Supplementation with live Saccharomyces cerevisiae boulardii during the initial 42 days of the feedlot phase in Nellore beef cattle.

This study aimed to assess the effect of Saccharomyces cerevisiae boulardii CNCM I-1079 supplementation during the initial feeding period on the performance of Nellore bulls in a feedlot system. One hundred ninety-eight Nellore bulls were used in a completely randomized block design, with blocking based on weight within each treatment group: light (331.4 kg; 4 pens), medium (349.7 kg; 4 pens), and heavy (362.5 kg; 3 pens). The treatments included CON-a basal diet, and SCB-basal diet plus a probiotic (Saccharomyces cerevisiae boulardii CNCM I-1079; 1.0 × 1010 CFU/head/d). Experimental diets were administered for the first 42 d (21 d in the step-up phase and 21 d in the finishing diet -870 g concentrate/kg dry matter [DM]). Subsequently, both treatment groups were transitioned to the same basal diet for an additional 76 d, completing 118 d on feed. Linear regression analysis was conducted for dry matter intake (DMI) data. During the initial 42 d, DMI tended to be higher for SCB (P = 0.09); also bulls fed SCB reached the plateau of the curve at 9.17 kg DMI/d earlier (39 d, R2 = 0.97) than those fed CON (43 d; R2 = 0.96) diets. For the first 42 d, the SCB treatment exhibited higher final weight (393.0 vs. 401.4 kg, P = 0.02), total gain (49.3 vs. 53.5 kg, P = 0.02), daily weight gain (1.124 vs. 1.274 kg, P = 0.02), and G:F (0.174 vs. 0.188, P = 0.04). Over the entire 118-d period, SCB-fed bulls had greater final body weight (509.5 vs. 518.0 kg, P = 0.02), total body weight gain (163.7 vs. 170.3 kg, P = 0.01), and average daily gain (1.366 vs. 1.420 kg, P = 0.01). The feed efficiency of SCB-supplemented bulls was 8.05% higher than CON (P = 0.04), and the final carcass weight was 1.69% greater for animals fed SCB (283.8 vs. 288.6 kg, P = 0.04). Total carcass weight gain (110.9 vs. 114.7 kg) and daily carcass weight gain (0.924 vs. 0.956 kg) tended (P = 0.06) to increase by 3.46% in SCB-fed animals compared with those fed CON. Gain yield, carcass conversion, and carcass yield did not differ between treatments. There were no significant differences in the apparent digestibility of DM, crude protein, neutral detergent fiber, and ether extract between treatments. However, starch digestibility (92.7% vs. 88%) was greater for the control treatment (P < 0.001). Including live Saccharomyces cerevisiae boulardii yeast as a probiotic supplement during the initial 42 d in the feedlot enhanced early-stage growth performance in Nellore bulls. Notably, this supplementation carried over carcass gain over the entire feedlot period.

Long-term exposure of Allonais inaequalis to a mixture of antibiotics in freshwater and synthetic wastewater matrices: Reproduction, recovery, and swimming responses.

Antibiotics from sulfonamide, fluoroquinolone, and diaminopyrimidine classes are widely used in human and veterinary medicine, and their combined occurrence in the aquatic environment is increasing around the world. In parallel, the understanding of how mixtures of these compounds affect non-target species from tropical freshwaters is scarce. Thus, this work aimed to study the long-term reproductive, recovery, and swimming effects of mixtures of 12 antibiotics from three different classes (up to 10 µg L-1 ) added to freshwater (FWM) and synthetic wastewater (SWM) matrices on freshwater worm Allonais inaequalis. Results revealed that at the reproduction level, the exposure to antibiotics in the SWM matrix does not cause a significant toxic effect on species after 10 days. On the other hand, exposures to initial dose mixtures (10 µg L-1 each) in FWM caused a significant reduction of offspring by 19.2%. In addition, recovery bioassays (10 days in an antibiotic-free environment) suggested that A. inaequalis has reduced offspring production due to previous exposure to antibiotic mixtures in both matrices. Furthermore, despite slight variation in swimming speed over treatments, no significant differences were pointed out. Regarding antibiotics in the water matrices after 10-day exposures, the highest concentrations were up to 2.7, 7.8, and 4.2 µg L-1 for antibiotics from sulfonamide, fluoroquinolone, and diaminopyrimidine classes, respectively. These findings suggest that a species positioned between primary producers and secondary consumers may experience late reproductive damage even in an antibiotic-free zone, after previous 10-day exposure to antibiotic mixtures. PRACTITIONER POINTS: A mixture of sulfonamide, fluoroquinolone, and diaminopyrimidine antibiotics in freshwater affects the offspring production of A. inaequalis after 10 days. After the 10-day antibiotic exposure, the reproduction of A. inaequalis remains affected in an antibiotic-free environment over the recovery period. The swimming speed of the worms does not change after 10 days of exposure to the antibiotic mixture. The concentration of dissolved solids can limit the natural degradation of sulfonamide, fluoroquinolone, and diaminopyrimidine antibiotics in the aquatic environment.

Compatibility of pesticides with the predatory mite Neoseiulus barkeri.

Multiple arthropod pests can affect the same crop in agricultural systems, requiring the integration of control methods. In the present study, the effects of residual exposure to four broad-spectrum insecticides/acaricides (azadiractin, abamectin, chlorfenapyr, and fenpyroximate) on immature (development and survival time) and adult females (longevity, fecundity, and fertility life table parameters) of the predatory mite Neoseiulus barkeri were evaluated. Additionally, the insecticides/acaricides were categorized according to their selectivity based on the classification proposed by the International Organization for Biological Control (IOBC) for assessing the susceptibility of arthropods in laboratory experiments. Method 004, proposed by the Insecticide Resistance Action Committee (IRAC), was adopted for the bioassays with predators exposed to insecticide-acaricide residues. Among the insecticides/acaricides studied, azadirachtin had minimal effects on immature and adult N. barkeri (all non-significant) and was considered harmless based on the classification of toxicity according to the standards/categories proposed by the IOBC. All other insecticides/acaricides affected immature and adult N. barkeri and were considered slightly harmful in terms of toxicity, according to the IOBC.

Not so much a sea bass: Divergent European sea bass (Dicentrarchus labrax L.) freshwater incursions.

European sea bass (Dicentrarchus labrax [Linnaeus, 1758]) is a euryhaline marine migrant fish highly valuable for fisheries and aquaculture. Although juveniles are known to use estuaries and occasionally move to freshwater environments, these freshwater incursions had not been reported for adults. Recently, this behavior was observed in the Tagus River (Portugal) for adults occurring up to 150 km from the river mouth, about 80 km upstream from the tidal influence, suggesting the existence of a freshwater contingent. Fisheries management of sea bass should consider the putative existence of a freshwater contingent.

Discovery of Novel miRNAs in Colorectal Cancer: Potential Biological Roles and Clinical Utility.

Deregulated miRNAs are associated with colorectal cancer (CRC), with alterations depending on the tumor location. Novel tissue-specific miRNAs have been identified in different tumors and are associated with cancer. We used miRMaster to identify novel miRNAs in CRC from the TCGA and GEO data (discovery and validation groups). We used TCGA data from five tissues to analyze miRNA tissue specificity. miRDB was used to predict miRNA targets, and the UCSC Xena Browser was used to evaluate target expression. After successive analyses, we identified 15 novel miRNAs with the same expression patterns in CRC in both the discovery and validation groups. Four molecules (nov-miR-13844-5p, nov-miR-7154-5p, nov-miR-5035-3p, and nov-miR-590-5p) were differentially expressed in proximal and distal CRC. The nov-miR-3345-5p and nov-miR-13172-3p, which are upregulated in tumors, are only expressed in colorectal tissues. These molecules have been linked to a worse prognosis in right-sided colon and rectal carcinomas. An analysis revealed an association between eight novel miRNAs and 81 targets, mostly cancer-related genes, with varying expression based on tumor location. These findings provide new miRNAs with potential biological relevance, molecular biomarkers, and therapeutic targets for CRC treatment.

Dimerization of melanocortin 4 receptor controls puberty onset and body size polymorphism.

Xiphophorus fish exhibit a clear phenotypic polymorphism in puberty onset and reproductive strategies of males. In X. nigrensis and X. multilineatus, puberty onset is genetically determined and linked to a melanocortin 4 receptor (Mc4r) polymorphism of wild-type and mutant alleles on the sex chromosomes. We hypothesized that Mc4r mutant alleles act on wild-type alleles by a dominant negative effect through receptor dimerization, leading to differential intracellular signaling and effector gene activation. Depending on signaling strength, the onset of puberty either occurs early or is delayed. Here, we show by Förster Resonance Energy Transfer (FRET) that wild-type Xiphophorus Mc4r monomers can form homodimers, but also heterodimers with mutant receptors resulting in compromised signaling which explains the reduced Mc4r signaling in large males. Thus, hetero- vs. homo- dimerization seems to be the key molecular mechanism for the polymorphism in puberty onset and body size in male fish.

Green synthesis of gold nanoparticles modulates lipopolysaccharide-induced lung inflammation in Wistar rats.

This study aimed to investigate the effect of intranasal treatment of gold nanoparticles (GNPs) and Curcumin (Cur) on the lipopolysaccharide (LPS)-induced acute pulmonary inflammatory response. A single intraperitoneal injection of LPS (0.5 mg/Kg) was performed, and the animals in the Sham group were injected with 0.9% saline. Treatment was daily intranasally with GNPs (2.5 mg/L), Cur (10 mg/kg) and GNP-Cur started 12 h after LPS administration and ended on the seventh day. The results show that the treatment performed with GNP-Cur was the most effective to attenuate the action of pro-inflammatory cytokines, and a lower leukocyte count in the bronchoalveolar lavage, in addition to positively regulating anti-inflammatory cytokines in relation to other groups. As a result, it promoted an oxirreductive balanced environment in the lung tissue, providing a histological outcome with a reduction in inflammatory cells and greater alveolar area. The group treated with GNPs-Cur was superior to the other groups, with better anti-inflammatory activity and reduced oxidative stress, resulting in less morphological damage to lung tissue. In conclusion, the use of reduced GNPs with curcumin demonstrates promising effects in the control of the acute inflammatory response, helping to protect the lung tissue at the biochemical and morphological levels.

The APSY-SED study: protocol of an observational, longitudinal, mixed methods and multicenter study exploring the psychological adjustment of relatives and healthcare providers of patients with cancer with continuous deep sedation until death.

BACKGROUND: Since 2016, France is the only country in the World where continuous deep sedation until death (CDSUD) is regulated by law. CDSUD serves as a response to refractory suffering in palliative situations where the patients' death is expected to occur in the following hours or days. Little is known on the psychological adjustment surrounding a CDSUD procedure for healthcare providers (HCPs) and relatives. Our study aims to gather qualitative and quantitative data on the specific processes behind the psychological adjustment of both relatives and HCPs, after the administration of CDSUD for patients with cancer. METHODS: The APSY-SED study is a prospective, longitudinal, mixed-methods and multicenter study. Recruitment will involve any French-speaking adult cancer patient for who a CDSUD is discussed, their relatives and HCPs. We plan to include 150 patients, 150 relatives, and 50 HCPs. The evaluation criteria of this research are: 1/ Primary criterion: Psychological adjustment of relatives and HCPs 6 and 13 months after the death of the patient with cancer (psychological adjustment = intensity of anxiety, depression and grief reactions, CDSUD-related distress, job satisfaction, Professional Stress and Professional experience). Secondary criteria: a)occurrence of wish for a CDSUD in patients in palliative phase; b)occurrence of wish for hastened death in patients in palliative phase; c)potential predictors of adjustment assessed after the discussion concerning CDSUD as an option and before the setting of the CDSUD; d) Thematic analysis and narrative account of meaning-making process concerning the grief experience. DISCUSSION: The APSY-SED study will be the first to investigate the psychological adjustment of HCPs and relatives in the context of a CDSUD procedure implemented according to French law. Gathering data on the grief process for relatives can help understand bereavement after CDSUD, and participate in the elaboration of specific tailored interventions to support HCPs and relatives. Empirical findings on CDSUD among patients with cancer in France could be compared with existing data in other countries and with results related to other medical fields where CDSUD is also conducted. TRIAL REGISTRATION: This protocol received the National Registration Number: ID-RCB2021-A03042-39 on 14/12/2021.

Adaptations in dental public health services during the COVID-19 pandemic in municipalities of Southern Brazil: a grounded theory and collaborative research.

The aim of this study was to analyze oral health actions in four municipalities in Brazil during the COVID-19 pandemic according to a theoretical framework model on oral healthcare management. It was a qualitative study carried out in two stages. A theoretical-empirical model on the significance of oral healthcare management was developed, following the Grounded Theory method. Fourteen dentists and five healthcare managers participated, through open interview. Subsequently, collaborative research was performed, and the model was applied to analyze the documents produced to address the pandemic by each of four municipalities in Santa Catarina State. The model provided a framework for analyzing actions for coping with the pandemic regarding oral health services. Actions were identified in all dimensions of the model: reduction in supply of dental care due to restricted access to elective services; search for biosafety care standards; dissemination of standardized science-based guidelines; attempt to maintain comprehensive dental assistance through re-adaptation of specialized services and collective actions; and relocation of oral health professionals to assist in other sectors. The oral health care management framework can serve as a reference for redesigning oral health actions and services in other municipalities during the COVID-19 pandemic, in a broader perspective.

Analisar as ações de saúde bucal em quatro municípios brasileiros durante a pandemia de COVID-19, segundo um modelo de referencial teórico sobre gestão da atenção à saúde bucal. Estudo qualitativo realizado em dois momentos. Foi desenvolvido um modelo teórico-empírico sobre o significado da gestão do cuidado em saúde bucal, seguindo o método da Teoria Fundamentada nos Dados. Participaram 14 dentistas e cinco gestores de saúde, por meio de entrevista aberta. Posteriormente, no segundo momento, foi realizada uma pesquisa colaborativa, e o modelo foi aplicado para analisar os documentos produzidos em cada município para o enfrentamento local da pandemia, em quatro municípios do Estado de Santa Catarina, sul do Brasil. O modelo forneceu uma estrutura para analisar as ações de enfrentamento da pandemia nos serviços de saúde bucal. Foram identificadas ações em todas as dimensões do modelo: redução da oferta de atendimento odontológico devido à restrição de acesso aos serviços eletivos; a busca por padrões de assistência à biossegurança; disseminação de diretrizes padronizadas e com base científica; a tentativa de manter a assistência odontológica integral por meio da readaptação de serviços especializados e ações coletivas; e realocação de profissionais de saúde bucal para atendimento em outros setores. O referencial de gestão da atenção à saúde bucal pode servir de referência para redesenhar as ações e serviços de saúde bucal em outros municípios em período de pandemia de COVID-19, em uma perspectiva mais ampla.

Adaptations in dental public health services during the COVID-19 pandemic in municipalities of Southern Brazil: a grounded theory and collaborative research / Adaptações nos serviços públicos odontológicos durante o Pandemia COVID-19 em municípios do Sul do Brasil: uma teoria fundamentada e pesquisa colaborativa

ABSTRACT The aim of this study was to analyze oral health actions in four municipalities in Brazil during the COVID-19 pandemic according to a theoretical framework model on oral healthcare management. It was a qualitative study carried out in two stages. A theoretical-empirical model on the significance of oral healthcare management was developed, following the Grounded Theory method. Fourteen dentists and five healthcare managers participated, through open interview. Subsequently, collaborative research was performed, and the model was applied to analyze the documents produced to address the pandemic by each of four municipalities in Santa Catarina State. The model provided a framework for analyzing actions for coping with the pandemic regarding oral health services. Actions were identified in all dimensions of the model: reduction in supply of dental care due to restricted access to elective services; search for biosafety care standards; dissemination of standardized science-based guidelines; attempt to maintain comprehensive dental assistance through re-adaptation of specialized services and collective actions; and relocation of oral health professionals to assist in other sectors. The oral health care management framework can serve as a reference for redesigning oral health actions and services in other municipalities during the COVID-19 pandemic, in a broader perspective.

RESUMO Analisar as ações de saúde bucal em quatro municípios brasileiros durante a pandemia de COVID-19, segundo um modelo de referencial teórico sobre gestão da atenção à saúde bucal. Estudo qualitativo realizado em dois momentos. Foi desenvolvido um modelo teórico-empírico sobre o significado da gestão do cuidado em saúde bucal, seguindo o método da Teoria Fundamentada nos Dados. Participaram 14 dentistas e cinco gestores de saúde, por meio de entrevista aberta. Posteriormente, no segundo momento, foi realizada uma pesquisa colaborativa, e o modelo foi aplicado para analisar os documentos produzidos em cada município para o enfrentamento local da pandemia, em quatro municípios do Estado de Santa Catarina, sul do Brasil. O modelo forneceu uma estrutura para analisar as ações de enfrentamento da pandemia nos serviços de saúde bucal. Foram identificadas ações em todas as dimensões do modelo: redução da oferta de atendimento odontológico devido à restrição de acesso aos serviços eletivos; a busca por padrões de assistência à biossegurança; disseminação de diretrizes padronizadas e com base científica; a tentativa de manter a assistência odontológica integral por meio da readaptação de serviços especializados e ações coletivas; e realocação de profissionais de saúde bucal para atendimento em outros setores. O referencial de gestão da atenção à saúde bucal pode servir de referência para redesenhar as ações e serviços de saúde bucal em outros municípios em período de pandemia de COVID-19, em uma perspectiva mais ampla.

Double heterozygous pathogenic variants prevalence in a cohort of patients with hereditary breast cancer.

Hereditary breast cancer (BC) corresponds to 5% of all BC and a larger parcel of early-onset disease. The incorporation of next-generation sequencing (NGS) techniques reduced the cost of molecular testing and allowed the inclusion of additional cancer predisposition genes in panels that are more comprehensive. This enabled the identification of germline pathogenic variants in carriers and the introduction of risk-reducing strategies. It also resulted in the identification of the co-occurrence of more than one germline pathogenic variant in BC genes in some families. This is a rare event, and there are few reports on its impact on cancer risk. We conducted a single-institution retrospective study in which 1,156 women with early onset BC and/or a family history of cancer were tested by a germline multi-gene hereditary cancer panel. Germline pathogenic variants in high- and/or moderate-penetrance BC genes were identified in 19.5% of the individuals (n = 226). The most frequent variants were found in TP53 (69 of 226; 55 of them represented by p.R337H), BRCA1 (47 of 226), and BRCA2 (41 of 226). Double heterozygous (DH) variants were detected in 14 cases, representing 1.2% of all individuals assessed. There were no significant differences in age of BC onset and risk for bilateral BC in DH carriers when compared with those with one germline variant.

MeHg exposure impairs both the catecholaminergic and cholinergic systems resulting in motor and non-motor behavioral changes in Drosophila melanogaster.

Human exposure to the natural environmental contaminant methylmercury (MeHg) has been associated to adverse health effects. Importantly, the mechanisms by which this organomercurial exerts its neurotoxicity have yet to be fully clarified. Therefore, the aim of this study was to evaluate whether exposure to MeHg alters dopamine (DA) and octopamine (OA) levels, acetylcholinesterase (AChE) activity and impacts both motor and non-motor behaviours. We studied the effect of MeHg by feeding 1-2 d old flies (male and females) with 25 and 50 µM MeHg for 4 d and determined effects on survival, motor and non-motor behaviours, oxidative stress, AChE and tyrosine hydroxylase (TH) activities, as well as DA and OA levels. We found that Drosophila melanogaster (D. melanogaster) exposed to MeHg showed a reduction in survival rate, associated with the inhibition of AChE and TH activities in head of flies and decreased DA and OA levels. These changes were accompanied by behavioural alterations, such as locomotor deficit and increased grooming behaviour, in addition to an increase in oxidative stress markers both in head and in body of flies, and an increase in glutathione-S-transferase (GST) activity in head of flies. Collectively, our data support the hypothesis that MeHg neurotoxicity is associated with altered OA and DA levels, AChE inhibition, which may serve, at least in part, as the underpinnings of both motor and non-motor behavioural changes.

Genetic and Epigenetic Mechanisms Deregulate the CRL2pVHL Complex in Hepatocellular Carcinoma.

Dysregulation of ubiquitin-proteasome pathway genes through copy number alteration, promoter hypomethylation, and miRNA deregulation is involved in cancer development and progression. Further characterizing alterations in these genes may uncover novel drug targets across a range of diseases in which druggable alterations are uncommon, including hepatocellular carcinoma (HCC). We analyzed 377 HCC and 59 adjacent non-malignant liver tissue samples, focusing on alterations to component genes of the widely studied CRL2pVHL E3 ubiquitin ligase complex. mRNA upregulation of the component genes was common, and was correlated with DNA hypomethylation and copy number increase, but many tumours displayed overexpression that was not explained by either mechanism. Interestingly, we found 66 miRNAs, including 39 previously unannotated miRNAs, that were downregulated in HCC and predicted to target one or more CRL2pVHL components. Several miRNAs, including hsa-miR-101-3p and hsa-miR-139-5p, were negatively correlated with multiple component genes, suggesting that miRNA deregulation may contribute to CRL2pVHL overexpression. Combining miRNA and mRNA expression, DNA copy number, and methylation status into one multidimensional survival analysis, we found a significant association between greater numbers of alterations and poorer overall survival for multiple component genes. While the intricacies of CRL2pVHL complex gene regulation require additional research, it is evident that multiple causes for the deregulation of these genes must be considered in HCC, including non-traditional mechanisms.

The effect of different resistance training protocols equalized by time under tension on the force-position relationship after 10 weeks of training period.

This study investigated the impact of performing two equalized resistance training (RT) protocols for 10 weeks that differ only by repetition duration and number in the force-position and EMG-position relationship. Participants performed an equalized (36 s of time under tension; 3-4 sets; 3 min between sets; 50-55% of one-repetition maximum; 3× week) RT intervention on the bench press and the only different change between protocols was repetition number (RN; 12 vs.6) or duration (RD; 3 s vs. 6 s). Two experimental groups (RN12RD3, n = 12; and RN6RD6, n = 12) performed the RT, while one group was the control (Control, n = 11). Maximal isometric contractions at 10%, 50% and 90% of total bench press range of motion were performed pre- and post-RT, while electromyography was recorded. It demonstrated an increase in isometric force (+14% to 24%, P < 0.001) shifting up the force-position relationship of the training groups after RT, although no difference was between training groups compared to the Control. Neuromuscular activation from pectoralis major presented an increase after training for both RT groups (+44%; P < 0.001) compared to the Control. However, although not significantly different, triceps brachii also presented an increase depending on the protocol (+25%). In conclusion, 10 weeks of an equalized RT with longer RN and shorter RD (or opposite) similarly increases the ability to produce maximal isometric force during the bench exercise across different angles, while neuromuscular activation of the pectoralis major partially explained the shift-up of the force-position relationship after training.

Equalization of Training Protocols by Time Under Tension Determines the Magnitude of Changes in Strength and Muscular Hypertrophy.

ABSTRACT: Martins-Costa, HC, Lacerda, LT, Diniz, RCR, Lima, FV, Andrade, AGP, Peixoto, GH, Gomes, MC, Lanza, MB, Bemben, MG, and Chagas, MH. Equalization of training protocols by time under tension determines the magnitude of changes in strength and muscular hypertrophy. J Strength Cond Res 36(7): 1770-1780, 2022-The aim of this study was to investigate the effects of 2 training protocols equalized by tension (TUT) on maximal strength (1 repetition maximum [RM]), regional cross-sectional areas (proximal, middle, and distal), and total cross-sectional areas (sum of the regional cross-sectional areas) of the pectoralis major and triceps brachii muscles. Thirty-eight men untrained in resistance training participated in the study and were allocated under 3 conditions: Protocol 3s (n = 11; 12 repetitions; 3s repetition duration), Protocol 6s (n = 11; 6 repetitions; 6s repetition duration), and Control (n = 11; no training). Training protocols (10 weeks; bench press exercise) were equated for TUT (36 seconds per set), number of sets (3-4), intensity (50-55% of 1RM), and rest between sets (3 minutes). Analysis of variance was used to examine a percentage change in variables of interest across the 3 groups with an alpha level of 0.05 used to establish statistical significance. Protocols 3s and 6s showed no differences in the increase of total and regional muscle cross-sectional areas. There were no differences in regional hypertrophy of the pectoralis major muscle. In the triceps brachii muscle, the increase in distal cross-sectional area was greater when compared with the middle and proximal regions. Both experimental groups had similar increases in the 1RM test. In conclusion, training protocols with the same TUT promote similar strength gains and muscle hypertrophy. Moreover, considering that the protocols used different numbers of repetitions, the results indicate that training volumes cannot be considered separately from TUT when evaluating neuromuscular adaptations.

A duplicated copy of id2b is an unusual sex-determining candidate gene on the Y chromosome of arapaima (Arapaima gigas).

Arapaima gigas is one of the largest freshwater fish species of high ecological and economic importance. Overfishing and habitat destruction are severe threats to the remaining wild populations. By incorporating a chromosomal Hi-C contact map, we improved the arapaima genome assembly to chromosome-level, revealing an unexpected high degree of chromosome rearrangements during evolution of the bonytongues (Osteoglossiformes). Combining this new assembly with pool-sequencing of male and female genomes, we identified id2bbY, a duplicated copy of the inhibitor of DNA binding 2b (id2b) gene on the Y chromosome as candidate male sex-determining gene. A PCR-test for id2bbY was developed, demonstrating that this gene is a reliable male-specific marker for genotyping. Expression analyses showed that this gene is expressed in juvenile male gonads. Its paralog, id2ba, exhibits a male-biased expression in immature gonads. Transcriptome analyses and protein structure predictions confirm id2bbY as a prime candidate for the master sex-determiner. Acting through the TGFß signaling pathway, id2bbY from arapaima would provide the first evidence for a link of this family of transcriptional regulators to sex determination. Our study broadens our current understanding about the evolution of sex determination genetic networks and provide a tool for improving arapaima aquaculture for commercial and conservation purposes.

Multi-analytical test based on serum miRNAs and proteins quantification for ovarian cancer early detection.

Advanced ovarian cancer is one of the most lethal gynecological tumor, mainly due to late diagnoses and acquired drug resistance. MicroRNAs (miRNAs) are small-non coding RNA acting as tumor suppressor/oncogenes differentially expressed in normal and epithelial ovarian cancer and has been recognized as a new class of tumor early detection biomarkers as they are released in blood fluids since tumor initiation process. Here, we evaluated by droplet digital PCR (ddPCR) circulating miRNAs in serum samples from healthy (N = 105) and untreated ovarian cancer patients (stages I to IV) (N = 72), grouped into a discovery/training and clinical validation set with the goal to identify the best classifier allowing the discrimination between earlier ovarian tumors from health controls women. The selection of 45 candidate miRNAs to be evaluated in the discovery set was based on miRNAs represented in ovarian cancer explorative commercial panels. We found six miRNAs showing increased levels in the blood of early or late-stage ovarian cancer groups compared to healthy controls. The serum levels of miR-320b and miR-141-3p were considered independent markers of malignancy in a multivariate logistic regression analysis. These markers were used to train diagnostic classifiers comprising miRNAs (miR-320b and miR-141-3p) and miRNAs combined with well-established ovarian cancer protein markers (miR-320b, miR-141-3p, CA-125 and HE4). The miRNA-based classifier was able to accurately discriminate early-stage ovarian cancer patients from health-controls in an independent sample set (Sensitivity = 80.0%, Specificity = 70.3%, AUC = 0.789). In addition, the integration of the serum proteins in the model markedly improved the performance (Sensitivity = 88.9%, Specificity = 100%, AUC = 1.000). A cross-study validation was carried out using four data series obtained from Gene Expression Omnibus (GEO), corroborating the performance of the miRNA-based classifier (AUCs ranging from 0.637 to 0.979). The clinical utility of the miRNA model should be validated in a prospective cohort in order to investigate their feasibility as an ovarian cancer early detection tool.

Chronic Antidiabetic Actions of Leptin: Evidence From Parabiosis Studies for a CNS-Derived Circulating Antidiabetic Factor.

We used parabiosis to determine whether the central nervous system (CNS)-mediated antidiabetic effects of leptin are mediated by release of brain-derived circulating factors. Parabiosis was surgically induced at 4 weeks of age, and an intracerebroventricular (ICV) cannula was placed in the lateral cerebral ventricle at 12 weeks of age for ICV infusion of leptin or saline vehicle. Ten days after surgery, food intake, body weight, and blood glucose were measured for 5 consecutive days, and insulin-deficiency diabetes was induced in all rats by a single streptozotocin (STZ) injection (40 mg/kg). Five days after STZ injection, leptin or vehicle was infused ICV for 7 days, followed by 5-day recovery period. STZ increased blood glucose and food intake. Chronic ICV leptin infusion restored normoglycemia in leptin-infused rats while reducing blood glucose by ∼27% in conjoined vehicle-infused rats. This glucose reduction was caused mainly by decreased hepatic gluconeogenesis. Chronic ICV leptin infusion also reduced net cumulative food intake and increased GLUT4 expression in skeletal muscle in leptin/vehicle compared with vehicle/vehicle conjoined rats. These results indicate that leptin's CNS-mediated antidiabetic effects are mediated, in part, by release into the systemic circulation of leptin-stimulated factors that enhance glucose utilization and reduce liver gluconeogenesis.