Characterization and control of oocyte large-scale chromatin configuration in different cattle breeds.

Made available in DSpace on 2020-12-11T01:57:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Differences in reproductive physiology between cattle breeds may help to explain distinct responses to assisted reproductive techniques and to define breed-specific protocols with improved efficiency. Germinal vesicle (GV) stage oocytes are characterized by increasing levels of chromatin compaction enclosed within the nucleus (graded from GV0 to GV3), associated with different developmental competence. The first objective of this study was to characterize chromatin configuration of GV stage oocytes recovered by OPU at random days of the estrous cycle from Nelore (Bos indices) and Holstein (Bos taurus) cows. In Nelore 90% of the oocytes presented advanced stages of chromatin compaction associated with higher developmental competence (GV2 and GV3), while in Holstein, only 65% of the oocytes were at these stages. Then, aiming to obtain a more homogeneous population of oocytes in Holstein, we tested two synchronization protocols combining aspiration of all visible follicles at a random day (day 0), two IM injections of FSH 12 h apart on day 2, and OPU on day 4 (OPU/D4) or 5 (OPU/D5). The protocol OPU/D4 provided around 45% of the oocytes with low chromatin compaction (GV1), while the protocol OPU/D5 provided 70% of the oocytes at GV2 and 20% at GV3. Finally, we assessed the effects of a culture system known to prevent meiotic resumption on chromatin configuration of the GV2 enriched oocyte population obtained with the protocol OPU/D5. After 9 h of culture most oocytes transited from GV2 to GV3, with 90% of the oocytes at GV3 stage. This study demonstrates differences between Nelore and Holstein cows regarding patterns of chromatin configuration that may account for their different performance in IVM/IVF. In addition, it provides novel references for the design of protocols aiming to regulate oocyte quality before IVM for the optimization of IVF outcomes. (C) 2019 Elsevier Inc. All rights reserved. Sao Paulo State Univ, Inst Biosci, Dept Physiol, Ovarian Mol Physiol Lab, Sao Paulo, Brazil Univ Sao Paulo, Dept Anim Reprod, Sao Paulo, Brazil Univ Milan, Dept Hlth Anim Sci & Food Safety, Reprod & Dev Biol Lab, Milan, Italy Sao Paulo State Univ, Inst Biosci, Dept Physiol, Ovarian Mol Physiol Lab, Sao Paulo, Brazil CAPES: 001 FAPESP: 2017/07588-4 FAPESP: 2016/21671-

Glyceraldehyde-3-phosphate dehydrogenase of Paracoccidioides brasiliensis is a cell surface protein involved in fungal adhesion to extracellular matrix proteins and interaction with cells

The pathogenic fungus Paracoccidioides brasiliensis causes paracoccidioidomycosis, a pulmonary mycosis acquired by inhalation of fungal airborne propagules, which may disseminate to several organs and tissues, leading to a severe form of the disease. Adhesion to and invasion of host cells are essential steps involved in the infection and dissemination of pathogens. Furthermore, pathogens use their surface molecules to bind to host extracellular matrix components to establish infection. Here, we report the characterization of the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of P. brasiliensis as an adhesin, which can be related to fungus adhesion and invasion. the P. brasiliensis GAPDH was overexpressed in Escherichia coli, and polyclonal antibody against this protein was obtained. By immunoelectron microscopy and Western blot analysis, GAPDH was detected in the cytoplasm and the cell wall of the yeast phase of P. brasiliensis. the recombinant GAPDH was found to bind to fibronectin, laminin, and type I collagen in ligand far-Western blot assays. of special note, the treatment of P. brasiliensis yeast cells with anti-GAPDH polyclonal antibody and the incubation of pneumocytes with the recombinant protein promoted inhibition of adherence and internalization of P. brasiliensis to those in vitro-cultured cells. These observations indicate that the cell wall-associated form of the GAPDH in P. brasiliensis could be involved in mediating binding of fungal cells to fibronectin, type I collagen, and laminin, thus contributing to the adhesion of the microorganism to host tissues and to the dissemination of infection.Univ Fed Goias, Inst Ciencias Biol, Mol Biol Lab, BR-74001970 Goiania, Go, BrazilUniv Brasilia, BR-70910900 Brasilia, DF, BrazilUniv Estadual Julio Mesquita Filho, Araraquara, SP, BrazilUniversidade Federal de São Paulo, São Paulo, BrazilUniversidade Federal de São Paulo, São Paulo, BrazilWeb of Scienc

Desempenho da solicitação do perfil lipídico entre os setores público e privado

OBJECTIVE: To evaluate the performance of lipid profile screening according to the mode of financing of medical appointments - public or private. METHODS: A population-based cross-sectional study was carried out with a multi-stage sampling strategy. The study included 3,136 adults (>; 20 years old) from the city of Pelotas, Southern Brazil, in 2006. The following indicators were calculated: coverage (proportion of screened individuals among those meeting screening criteria), focus (proportion of individuals meeting screening criteria among those who were tested), screening errors (tests in individuals not meeting screening criteria - overscreening - and no screening of individuals meeting the screening criteria - underscreening), and screening ratio (ratio between number of individuals who met screening criteria and the number who failed to meet screening criteria among all individuals tested). Bivariate analyses were performed using Chi-squared tests. 95% confidence intervals (95% CI) were calculated for all parameters assessed. RESULTS: General coverage was 73.0% (95%CI: 70.8;75.2), and focus was 67.2% (95% CI: 64.7;69.3). In the public sector, compared to the private/health plan sector, coverage was lower (65.2% vs. 82.2%; pOBJETIVO: Avaliar o desempenho do rastreio do perfil lipídico conforme o financiamento da consulta médica - setores público e privado. MÉTODOS: Estudo transversal de base populacional com amostragem em múltiplos estágios, incluindo 3.136 adultos (>; 20 anos) de Pelotas, Brasil, 2006. Foram calculados: cobertura (proporção de indivíduos rastreados entre aqueles com indicação de rastreio), foco (proporção de indivíduos com indicação entre os rastreados), erros de rastreio (sobre-rastreio: exames em indivíduos sem indicação, e sub-rastreio: não solicitação entre indivíduos com risco) e razão de rastreio (razão entre número de rastreados que atendiam aos critérios e número dos que não atendiam). Análises bivariadas foram realizadas por meio do teste qui-quadrado e intervalos de confiança de 95% foram estimados para os parâmetros avaliados. RESULTADOS: A cobertura geral foi 73,0% (IC 95%: 70,8;75,2) e o foco 67,2% (IC 95%: 64,7;69,3). O setor público teve, em relação ao privado/convênio, menor cobertura (65,2% versus 82,2%;

The emergence of arboviruses changes the profile of viral meningitis in Salvador, Bahia: A case series

Background: Recently, different arboviruses became endemic in Brazil mostlycausing acute febrile illnesses, however, neurological manifestations have alsobeen reported. This study aimed to investigate which viruses were involved inthe meningitis etiology and the contribution of the circulating arboviruses inSalvador, Bahia, Brazil.Methods: From June 2014 to February 2016, 170 patients with suspected viralmeningitis were identified in Couto Maia Hospital, Salvador-BA, Brazil. TheirCSF samples were investigated for possible viral etiology by reversetranscription-PCR (RT-PCR) for different arboviruses: DENV, ZIKV and CHIKV;and for the EV; and by PCR for the HHV1-5 complex (HSV1-2, VZV, EBV andCMV). Also, ELISA was carried out in a subgroup of remaining samples fordetection of DENV IgM and NS1 antigen, CHIKV IgM and ZIKV IgM.Results: Thirty-seven patients were PCR or ELISA positive for at least one of thestudied viruses (overall positivity 21.8%). EV was the agent most frequentlydetected (10 cases; 27.0%), along with all four DENV serotypes (10 cases;27.0%); followed by CHIKV (6 cases; 16.2%), ZIKV (6 cases; 16.2%), and Varicellazoster virus (VZV) (1 case; 2.7%). Four cases (10.8%) presented viral co-infectiondetected: DENV1 + CHIKV, DENV1 + EV, DENV4 + ZIKV, and CHIKV + ZIKV.Arboviruses (DENV, CHIKV and ZIKV) accounted for the great majority of cases(26 cases; 70.3%) of all single and co-infections: DENV has been the mostfrequently detected arbovirus (13 cases; 35.1%). Among non-arboviralmeningitis,the most common etiology was the EV (11 cases; 29.7%).Conclusions: Arboviruses accounted for the majority of identified virusesamong patients with suspected viral meningitis. In areas where they areendemic it is crucial to increase viral surveillance and consider them in thedifferential diagnosis of meningitis.Fil: Dias, Tamiris T.. Fundación Oswaldo Cruz; BrasilFil: Tauro, Laura Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Nordeste. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú | Universidad Nacional de Misiones. Instituto de Biología Subtropical. Instituto de Biología Subtropical - Nodo Puerto Iguazú; ArgentinaFil: Macêdo, Lara E. N.. Fundación Oswaldo Cruz; BrasilFil: Brito, Liz O.. Fundación Oswaldo Cruz; BrasilFil: Ribeiro, Victor H. O.. Fundación Oswaldo Cruz; BrasilFil: Santos, Cleiton S.. Fundación Oswaldo Cruz; BrasilFil: Jacob-Nascimento, Leile C.. Fundación Oswaldo Cruz; BrasilFil: Vilas-Boas, Letícia S.. Fundación Oswaldo Cruz; BrasilFil: Amado, Caio. Fundación Oswaldo Cruz; BrasilFil: Barbosa, Paula S.. Fundación Oswaldo Cruz; BrasilFil: Reis, Joice N.. Fundación Oswaldo Cruz; BrasilFil: Soares Campos, Gubio. Fundación Oswaldo Cruz; BrasilFil: Ribeiro, Guilherme S.. Fundación Oswaldo Cruz; BrasilFil: Siqueira, Isadora C.. Fundación Oswaldo Cruz; BrasilFil: Silva, Luciano K.. Fundación Oswaldo Cruz; BrasilFil: Reis, Mitermayer G.. Fundación Oswaldo Cruz; Brasi

Geographic patterns of genetic variation in a broadly distributed marine vertebrate: new insights into loggerhead turtle stock structure from expanded mitochondrial DNA sequences

Previous genetic studies have demonstrated that natal homing shapes the stock structure of marine turtle nesting populations. However, widespread sharing of common haplotypes based on short segments of the mitochondrial control region often limits resolution of the demographic connectivity of populations. Recent studies employing longer control region sequences to resolve haplotype sharing have focused on regional assessments of genetic structure and phylogeography. Here we synthesize available control region sequences for loggerhead turtles from the Mediterranean Sea, Atlantic, and western Indian Ocean basins. These data represent six of the nine globally significant regional management units (RMUs) for the species and include novel sequence data from Brazil, Cape Verde, South Africa and Oman. Genetic tests of differentiation among 42 rookeries represented by short sequences (380 bp haplotypes from 3,486 samples) and 40 rookeries represented by long sequences (~800 bp haplotypes from 3,434 samples) supported the distinction of the six RMUs analyzed as well as recognition of at least 18 demographically independent management units (MUs) with respect to female natal homing. A total of 59 haplotypes were resolved. These haplotypes belonged to two highly divergent global lineages, with haplogroup I represented primarily by CC-A1, CC-A4, and CC-A11 variants and haplogroup II represented by CC-A2 and derived variants. Geographic distribution patterns of haplogroup II haplotypes and the nested position of CC-A11.6 from Oman among the Atlantic haplotypes invoke recent colonization of the Indian Ocean from the Atlantic for both global lineages. The haplotypes we confirmed for western Indian Ocean RMUs allow reinterpretation of previous mixed stock analysis and further suggest that contemporary migratory connectivity between the Indian and Atlantic Oceans occurs on a broader scale than previously hypothesized. This study represents a valuable model for conducting comprehensive international cooperative data management and research in marine ecology