Generation and validation of genetic markers for the selection of carioca dry bean genotypes with the slow-darkening seed coat trait

Slow darkening (SD) is a trait that helps to maintain a brighter seed coat appearance in certain market classes of dry beans. The aim of this study was to generate new fluorescence-based markers and validate previously identified microsatellite markers for linkage to the SD trait in lines of the carioca market class. Four segregating populations were generated by Embrapa, the Brazilian Agricultural Research Corporation, from crosses between the SD cultivar BRSMG Madrepe´rola and the regular-darkening cultivars BRS Estilo, BRS Cometa, BRS Nota´vel and BRS Sublime. These populations were screened with the simple-sequence markers Pvsd- 1158 and PVM02TC116 and with a TaqManTM marker designed for the single-nucleotide polymorphism (SNP) PvbHLHp12804. A KASP marker was also designed for the PvbHLHp12804 marker for testing on advanced carioca lines developed by the University of Saskatchewan. In the carioca lines developed by Embrapa, PVM02TC116 proved unsuitable for marker-assisted selection (MAS). Both the Pvsd-1158 and PvbHLHp12804 markers were found to be tightly linked to the gene responsible for the SD trait, with genetic distances calculated at 2.8 cM for Pvsd-1158 and 2.0 and 3.1 cM for PvbbHLHp12804, respectively. These markers presented more than 97% of selection efficiency. The genotypic scoring using the PvbHLHp12804 KASP marker was perfectly correlated with the phenotype in all lines of the University of Saskatchewan. The results of this study validates the use of Pvsd-1158 as a gel-based marker for SD in carioca beans. The new fluorescence-based SNP PvbHLHp12804 markers exhibited very tight linkage to SD in carioca and pinto bean lines. These markers will be ideal for MAS for the SD trait in these market classes

Lack of effects of typical and atypical antipsychotics in DARPP-32 and NCS-1 levels in PC12 cells overexpressing NCS-1

<p>Abstract</p> <p>Background</p> <p>Schizophrenia is the major psychiatry disorder, which the exact cause remains unknown. However, it is well known that dopamine-mediated neurotransmission imbalance is associated with this pathology and the main target of antipsychotics is the dopamine receptor D₂. Recently, it was described alteration in levels of two dopamine signaling related proteins in schizophrenic prefrontal cortex (PFC): Neuronal Calcium Sensor-1 (NCS-1) and DARPP-32. NCS-1, which is upregulated in PFC of schizophrenics, inhibits D₂internalization. DARPP-32, which is decreased in PFC of schizophrenics, is a key downstream effector in transducing dopamine signaling. We previously demonstrated that antipsychotics do not change levels of both proteins in rat's brain. However, since NCS-1 and DARPP-32 levels are not altered in wild type rats, we treated wild type PC12 cells (PC12 WT) and PC12 cells stably overexpressing NCS-1 (PC12 Clone) with antipsychotics to investigate if NCS-1 upregulation modulates DARPP-32 expression in response to antipsychotics treatment.</p> <p>Results</p> <p>We chronically treated both PC12 WT and PC12 Clone cells with typical (Haloperidol) or atypical (Clozapine and Risperidone) antipsychotics for 14 days. Using western blot technique we observed that there is no change in NCS-1 and DARPP-32 protein levels in both PC12 WT and PC12 Clone cells after typical and atypical antipsychotic treatments.</p> <p>Conclusions</p> <p>Because we observed no alteration in NCS-1 and DARPP-32 levels in both PC12 WT and Clone cells treated with typical or atypical antipsychotics, we suggest that the alteration in levels of both proteins in schizophrenic's PFC is related to psychopathology but not with antipsychotic treatment.</p

TRANSPORTE TRANSDÉRMICO DE NANOPARTÍCULAS METÁLICAS UTILIZANDO FONOFORESE: ESTUDOS ELETROQUÍMICOS

Existem diversos métodos de tratamento de doenças. Dentre eles, adesivos poliméricos contendo fármacos vêm sendo muito explorados por serem menos invasivos se comparados a outros métodos[1][2]. Nanopartículas de prata também são alvo de estudos devido a atividade antimicrobiana e cicatrizante que apresentam[3]. O trabalho teve como objetivo a avaliação da permeação de nanopartículas de prata através de uma membrana polimérica por meios eletroquímicos

The Brazilian consensus for the diagnosis and treatment of hyperthyroidism: recommendations by the Thyroid Department of the Brazilian Society of Endocrinology and Metabolism

INTRODUCTION: Hyperthyroidism is characterized by increased synthesis and release of thyroid hormones by the thyroid gland. Thyrotoxicosis refers to the clinical syndrome resulting from excessive circulating thyroid hormones, secondary to hyperthyroidism or due to other causes. This article describes evidence-based guidelines for the clinical management of thyrotoxicosis. OBJECTIVE: This consensus, developed by Brazilian experts and sponsored by the Department of Thyroid Brazilian Society of Endocrinology and Metabolism, aims to address the management, diagnosis and treatment of patients with thyrotoxicosis, according to the most recent evidence from the literature and appropriate for the clinical reality of Brazil. MATERIALS AND METHODS: After structuring clinical questions, search for evidence was made available in the literature, initially in the database MedLine, PubMed and Embase databases and subsequently in SciELO - Lilacs. The strength of evidence was evaluated by Oxford classification system was established from the study design used, considering the best available evidence for each question. RESULTS: We have defined 13 questions about the initial clinical approach for the diagnosis and treatment that resulted in 53 recommendations, including the etiology, treatment with antithyroid drugs, radioactive iodine and surgery. We also addressed hyperthyroidism in children, teenagers or pregnant patients, and management of hyperthyroidism in patients with Graves' ophthalmopathy and various other causes of thyrotoxicosis. CONCLUSIONS: The clinical diagnosis of hyperthyroidism usually offers no difficulty and should be made with measurements of serum TSH and thyroid hormones. The treatment can be performed with antithyroid drugs, surgery or administration of radioactive iodine according to the etiology of thyrotoxicosis, local availability of methods and preferences of the attending physician and patient.INTRODUÇÃO: O hipertireoidismo é caracterizado pelo aumento da síntese e liberação dos hormônios tireoidianos pela glândula tireoide. A tireotoxicose refere-se à síndrome clínica decorrente do excesso de hormônios tireoidianos circulantes, secundário ao hipertireoidismo ou não. Este artigo descreve diretrizes baseadas em evidências clínicas para o manejo da tireotoxicose. OBJETIVO: O presente consenso, elaborado por especialistas brasileiros e patrocinado pelo Departamento de Tireoide da Sociedade Brasileira de Endocrinologia e Metabologia, visa abordar o manejo, diagnóstico e tratamento dos pacientes com tireotoxicose, de acordo com as evidências mais recentes da literatura e adequadas para a realidade clínica do país. MATERIAIS E MÉTODOS: Após estruturação das questões clínicas, foi realizada busca das evidências disponíveis na literatura, inicialmente na base de dados do MedLine-PubMed e posteriormente nas bases Embase e SciELO - Lilacs. A força das evidências, avaliada pelo sistema de classificação de Oxford, foi estabelecida a partir do desenho de estudo utilizado, considerando-se a melhor evidência disponível para cada questão. RESULTADOS: Foram definidas 13 questões sobre a abordagem clínica inicial visando ao diagnóstico e ao tratamento que resultaram em 53 recomendações, incluindo investigação etiológica, tratamento com drogas antitireoidianas, iodo radioativo e cirurgia. Foram abordados ainda o hipertireoidismo em crianças, adolescentes ou pacientes grávidas e o manejo do hipertireoidismo em pacientes com oftalmopatia de Graves e com outras causas diversas de tireotoxicose. CONCLUSÕES: O diagnóstico clínico do hipertireoidismo, geralmente, não oferece dificuldade e a confirmação diagnóstica deverá ser feita com as dosagens das concentrações séricas de TSH e hormônios tireoidianos. O tratamento pode ser realizado com drogas antitireoidianas, administração de radioiodoterapia ou cirurgia de acordo com a etiologia da tireotoxicose, as características clínicas, disponibilidade local de métodos e preferências do médico-assistente e paciente.20523

Protective Human Leucocyte Antigen Haplotype, HLA-DRB1*01-B*14, against Chronic Chagas Disease in Bolivia

Chronic Chagas disease consists of four different forms categorized on the basis of their clinical manifestations, namely; cardiac, digestive, cardiodigestive and indeterminate. In Latin America, there are 8–10 million seropositive persons who are at risk of, or have already developed serious clinical complications and who have limited access to effective treatment. The cardiac and digestive forms are characterized by tissue damage caused by persistent infection of Trypanosoma cruzi and are thought to be modulated by host immunity. In our large scale screening for chronic Chagas disease in Santa Cruz, Bolivia, hearts and colons of 229 seropositive patients were examined. We found 31.4% of patients had abnormal electrocardiograms (ECGs), 15.7% presented with megacolon, 5.2% had a combination of abnormal ECG and megacolon, and 58.1% were of indeterminate status. Previously, we attempted to ascertain whether parasite genetic polymorphism might account for the differences in clinical manefestations, by analyzing parasite DNA taken from the same study group (with the addition of a further 62 megacolon post-operational patients). We found no relationships between parasite lineages and clinical disease form. The present study reveals that host HLA polymorphisms associate with clinical manifestations of Chagas

Regulatory T Cells Phenotype in Different Clinical Forms of Chagas' Disease

CD25High CD4+ regulatory T cells (Treg cells) have been described as key players in immune regulation, preventing infection-induced immune pathology and limiting collateral tissue damage caused by vigorous anti-parasite immune response. In this review, we summarize data obtained by the investigation of Treg cells in different clinical forms of Chagas' disease. Ex vivo immunophenotyping of whole blood, as well as after stimulation with Trypanosoma cruzi antigens, demonstrated that individuals in the indeterminate (IND) clinical form of the disease have a higher frequency of Treg cells, suggesting that an expansion of those cells could be beneficial, possibly by limiting strong cytotoxic activity and tissue damage. Additional analysis demonstrated an activated status of Treg cells based on low expression of CD62L and high expression of CD40L, CD69, and CD54 by cells from all chagasic patients after T. cruzi antigenic stimulation. Moreover, there was an increase in the frequency of the population of Foxp3+ CD25HighCD4+ cells that was also IL-10+ in the IND group, whereas in the cardiac (CARD) group, there was an increase in the percentage of Foxp3+ CD25High CD4+ cells that expressed CTLA-4. These data suggest that IL-10 produced by Treg cells is effective in controlling disease development in IND patients. However, in CARD patients, the same regulatory mechanism, mediated by IL-10 and CTLA-4 expression is unlikely to be sufficient to control the progression of the disease. These data suggest that Treg cells may play an important role in controlling the immune response in Chagas' disease and the balance between regulatory and effector T cells may be important for the progression and development of the disease. Additional detailed analysis of the mechanisms on how these cells are activated and exert their function will certainly give insights for the rational design of procedure to achieve the appropriate balance between protection and pathology during parasite infections

Generation and validation of genetic markers for the selection of carioca dry bean genotypes with the slow-darkening seed coat trait

Slow darkening (SD) is a trait that helps to maintain a brighter seed coat appearance in certain market classes of dry beans. The aim of this study was to generate new fluorescence-based markers and validate previously identified microsatellite markers for linkage to the SD trait in lines of the carioca market class. Four segregating populations were generated by Embrapa, the Brazilian Agricultural Research Corporation, from crosses between the SD cultivar BRSMG Madrepe´rola and the regular-darkening cultivars BRS Estilo, BRS Cometa, BRS Nota´vel and BRS Sublime. These populations were screened with the simple-sequence markers Pvsd- 1158 and PVM02TC116 and with a TaqManTM marker designed for the single-nucleotide polymorphism (SNP) PvbHLHp12804. A KASP marker was also designed for the PvbHLHp12804 marker for testing on advanced carioca lines developed by the University of Saskatchewan. In the carioca lines developed by Embrapa, PVM02TC116 proved unsuitable for marker-assisted selection (MAS). Both the Pvsd-1158 and PvbHLHp12804 markers were found to be tightly linked to the gene responsible for the SD trait, with genetic distances calculated at 2.8 cM for Pvsd-1158 and 2.0 and 3.1 cM for PvbbHLHp12804, respectively. These markers presented more than 97% of selection efficiency. The genotypic scoring using the PvbHLHp12804 KASP marker was perfectly correlated with the phenotype in all lines of the University of Saskatchewan. The results of this study validates the use of Pvsd-1158 as a gel-based marker for SD in carioca beans. The new fluorescence-based SNP PvbHLHp12804 markers exhibited very tight linkage to SD in carioca and pinto bean lines. These markers will be ideal for MAS for the SD trait in these market classes