osteopenia from osteoporosis in postmenopausal women, identifying the main metabolites associated to the separation between the groups. The Assays were performed using seventy-eight samples, being twenty-eight healthy volunteers, twenty-six osteopenia patients and twenty-four osteoporosis patients. PCA, LDA, PLS-DA and OPLS-DA formalisms were used. PCA discriminated the samples from healthy volunteers from diseased patient samples. Osteopenia-osteoporosis discrimination was only obtained using Analysis Discriminants formalisms, as LDA, PLS-DA and OPLS-DA. The metabonomics model using LDA formalism presented 88.0% accuracy, 88.5% specificity and 88.0% sensitivity. Cross-Validation, however, presented some problems as the accuracy of modeling decreased. LOOCV resulted in 78.0% accuracy. The OPLS-DA based model was better: R2Y and Q2 values equal to 0.871 (p<0.001) and 0.415 (p<0.001). LDA and OPLS-DA indicated the important spectral regions for discrimination, making possible to assign the metabolites involved in the skeletal system homeostasis, as follows: VLDL, LDL, leucine, isoleucine, allantoin, taurine and unsaturated lipids. These results indicate that 1H NMR-based metabonomics can be used as a diagnosis tool to discriminate osteoporosis from osteopenia using a single serum sample. [ABSTRACT FROM AUTHOR]

This cross-sectional study aims to describe doubts regarding the diagnosis and treatment of syphilis in pregnancy among primary care professionals in a telehealth service. All teleconsultations (TCs) offered through TelessaúdeRS-UFRGS to primary health care (PHC) services in the state of Rio Grande do Sul between 2018 and 2021 involving syphilis in pregnancy were included. A total of 356 (TCs) were analyzed. The main doubts about syphilis during pregnancy raised by primary care professionals were related to the need for retreatment (35%), diagnostic definition (23%) and initial treatment (16%). In addition, 95% of TCs were suitable for diagnosing and treating syphilis based on the 2020 Brazilian Ministry of Health guideline. This study suggests that TCs can identify failures in the diagnosis and treatment of public health problems and support decision making in PHC involving syphilis in pregnancy., Competing Interests: I have read the journal’s policy and the authors of this manuscript have the following competing interests: The authors declare that being collaborators of TelessaúdeRS-UFRGS raises a potential conflict of interest., (Copyright: © 2024 de Carvalho 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.)

Robust prognostic and predictive factors for hepatocellular carcinoma, a leading cause of cancer-related deaths worldwide, have not yet been identified. Previous studies have identified potential HCC determinants such as genetic mutations, epigenetic alterations, and pathway dysregulation. However, the clinical significance of these molecular alterations remains elusive. MicroRNAs are major regulators of protein expression. MiRNA functions are frequently altered in cancer. In this study, we aimed to explore the prognostic value of differentially expressed miRNAs in HCC, to elucidate their associated pathways and their impact on treatment response. To this aim, bioinformatics techniques and clinical dataset analyses were employed to identify differentially expressed miRNAs in HCC compared to normal hepatic tissue. We validated known associations and identified a novel miRNA signature with potential prognostic significance. Our comprehensive analysis identified new miRNA-targeted pathways and showed that some of these protein coding genes predict HCC patients' response to the tyrosine kinase inhibitor sorafenib., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Smith 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.)

The objective of this study was to investigate how Brazilian physical therapists (PTs) use therapeutic exercises in the rehabilitation of individuals with rotator cuff (RC) tendinopathy. The study used an online survey with a mix of 62 open- and closed-ended questions divided into three sections: participant demographics, professional experience, and clinical practice in the rehabilitation of patients with RC tendinopathy. One hundred and fifty-nine Brazilian physical therapists completed the survey. Most of our sample recommended isometric exercises (69.9%) in the initial phase of rehabilitation and eccentric exercises (47.4%) in the advanced phase. However, there was a wide variability in determining the volume of exercises, particularly with isometric exercises. Most of our sample considered patient comfort and pain levels when adjusting exercise intensity, regardless of exercise type. The majority (48.40%) recommended weekly reassessment and modification of exercises. Additionally, despite pain being a key factor for discharge and the primary adverse effect of exercise, most of our sample would not discontinue exercises in case of pain during the early and late phases of rehabilitation. Despite the lack of consensus on some aspects, the clinical practice of our sample is in line with the current literature and practice in other countries. However, further research and implementation are crucial to enhance future rehabilitation outcomes, including exploring the exercise training volume, the safety and effectiveness of exercising with pain and identifying the optimal pain level for best results., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Augusto 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.)

Background: Violence against women has a negative impact on multiple dimensions of women's health. During the Covid-19 pandemic, intimate partner violence against women has continued, and in some contexts has intensified. The aim of this study was to identify the prevalence of intimate partner violence against women during covid-19 pandemic and its association with socioeconomic, behavioral, and life-experience factors., Methods and Findings: Cross-sectional, population-based study conducted in the municipality of Vitória, state of Espírito Santo, from January to May 2022, where 1,086 women aged 18 years and over were interviewed. The World Health Organization (WHO) instrument on violence against women was used to screen outcomes. The prevalence of violence during the pandemic (psychological, physical, and sexual) and bivariate analysis with sociodemographic, behavioral, family, and life history characteristics of women were estimated. The multivariate analysis was carried out for each type of violence, the Poisson regression model was performed with an estimate of robust variance, inserting the variables of interest with (p<0.20). Those with p<0.05 remained in the adjusted model., Results: The prevalence of violence psychological against women perpetrated by an intimate partner during the pandemic was the most frequent (20.2%), followed by physical (9.0%) and sexual violence (6.5%). Women with less schooling and who were single had a higher prevalence of physical and psychological violence, as did those with a history of sexual abuse in childhood and whose mothers had been beaten by their intimate partners. Sexual violence was more prevalent among non-white, with up to eight years of schooling, whose mothers had a history of intimate partner violence, and who consumed alcohol during four days or more (p<0.01)., Conclusion: Psychological, physical, and sexual violence perpetrated by the intimate partner during the pandemic presented high magnitude among women living in Vitória. Sociodemographic, behavioral factors, and personal and maternal experiences of violence were associated with the phenomenon., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Leite 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.)

The aim of this study was to analyze levels of fatigue and resilience of Brazilian graduate students during the COVID-19 pandemic and to determine whether there is an association between fatigue and resilience and sociodemographic and academic factors. Data were analyzed using descriptive and inferential statistics, and it was discovered that the variables associated with higher levels of resilience were age; having children; being retired; receiving income above five minimum wages; having had greater problems in other phases of the research schedule; coming from private universities; being from the north of Brazil; studying the area of Health; and having their research schedule unaffected during the pandemic. On the other hand, lack of resilience was associated with not having children; being less well-off financially; being younger; being a woman; studying in a public university; and having to postpone part of the research during the pandemic. The conclusion of the study indicated the need for graduate programs to design strategies to deal with fatigue and promote resilience in Master's and PhD students., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Valóta 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.)

The efficiency of the DNA barcoding relies on sequencing fragment of the Cytochrome C Subunit I (COI) gene, which has been claimed as a tool to biodiversity identification from distinct groups. Accordingly, the goal of this study was to identify juvenile fish species along an estuary of Caeté River in the Brazilian Blue Amazon based on. For this purpose, we applied the DNA barcoding and discuss this approach as a tool for discrimination of species in early ontogenetic stages. A 500-bp fragment was obtained from 74 individuals, belonging to 23 species, 20 genera, 13 families and seven orders. About 70% of the 46 haplotypes revealed congruence between morphological and molecular species identification, while 8% of them failed in identification of taxa and 22% demonstrated morphological misidentification. These results proved that COI fragments were effective to diagnose fish species at early life stages, allowing identifying all samples to a species-specific status, except for some taxa whose COI sequences remain unavailable in public databases. Therefore, we recommend the incorporation of DNA barcoding to provide additional support to traditional identification, especially in morphologically controversial groups. In addition, periodic updates and comparative analyses in public COI datasets are encouraged., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Lutz 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.)

Psychopathy is a personality disorder characterized by a mix of traits belonging to four facets: affective (e.g., callous/lack of empathy), interpersonal (e.g., grandiosity), behavioral instability (e.g., impulsivity, poor behavioral controls), and social deviance (e.g., juvenile delinquency, criminal versatility). Several scholars have argued that early childhood maltreatment impacts the development of psychopathy, although views regarding its role in the four facets differ. We conducted a meta-analysis including 47 studies comprising a total of 389 effect sizes and 12,737 participants, to investigate the association between psychopathy and four types of child maltreatment: physical abuse, emotional abuse, neglect, and sexual abuse. We found support for a moderate link between overall psychopathy and childhood physical abuse, emotional abuse, and neglect, as well as overall childhood maltreatment. The link between psychopathy and childhood sexual abuse was small, but still statistically significant. These associations were stronger for the behavioral and antisocial facets than for the affective and interpersonal facets of psychopathy, but nearly all associations were statistically significant. Our findings are consistent with recently developed theories on the role of complex trauma in the development of severe personality disorders. Trauma-focused preventive and therapeutic interventions can provide further tests of the trauma-psychopathy hypothesis., Competing Interests: All authors have read the journal’s policy and the authors of this manuscript have the following competing interests. The first author has provided training in the use of the Psychopathy Checklist-Revised and Psychopathy Checklist:Youth Version. The other authors have declared that no competing interests exist. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

The melanocortin-1 receptor (MC1R) is one of the key proteins involved in the regulation of melanin production and several polymorphisms have been associated with different phenotypes of skin and hair color in human and nonhuman species. Most of the knowledge is centered on more homogeneous populations and studies involving an admixed group of people should be encouraged due to the great importance of understanding the human color variation. This work evaluates the MC1R diversity and the possible impacts of MC1R variants in an admixed sample population of Rio de Janeiro, Brazil, which is a product of Native American, African, and European miscegenation. Sequencing of complete coding region and part of the 3´UTR of MC1R gene identified 31 variants including one insertion and three novel synonymous substitutions in sample population grouped according to skin, hair and eye pigmentation levels. In nonmetric multidimensional scaling analysis (NMDS), three main clusters were identified, in which the Brazilian dark skin group remained in the African cluster whereas the intermediate and the light skin color phenotype in the European one. None gathered with Asians since their immigration to Brazil was a recent event. In silico analyses demonstrated that Cys35Tyr, Ile155Thr and Pro256Ser, found in our population, have a negative effect on receptor function probably due to changes on the receptor structure. Notably, Cys35Tyr mutation could potentially impair agonist binding. Altogether, this work contributes to the understanding of the genetic background of color variation on an admixed population and gives insights into the damaging effects of MC1R variants., Competing Interests: The authors have declared that no competing interests exist.

Background: Identifying low skeletal muscle strength (SMS), skeletal muscle mass (SMM) and skeletal muscle quality (SMQ) is pivotal for diagnosing sarcopenia cases. Age-related declines in SMS, SMM, and SMQ are dissimilar between the upper (UL) and lower limbs (LL). Despite this, both UL and LL measures have been used to assess SMS, SMM and SMQ in older adults. However, it is not clear whether there is agreement between UL and LL measures to identify older adults with low SMS, SMM and SMQ., Objective: To investigate the agreement between UL and LL measures to identify older adults with low SMS, SMM and SMQ., Methods: Participants (n = 385; 66.1 ± 5.1 years; 75,4% females) performed the handgrip strength test (HGS) and the 30-s chair stand test (CST) to assess UL- and LL-SMS, respectively. The SMM was assessed by dual-energy X-ray absorptiometry (DXA). The UL-SMQ was determined as: handgrip strength (kgf) ÷ arm SMM (kg). LL-SMQ was determined as: 30-s CST performance (repetitions) ÷ leg SMM (kg). Results below the 25th percentile stratified by sex and age group (60-69 and 70-80 years) were used to determine low SMS, SMM and SMQ. Cohen's kappa coefficient (κ) was used for the agreement analyses., Results: There was a slight and non-significant agreement between UL and LL measures to identify older adults with low SMS (κ = 0.046; 95% CI 0.093-0.185; p = 0.352). There was a moderate agreement to identify low SMM (κ = 0.473; 95% CI 0.371-0.574; p = 0.001) and a fair agreement to identify low SMQ (κ = 0.206; 95% CI 0.082 to 0.330; p = 0.005)., Conclusion: The agreement between UL and LL measures to identify older adults with low SMS, SMM and SMQ is limited, which might generate different clinical interpretations for diagnosing sarcopenia cases., Competing Interests: The authors have declared that no competing interests exist.

Glycation process refers to reactions between reduction sugars and amino acids that can lead to formation of advanced glycation end products (AGEs) which are related to changes in chemical and functional properties of biological structures that accumulate during aging and diseases. The aim of this study was to perform and analyze in vitro glycation by fructose and methylglyoxal (MGO) using salivary fluid, albumin, lysozyme, and salivary α-amylase (sAA). Glycation effect was analyzed by biochemical and spectroscopic methods. The results were obtained by fluorescence analysis, infrared spectroscopy (total attenuated reflection-Fourier transform, ATR-FTIR) followed by multivariate analysis of principal components (PCA), protein profile, immunodetection, enzymatic activity and oxidative damage to proteins. Fluorescence increased in all glycated samples, except in saliva with fructose. The ATR-FTIR spectra and PCA analysis showed structural changes related to the vibrational mode of glycation of albumin, lysozyme, and salivary proteins. Glycation increased the relative molecular mass (Mr) in protein profile of albumin and lysozyme. Saliva showed a decrease in band intensity when glycated. The analysis of sAA immunoblotting indicated a relative reduction in intensity of its correspondent Mr after sAA glycation; and a decrease in its enzymatic activity was observed. Carbonylation levels increased in all glycated samples, except for saliva with fructose. Thiol content decreased only for glycated lysozyme and saliva with MGO. Therefore, glycation of salivary fluid and sAA may have the potential to identify products derived by glycation process. This opens perspectives for further studies on the use of saliva, an easy and non-invasive collection fluid, to monitor glycated proteins in the aging process and evolution of diseases., Competing Interests: The authors have declared that no competing interests exist.

This article aims to identify networks of constitutional text structured in an imperceptible way. Constitutions has the power to reveal how its devices "dialogue" with each other, forming their own communicative communities. With the help from the network analysis methodology, an interpretative model of textual mapping is proposed based on internal references between the provisions of one Constitution. We use the Brazilian Constitution, whose text is very detailed, analytical and deals with various issues. As a result, a network was identified that included 174 connections between 95 articles of the Constitution and the existence of normative communities that share codes or communicative standards. The study demonstrates the strong possibility to found structuring axis of this communities and relates to the other issues that have reached constitutional stature, showing that the complexity of some Constitutions can be observed beyond its logical sequential structure., Competing Interests: The authors have declared that no competing interests exist.

Adaptive memory is the propensity of human memory to easily store and retrieve important information to deal with challenges related to the Pleistocene. Recent evidence shows that humans have had a multiregional evolution across the African continent, including the rainforests and deciduous forests; however, there is little evidence regarding the implications of these origins and the relevant and recurring challenges of these environments on survival processing advantage in memory. In this study, we conducted an experiment with volunteers to analyze whether adaptive memory operates in the retrieval of important information to solve challenges of using medicinal plants to treat diseases in the ancestral environments of the savanna, rainforests, and deciduous forests compared to the modern environments of desert, tundra, coniferous forest, and urban areas. We used simulated survival environments and asked volunteers (30 per simulated scenario) to imagine themselves sick in one of these environments, and needing to find medicinal plants to treat their disease. The volunteers rated the relevance of 32 words to solve this challenge, followed by a surprise memory test. Our results showed no ancestral priority in recalling relevant information, as both ancestral and modern environments showed a similar recall of relevant information. This suggests that the evolved cognitive apparatus allows human beings to survive and can create survival strategies to face challenges imposed in various environments. We believe that this is only possible if the human mind operates through a flexible cognitive mechanism. This flexibility can reflect, for example, the different environments that the first hominids inhabited and the different dangerous situations that they faced., Competing Interests: The authors have declared that no competing interests exist.

The family Arecaceae is distributed throughout tropical and subtropical regions of the world. Among the five subfamilies, Arecoideae is the most species-rich and still contains some ambiguous inter-generic relationships, such as those within subtribes Attaleinae and Bactridineae. The hypervariable regions of plastid genomes (plastomes) are interesting tools to clarify unresolved phylogenetic relationships. We sequenced and characterized the plastome of Bactris gasipaes (Bactridinae) and compared it with eight species from the three Cocoseae sub-tribes (Attaleinae, Bactridinae, and Elaeidinae) to perform comparative analysis and to identify hypervariable regions. The Bactris gasipaes plastome has 156,646 bp, with 113 unique genes. Among them, four genes have an alternative start codon (cemA, rps19, rpl2, and ndhD). Plastomes are highly conserved within tribe Cocoseae: 97.3% identity, length variation of ~2 kb, and a single ~4.5 kb inversion in Astrocaryum plastomes. The LSC/IR and IR/SSC junctions vary among the subtribes: in Bactridinae and Elaeidinae the rps19 gene is completely contained in the IR region; in the subtribe Attaleinae the rps19 gene is only partially contained in the IRs. The hypervariable regions selected according to sequence variation (SV%) and frequency of parsimony informative sites (PIS%) revealed plastome regions with great potential for molecular analysis. The ten regions with greatest SV% showed higher variation than the plastid molecular markers commonly used for phylogenetic analysis in palms. The phylogenetic trees based on the plastomes and the hypervariable regions (SV%) datasets had well-resolved relationships, with consistent topologies within tribe Cocoseae, and confirm the monophyly of the subtribes Bactridinae and Attaleinae., Competing Interests: The authors have declared that no competing interests exist.

In Dec 2020 Brazil became one of the worldwide epicenters of the COVID-19 pandemic with more than 7.2M reported cases. Brazil has a large territory with unequal distribution of healthcare resources including physicians. Resource limitation has been one of the main factors hampering Brazil's response to the COVID-19 crisis. Telemedicine has been an effective approach for COVID-19 management as it allows to reduce the risk of cross-contamination and provides support to remote rural locations. Here we present the analyses of teleconsultations from a countrywide telemedicine service (TelessáudeRS-UFRGS, TRS), that provides physician-to-physician remote support during the COVID-19 pandemic in Brazil. We performed a descriptive analysis of the teleconsultation incoming calls and a text analysis from the call transcripts. Our findings indicate that TRS teleconsultations in Brazil experienced an exponential increment of 802.% during a period of 6 days, after the first death due to COVID-19 was reported. However, the number of teleconsultations cases decreased over time, despite the number of reported COVID-19 cases continuously increasing. The results also showed that physicians in low-income municipalities, based on GDP per capita, are less likely to consult the telemedicine service despite facing higher rates of COVID-19 cases. The text analysis of call transcripts from medical teleconsultations showed that the main concern of physicians were "asymptomatic" patients. We suggest an immediate reinforcement of telehealth services in the regions of lower income as a strategy to support COVID-19 management., Competing Interests: The authors have declared that no competing interests exist.

This study aimed to elucidate the effects of selenium-loaded chitosan nanoparticles used as a dietary supplement on Nile tilapia (Oreochromis niloticus) antioxidant and growth responses. First, chitosan-based nanoparticles containing selenium (Se) were synthesized using the ionotropic gelation method and their physicochemical characteristics, controlled release profile, and antioxidant activity properties were investigated. Thereafter, the effects on glutathione peroxidase and antioxidant activities (by radical scavenging activity), growth, and whole-body composition of Nile tilapia were evaluated when they were fed with Se-loaded chitosan nanoparticles and compared with other selenium dietary supplements. Se-loaded chitosan nanoparticles showed high entrapment efficiency (87%), spherical shape, smooth surface, and broad size distribution. The controlled release of Se consisted of an initial burst followed by a gradual release over 48 h. Se-loaded nanoparticles presented significantly higher antioxidant activity compared to free Se. A 60-day feeding trial was conducted to compare the effects of supplementing different dietary Se sources, including selenomethionine (as organic source), sodium selenite (as inorganic source), and Se-loaded chitosan nanoparticles (Se-Nano and Se-Nano x1.5) on antioxidant and growth responses of Nile tilapia. A basal diet without Se supplementation was used as the control. The dietary supplementations with different Se sources (free and encapsulated selenium) lead to significant improvements in final weight and feed efficiency of Nile tilapia fingerlings. However, dietary treatments did not affect whole-body protein and lipid content. Diets containing Se-Nano and Se-Nano x1.5 were more effective than sodium selenite and selenomethionine in preventing oxidative stress and improving antioxidant activity in Nile tilapia. Overall, Se-loaded nanoparticles presented a great potential as an efficient source for delivering dietary Se to Nile tilapia, directly affecting the growth performance, feed efficiency, oxidative stress, and antioxidant activity of this species., Competing Interests: The authors have declared that no competing interests exist.

Objectives: The prevalence of excess body weight (EBW) has increased over the last decades in Brazil, where 55.4% of the adult population was overweight in 2019. EBW is a well-known risk factor for several types of cancer. We estimated the federal cost of EBW-related cancers in adults, considering the medical expenditures in the Brazilian Public Health System., Methods: We calculated the costs related to 11 types of cancer considering the procedures performed in 2018 by all organizations that provide cancer care in the public health system. We obtained data from the Hospital and Ambulatory Information Systems of the Brazilian Public Health System. We calculated the fractions of cancer attributable to EBW using the relative risks from the literature and prevalence from a nationally representative survey. We converted the monetary values in Reais (R$) to international dollars (Int$), considering the purchasing power parity (PPP) of 2018., Results: In Brazil, the 2018 federal cost for all types of cancers combined was Int$ 1.73 billion, of which nearly Int$ 710 million was spent on EBW-related cancer care and Int$ 30 million was attributable to EBW. Outpatient and inpatient expenditures reached Int$ 20.41 million (of which 80% was for chemotherapy) and Int$ 10.06 million (of which 82% was for surgery), respectively. Approximately 80% of EBW-attributable costs were due to breast, endometrial and colorectal cancers., Conclusion: A total of 1.76% of all federal cancer-related costs could be associated with EBW, representing a substantial economic burden for the public health system. We highlight the need for integrated policies for excess body weight control and cancer prevention., Competing Interests: Two authors, LRB and MQMR, received fee from consulting from Pan- American Health Organization (PAHO). This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Antigen cross presentation, whereby exogenous antigens are presented by MHC class I molecules to CD8+ T cells, is essential for generating adaptive immunity to pathogens and tumor cells. Following endocytosis, it is widely understood that protein antigens must be transferred from endosomes to the cytosol where they are subject to ubiquitination and proteasome degradation prior to being translocated into the endoplasmic reticulum (ER), or possibly endosomes, via the TAP1/TAP2 complex. Revealing how antigens egress from endocytic organelles (endosome-to-cytosol transfer, ECT), however, has proved vexing. Here, we used two independent screens to identify the hydrogen peroxide-transporting channel aquaporin-3 (AQP3) as a regulator of ECT. AQP3 overexpression increased ECT, whereas AQP3 knockout or knockdown decreased ECT. Mechanistically, AQP3 appears to be important for hydrogen peroxide entry into the endosomal lumen where it affects lipid peroxidation and subsequent antigen release. AQP3-mediated regulation of ECT was functionally significant, as AQP3 modulation had a direct impact on the efficiency of antigen cross presentation in vitro. Finally, AQP3-/- mice exhibited a reduced ability to mount an anti-viral response and cross present exogenous extended peptide. Together, these results indicate that the AQP3-mediated transport of hydrogen peroxide can regulate endosomal lipid peroxidation and suggest that compromised membrane integrity and coordinated release of endosomal cargo is a likely mechanism for ECT., Competing Interests: The authors SCN, RBS, MD, CC, MLA and IM are employed by Genentech. The author ABM is employed by CSL Ltd.. The author EST is employed by Boehringer Ingelheim. The commercial affiliation of the authors does not alter our adherence to PLOS ONE policies on sharing data and materials. There are no patents, products in development or marketed products to declare.

In 2015, UN member states committed to eliminate female genital mutilation (FGM) by 2030 as part of the Sustainable Development Agenda. To reach this goal, interventions need to be targeted and guided by the best available evidence. To date, however, estimates of the number of girls and women affected by FGM and their trends over time and geographic space have been limited by the availability, specificity and quality of population-level data. We present new estimates based on all publicly available nationally representative surveys collected since the 1990s that contain both information on FGM status and on the age at which FGM occurred. Using survival analysis, we generate estimates of FGM risk by single year of age for all countries with available data, and for rural and urban areas separately. The likelihood of experiencing FGM has decreased at the global level, but progress has been starkly uneven between countries. The available data indicate no progress in reducing FGM risk in Gambia, Guinea-Bissau, Mali and Guinea. In addition, rural and urban areas have diverged over the last two decades, with FGM declining more rapidly in urban areas. We describe limitations in the availability and quality of data on FGM occurrence and age-at-FGM. Based on current trends, the SDG goal of eliminating FGM by 2030 is out of reach, and the pace at which the practice is being abandoned would need to accelerate to eliminate FGM by 2030. The heterogeneity in trends between countries and rural vs urban areas offers an opportunity to contrast countries where FGM is in rapid decline and explore potential policy lessons and programmatic implications for countries where the practice of FGM appears to remain entrenched., Competing Interests: The authors have declared that no competing interests exist.

During alcohol consumption, the esophageal mucosa is directly exposed to high concentrations of ethanol (EtOH). We therefore investigated the response of normal human esophageal epithelial cell lines EPC1, EPC2 and EPC3 to acute EtOH exposure. While these cells were able to tolerate 2% EtOH for 8 h in both three-dimensional organoids and monolayer culture conditions, RNA sequencing suggested that EtOH induced mitochondrial dysfunction. With EtOH treatment, EPC1 and EPC2 cells also demonstrated decreased mitochondrial ATPB protein expression by immunofluorescence and swollen mitochondria lacking intact cristae by transmission electron microscopy. Mitochondrial membrane potential (ΔΨm) was decreased in a subset of EPC1 and EPC2 cells stained with ΔΨm-sensitive dye MitoTracker Deep Red. In EPC2, EtOH decreased ATP level while impairing mitochondrial respiration and electron transportation chain functions, as determined by ATP fluorometric assay, respirometry, and liquid chromatography-mass spectrometry. Additionally, EPC2 cells demonstrated enhanced oxidative stress by flow cytometry for mitochondrial superoxide (MitoSOX), which was antagonized by the mitochondria-specific antioxidant MitoCP. Concurrently, EPC1 and EPC2 cells underwent autophagy following EtOH exposure, as evidenced by flow cytometry for Cyto-ID, which detects autophagic vesicles, and immunoblots demonstrating induction of the lipidated and cleaved form of LC3B and downregulation of SQSTM1/p62. In EPC1 and EPC2, pharmacological inhibition of autophagy flux by chloroquine increased mitochondrial oxidative stress while decreasing cell viability. In EPC2, autophagy induction was coupled with phosphorylation of AMP activated protein kinase (AMPK), a cellular energy sensor responding to low ATP levels, and dephosphorylation of downstream substrates of mechanistic Target of Rapamycin Complex (mTORC)-1 signaling. Pharmacological AMPK activation by AICAR decreased EtOH-induced reduction of ΔΨm and ATP in EPC2. Taken together, acute EtOH exposure leads to mitochondrial dysfunction and oxidative stress in esophageal keratinocytes, where the AMPK-mTORC1 axis may serve as a regulatory mechanism to activate autophagy to provide cytoprotection against EtOH-induced cell injury., Competing Interests: The authors have declared that no competing interests exist.

Purpose: To determine whether teleophthalmology can help physicians in assessing and managing eye conditions and to ascertain which clinical conditions can be addressed by teleophthalmology in primary care setting., Methods: We evaluated the resolution capacity of TeleOftalmo, strategy implemented in the public health system of southern Brazil. Resolution capacity was defined as the ability to fully address patients' eye complaints in primary care with remote assistance from ophthalmologists. Data from tele-eye reports were collected over 14 months. Resolution capacity was compared across different age groups and different ocular conditions., Results: Overall, 8,142 patients had a tele-eye report issued in the study period. Resolution capacity was achieved in 5,748 (70.6%) patients. When stratified into age groups, the lowest capacity was 43.1% among subjects aged ≥65 years, while the highest was 89.7% among subjects aged 13-17 years (p<0.001). Refractive error (70.3%) and presbyopia (56.3%) were the most prevalent conditions followed by cataract (12.4%) and suspected glaucoma (7.6%). Resolution capacity was higher in cases of refractive error, presbyopia, spasm of accommodation and lid disorders than in patients diagnosed with other condition (p<0.001)., Conclusions: With telemedicine support, primary care physicians solved over two-thirds of patients' eye or vision complaints. Refractive errors had high case resolution rates, thus having a great impact on reducing the number of referrals to specialty care. Teleophthalmology adoption in primary-care settings as part of the workup of patients with eye or vision complaints promotes a more effective use of specialty centers and will hopefully reduce waiting times for specialty referral., Competing Interests: The authors have declared that no competing interests exist.

Monitoring of blood glucose is an invasive, painful and costly practice in diabetes. Consequently, the search for a more cost-effective (reagent-free), non-invasive and specific diabetes monitoring method is of great interest. Attenuated total reflectance Fourier transform infrared (ATR-FTIR) spectroscopy has been used in diagnosis of several diseases, however, applications in the monitoring of diabetic treatment are just beginning to emerge. Here, we used ATR-FTIR spectroscopy to evaluate saliva of non-diabetic (ND), diabetic (D) and insulin-treated diabetic (D+I) rats to identify potential salivary biomarkers related to glucose monitoring. The spectrum of saliva of ND, D and D+I rats displayed several unique vibrational modes and from these, two vibrational modes were pre-validated as potential diagnostic biomarkers by ROC curve analysis with significant correlation with glycemia. Compared to the ND and D+I rats, classification of D rats was achieved with a sensitivity of 100%, and an average specificity of 93.33% and 100% using bands 1452 cm-1 and 836 cm-1, respectively. Moreover, 1452 cm-1 and 836 cm-1 spectral bands proved to be robust spectral biomarkers and highly correlated with glycemia (R2 of 0.801 and 0.788, P < 0.01, respectively). Both PCA-LDA and HCA classifications achieved an accuracy of 95.2%. Spectral salivary biomarkers discovered using univariate and multivariate analysis may provide a novel robust alternative for diabetes monitoring using a non-invasive and green technology., Competing Interests: No competing interests.

Cyclo-Gly-Pro (CGP) attenuates nociception, however its effects on salivary glands remain unclear. In this study, we investigated the acute effects of CGP on salivary flow and composition, and on the submandibular gland composition, compared with morphine. Besides, we characterized the effects of naloxone (a non-selective opioid receptor antagonist) on CGP- and morphine-induced salivary and glandular alterations in mice. After that, in silico analyses were performed to predict the interaction between CGP and opioid receptors. Morphine and CGP significantly reduced salivary flow and total protein concentration of saliva and naloxone restored them to the physiological levels. Morphine and CGP also reduced several infrared vibrational modes (Amide I, 1687-1594cm-1; Amide II, 1594-1494cm-1; CH2/CH3, 1488-1433cm-1; C = O, 1432-1365cm-1; PO2 asymmetric, 1290-1185cm-1; PO2 symmetric, 1135-999cm-1) and naloxone reverted these alterations. The in silico docking analysis demonstrated the interaction of polar contacts between the CGP and opioid receptor Cys219 residue. Altogether, we showed that salivary hypofunction and glandular changes elicited by CGP may occur through opioid receptor suggesting that the blockage of opioid receptors in superior cervical and submandibular ganglions may be a possible strategy to restore salivary secretion while maintaining antinociceptive action due its effects on the central nervous system., Competing Interests: The authors have declared that no competing interests exist.

Background: Kidney transplantation is the treatment of choice in patient with end stage chronic kidney disease, offering the best long term survival and greater Quality of Life in this group of patients. Graft volume was correlated with improved renal function in living donor transplantations. The primary aim of this study was to correlate renal volume adjusted to body surface area with renal function one year (estimated glomerular filtration rate; eGFR) after kidney transplantation., Methods: This single-center, prospective cohort study included 256 patients who underwent kidney transplantation from January 2011 through December 2015 at Hospital das Clínicas de Botucatu-UNESP. We evaluated three kidney measurements during the bench surgery; the final graft volume was calculated using the ellipsoid formula and adjusted to body surface area., Results: In the living donors there was positive correlation between adjusted graft volume and eGFR (r = 0.311, p = 0.008). Multivariate analysis revealed that low rejection rate and increased adjusted graft volume were independent factors correlated with eGFR. In deceased donors, there was no correlation between adjusted kidney volume and eGFR (r = 0.08, p = 0.279) in univariate analysis, but a multivariate analysis indicated that lower kidney donor profile index (KDPI), absence of rejection and high adjusted kidney volume were independent factors for better eGFR., Conclusion: Adjusted kidney volume was positively correlated with a satisfactory eGFR at one year after living donor and deceased donor transplantations., Competing Interests: The authors have declared that no competing interests exist.

Respiratory infection can be exacerbated by the high glucose concentration in the airway surface liquid (ASL). We investigated the effects of salbutamol and phlorizin on the pulmonary function, oxidative stress levels and SGLT1 activity in lung, pulmonary histopathological damages and survival rates of rats with sepsis. Sepsis was induced by cecal ligation and puncture surgery (CLP). Twenty-four hours after surgery, CLP rats were intranasally treated with saline, salbutamol or phlorizin. After 2 hours, animals were anesthetized and sacrificed. Sepsis promoted atelectasis and bronchial inflammation, and led to increased expression of SGLT1 on cytoplasm of pneumocytes. Salbutamol treatment reduced bronchial inflammation and promoted hyperinsuflation in CLP rats. The interferon-ɤ and Interleucin-1β concentrations in bronchoalveolar lavage (BAL) were closely related to the bronchial inflammation regulation. Salbutamol stimulated SGLT1 in plasma membrane; whereas, phlorizin promoted the increase of SGLT1 in cytoplasm. Phlorizin reduced catalase activity and induced a significant decrease in the survival rate of CLP rats. Taken together, sepsis promoted atelectasis and lung inflammation, which can be associated with SGLT1 inhibition. The loss of function of SGLT1 by phlorizin are related to the augmented disease severity, increased atelectasis, bronchial inflammation and a significant reduction of survival rate of CLP rats. Alternatively salbutamol reduced BAL inflammatory cytokines, bronchial inflammation, atelectasis, and airway damage in sepsis. These data suggest that this selective β2-adrenergic agonist may protect lung of septic acute effects., Competing Interests: The authors have declared that no competing interests exist.

Coffee farms have been adopting the microterraces system, a technique that reduces the effect of the slope by moving the soil between the crop lines. In this way, all the mechanized operations can be carried out normally, except for harvesting, due to the work limitation of the harvesters, who work in areas with a maximum slope of 20%. One option is to use unilateral harvesters, which crop one side at a time; however, there has been no research on these microterrace machines to evaluate their performance and to compare it with those of the other harvesting methods in those regions. This study aimed to compare the mechanized harvest performance in the microterraces with the manual and semimechanized harvesting methods. The study was carried out in an agricultural area of the municipality of Ouro Fino / MG, Brazil, in a crop production site where the microterraces were built six years before the experiment. The treatments were assigned to a split-block design with seven repetitions and consisted of mechanized harvest-unilateral harvester with bag storage; manual harvest-regionally experienced workers; and semimechanized harvest-with portable breakers. Through an analysis of the times and movements, the operational efficiency and operational and effective field capabilities were measured. The adoption of microterraces allows the efficient mechanization of areas previously impossible to mechanize. The unilateral harvester is a potential tool for the partial replacement of manual labor in the harvest, performing a service equivalent to that of 23.68 manual workers and 10.55 manual workers in the semimechanized system., Competing Interests: The authors have declared that no competing interests exist.

Throughout evolutionary history, humans have been exposed to a wide variety of diseases, some of which have serious and even lethal consequences. Memorizing medicinal plants for the treatment of serious diseases likely maximized the chances of survival and reproduction and was instrumental in the evolutionary success of our species. In the present study, we used the idea of adaptive memory to understand whether human memory evolved to recall information about medicinal plants for the treatment of serious diseases. We considered plant-disease pairs of words as units of information available in a medical system based on the use of medicinal plants. The pairs included in the categories of chronic infectious diseases and transmissible infectious diseases were considered to be of higher adaptive value, whereas those included in the category of common conditions were considered to be of lower adaptive value. Pairs grouped into the category of emerging and reemerging diseases were employed to investigate conformity bias; pairs belonging to the category esthetic uses were considered to be of little adaptive relevance and utilized as an experimental control. Our results revealed that plant-disease pairs associated with the category of common conditions, considered by us to be of lower severity and less adaptive relevance for humans, were better remembered and retained in the participants' memory. We believe that prior experience with common conditions and the frequency of these conditions in the population may have intensified the ability to remember the plant-disease pairs associated with this group of diseases., Competing Interests: The authors have declared that no competing interests exist.

The galling habit represents a complex type of interaction between insects and plants, ranging from antagonism to mutualism. The obligate pollination mutualism between Ficus and fig wasps relies strongly on the induction of galls in Ficus flowers, where wasps' offspring develop. Even though gall induction plays an important role in many insect-plant interactions, the mechanisms that trigger gall formation are still not completely known. Using a fingerprinting approach, we show here that venom protein profiles from galling fig wasps differ from the venom profiles of non-galling species, suggesting the secretion plays different roles according to the type of interaction it is involved in. Each studied cleptoparasitic species had a distinct venom profile, suggesting that cleptoparasitism in fig wasps covers a vast diversity of molecular interactions. Fig wasp venoms are mainly composed of peptides. No low molecular weight compounds were detected by UPLC-DAD-MS, suggesting that such compounds (e.g., IAA and cytokinines) are not involved in gall induction. The differences in venom composition observed between galling and non-galling fig wasp species bring new perspectives to the study of gall induction processes and the role of insect secretions., Competing Interests: The authors have declared that no competing interests exist.

Microorganisms developing in the liner of the spent fuel pool (SFP) and the fuel transfer channel (FTC) of a Nuclear Power Plant (NPP) can form high radiation resistant biofilms and cause corrosion. Due to difficulties and limitations to obtain large samples from SFP and FTC, cotton swabs were used to collect the biofilm from the wall of these installations. Molecular characterization was performed using massively parallel sequencing to obtain a taxonomic and functional gene classification. Also, samples from the drainage system were evaluated because microorganisms may travel over the 12-meter column of the pool water of the Brazilian Nuclear Power Plant (Angra1), which has been functioning since 1985. Regardless of the treatment of the pool water, our data reveal the unexpected presence of Fungi (Basidiomycota and Ascomycota) as the main contaminators of the SFP and FTC. Ustilaginomycetes (Basidiomycota) was the major class contributor (70%) in the SFP and FTC reflecting the little diversity in these sites; nevertheless, Proteobacteria, Actinobacteria, Firmicutes (Bacilli) were present in small proportions. Mapping total reads against six fungal reference genomes indicate that there is, in fact, a high abundance of fungal sequences in samples collected from SFP and FTC. Analysis of the ribosomal internal transcribed spacer (ITS) 1 and 2 regions and the protein found in the mitochondria of eukaryotic cells, cytochrome b (cytb) grouped our sample fungi in the clade 7 as Ustilago and Pseudozyma. In contrast, in the drainage system, Alphaproteobacteria were present in high abundances (55%). The presence of Sphingopyxis, Mesorhizobium, Erythrobacter, Sphingomonas, Novosphingobium, Sphingobium, Chelativorans, Oceanicaulis, Acidovorax, and Cyanobacteria was observed. Based on genomic annotation data, the assessment of the biological function found a higher proportion of protein-coding sequences related to respiration and protein metabolism in SFP and FTC samples. The knowledge of this biological inventory present in the system may contribute to further studies of potential microorganisms that might be useful for bioremediation of nuclear waste., Competing Interests: The author has declared that no competing interests exist.

Background: Given the relative importance of cognitive impairment, there was considerable interest in identifying the cognitive profile of PD patients, in order to ensure specific and appropriate therapeutic interventions., Purpose: To determine the effects of physical exercise programs on cognitive function in PD patients, compared with the control group., Data Sources: Medline, Cochrane, Scopus, PEDro and Web of Science (last searched in September 2016)., Study Selection: Randomized clinical trials examining the effects of physical exercise programs and cognitive function in PD patients. Nine studies fulfilled the selection criteria and were included in this review., Data Extraction: Characteristics of the publication, characteristics of the participants, test used for cognitive screening, cognitive domain assessed, tools used to assess cognitive function, characteristics of the experimental intervention, characteristics of the control group, mean results and standard deviation of function cognitive. The PEDro score was used to evaluate methodological quality., Data Synthesis: Most eligible studies showed good methodological quality based on the PEDro scale. Studies have shown that adapted tango for PD patients, cognitive training combined with motor training, and treadmill training promote the preservation or improvement of cognitive function in PD patients., Limitations: The diversity of cognitive tests used to assess cognitive function and the high heterogeneity identified between the physical exercise programs., Conclusions: Physical exercise programs promote positive and significant effects on global cognitive function, processing speed, sustained attention and mental flexibility in PD patients, at a mild to moderate stage for patients with a 6-year clinical diagnosis of PD. However, treadmill training performed 3 times a week for about 60 minutes and for a period of 24 weeks produced larger improvements in cognition.

Theta-burst stimulation (TBS) can be a non-invasive technique to modulate cognitive functions, with promising therapeutic potential, but with some contradictory results. Event related potentials are used as a marker of brain deterioration and can be used to evaluate TBS-related cognitive performance, but its use remains scant. This study aimed to study bilateral inhibitory and excitatory TBS effects upon neurocognitive performance of young healthy volunteers, using the auditory P300' results. Using a double-blind sham-controlled study, 51 healthy volunteers were randomly assigned to five different groups, two submitted to either excitatory (iTBS) or inhibitory (cTBS) stimulation over the left dorsolateral pre-frontal cortex (DLPFC), two other actively stimulated the right DLPFC and finally a sham stimulation group. An oddball based auditory P300 was performed just before a single session of iTBS, cTBS or sham stimulation and repeated immediately after. P300 mean latency comparison between the pre- and post-TBS stimulation stages revealed significantly faster post stimulation latencies only when iTBS was performed on the left hemisphere (p = 0.003). Right and left hemisphere cTBS significantly delayed P300 latency (right p = 0.026; left p = 0.000). Multiple comparisons for N200 showed slower latencies after iTBS over the right hemisphere. No significant difference was found in amplitude variation. TBS appears to effectively influence neural networking involved in P300 formation, but effects seem distinct for iTBS vs cTBS and for the right or the left hemisphere. P300 evoked potentials can be an effective and practical tool to evaluate transcranial magnetic stimulation related outcomes.

In trypanosomatids, the kinetoplast is the portion of the single mitochondrion that is connected to the basal body and contains the kDNA, a network composed by circular and interlocked DNA. The kDNA packing is conducted by Kinetoplast Associated Proteins (KAPs), which are similar to eukaryotic histone H1. In symbiont-harboring trypanosomatids (SHTs) such as Angomonas deanei and Strigomonas culicis, a ß-proteobacterium co-evolves with the host in a mutualistic relationship. The prokaryote confers nutritional benefits to the host and affects its cell structure. Atomic force microscopy showed that the topology of isolated kDNA networks is quite similar in the two SHT species. Ultrastructural analysis using high-resolution microscopy techniques revealed that the DNA fibrils are more compact in the kinetoplast region that faces the basal body and that the presence of the symbiotic bacterium does not interfere with kDNA topology. However, RT-PCR data revealed differences in the expression of KAPs in wild-type protozoa as compared to aposymbiotic cells. Immunolocalization showed that different KAPs present distinct distributions that are coincident in symbiont-bearing and in symbiont-free cells. Although KAP4 and KAP7 are shared by all trypanosomatid species, the expanded repertoire of KAPs in SHTs can be used as phylogenetic markers to distinguish different genera.

Global Positioning System (GPS) tags are nowadays widely used in wildlife tracking. This geolocation technique can suffer from fix loss biases due to poor satellite GPS geometry, that result in tracking data gaps leading to wrong research conclusions. In addition, new solar-powered GPS tags deployed on birds can suffer from a new "battery drain bias" currently ignored in movement ecology analyses. We use a GPS tracking dataset of bearded vultures (Gypaetus barbatus), tracked for several years with solar GPS tags, to evaluate the causes and triggers of fix and data retrieval loss biases. We compare two models of solar GPS tags using different data retrieval systems (Argos vs GSM-GPRS), and programmed with different duty cycles. Neither of the models was able to accomplish the duty cycle programed initially. Fix and data retrieval loss rates were always greater than expected, and showed non-random gaps in GPS locations. Number of fixes per month of tracking was a bad criterion to identify tags with smaller biases. Fix-loss rates were four times higher due to battery drain than due to poor GPS satellite geometry. Both tag models were biased due to the uneven solar energy available for the recharge of the tag throughout the annual cycle, resulting in greater fix-loss rates in winter compared to summer. In addition, we suggest that the bias found along the diurnal cycle is linked to a complex three-factor interaction of bird flight behavior, topography and fix interval. More fixes were lost when vultures were perching compared to flying, in rugged versus flat topography. But long fix-intervals caused greater loss of fixes in dynamic (flying) versus static situations (perching). To conclude, we emphasize the importance of evaluating fix-loss bias in current tracking projects, and deploying GPS tags that allow remote duty cycle updates so that the most appropriate fix and data retrieval intervals can be selected.

Aedes aegypti is one of the species most favored by changes in the environment caused by urbanization. Its abundance increases rapidly in the face of such changes, increasing the risk of disease transmission. Previous studies have shown that mosquito species that have adapted to anthropogenic environmental changes benefit from urbanization and undergo population expansion. In light of this, we used microsatellite markers to explore how urbanization processes may be modulating Ae. aegypti populations collected from three areas with different levels of urbanization in the city of São Paulo, Brazil. Specimens were collected at eleven sites in three areas with different degrees of urbanization in the city of São Paulo: conserved, intermediate and urbanized. Ten microsatellite loci were used to characterize the populations from these areas genetically. Our findings suggest that as urbanized areas grow and the human population density in these areas increases, Ae. aegypti populations undergo a major population expansion, which can probably be attributed to the species' adaptability to anthropogenic environmental changes. Our findings reveal a robust association between, on the one hand, urbanization processes and densification of the human population and, on the other, Ae. aegypti population structure patterns and population expansion. This indicates that this species benefits from anthropogenic effects, which are intensified by migration of the human population from rural to urban areas, increasing the risk of epidemics and disease transmission to an ever-increasing number of people.

Purpose: To differentiate the plasma metabolomic profile of patients with age related macular degeneration (AMD) from that of controls, by Nuclear Magnetic Resonance (NMR) spectroscopy., Methods: Two cohorts (total of 396 subjects) representative of central Portugal and Boston, USA phenotypes were studied. For each cohort, subjects were grouped according to AMD stage (early, intermediate and late). Multivariate analysis of plasma NMR spectra was performed, followed by signal integration and univariate analysis., Results: Small changes were detected in the levels of some amino acids, organic acids, dimethyl sulfone and specific lipid moieties, thus providing some biochemical information on the disease. The possible confounding effects of gender, smoking history and age were assessed in each cohort and found to be minimal when compared to that of the disease. A similar observation was noted in relation to age-related comorbidities. Furthermore, partially distinct putative AMD metabolite fingerprints were noted for the two cohorts studied, reflecting the importance of nutritional and other lifestyle habits in determining AMD metabolic response and potential biomarker fingerprints. Notably, some of the metabolite changes detected were noted as potentially differentiating controls from patients diagnosed with early AMD., Conclusion: For the first time, this study showed metabolite changes in the plasma of patients with AMD as compared to controls, using NMR. Geographical origins were seen to affect AMD patients´ metabolic profile and some metabolites were found to be valuable in potentially differentiating controls from early stage AMD patients. Metabolomics has the potential of identifying biomarkers for AMD, and further work in this area is warranted.

The purpose of this study was to identify genomic regions associated with carcass traits in an experimental Nelore cattle population. The studied data set contained 2,306 ultrasound records for longissimus muscle area (LMA), 1,832 for backfat thickness (BF), and 1,830 for rump fat thickness (RF). A high-density SNP panel (BovineHD BeadChip assay 700k, Illumina Inc., San Diego, CA) was used for genotyping. After genomic data quality control, 437,197 SNPs from 761 animals were available, of which 721 had phenotypes for LMA, 669 for BF, and 718 for RF. The SNP solutions were estimated using a single-step genomic BLUP approach (ssGWAS), which calculated the variance for windows of 50 consecutive SNPs and the regions that accounted for more than 0.5% of the additive genetic variance were used to search for candidate genes. The results indicated that 12, 18, and 15 different windows were associated to LMA, BF, and RF, respectively. Confirming the polygenic nature of the studied traits, 43, 65, and 53 genes were found in those associated windows, respectively for LMA, BF, and RF. Among the candidate genes, some of them, which already had their functions associated with the expression of energy metabolism, were found associated with fat deposition in this study. In addition, ALKBH3 and HSD17B12 genes, which are related in fibroblast death and metabolism of steroids, were found associated with LMA. The results presented here should help to better understand the genetic and physiologic mechanism regulating the muscle tissue deposition and subcutaneous fat cover expression of Zebu animals. The identification of candidate genes should contribute for Zebu breeding programs in order to consider carcass traits as selection criteria in their genetic evaluation., Competing Interests: The authors have declared that no competing interests exist.

Combined physiological and behavioural responses to salt loads during development have rarely been studied in air-breathing vertebrates able to inhabit hypersaline habitats, but they may be of particular importance in understanding, for example, the differences among species in patterns of habitat use or ontogenetic diet switches. Here, we compared the physiological and behavioural responses of self-feeding precocial chicks developed in contrasting levels of water salinity. The model species was the Black-winged Stilt (Himantopus himantopus) a precocial shorebird that breeds in a range of habitats from freshwater to hypersaline wetlands. Specifically, we compared resting metabolic rate (RMR), heat shock proteins (Hsp70), plasma ions, hematocrit, body mass, body size, growth rate and head-shaking behaviour of captive-reared Black-winged Stilt fledglings developed under fresh (0 ‰), saline (20 ‰), and hypersaline (60 ‰) water. Contrary to expectations, none of the physiological and morphological variables measured differed significantly among treatments. Likewise, the RMR of wild and captive-reared fledglings was similar. Surprisingly, the saltgland mass of wild fledglings from freshwater and those from hypersaline habitats was also similar. However, head-shaking, a behavioural response associated to minimize salt intake and to expel the secretions of salt glands, differed according to salinity source: head-shaking rate increased with increasing salinity. The results of this study support the key role of behavioural osmoregulation in avoiding salt stress during development., Competing Interests: The authors have declared that no competing interests exist.

Habitat structure plays an important mediating role in predator-prey interactions. However the effects are strongly dependent on regional predator pools, which can drive predation risk in habitats with very similar structure in opposite directions. In the Mediterranean Sea predation on juvenile sea urchins is commonly known to be regulated by seagrass structure. In this study we test whether the possibility for juvenile Paracentrotus lividus to be predated changes in relation to the fragmentation of the seagrass Posidonia oceanica (four habitat classes: continuous, low-fragmentation, high-fragmentation and rocks), and to the spatial arrangement of such habitat classes at a landscape scale. Sea urchin predation risk was measured in a 20-day field experiment on tethered individuals placed in three square areas 35×35 m2 in size. Variability of both landscape and habitat structural attributes was assessed at the sampling grain 5×5 m2. Predation risk changed among landscapes, as it was lower where more 'rocks', and thus less seagrass, were present. The higher risk was found in the 'continuous' P. oceanica rather than in the low-fragmentation, high-fragmentation and rock habitats (p-values = 0.0149, 0.00008, and 0.0001, respectively). Therefore, the expectation that juvenile P. lividus survival would have been higher in the 'continuous' seagrass habitat, which would have served as shelter from high fish predation pressure, was not met. Predation risk changed across habitats due to different success between attack types: benthic attacks (mostly from whelks) were overall much more effective than those due to fish activity, the former type being associated with the 'continuous' seagrass habitat. Fish predation on juvenile sea urchins on rocks and 'high-fragmentation' habitat was less likely than benthic predation in the 'continuous' seagrass, with the low seagrass patch complexity increasing benthic activity. Future research should be aimed at investigating, derived from the complex indirect interactions among species, how top-down control in marine reserves can modify seagrass habitat effects., Competing Interests: The authors have declared that no competing interests exist.

The Caribbean snapper Lutjanus purpureus is a marine species fish commonly found associated with rocky seabeds and is widely distributed along of Western Atlantic. Data on stock delineation and stock recognition are essential for establishing conservation measures for commercially fished species. However, few studies have investigated the population genetic structure of this economically valuable species, and previous studies (based on only a portion of the mitochondrial DNA) provide an incomplete picture. The present study used a multi-locus approach (12 segments of mitochondrial and nuclear DNA) to elucidate the levels of genetic diversity and genetic connectivity of L. purpureus populations and their demographic history. L. purpureus has high levels of genetic diversity, which probably implies in high effective population sizes values for the species. The data show that this species is genetically homogeneous throughout the geographic region analyzed, most likely as a result of dispersal during larval phase. Regarding demographic history, a historical population growth event occurred, likely due to sea level changes during the Pleistocene., Competing Interests: The authors have declared that no competing interests exist.

Mosquitoes are responsible for the transmission of important infectious diseases, causing millions of deaths every year and endangering approximately 3 billion people around the world. As such, precise identification of mosquito species is crucial for an understanding of epidemiological patterns of disease transmission. Currently, the most common method of mosquito identification relies on morphological taxonomic keys, which do not always distinguish cryptic species. However, wing geometric morphometrics is a promising tool for the identification of vector mosquitoes, sibling and cryptic species included. This study therefore sought to accurately identify mosquito species from the three most epidemiologically important mosquito genera using wing morphometrics. Twelve mosquito species from three epidemiologically important genera (Aedes, Anopheles and Culex) were collected and identified by taxonomic keys. Next, the right wing of each adult female mosquito was removed and photographed, and the coordinates of eighteen digitized landmarks at the intersections of wing veins were collected. The allometric influence was assessed, and canonical variate analysis and thin-plate splines were used for species identification. Cross-validated reclassification tests were performed for each individual, and a Neighbor Joining tree was constructed to illustrate species segregation patterns. The analyses were carried out and the graphs plotted with TpsUtil 1.29, TpsRelw 1.39, MorphoJ 1.02 and Past 2.17c. Canonical variate analysis for Aedes, Anopheles and Culex genera showed three clear clusters in morphospace, correctly distinguishing the three mosquito genera, and pairwise cross-validated reclassification resulted in at least 99% accuracy; subgenera were also identified correctly with a mean accuracy of 96%, and in 88 of the 132 possible comparisons, species were identified with 100% accuracy after the data was subjected to reclassification. Our results showed that Aedes, Culex and Anopheles were correctly distinguished by wing shape. For the lower hierarchical levels (subgenera and species), wing geometric morphometrics was also efficient, resulting in high reclassification scores., Competing Interests: The authors have declared that no competing interests exist.

Objectives: To assess the prospective associations of physical activity behaviors and screen time with early sexual intercourse initiation (i.e., before 15 years) in a large sample of adolescents., Methods: We used two waves of data from the Rotterdam Youth Monitor, a longitudinal study conducted in the Netherlands. The analysis sample consisted of 2,141 adolescents aged 12 to 14 years (mean age at baseline = 12.2 years, SD = 0.43). Physical activity (e.g., sports outside school), screen time (e.g., computer use), and early sexual intercourse initiation were assessed by means of self-report questionnaires. Logistic regression models were tested to assess the associations of physical activity behaviors and screen time (separately and simultaneously) with early sexual intercourse initiation, controlling for confounders (i.e., socio-demographics and substance use). Interaction effects with gender were tested to assess whether these associations differed significantly between boys and girls., Results: The only physical activity behavior that was a significant predictor of early sexual intercourse initiation was sports club membership. Adolescent boys and girls who were members of a sports club) were more likely to have had early sex (OR = 2.17; 95% CI = 1.33, 3.56. Significant gender interaction effects indicated that boys who watched TV ≥2 hours/day (OR = 2.00; 95% CI = 1.08, 3.68) and girls who used the computer ≥2 hours/day (OR = 3.92; 95% CI = 1.76, 8.69) were also significantly more likely to have engaged in early sex., Conclusion: These findings have implications for professionals in general pediatric healthcare, sexual health educators, policy makers, and parents, who should be aware of these possible prospective links between sports club membership, TV watching (for boys), and computer use (for girls), and early sexual intercourse initiation. However, continued research on determinants of adolescents' early sexual initiation is needed to further contribute to the strategies for improving adolescents' healthy sexual development and behaviors.

Background Context: Low back pain (LBP) is one of the leading causes of disability worldwide and among the most common reasons for seeking primary sector care. Chiropractors, physical therapists and general practitioners are among those providers that treat LBP patients, but there is only limited evidence regarding the effectiveness and economic evaluation of care offered by these provider groups., Purpose: To estimate the clinical effectiveness and to systematically review the literature of full economic evaluation of chiropractic care compared to other commonly used care approaches among adult patients with non-specific LBP., Study Design: Systematic reviews of interventions and economic evaluations., Methods: A comprehensive search strategy was conducted to identify 1) pragmatic randomized controlled trials (RCTs) and/or 2) full economic evaluations of chiropractic care for low back pain compared to standard care delivered by other healthcare providers. Studies published between 1990 and 4th June 2015 were considered. Primary outcomes included pain, functional status and global improvement. Study selection, critical quality appraisal and data extraction were conducted by two independent reviewers. Data from RCTs with low risk of bias were included in a meta-analysis to determine effect estimates. Cost estimates of full economic evaluations were converted to 2015 USD and results summarized using Slavin's qualitative best-evidence synthesis., Results: Six RCTs and three full economic evaluations were scientifically admissible. Five RCTs with low risk of bias compared chiropractic care to exercise therapy (n = 1), physical therapy (n = 3) and medical care (n = 1). Overall, we found similar effects for chiropractic care and the other types of care and no reports of serious adverse events. Three low to high quality full economic evaluations studies (one cost-effectiveness, one cost-minimization and one cost-benefit) compared chiropractic to medical care. Given the divergent conclusions (favours chiropractic, favours medical care, equivalent options), mixed-evidence was found for economic evaluations of chiropractic care compared to medical care., Conclusion: Moderate evidence suggests that chiropractic care for LBP appears to be equally effective as physical therapy. Limited evidence suggests the same conclusion when chiropractic care is compared to exercise therapy and medical care although no firm conclusion can be reached at this time. No serious adverse events were reported for any type of care. Our review was also unable to clarify whether chiropractic or medical care is more cost-effective. Given the limited available evidence, the decision to seek or to refer patients for chiropractic care should be based on patient preference and values. Future studies are likely to have an important impact on our estimates as these were based on only a few admissible studies.

Background: Malawi ratified a compulsory birth and death registration system in 2009. Until it captures complete coverage of vital events, Malawi relies on other data sources to calculate mortality estimates. We tested a community-based method to estimate annual under-five mortality rates (U5MR) through the Real-Time Monitoring of Under-Five Mortality (RMM) project in Malawi. We implemented RMM in two phases, and conducted an independent evaluation of phase one after 21 months of implementation. We present results of the phase two validation that covers the full project time span, and compare the results to those of the phase one validation., Methods and Findings: We assessed the completeness of the counts of births and deaths and the accuracy of disaggregated U5MR from the community-based method against a retrospective full pregnancy history for rolling twelve-month periods after the independent evaluation. We used full pregnancy histories collected through household interviews carried out between November 2013 and January 2014 as the validation data source. Health Surveillance Agents (HSAs) across the 160 catchment areas submitted routine reports on pregnancies, births, and deaths consistently. However, for the 15-month implementation period post-evaluation, average completeness of birth event reporting was 76%, whereas average completeness of death event reporting was 67% relative to that expected from a comparable pregnancy history. HSAs underestimated the U5MR by an average of 21% relative to that estimated from a comparable pregnancy history., Conclusions: On a medium scale, the community-based RMM method in Malawi produced substantial underestimates of annualized U5MR relative to those obtained from a full pregnancy history, despite the additional incentives and quality-control activities. We were not able to achieve an optimum level of incentive and support to make the system work while ensuring sustainability. Lessons learned from the implementation of RMM can inform programs supporting community-based interventions through HSAs in Malawi.

Introduction: Most low-income countries lack complete and accurate vital registration systems. As a result, measures of under-five mortality rates rely mostly on household surveys. In collaboration with partners in Ethiopia, Ghana, Malawi, and Mali, we assessed the completeness and accuracy of reporting of births and deaths by community-based health workers, and the accuracy of annualized under-five mortality rate estimates derived from these data. Here we report on results from Ethiopia, Malawi and Mali., Method: In all three countries, community health workers (CHWs) were trained, equipped and supported to report pregnancies, births and deaths within defined geographic areas over a period of at least fifteen months. In-country institutions collected these data every month. At each study site, we administered a full birth history (FBH) or full pregnancy history (FPH), to women of reproductive age via a census of households in Mali and via household surveys in Ethiopia and Malawi. Using these FBHs/FPHs as a validation data source, we assessed the completeness of the counts of births and deaths and the accuracy of under-five, infant, and neonatal mortality rates from the community-based method against the retrospective FBH/FPH for rolling twelve-month periods. For each method we calculated total cost, average annual cost per 1,000 population, and average cost per vital event reported., Results: On average, CHWs submitted monthly vital event reports for over 95 percent of catchment areas in Ethiopia and Malawi, and for 100 percent of catchment areas in Mali. The completeness of vital events reporting by CHWs varied: we estimated that 30%-90% of annualized expected births (i.e. the number of births estimated using a FPH) were documented by CHWs and 22%-91% of annualized expected under-five deaths were documented by CHWs. Resulting annualized under-five mortality rates based on the CHW vital events reporting were, on average, under-estimated by 28% in Ethiopia, 32% in Malawi, and 9% in Mali relative to comparable FPHs. Costs per vital event reported ranged from $21 in Malawi to $149 in Mali., Discussion: Our findings in Mali suggest that CHWs can collect complete and high-quality vital events data useful for monitoring annual changes in under-five mortality rates. Both the supervision of CHWs in Mali and the rigor of the associated field-based data quality checks were of a high standard, and the size of the pilot area in Mali was small (comprising of approximately 53,205 residents in 4,200 households). Hence, there are remaining questions about whether this level of vital events reporting completeness and data quality could be maintained if the approach was implemented at scale. Our experience in Malawi and Ethiopia suggests that, in some settings, establishing and maintaining the completeness and quality of vital events reporting by CHWs over time is challenging. In this sense, our evaluation in Mali falls closer to that of an efficacy study, whereas our evaluations in Ethiopia and Malawi are more akin to an effectiveness study. Our overall findings suggest that no one-size-fits-all approach will be successful in guaranteeing complete and accurate reporting of vital events by CHWs.

Background: While community health workers are being recognized as an integral work force with growing responsibilities, increased demands can potentially affect motivation and performance. The ubiquity of mobile phones, even in hard-to-reach communities, has facilitated the pursuit of novel approaches to support community health workers beyond traditional modes of supervision, job aids, in-service training, and material compensation. We tested whether supportive short message services (SMS) could improve reporting of pregnancies and pregnancy outcomes among community health workers (Health Surveillance Assistants, or HSAs) in Malawi., Methods and Findings: We designed a set of one-way SMS that were sent to HSAs on a regular basis during a 12-month period. We tested the effectiveness of the cluster-randomized intervention in improving the complete documentation of a pregnancy. We defined complete documentation as a pregnancy for which a specific outcome was recorded. HSAs in the treatment group received motivational and data quality SMS. HSAs in the control group received only motivational SMS. During baseline and intervention periods, we matched reported pregnancies to reported outcomes to determine if reporting of matched pregnancies differed between groups and by period. The trial is registered as ISCTRN24785657., Conclusions: Study results show that the mHealth intervention improved the documentation of matched pregnancies in both the treatment (OR 1.31, 95% CI: 1.10-1.55, p<0.01) and control (OR 1.46, 95% CI: 1.11-1.91, p = 0.01) groups relative to the baseline period, despite differences in SMS content between groups. The results should be interpreted with caution given that the study was underpowered. We did not find a statistically significant difference in matched pregnancy documentation between groups during the intervention period (OR 0.94, 95% CI: 0.63-1.38, p = 0.74). mHealth applications have the potential to improve the tracking and data quality of pregnancies and pregnancy outcomes, particularly in low-resource settings.

Background: Ethiopia has scaled up its community-based programs over the past decade by training and deploying health extension workers (HEWs) in rural communities throughout the country. Consequently, child mortality has declined substantially, placing Ethiopia among the few countries that have achieved the United Nations' fourth Millennium Development Goal. As Ethiopia continues its efforts, results must be assessed regularly to provide timely feedback for improvement and to generate further support for programs. More specifically the expansion of HEWs at the community level provides a unique opportunity to build a system for real-time monitoring of births and deaths, linked to a civil registration and vital statistics system that Ethiopia is also developing. We tested the accuracy and completeness of births and deaths reported by trained HEWs for monitoring child mortality over 15 -month periods., Methods and Findings: HEWs were trained in 93 randomly selected rural kebeles in Jimma and West Hararghe zones of the Oromia region to report births and deaths over a 15-month period from January, 2012 to March, 2013. Completeness of number of births and deaths, age distribution of deaths, and accuracy of resulting under-five, infant, and neonatal mortality rates were assessed against data from a large household survey with full birth history from women aged 15-49. Although, in general HEWs, were able to accurately report events that they identified, the completeness of number of births and deaths reported over twelve-month periods was very low and variable across the two zones. Compared to household survey estimates, HEWs reported only about 30% of births and 21% of under-five deaths occurring in their communities over a twelve-month period. The under-five mortality rate was under-estimated by around 30%, infant mortality rate by 23% and neonatal mortality by 17%. HEWs reported disproportionately higher number of deaths among the very young infants than among the older children., Conclusion: Birth and death data reported by HEWs are not complete enough to support the monitoring of changes in childhood mortality. HEWs can significantly contribute to the success of a CRVS in Ethiopia, but cannot be relied upon as the sole source for identification of vital events. Further studies are needed to understand how to increase the level of completeness.