Myostatin gene knockdown through lentiviral-mediated delivery of shRNA for in vitro production of transgenic bovine embryos

Myostatin is described as a negative regulator of the skeletal muscle growth. Genetic engineering, in order to produce animals with double the muscle mass and that can transmit the characteristic to future progeny, may be useful. In this context, the present study aimed to analyse the feasibility of lentiviral-mediated delivery of short hairpin RNA (shRNA) targeting of myostatin into in vitro produced transgenic bovine embryos. Lentiviral vectors were used to deliver a transgene that expressed green fluorescent protein (GFP) and an shRNA that targeted myostatin. Vector efficiency was verified through in vitro murine myoblast (C2C12) cell morphology after inductive differentiation and by means of real-time PCR. The lentiviral vector was microinjected into the perivitellinic space of in vitro matured oocytes. Non-microinjected oocytes were used as the control. After injection, oocytes were fertilized and cultured in vitro. Blastocysts were evaluated by epifluorescence microscopy. Results demonstrated that the vector was able to inhibit myostatin mRNA in C2C12 cells, as the transducted group had a less amount of myostatin mRNA after 72 h of differentiation (p < 0.05) and had less myotube formation than the non-transduced group (p < 0.05). There was no difference in cleavage and blastocyst rates between the microinjected and control groups. After hatching, 3.07% of the embryos exhibited GFP expression, indicating that they expressed shRNA targeting myostatin. In conclusion, we demonstrate that a lentiviral vector effectively performed shRNA myostatin gene knockdown and gene delivery into in vitro produced bovine embryos. Thus, this technique can be considered a novel option for the production of transgenic embryos and double muscle mass animals.

Detecção de salmonella sp. em psitacídeos de cativeiro através da reação em cadeia da polimerase (PCR)

O interesse da Medicina Veterinária nas espécies silvestres tem aumentado gradativamente, principalmente no estudo dos contextos ecológicos de saúde. Dentro desse contexto, autores realizaram estudos com o objetivo de conhecer a importância de Salmonella sp. na saúde das aves silvestres e seu potencial de transmissão para humanos e outros animais. Informações sobre a prevalência e distribuição dos sorovares de salmonelas na população de animais silvestres e domésticos são essenciais para relacionar os possíveis reservatórios que possam ser responsáveis pela transmissão dessa zoonose. Este trabalho teve como objetivo a detecção de Salmonella sp. em psitacídeos clinicamente sadios por Reação em Cadeia da Polimerase (PCR). Foram coletados suabes cloacais de 280 psitacídeos mantidos em cativeiro no Estado do Rio Grande do Sul, pertencentes a treze espécies, provenientes de um zoológico, um criadouro conservacionista e um criadouro comercial. O DNA das amostras foi extraído pelo método de fenol-clorofórmio e examinados pela PCR com a utilização de um par de iniciadores que amplifica um fragmento de 284 pb do gene invA pertencente ao gênero Salmonella, resultando em 37 amostras positivas. Não houve diferença na prevalência de salmonela entre os três plantéis nem entre as 13 espécies analizadas. Não foi possível a detecção desse patógeno pela PCR com iniciadores para a identificação de S. Typhimurium, S. Enteritidis, S. Pullorum e S. Gallinarum, nem através da Técnica Microbiológica Convencional nas amostras detectadas pela PCR genérica, provavelmente devido a maior sensibilidade e especificidade da PCR genérica. De acordo com a revisão bibliográfica realizada, este foi o primeiro trabalho de detecção direta de Salmonella em psitacídeos utilizando a PCR. Os resultados indicaram que aproximadamente 13,2% dos psitacídeos mantidos em cativeiro eram portadores assintomáticos ou eram transientemente infectados pelo gênero Salmonella.

Crises de liquidez e o seguro de depósitos bancários

A definição de um contrato de seguro depósito eficiente é analisada face à rede de segurança constituída nos sistemas financeiros. Busca-se, portanto, a harmonização do redesconto, o controle prudencial, o seguro depósito e eventuais programas de estímulos a fusões e aquisições de bancos em dificuldades (o PROER), de forma a equacionar o risco de crises de liquidez, ao mesmo tempo, em que minimizando o problema de assimetria de informações e os efeitos do elevado nível de alavancagem financeira dos bancos.

O impacto da afiliação a grupos econômicos no desempenho empresarial pós-aquisição no Brasil

Os grupos econômicos representam a internalização das falhas de mercado por empreendedores buscando superar as dificuldades de obtenção de recursos produtivos em mercados emergentes. A proposta deste trabalho é verificar se o acesso a um mercado interno de recursos permite a uma empresa afiliada a um grupo econômico obter um desempenho superior em relação a empresas independentes no caso de aquisições. Com base nas perspectivas teóricas da economia e da visão baseada em recursos, analisou-se o desempenho pós-aquisição de empresas de capital aberto afiliadas e não-afiliadas a grupos econômicos. Por meio de regressões com dados em painel, foram encontrados indícios de que a afiliação a grupos econômicos afeta negativamente o desempenho financeiro das empresas pós-aquisições. A principal implicação deste resultado para a prática refere-se à necessidade de revisão das estratégias de crescimento inorgânico e de seleção de empresas-alvo para aquisição por parte de afiliadas a grupos econômicos.

Pontas de pulverização e eletrificação das gotas na deposição da calda em plantas de crisântemo

O objetivo deste trabalho foi avaliar a deposição das gotas de pulverização, dotadas de carga elétrica (eletrostática), em comparação à técnica de pulverização convencional em crisântemo, com uso de diferentes pontas de pulverização. O experimento foi conduzido em delineamento inteiramente ao acaso, com oito tratamentos: combinação das pontas TXVK-3, AXI 110015, AXI 12002 TWIN e AXI 11003, com duas técnicas de pulverização (com e sem eletrostática), e quatro repetições. Cada repetição foi representada por 12 plantas, às quais foram afixados papéis do tipo mata-borrão na superfície abaxial e adaxial dos folíolos, e em duas posições da planta: ápice e base. Um corante marcador (Rodamina B) foi pulverizado na proporção de 5 g por 100 L d'água em cada um dos tratamentos. Os depósitos do marcador foram quantificados por fluorometria. As pontas com gotas de menor diâmetro mediano volumétrico (TXVK-3 e AXI 110015) apresentaram maiores depósitos na superfície abaxial da folha, quando se utilizou a pulverização eletrostática. Esse fato não foi observado, quando foram pulverizadas gotas com maior diâmetro mediano volumétrico e desprovidas de carga elétrica, nas diferentes partes da planta.

Mapping the knowledge covered by library classification systems

This study explores, in 3 steps, how the 3 main library classification systems, the Library of Congress Classification, the Dewey Decimal Classification, and the Universal Decimal Classification, cover human knowledge. First, we mapped the knowledge covered by the 3 systems. We used the 10 Pillars of Knowledge: Map of Human Knowledge, which comprises 10 pillars, as an evaluative model. We mapped all the subject-based classes and subclasses that are part of the first 2 levels of the 3 hierarchical structures. Then, we zoomed into each of the 10 pillars and analyzed how the three systems cover the 10 knowledge domains. Finally, we focused on the 3 library systems. Based on the way each one of them covers the 10 knowledge domains, it is evident that they failed to adequately and systematically present contemporary human knowledge. They are unsystematic and biased, and, at the top 2 levels of the hierarchical structures, they are incomplete.

Prevalence of metabolic syndrome in elderly Japanese-Brazilians

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Prevalence of Metabolic Syndrome in Japanese-Brazilians According to Specific Definitions for Ethnicity

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Crystallization and preliminary X-ray diffraction analysis of a class II phospholipase D from Loxosceles intermedia venom

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Project and Development of ErgoCoIn Version 2.0

Usability evaluation (UE) methods may have several purposes: identify and diagnose usability problems; evaluate the implementation, comparing the one under evaluation with other systems, and also perform acceptance test. These methods are usually expensive, time-consuming and complex. In this context, the ErgoCoIn 1.0 has being used as an accessible system that can be used by several people all over the world, even if they are not usability experts or evaluators. This paper aims to present a new version of this environment considering that its basic objective is to propose a higher quality for the questions. Additionally, a new presentation questionnaire window small enough to be used both in web browsers and mobile devices is proposed.

Independent impact of glycemia ad blood pressure in albuminuria on high-risk subjects for metabolic syndrome

Background: Microalbuminuria may reflect diffuse endothelial damage. Considering that diabetes and hypertension cause vasculopathy, we investigated associations of albumin-to-creatinine ratio (ACR) with plasma glucose and blood pressure levels in high-risk subjects for metabolic syndrome. Methods: A sample of 519 (246 men) Japanese-Brazilians (aged 60 ± 11 years), who participated in a population-based study, had their ACR determined in a morning urine specimen. Backward models of multiple linear regression were created for each gender including log-transformed values of ACR as dependent variable; an interaction term between diabetes and hypertension was included. Results: Macroalbuminuria was found in 18 subjects. ACR mean values for subjects with normal glucose tolerance, impaired fasting glycemia, impaired glucose tolerance and diabetes were 9.9 ± 6.0, 19.0 ± 35.4, 20.7 ± 35.4, and 33.9 ± 55.0 mg/g, respectively. Diabetic subjects showed higher ACR than the others (p < 0.05). An increase in the proportion of albuminuric subjects was observed as glucose metabolism deteriorated (4.9, 17.0, 23.0 and 36.0%). Stratifying into 4 groups according to postchallenge glycemia (< 7.8 mmol/l, n = 9 1; ≥ 7.8 mmol/l, n = 4 10) and hypertension, hypertensive and glucose-intolerant subgroups showed higher ACR values. ACR was associated with gender, waist circumference, blood pressure, plasma glucose and triglyceride (p < 0.05); albuminuric subjects had significantly higher levels of such variables than the normoalbuminuric ones. In the final models of linear regression, systolic blood pressure and 2-hour glycemia were shown to be independent predictors of ACR for both genders (p < 0.05). In men, also waist was independently associated with ACR. No interaction was detected between diabetes and hypertension. Conclusions: These findings suggest that both glucose intolerance and hypertension could have independent but not synergistic effects on endothelial function - reflected by albumin loss in urine. Such hypothesis needs to be confirmed in prospective studies. © 2004 Dustri-Verlag Dr. K. Feistle.

Autoimmunity does not contribute to the highly prevalent glucose metabolism disturbances in a Japanese Brazilian population

The Japanese Brazilian population has one of the highest prevalences of diabetes worldwide. Despite being non-obese according to standard definitions, their body fat distribution is typically central. We investigated whether a subset of these subjects had autoantibodies that would suggest a slowly progressive form of type 1 diabetes. A total of 721 Japanese Brazilians (386 men) in the 30- to 60-year age group underwent clinical examination and laboratory procedures, including a 75-g oral glucose tolerance test and determinations of serum autoantibodies. Antibodies to glutamic acid decarboxylase (GADab) were determined by radioimmunoassay and to thyroglobulin (TGab) and thyroperoxidase (TPOab) by flow-cytometry assays. Mean body mass index was 25.2 ± 3.8 kg/m2, but waist circumference was elevated according to the Asian standards. Diabetes, impaired glucose tolerance, and impaired fasting glycemia were found in 31%, 22%, and 22%, respectively, and 53% of the subjects had metabolic syndrome. Glutamic acid decarboxylase (GADab) was positive in 4.72%, TGab in 9.6%, and TPOab in 10% of the whole sample. When participants were stratified according to the presence of thyroid antibodies, similar frequencies of GADab were found in positive and negative groups. The prevalence rates of glucose metabolism disturbances did not differ between GADab positive and negative groups. Our data did not support the view that autoimmune injury could contribute to the high prevalence of diabetes seen in Japanese Brazilians, and the presence of co-morbidities included in the spectrum of metabolic syndrome favors the classification as type 2 diabetes.

Structure of a novel class II phospholipase D: Catalytic cleft is modified by a disulphide bridge

Phospholipases D (PLDs) are principally responsible for the local and systemic effects of Loxosceles envenomation including dermonecrosis and hemolysis. Despite their clinical relevance in loxoscelism, to date, only the SMase I from Loxosceles laeta, a class I member, has been structurally characterized. The crystal structure of a class II member from Loxosceles intermedia venom has been determined at 1.7. Å resolution. Structural comparison to the class I member showed that the presence of an additional disulphide bridge which links the catalytic loop to the flexible loop significantly changes the volume and shape of the catalytic cleft. An examination of the crystal structures of PLD homologues in the presence of low molecular weight compounds at their active sites suggests the existence of a ligand-dependent rotamer conformation of the highly conserved residue Trp230 (equivalent to Trp192 in the glycerophosphodiester phosphodiesterase from Thermus thermophofilus, PDB code: 1VD6) indicating its role in substrate binding in both enzymes. Sequence and structural analyses suggest that the reduced sphingomyelinase activity observed in some class IIb PLDs is probably due to point mutations which lead to a different substrate preference. © 2011 Elsevier Inc.

O riso pela fama: análise dos filmes publicitários da topper e Confederação Brasileira de Rugby

Pós-graduação em Comunicação - FAAC

Primeiro Colóquio Brasileiro de Matemática: identificação de um registro e pequenas biografias de seus participantes

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)