RESUMO
The venom color variation of Crotalus durissus terrificus (Cdt) is attributed to the presence of the toxin L-amino acid oxidase (LAAO). During the venom milking routine of Instituto Butantan, we have noticed that most venoms of captive Cdt specimens show a yellowish color, while most venoms of wild specimens are white. Here we describe a comparative analysis of long-term captive (LTC) and recently wild-caught (RWC) Cdt, focusing on LAAO variation. For the identification of LAAO in individual venoms, four different approaches were employed: evaluation of the enzymatic activity, SDS-PAGE, Western blotting, and ELISA. In addition, mass spectrometry analysis was performed using pooled samples. Although some variation among these methodologies was observed, it was possible to notice that the presence of LAAO was significantly higher in the venom of LTC individuals. LAAO was identified in 60-80% LTC specimens and in only 10-12% of RWC specimens. Furthermore, this enzyme accounts for 5.6% of total venom proteins of LTC Cdt pooled venom, while it corresponds to only 0.7% of RWC Cdt pooled venom. These findings strongly suggest that captive maintenance increases the expression of LAAO in Cdt venom.
Assuntos
Venenos de Crotalídeos , Crotalus , L-Aminoácido Oxidase/metabolismo , Animais , Eletroforese em Gel de Poliacrilamida , Humanos , Venenos de SerpentesRESUMO
Plasma in several organisms has components that promote resistance to envenomation by inhibiting specific proteins from snake venoms, such as phospholipases A2 (PLA2s). The major hypothesis for inhibitor's presence would be the protection against self-envenomation in venomous snakes, but the occurrence of inhibitors in non-venomous snakes and other animals has opened new perspectives for this molecule. Thus, this study showed for the first time the structural and functional characterization of the PLA2 inhibitor from the Boa constrictor serum (BoaγPLI), a non-venomous snake that dwells extensively the Brazilian territory. Therefore, the inhibitor was isolated from B. constrictor serum, with 0.63% of recovery. SDS-PAGE showed a band at ~25 kDa under reducing conditions and ~20 kDa under non-reducing conditions. Chromatographic analyses showed the presence of oligomers formed by BoaγPLI. Primary structure of BoaγPLI suggested an estimated molecular mass of 22 kDa. When BoaγPLI was incubated with Asp-49 and Lys-49 PLA2 there was no severe change in its dichroism spectrum, suggesting a non-covalent interaction. The enzymatic assay showed a dose-dependent inhibition, up to 48.2%, when BoaγPLI was incubated with Asp-49 PLA2, since Lys-49 PLA2 has a lack of enzymatic activity. The edematogenic and myotoxic effects of PLA2s were also inhibited by BoaγPLI. In summary, the present work provides new insights into inhibitors from non-venomous snakes, which possess PLIs in their plasma, although the contact with venom is unlikely.
Assuntos
Boidae/sangue , Fosfolipases A2 do Grupo IV/antagonistas & inibidores , Inibidores de Fosfolipase A2/sangue , Sequência de Aminoácidos , Animais , Bothrops/metabolismo , Brasil , Venenos de Crotalídeos/antagonistas & inibidores , Venenos de Crotalídeos/química , Fosfolipases A2 do Grupo IV/química , Peso Molecular , Inibidores de Fosfolipase A2/química , Domínios e Motivos de Interação entre Proteínas , Venenos de Serpentes/antagonistas & inibidores , Venenos de Serpentes/química , Espectrometria de Massas em TandemRESUMO
O objetivo é analisar os dados obtidos sobre curso de capacitação de recursos digitais para empreendedoras com idade acima de 50 anos, em termos de aprendizagem, ganhos pessoais e profissionais. Amostra formada por 28 empresárias, com idade igual ou superior a 50 anos, residentes na cidade de São Paulo. Como resultados e discussão, destaca-se que 80% das alunas afirmaram que iriam aplicar os conhecimentos obtidos no dia a dia de suas empresas, demonstrando a pertinência do conteúdo com a prática das empreendedoras.
Objective: to analyze the data obtained on the training course of digital resources for entrepreneurs aged over 50, in terms of learning, personal and professional gain. Methods: a sample of 28 women entrepreneurs, aged 50 years or over, living in the city of São Paulo. Results and discussion: 80% of the students affirmed that they would apply the knowledge obtained in their companies' day-to-day activities, demonstrating the relevance of the content to the practice of entrepreneurs.
El objetivo es analizar los datos obtenidos sobre el curso de capacitación en recursos digitales para emprendedores mayores de 50 años, en términos de aprendizaje, ganancias personales y profesionales. Una muestra de 28 mujeres de negocios, de 50 años o más, que residen en la ciudad de São Paulo. Como resultados y discusión, es digno de mención que el 80% de los estudiantes dijeron que aplicarían los conocimientos obtenidos en la vida diaria de sus empresas, lo que demuestra la relevancia del contenido con la práctica de los emprendedores.
Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Mulheres , Mulheres Trabalhadoras/educação , Informática/educação , Empreendedorismo , Aprendizagem , Pessoa de Meia-IdadeRESUMO
O estudo objetivou descrever o curso Mulheres 50+ em Rede pela ótica das participantes. A amostra foi formada por 28 empresárias, com idade igual ou superior a 50 anos, residentes na cidade de São Paulo. O curso foi desenvolvido em dez módulos, no segundo semestre de 2015. Utilizado questionário com perguntas abertas e fechadas para avaliar a satisfação. Resultados e Discussão: idade média 57 anos, 62% com graduação completa. A avaliação do curso foi positiva, com ganhos pessoais e trocas de experiências entre as participantes.
The study aimed to describe the 50+ Women Network course from the participants' perspective. Methods: A sample of 28 women entrepreneurs, aged 50 years or over, living in the city of São Paulo. The course was developed in modules in the second half of 2015. A questionnaire was used with open and closed questions to evaluate satisfaction. Results and Discussion: mean age 57 years, 62% with full graduation. Evaluation of the course was positive, with gains and exchanges of experiences among participants.
El estudio objetivó describir el curso Mujeres 50+ en Red por la óptica de las participantes. La muestra fue formada por 28 empresarias, con edad igual o superior a 50 años, residentes en la ciudad de São Paulo. El curso fue desarrollado en diez módulos, en el segundo semestre de 2015. Utilizado cuestionario con preguntas abiertas y cerradas para evaluar la satisfacción. Resultados y Discusión: edad media 57 años, 62% con graduación completa. La evaluación del curso fue positiva, con ganancias personales e intercambios de experiencias entre las participantes.