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Lima, Walter dos Santos
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Id: lil-679429
Autor: Soares, Filippe Elias de Freitas; Braga, Fabio Ribeiro; Araújo, Jackson Victor de; Lima, Walter dos Santos; Mozzer, Lanuze Rose; Queiroz, José Humberto de.
Título: Optimization of protease production by the fungus Monacrosporium thaumasium and its action against Angiostrongylus vasorum larvae / Otimização da produção de protease pelo fungo Monacrosporium thaumasium e sua ação contra larvas de Angiostrongylus vasorum
Fonte: Rev. bras. parasitol. vet;22(2):285-288, Apr.-June 2013. tab.
Idioma: en.
Resumo: The objectives of this study were to optimize protease production from the nematophagous fungus Monacrosporium thaumasium (NF34a) and evaluate its larvicidal activity and biological stability. An isolate of the nematophagous fungus Monacrosporium thaumasium (NF34a) was used to produce the enzyme. The Plackett-Burman design was used in order to scan which components of the culture medium could have a significant influence on protease production by the fungus NF34a. An in vitro assay was also performed to evaluate the larvicidal activity of NF34a. It was observed that only one component of the culture medium (yeast extract), at the levels studied, had any significant effect (p < 0.05) on protease production. There was a reduction (p < 0.01) in the mean number of larvae recovered from the treated groups, compared with the control groups. The results confirm previous reports on the efficiency of nematophagous fungi for controlling nematode larvae that are potentially zoonotic. Thus, given the importance of biological control, we suggest that further studies should be conducted on the protease produced by the fungus Monacrosporium thaumasium.

O objetivo deste trabalho foi otimizar a produção de proteases do fungo nematófago Monacrosporium thaumasium (NF34a), avaliar sua atividade larvicida e sua estabilidade biológica. Foi utilizado um isolado do fungo nematófago Monacrosporium thaumasium (NF34a) para a produção de enzima. O delineamento Plackett-Burman foi utilizado com o objetivo de se escanear quais componentes do meio de cultura, poderiam ter influência significava na produção de protease pelo fungo NF34a. Foi realizado um ensaio in vitro para avaliar a atividade larvicida de NF34a. Observou-se que apenas um dos componentes do meio de cultura (extrato de levedura), nos níveis avaliados, apresentou um efeito significativo (p < 0,05) sobre a produção de protease. Houve redução (p < 0,01) entre as médias de larvas recuperadas dos grupos tratados em relação às dos grupos controle. Os resultados confirmam trabalhos anteriores da eficiência de fungos nematófagos no controle de larvas de nematóides potencialmente zoonóticos. Assim, devido a importância do controle biológico, os autores sugerem estudos mais aprofundados sobre a protease produzida pelo fungo Monacrosporium thaumasium.
Descritores: Angiostrongylus
Ascomicetos/enzimologia
Peptídeo Hidrolases/biossíntese
-Larva
Limites: Animais
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Id: lil-795230
Autor: Andreola, Patrícia; Demathé, Adriana; Galafassi, Daniel; Elsemann, Estelamari Barbieri; Elsemann, Rogério Brasiliense; Gazzoni, Alexandra Flávia.
Título: Estudo comparativo entre a produção de fosfolipases extracelulares e proteinases do gênero Candida isoladas a partir de infecções de cavidade oral / Comparative study between extracellular phospholipase and proteinase production in clinically important of the genus Candida isolated in oral candidiasis patients
Fonte: Rev. odontol. UNESP (Online);45(4):219-226, July-Aug. 2016. tab, ilus.
Idioma: pt.
Projeto: CNPq.
Resumo: Introdução: A habilidade da Candida spp. em produzir enzimas proteolíticas, tais como fosfolipase e proteinases, tem um papel importante na patogenicidade destas leveduras. Objetivo: Determinar as espécies causadoras das infecções orais por Candida spp., além de investigar a atividade in vitro das fosfolipases e proteinases em isolados clínicos do gênero Candida, provenientes de pacientes com candidíase oral. Material e método: Isolados de Candida spp., pertencentes à Coleção de Cultivos Fúngicos do Laboratório de Microbiologia e Patologia Oral do Departamento de Odontologia da Faculdade da Serra Gaúcha, foram analisados. Produção de fosfolipases foi analisada utilizando-se Ágar gema de ovo. Liberação de proteinases foi medida utilizando-se extrato de levedura adicionado à albumina bovina. Resultado: Um total de 35 isolados clínicos do gênero Candida foi testado. C. albicans foi a espécie predominante (77%). Os demais isolados identificados foram: C. parapsilosis (20%) e C. tropicalis (2%). Ao comparar a atividade de fosfolipase do grupo C. albicans com o grupo Candida não-albicans, foi encontrada diferença significativa (P=0,04). Não foi encontrada diferença significativa entre a C. albicans e a C. não-albicans, para a produção de proteinase. A liberação de proteinase foi significativamente maior quando comparada à produção de fosfolipase para o gênero Candida (P=0,04). Diferença estatisticamente significativa foi encontrada quando a atividade de fosfolipase e proteinase da C. albicans foi comparada à atividade das espécies de C. não-albicans (P=0,02). Conclusão: Diferentes quantificações de fosfolipase extracelular e atividade de proteinase têm sido atribuídas aos isolados clínicos de C. albicans quando comparados a outras espécies de Candida.

Introduction: The ability of the genus Candida to produce secretory enzimes such as proteinase and phospholipases to play an important role in the patogenicity of these yeasts. Objective: To determine the Candida species isolates from oral cavity infections and to investigate in vitro phospholipase and protease activities in clinically important of the genus Candida isolated in oral candidiasis patients. Material and method: Candida species isolated of the Fungal Culture Collection of Oral Microbiology and Pathology Testing Service Laboratory of the Dentistry Departament of Faculdade da Serra Gaúcha were evaluated. The phospholipases detection was assayed using the egg yolk agar plate method. Determination of protease production was performed in agar plates containing bovine serum albumine and yeasts extract. Result: A total of 35 isolates of the genus Candida were tested. C. albicans was the species predominant (77% of isolates), followed by C. parapsilosis (20%) and C. tropicalis (2%). When compared the phospholipase activity of the C. albicans group with the non-albicans Candida species types group was observed significant difference among of this groups (P=0.04). No statiscally significant difference between the C.albicans and non-albicans Candida species types when was compared to proteinase production. Proteinase production was higher and statiscally significant when compared to phospholipase activity in the genus Candida isolates (P=0.04). Statiscally significant differences were found between phosphoplipases and proteases activity for C. albicans when compared to non-albicans Candida species types (P=0.02). Conclusion: Differences in the secretion rates of phospholipase extracellular and proteases activity have been attributed to clinical strains of C. albicans when compared to others Candida species types.
Descritores: Peptídeo Hidrolases
Fosfolipases
Técnicas In Vitro
Candida
Candida albicans
Candidíase Bucal
Soroalbumina Bovina
Fatores de Virulência
Enzimas
-Leveduras
Infecções
Boca
Responsável: BR39.2 - Biblioteca Professora Maria Dilma de Oliveira Gonçalves


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Id: lil-314887
Autor: Sepúlveda Carvajal, Cecilia.
Título: Inflamación musculoesquelética: estado del arte / Musculoskeletal inflammation: state of the art
Fonte: Rev. chil. reumatol;17(4):147-155, 2001. ilus, tab.
Idioma: es.
Descritores: Doenças Musculoesqueléticas/fisiopatologia
-Linfócitos
Histamina
Serotonina
Adenosina
Ativação Plaquetária/fisiologia
Células Matadoras Naturais/fisiologia
Macrófagos/fisiologia
Mediadores da Inflamação/fisiologia
Neutrófilos/fisiologia
Óxido Nítrico/fisiologia
Peptídeo Hidrolases/fisiologia
Limites: Humanos
Responsável: CL1.1 - Biblioteca Central


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Id: lil-736980
Autor: M.M. Silva, Germana; Bezerra, Raquel P; Teixeira, José A; Porto, Tatiana S; Lima-Filho, José L; Porto, Ana Lúcia F.
Título: Fibrinolytic protease production by new Streptomyces sp: DPUA 1576 from Amazon lichens
Fonte: Electron. j. biotechnol;18(1):16-19, Jan. 2015. ilus, graf, tab.
Idioma: en.
Projeto: National Counsel of Technological and Scientific Development; . Fundação de Amparo à Ciência e Tecnologia do Estado de Pernambuco.
Resumo: Background Streptomyces sp. DPUA 1576 from Amazon lichens was studied to protease and fibrinolytic production. A 2² factorial experimental design was applied to optimize its protease enzyme production using two independent variables, namely soybean flour and glucose concentrations. Results The optimal conditions to obtain high protease production (83.42 U/mL) were 1.26% soybean flour and 1.23% glucose concentration. A polynomial model was fitted to correlate the relationship between the two variables and protease activity. In relation to fibrinolytic activity, the highest activity of 706.5 mm² was obtained at 1.7% soybean flour and 1.0% glucose concentration, which was 33% higher than plasmin. Fibrinolytic production was not optimized in the studied conditions. Conclusions These results show that the optimization of the culture medium can enhance protease production, thus becoming a good process for further research. In addition, Streptomyces sp. DPUA 1576, isolated from Amazon lichens, might be a potential strain for fibrinolytic protease production.
Descritores: Peptídeo Hidrolases/biossíntese
Streptomyces/enzimologia
Fibrinolíticos/metabolismo
-Soja
Modelos Estatísticos
Actinobacteria
Farinha
Glucose/análise
Líquens
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-830618
Autor: Neira, Víctor; Corbalán, Ramón; Pereira, Jaime; Panes, Olga; Garayar, Bernardita; Aizman, Andrés; Llevaneras, Silvana; Villarroel, Luis.
Título: Evaluación de la anticoagulación con rivaroxaban, en pacientes con fibrilación auricular no valvular de reciente diagnóstico / Evaluation of oral anticoagulation with rivaroxaban, in patients with new onset non valvular atrial fibrillation
Fonte: Rev. méd. Chile;144(9):1103-1111, set. 2016. graf, tab.
Idioma: es.
Projeto: Pontificia Universidad Católica de Chile. Dirección de Investigación; . Pontificia Universidad Católica de Chile. División de Enfermedades Cardiovasculares.
Resumo: Background: Atrial fibrillation (AF) generates a hypercoagulable state with an increased thrombin generation and raised levels of thrombin-antithrombin complexes, which results in a high risk of stroke and thromboembolism. Aim: To evaluate the anticoagulant effect of rivaroxaban by anti-Xa factor activity and its correlation with thrombin-antithrombin complexes, thrombin generation and prothrombin time in patients newly diagnosed with non-valvular AF. Patients and Methods: Prospective study in patients with indication of anticoagulation. Demographic variables, cardiovascular risk factors, CHA2DS2-VASc and HAS-BLED scores were recorded. Blood samples were taken at baseline, at 3 and 24 hours after the administration of the drug and at 30 days. Rivaroxaban levels, anti-Xa activity, prothrombin time, thrombin generation and plasma levels of thrombin-antithrombin complexes were determined. Results: We studied 20 patients aged 76.3 ± 8.0 years (60% female) with a CHA2DS2-VASc score > 2 points. The anti-Xa factor activity correlated with rivaroxaban plasma levels at 3 hours (r = 0.61, p < 0.01), at 24 hours (r = 0.85, p < 0.01) and at 30 days (r = 0.99, p < 0.01), with prothrombin time at 3 hours (r = -0.86, p = 0.019) and at 30 days (r = -0.63, p = 0.02) and with a sustained decrease in thrombin generation at 30 days of follow-up (r = -0.74, p < 0.01). There was no correlation with thrombin-antithrombin complexes (r = -0.02, p = 0.83). Conclusions: Rivaroxaban consistently inhibited the mild pro-coagulant state found in newly diagnosed non-valvular AF patients through the first 24 hours and this effect was maintained at 30 days. Plasma levels of the drug correlated with anti-Xa factor activity, thrombin generation and prothrombin time
Descritores: Peptídeo Hidrolases/efeitos dos fármacos
Fibrilação Atrial/sangue
Trombina/efeitos dos fármacos
Fator Xa/efeitos dos fármacos
Antitrombina III/efeitos dos fármacos
Inibidores do Fator Xa/farmacologia
Rivaroxabana/farmacologia
-Tempo de Protrombina
Fatores de Tempo
Trombina/metabolismo
Fator Xa/metabolismo
Administração Oral
Estudos Prospectivos
Limites: Humanos
Masculino
Feminino
Idoso
Idoso de 80 Anos ou mais
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1339188
Autor: Zhang, Yan; Yin, Wei-hua; Yang, Fan; An, Yun-qiang; Zhou, Wei; Yu, Hui; Xie, Hong; Zhang, Yan-ling; Zhu, Yue; Shen, Xiang-chun; Tian, Ruiqing.
Título: VEGF121 como Mediador de Efeitos Cardioprotetores Pós-Hipóxia via CaSR e via da Protease Dependente de Mitocôndria / VEGF121 Mediates Post-Hypoxia Cardioprotective Effects Via CaSR and Mitochondria-Dependent Protease Pathway
Fonte: Arq. bras. cardiol;117(3):476-483, Sept. 2021. graf.
Idioma: en; pt.
Projeto: The Joint Fund of Guizhou Province; . Science and Technology Department Academic New Seedling Project; . Department of Education Natural Science Youth Talent project of Guizhou Province.
Resumo: Resumo Fundamento: A doença cardiovascular é a principal causa de morte em todo o mundo. A apoptose mediada por hipóxia em cardiomiócitos é uma das principais causas de distúrbios cardiovasculares. O tratamento com a proteína do fator de crescimento endotelial vascular (VEGF, do inglês vascular endothelial growth factor) foi testado, mas as dificuldades operacionais limitaram seu uso. Entretanto, com os avanços da terapia gênica, aumentou o interesse na terapia gênica baseada no VEGF em doenças cardiovasculares. No entanto, o mecanismo preciso pelo qual a reposição de VEGF resgata os danos pós-hipóxia em cardiomiócitos não é conhecido. Objetivos: Investigar o efeito da expressão de VEGF121 pós-hipóxia utilizando cardiomiócitos de ratos neonatos. Métodos: Cardiomiócitos isolados de ratos neonatos foram utilizados para estabelecer um modelo in vitro de lesão cardíaca induzida por hipóxia. O efeito da superexpressão de VEGF, isolado ou em conjunto com inibidores de moléculas pequenas que têm como alvo os canais de cálcio, receptores sensíveis ao cálcio (CaSR, do inglês calcium-sensitive receptors) e calpaína, no crescimento e proliferação celular em lesão de cardiomiócitos induzidos por hipóxia, foram determinados com ensaio de MTT, coloração TUNEL, coloração com Anexina V/PI, lactato desidrogenase e atividade da caspase. Para análise estatística, um valor de p<0,05 foi considerado significativo. Resultados: Verificou-se que o efeito do VEGF121 foi mediado por CaSR e calpaína, mas não foi dependente dos canais de cálcio. Conclusões: Nossos resultados, mesmo em um ambiente in vitro, estabelecem as bases para uma validação futura e testes pré-clínicos da terapia gênica baseada em VEGF em doenças cardiovasculares.

Abstract Background: Cardiovascular disease is the major cause of death worldwide. Hypoxia-mediated apoptosis in cardiomyocytes is a major cause of cardiovascular disorders. Treatment with vascular endothelial growth factor (VEGF) protein has been tested but operational difficulties have limited its use. However, with the advancements of gene therapy, interest has risen in VEGF-based gene therapy in cardiovascular disorders. However, the precise mechanism by which VEGF replenishment rescues post-hypoxia damage in cardiomyocytes is not known. Objectives: To investigate the effect of post-hypoxia VEGF121 expression using neonatal rat cardiomyocytes. Methods: Cardiomyocytes isolated from neonatal rats were used to establish an in vitro model of hypoxia-induced cardiac injury. The effect of VEGF overexpression, alone or in combination with small-molecule inhibitors targeting calcium channel, calcium sensitive receptors (CaSR), and calpain on cell growth and proliferation on hypoxia-induced cardiomyocyte injury were determined using an MTT assay, TUNEL staining, Annexin V/PI staining, lactate dehydrogenase and caspase activity. For statistical analysis, a value of P<0.05 was considered to be significant. Results: The effect of VEGF121 was found to be mediated by CaSR and calpain but was not dependent on calcium channels. Conclusions: Our findings, even though using an in vitro setting, lay the foundation for future validation and pre-clinical testing of VEGF-based gene therapy in cardiovascular diseases.
Descritores: Fator A de Crescimento do Endotélio Vascular/metabolismo
Receptores de Detecção de Cálcio/metabolismo
-Peptídeo Hidrolases/metabolismo
Miócitos Cardíacos/metabolismo
Hipóxia
Mitocôndrias
Limites: Animais
Ratos
Responsável: BR1.1 - BIREME


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Id: biblio-893617
Autor: GOMES, Cinthya Cristina; GUIMARÃES, Ludmila Silva; PINTO, Larissa Christina Costa; CAMARGO, Gabriela Alessandra da Cruz Galhardo; VALENTE, Maria Isabel Bastos; SARQUIS, Maria Inêz de Moura.
Título: Investigations of the prevalence and virulence of Candida albicans in periodontal and endodontic lesions in diabetic and normoglycemic patients
Fonte: J. appl. oral sci;25(3):274-281, May-June 2017. tab, graf.
Idioma: en.
Projeto: Rio de Janeiro Research Foundation.
Resumo: Abstract Pulpal and periodontal tissues have similar microbiota that allows cross-contamination between the pulp and periodontal tissues. Objective The aim of this study was to investigate the prevalence of isolated Candida albicans from periodontal endodontic lesions in diabetic and normoglycemic patients, and the fungi's virulence in different atmospheric conditions. Material and Methods A case-control study was conducted on 15 patients with type 2 diabetes mellitus (G1) and 15 non-diabetics (G2) with periodontal endodontic lesions. Samples of root canals and periodontal pockets were plated on CHROMagar for later identification by polymerase chain reaction (PCR) and virulence test. Results C. albicans was identified in 79.2% and 20.8% of the 60 samples collected from diabetic and normoglycemic patients, respectively. Of the 30 samples collected from periodontal pockets, 13 showed a positive culture for C. albicans, with 77% belonging to G1 and 23% to G2. Of the 11 positive samples from root canals, 82% were from G1 and 18% from G2. Production of proteinase presented a precipitation zone Pz<0.63 of 100% in G1 and 72% in G2, in redox and negative (Pz=1), under anaerobic conditions in both groups. Hydrophobicity of the strains from G1 indicated 16.4% with low, 19.3% with moderate, and 64.3% with high hydrophobicity in redox. In G2, 42.2% had low, 39.8% had moderate, 18% had high hydrophobicity in redox. In anaerobic conditions, G1 showed 15.2% with low, 12.8% with moderate, and 72% with high hydrophobicity; in G2, 33.6% had low, 28.8% had moderate, and 37.6% had high hydrophobicity. There was statistical difference in the number of positive cultures between G1 and G2 (p<0.05) with predominance in G1. There was statistical difference for all virulence factors, except hemolysis (p=0.001). Conclusions Candida albicans was isolated more frequently and had higher virulence in diabetic patients.
Descritores: Doenças Periodontais/microbiologia
Candida albicans/isolamento & purificação
Candida albicans/patogenicidade
Doenças da Polpa Dentária/microbiologia
Diabetes Mellitus Tipo 2/microbiologia
-Oxirredução
Peptídeo Hidrolases/análise
Doenças Periodontais/fisiopatologia
Doenças Periodontais/diagnóstico por imagem
Bolsa Periodontal/microbiologia
Fosfolipases/análise
Virulência
DNA Fúngico
Radiografia Dentária
Estudos de Casos e Controles
Reação em Cadeia da Polimerase
Estatísticas não Paramétricas
Cavidade Pulpar/microbiologia
Doenças da Polpa Dentária/fisiopatologia
Doenças da Polpa Dentária/diagnóstico por imagem
Diabetes Mellitus Tipo 2/fisiopatologia
Eletroforese
Interações Hidrofóbicas e Hidrofílicas
Limites: Humanos
Masculino
Feminino
Adulto
Pessoa de Meia-Idade
Idoso
Responsável: BR1.1 - BIREME


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Id: biblio-954707
Autor: Lee, Chi-Chiu; Tsai, Wann-Sheng; Hsieh, Hernyi Justin; Hwang, Deng-Fwu.
Título: Hemolytic activity of venom from crown-of-thorns starfish Acanthaster planci spines
Fonte: J. venom. anim. toxins incl. trop. dis;19:22, maio 2013. tab, graf.
Idioma: en.
Resumo: Background : The crown-of-thorns starfish Acanthaster planci is a venomous species from Taiwan whose venom provokes strong hemolytic activity. To understand the hemolytic properties of A. planci venom, samples were collected from A. planci spines in the Penghu Islands, dialyzed with distilled water, and lyophilized into A. planci spine venom (ASV) powder. Results : Both crude venom and ASV cause 50% hemolysis at a concentration of 20 μg/mL. The highest hemolytic activity of ASV was measured at pH 7.0-7.4; ASV-dependent hemolysis was sharply reduced when the pH was lower than 3 or greater than 8. There was almost no hemolytic activity when the Cu2+ concentration was increased to 10 mM. Furthermore, incubation at 100°C for 30 to 60 minutes sharply decreased the hemolytic activity of ASV. After treatment with the protease α-chymotrypsin, the glycoside hydrolase cellulase, and the membrane component cholesterin, the hemolytic activity of ASV was significantly inhibited. Conclusions : The results of this study provide fundamental information about A. planci spine venom. The hemolytic activity was affected by pH, temperature, metal ions, EDTA, cholesterin, proteases, and glycoside hydrolases. ASV hemolysis was inhibited by Cu2+, cholesterin, α-chymotrypsin, and cellulose, factors that might prevent the hemolytic activity of venom and provide the medical treatment for sting.(AU)
Descritores: Peptídeo Hidrolases
Coluna Vertebral
Estrelas-do-Mar
Hemólise
Limites: Animais
Responsável: BR68.1 - Biblioteca Virginie Buff D'Ápice


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Id: biblio-954804
Autor: Campos, Fabiana V; Menezes, Thiago N; Malacarne, Pedro F; Costa, Fábio L. S; Naumann, Gustavo B; Gomes, Helena L; Figueiredo, Suely G.
Título: A review on the Scorpaena plumieri fish venom and its bioactive compounds
Fonte: J. venom. anim. toxins incl. trop. dis;22:35, 2016. tab, graf, ilus.
Idioma: en.
Projeto: FAPES/CNPq; . FAPES-Profix; . INCTTOX/CNPq; . CAPES; . CAPES; . AUXPE Toxinologia.
Resumo: The most poisonous fish species found along the Brazilian coast is the spotted scorpionfish Scorpaena plumieri. Though hardly ever life-threatening to humans, envenomation by S. plumieri can be quite hazardous, provoking extreme pain and imposing significant socioeconomic costs, as the victims may require days to weeks to recover from their injuries. In this review we will walk the reader through the biological features that distinguish this species as well as the current epidemiological knowledge related to the envenomation and its consequences. But above all, we will discuss the challenges involved in the biochemical characterization of the S. plumieri venom and its compounds, focusing then on the successful isolation and pharmacological analysis of some of the bioactive molecules responsible for the effects observed upon envenomation as well as on experimental models. Despite the achievement of considerable progress, much remains to be done, particularly in relation to the non-proteinaceous components of the venom. Therefore, further studies are necessary in order to provide a more complete picture of the venom's chemical composition and physiological effects. Given that fish venoms remain considerably less studied when compared to terrestrial venoms, the exploration of their full potential opens a myriad of possibilities for the development of new drug leads and tools for elucidating the complex physiological processes.(AU)
Descritores: Peptídeo Hidrolases
Venenos de Peixe/toxicidade
Peixes
-Inflamação
Limites: Animais
Responsável: BR68.1 - Biblioteca Virginie Buff D'Ápice


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Id: biblio-1153464
Autor: Silva, J A F; Silva, M K S; Silva, T A; Costa, L D A; Leal, M L E; Bezerra, R S; Costa, H M S; Freitas-Júnior, A C V.
Título: Obtainment and characterization of digestive aspartic proteases from the fish Caranx hippos (Linnaeus, 1766) / Obtenção e caracterização de proteases aspárticas digestórias do peixe Caranx hippos (Linnaeus, 1766)
Fonte: Braz. j. biol;82:e234500, 2022. tab, graf.
Idioma: en.
Resumo: Abstract This work aimed to obtain aspartic proteases of industrial and biotechnological interest from the stomach of the crevalle jack fish (Caranx hippos). In order to do so, a crude extract (CE) of the stomach was obtained and subjected to a partial purification by salting-out, which resulted in the enzyme extract (EE) obtainment. EE proteases were characterized physicochemically and by means of zymogram. In addition, the effect of chemical agents on their activity was also assessed. By means of salting-out it was possible to obtain a purification of 1.6 times with a yield of 49.4%. Two acid proteases present in the EE were observed in zymogram. The optimum temperature and thermal stability for EE acidic proteases were 55 ºC and 45 °C, respectively. The optimum pH and pH stability found for these enzymes were pH 1.5 and 7.0, respectively. Total inhibition of EE acid proteolytic activity was observed in the presence of pepstatin A. dithiothreitol (DTT) and Ca2+ did not promote a significant effect on enzyme activity. In the presence of heavy metals, such as Al3+, Cd2+ and Hg2+, EE acidic proteases showed more than 70% of their enzymatic activity. The results show that it is possible to obtain, from the stomach of C. hippos, aspartic proteases with high proteolytic activity and characteristics that demonstrate potential for industrial and biotechnological applications.

Resumo Este trabalho objetivou obter proteases aspárticas de interesse industrial e biotecnológico a partir do estômago do peixe xaréu (Caranx hippos). Para isso, foi obtido um extrato bruto do estômago, o qual foi submetido a uma purificação parcial por salting-out onde se obteve o extrato enzimático (EE). As proteases do EE foram caracterizadas físico-quimicamente e através de zimograma. Além disso, o efeito de agentes químicos sobre sua atividade também foi avaliado. Através de salting-out foi possível obter uma purificação de 1,6 vezes com rendimento de 49,4%. Foram observadas duas proteases ácidas presentes no EE através de zimograma. A temperatura ótima e a estabilidade térmica para as proteases ácidas do EE foram de 55 ºC e 45 °C, respectivamente. O pH ótimo e a estabilidade ao pH encontrados para estas enzimas foram o pH 1,5 e 7,0, respectivamente. Observou-se a inibição total da atividade proteolítica ácida do EE na presença de pepstatina A. O ditiotreitol (DTT) e o Ca2+ não promoveram efeito significativo na atividade enzimática. Na presença de metais pesados, como Al3+, Cd2+ e Hg2+, o EE manteve mais de 70% de atividade enzimática do EE. Os resultados mostram que é possível obter, a partir do estômago de C. hippos, proteases aspárticas com alta atividade proteolítica e características que demonstram potencial para aplicações industriais e biotecnológicas.
Descritores: Peptídeo Hidrolases
-Temperatura
Concentração de Íons de Hidrogênio
Limites: Animais
Responsável: BR1.1 - BIREME



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