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Id: biblio-950843
Autor: Negrón-Oyarzo, Ignacio; Lara-Vásquez, Ariel; Palacios-García, Ismael; Fuentealba, Pablo; Aboitiz, Francisco.
Título: Schizophrenia and reelin: a model based on prenatal stress to study epigenetics, brain development and behavior
Fonte: Biol. Res;49:1-10, 2016. ilus, graf.
Idioma: en.
Projeto: Millennium Center for the Neuroscience of Memory; . Ministry of Economy, Development and Tourism; . FONDECYT.
Resumo: Schizophrenia is a severe psychiatric disorder that results in a significant disability for the patient. The disorder is characterized by impairment of the adaptive orchestration of actions, a cognitive function that is mainly dependent on the prefrontal cortex. This behavioral deficit, together with cellular and neurophysiological alterations in the prefrontal cortex, as well as reduced density of GABAergic cells and aberrant oscillatory activity, all indicate structural and functional deficits of the prefrontal cortex in schizophrenia. Among the several risk factors for the development of schizophrenia, stress during the prenatal period has been identified as crucial. Thus, it is proposed that prenatal stress induces neurodevelopmental alterations in the prefrontal cortex that are expressed as cognitive impairment observed in schizophrenia. However, the precise mechanisms that link prenatal stress with the impairment of prefrontal cortex function is largely unknown. Reelin is an extracellular matrix protein involved in the development of cortical neural connectivity at embryonic stages, and in synaptic plasticity at postnatal stages. Interestingly, down-regulation of reelin expression has been associated with epigenetic changes in the reelin gene of the prefrontal cortex of schizophrenic patients. We recently showed that, similar to schizophrenic patients, prenatal stress induces down-expression of reelin associated with the methylation of its promoter in the rodent prefrontal cortex. These alterations were paralleled with altered prefrontal cortex functional connectivity and impairment in prefrontal cortex-dependent behavioral tasks. Therefore, considering molecular, cellular, physiological and behavioral evidence, we propose a unifying framework that links prenatal stress and prefrontal malfunction through epigenetic alterations of the reelin gene.
Descritores: Efeitos Tardios da Exposição Pré-Natal/fisiopatologia
Esquizofrenia/etiologia
Esquizofrenia/fisiopatologia
Estresse Fisiológico/fisiologia
Encéfalo/embriologia
Serina Endopeptidases/genética
Moléculas de Adesão Celular Neuronais/genética
Proteínas da Matriz Extracelular/genética
Epigênese Genética/fisiologia
Proteínas do Tecido Nervoso/genética
-Transtornos do Comportamento Social/fisiopatologia
Encéfalo/fisiopatologia
Expressão Gênica
Fatores de Risco
Transtornos Cognitivos/fisiopatologia
Metilação de DNA
Limites: Humanos
Feminino
Gravidez
Tipo de Publ: Research Support, Non-U.S. Gov't
Revisão
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-838089
Autor: Handa, Priya; Maliken, Bryan D; Nelson, James E; Hennessey, Kelly A; Vemulakonda, L. Akhila; Morgan-Stevenson, Vicki; Dhillon, Barjinder K; Gupta, Rohit; Yeh, Matthew M; Kowdley, Kris V.
Título: Differences in Hepatic Expression of Iron, Inflammation and Stress-Related Genes in Patients with Nonalcoholic Steatohepatitis
Fonte: Ann. hepatol;16(1):77-85, Jan.-Feb. 2017. graf.
Idioma: en.
Resumo: Abstract: Nonalcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease worldwide. We have previously shown that hepatic reticuloendothelial system (RES) iron deposition is associated with an advanced degree of nonalcoholic steatohepatitis (NASH) in humans. In this study, we aimed to determine differentially expressed genes related to iron overload, inflammation and oxidative stress pathways, with the goal of identifying factors associated with NASH progression. Seventy five patients with NAFLD were evaluated for their biochemical parameters and their liver tissue analyzed for NASH histological characteristics. Gene expression analysis of pathways related to iron homeostasis, inflammation and oxidative stress was performed using real-time PCR. Gene expression was compared between subjects based on disease status and presence of hepatic iron staining. We observed increased gene expression of hepcidin (HAMP) (2.3 fold, p = 0.027), transmembrane serine proteinase 6 (TMPRSS6) (8.4 fold, p = 0.003), signal transducer and activator of transcription 3 (STAT3) (5.5 fold, p = 0.004), proinflammatory cytokines; IL-1β (2.7 fold, p = 0.046) and TNF-α (3.8 fold, p = 0.001) in patients with NASH. TMPRSS6, a negative regulator of HAMP, is overexpressed in patients with NASH and HIF1α (hypoxia inducible factor-1) is downregulated. NAFLD patients with hepatic iron deposition exhibited higher hepcidin expression (3.1 fold, p = 0.04) but lower expression of cytokines. In conclusion, we observed elevated hepatic HAMP expression in patients with NASH and in NAFLD patients who had hepatic iron deposition, while proinflammatory cytokines displayed elevated expression only in patients with NASH, suggesting a regulatory role for hepcidin in NAFL to NASH transition and in mitigating inflammatory responses.
Descritores: Estresse Oxidativo/genética
Sobrecarga de Ferro/genética
Hepatopatia Gordurosa não Alcoólica/genética
Inflamação/genética
Ferro/análise
Fígado/química
-Serina Endopeptidases/genética
Regulação da Expressão Gênica
Fator de Necrose Tumoral alfa/genética
Fator de Necrose Tumoral alfa/sangue
Mediadores da Inflamação/sangue
Sobrecarga de Ferro/diagnóstico
Sobrecarga de Ferro/sangue
Fator de Transcrição STAT3/genética
Interleucina-1beta/genética
Interleucina-1beta/sangue
Reação em Cadeia da Polimerase em Tempo Real
Hepcidinas/genética
Hepatopatia Gordurosa não Alcoólica/diagnóstico
Hepatopatia Gordurosa não Alcoólica/sangue
Inflamação/diagnóstico
Inflamação/sangue
Fígado/patologia
Proteínas de Membrana/genética
Limites: Humanos
Masculino
Feminino
Pessoa de Meia-Idade
Responsável: BR1.1 - BIREME


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Id: biblio-888879
Autor: Candiotto, F B; Freitas-Júnior, A C V; Neri, R C A; Bezerra, R S; Rodrigues, R V; Sampaio, L A; Tesser, M B.
Título: Characterization of digestive enzymes from captive Brazilian flounder Paralichthys orbignyanus / Caracterização de enzimas digestivas do linguado brasileiro de cativeiro Paralichthys orbignyanus
Fonte: Braz. j. biol;78(2):281-288, May-Aug. 2018. tab, graf.
Idioma: en.
Resumo: Abstract Knowledge of specific enzyme activity, along with animal habits and digestive capacity is essential in formulating an appropriate diet for any species. In this study, we evaluated and characterized the activity of digestive enzymes present in the liver, intestine, and stomach of Paralichthys orbignyanus. The effects of pH and temperature on enzyme activity were also evaluated via the use of specific substrates. The use of specific substrates and inhibitors showed strong evidence of the presence of trypsin (BApNA= 0.51 ± 0.2 mU mg-1), chimotrypsin (SApNA= 2.62 ± 1.8 mU mg-1), and aminopeptidases (Leu-p-Nan =0.9709 ± 0.83 mU mg-1) in the intestine. Optimum pH for the activity of trypsin, chemotrypsin, leucino aminopeptidase, amilase, and pepsin were 9.5, 9.0, 8.0, 7.5, and 3.5, respectively, while optimum temperatures were 50, 50, 50, 40, and 45 °C, respectively. These results provide additional information regarding the biology of Brazilian flounder and can be used as a basis for further studies regarding fish feeding physiology.

Resumo O conhecimento da atividade enzimática é essencial para formular uma correta dieta específica para espécie, além de estarem correlacionadas com o hábito da alimentação e capacidade digestive. Neste estudo determinamos e caracterizamos a atividade enzimática presente no intestino, estômago e fígado do linguado Paralichthys orbignyanus. Os efeitos da temperatura e pH sobre a atividade enzimática também foram avaliados utilizando substratos específicos. O uso de substratos e inibidores específicos mostrou uma forte evidência da presença da tripsina (BApNA = 0,51 ± 0,2 mU mg-1), quimotripsina (SAPNA = 2,62 ± 1,8 mU mg-1), e as aminopeptidases (Leu-p-Nan = 0,97 ± 0,83 mU mg-1) no intestino. O pH ótimo observado para a atividade de tripsina, quimotripsina, leucino aminopeptidase, amilase e pepsina foi 9,5, 9,0, 8,0, 7,5 e 3,5, respectivamente. A temperatura ótima observada foi 50, 50, 50, 40 e 45 °C, respectivamente. Estes resultados fornecem informações adicionais sobre a biologia do linguado brasileiro e pode ser usado como base para novos estudos sobre fisiologia alimentar.
Descritores: Linguado/fisiologia
Proteínas de Peixes/metabolismo
Proteínas de Peixes/química
Trato Gastrointestinal/enzimologia
Aminopeptidases/metabolismo
Aminopeptidases/química
-Temperatura
Estabilidade Enzimática
Brasil
Serina Endopeptidases/metabolismo
Serina Endopeptidases/química
Concentração de Íons de Hidrogênio
Fígado/enzimologia
Limites: Animais
Responsável: BR1.1 - BIREME


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Tersariol, I. L. S
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Id: lil-303555
Autor: Tersariol, I. L. S; Pimenta, D. C; Chagas, J. R; Almeida, P. C.
Título: Proteinase activity regulation by glycosaminoglycans
Fonte: Braz. j. med. biol. res = Rev. bras. pesqui. méd. biol;35(2):135-144, Feb. 2002. tab.
Idioma: en.
Conferência: Apresentado em: SIMEC 2000 - International Symposium on Extracellular Matrix, Angra dos Reis, 24-27 Sept. 2000.
Resumo: There are few reports concerning the biological role and the mechanisms of interaction between proteinases and carbohydrates other than those involved in clotting. It has been shown that the interplay of enzymes and glycosaminoglycans is able to modulate the activity of different proteases and also to affect their structures. From the large number of proteases belonging to the well-known protease families and also the variety of carbohydrates described as widely distributed, only few events have been analyzed more deeply. The term "family" is used to describe a group of proteases in which every member shows an evolutionary relationship to at least one other protease. This relationship may be evident throughout the entire sequence, or at least in that part of the sequence responsible for catalytic activity. The majority of proteases belong to the serine, cysteine, aspartic or metalloprotease families. By considering the existing limited proteolysis process, in addition to the initial idea that the proteinases participate only in digestive processes, it is possible to conclude that the function of the enzymes is strictly limited to the cleavage of intended substrates since the destruction of functional proteins would result in normal tissue damage. In addition, the location as well as the eventual regulation of protease activity promoted by glycosaminoglycans can play an essential role in the development of several physiopathological conditions
Descritores: Endopeptidases
Glicosaminoglicanos
-Cisteína Endopeptidases
Serina Endopeptidases
Heparina
Inibidores de Serino Proteinase
Inibidores Teciduais de Metaloproteinases
Metaloproteinases da Matriz
Glicosaminoglicanos
Limites: Humanos
Animais
Tipo de Publ: Revisão
Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Texto completo
Id: lil-783800
Autor: Girgin, Mustafa; Binnetoglu, Kenan; Duman, Kazim; Kanat, Burhan Hakan; Cetinkaya, Ziya; Ayten, Refik; Ilhan, Yavuz Selim; Ilhan, Necip; Seker, Ibrahim; Timurkaan, Necati.
Título: Effects of platelet rich plasma on fascial healing in rats with fecal peritonitis
Fonte: Acta cir. bras;31(5):314-319, May 2016. tab, graf.
Idioma: en.
Resumo: ABSTRACT PURPOSE : To evaluate the effects of platelet rich plasma (PRP) on the healing of fascia wherein peritonitis has been created. METHODS: Twenty eight Wistar Albino rats were divided into four groups. Only a primary fascial repair following laparotomy was performed on Group 1, a primary fascial repair performed and PRP treatment applied following laparotomy on Group 2, and a fecal peritonitis created following laparotomy and a primary fascial repair carried out on Group 3. A fecal peritonitis was created following laparotomy and primary fascial repair and PRP treatment on the fascia was carried out on Group 4. RESULTS: TNF-α was found to be significantly lower in the control group (Group 1). It was detected at the highest level in the group in which fecal peritonitis was created and PRP applied (Group 4). TGF-β was determined as being significantly higher only in Group 4. Histopathologically, the differences between the groups in terms of cell infiltration and collagen deposition were not found to be significant. CONCLUSION: When platelet rich plasma was given histologically and biochemicaly as wound healing parameters cellular infiltration, collagen accumulation, and tissue hydroxyiproline levels were not increased but neovascularization, fibroblast activation and TNF Alfa levels were increased and PRP accelerated wound healing.
Descritores: Peritonite/complicações
Cicatrização
Plasma Rico em Plaquetas
Fáscia/fisiologia
-Peritonite/metabolismo
Serina Endopeptidases/metabolismo
Distribuição Aleatória
Fator de Crescimento Transformador beta/análise
Fator de Crescimento Transformador beta/metabolismo
Colágeno/efeitos dos fármacos
Colágeno/metabolismo
Fator de Necrose Tumoral alfa/análise
Fator de Necrose Tumoral alfa/metabolismo
Ratos Wistar
Gelatinases/metabolismo
Neovascularização Fisiológica
Modelos Animais
Fáscia/irrigação sanguínea
Hidroxiprolina/análise
Hidroxiprolina/metabolismo
Proteínas de Membrana/metabolismo
Limites: Animais
Responsável: BR1.1 - BIREME


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Id: biblio-1022642
Autor: Chen, Quangang; Chen, Renjin; Yuan, Honghua; Liu, Peng; Hu, Ankang; Wu, Lianlian; Liu, Jing.
Título: Immunosuppressive mechanism of Hypoderma lineatum secreted serine esterase, a potential modulatory method used to inhibit transplant rejection
Fonte: Electron. j. biotechnol;32:63-68, Mar. 2018. tab, graf.
Idioma: en.
Projeto: Natural Science Foundation of Jiangsu province; . Nature Science Foundation of Jiangsu Higher Education Institutions of China; . Natural Science Foundation of Jiangsu province; . China Postdoctoral Science Foundation funded project; . Jiangsu Planned Projects For Postdoctoral Research Funds; . Master's Innovation Team Project of Xuzhou Central Hospital.
Resumo: Background: Although immunosuppressive therapies have made organ transplantation a common medical procedure worldwide, chronic toxicity has a major issue for long-term treatment. One method to improve therapies and methods is the application of immunomodulatory agents from parasites such as Hypoderma lineatum. Hypodermin A (HA) is a serine esterase secreted by the larvae of Hypoderma lineatum, several studies demonstrated its immunosuppressive mechanism in vitro, and recently we discovered that HA inhibits the expression of interferon (IFN)-γ and interleukin (IL)-2 and activates IL-10 expression. Therefore, we hypothesized that it might be a potential agent used to block allograft rejections. However, most studies of the immunosuppressive mechanisms associated with HA were undertaken at the cellular level. In order to augment these studies, we evaluated the immunosuppressive effects of HA in vivo using an HA transgenic mouse model. Result: Our results revealed similar findings to those reported by in vitro studies, specifically that HA induced prostaglandin E2 expression, downregulated IFN-γ and IL-2 expression, and promoted IL-10 secretion via E-type prostanoid receptor 4. Additionally, we observed that HA overexpression inhibited lipopolysaccharide-induced TLR4 activation. These findings provide insight into a new potential agent capable of blocking graft rejection. Conclusion: Our founding suggested that HA-related treatment could be a promising option to improve the viability of grafts in human.
Descritores: Serina Endopeptidases/imunologia
Dípteros/enzimologia
Dípteros/imunologia
Rejeição de Enxerto/imunologia
-Ensaio de Imunoadsorção Enzimática
Serina Endopeptidases/metabolismo
Western Blotting
Citocinas
Imunossupressão
Interleucinas/antagonistas & inibidores
Interferons/antagonistas & inibidores
Interleucina-10/metabolismo
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Receptor 4 Toll-Like
Reação em Cadeia da Polimerase em Tempo Real
Rejeição de Enxerto/enzimologia
Rejeição de Enxerto/prevenção & controle
Limites: Animais
Camundongos
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1016095
Autor: Fitriani, Dewi; Rohman, M Saifur; Prijambada, Irfan D.
Título: Proteolytic activity of recombinant DegP from Chromohalobacter salexigens BKL5
Fonte: Electron. j. biotechnol;29:7-12, sept. 2017. ilus, graf, tab.
Idioma: en.
Resumo: Background: DegP is a serine protease that specifically cleaves and refolds unfolding proteins in the periplasmic space of the cells. To date, there is no information regarding DegP from halophilic bacteria. Chromohalobacter salexigens BKL5 is a moderately halophilic bacterium that has the ability to grow in a media containing more than 15% salt. Therefore, the objectives of this work were to clone and overexpress DegP-encoding gene from C. salexigens BKL5 and characterize its biochemical properties. Results: DegP-encoding gene was overexpressed in Escherichia coli BL21(DE3) CodonPlus in an active form. SDS-PAGE analysis showed that the molecular weight of the recombinant DegP was 45 kDa. Size-exclusion chromatography analysis suggested that recombinant DegP was present in two multimeric states, hexameric and dodecameric, with molecular weights of 297.9 and 579.12 kDa, respectively. Both conformations were enzymatically active when casein was used as substrate for enzymatic assay. Circular dichroism analysis showed that recombinant DegP was composed of 0.21­0.29 helical content, which was comparable to the helical content in the crystal structure of E. coli DegP. The basic/acidic residue ratio of recombinant DegP was 0.56, which was slightly higher than that of DegP from extreme halophiles (average, 0.45) but significantly lower than that of DegP from nonhalophiles (average, 0.94). Conclusions: Recombinant DegP from C. salexigens BKL5 showed proteolytic activity when ß-casein was used as a substrate. In silico analysis indicated that recombinant DegP had characteristics similar to those of halophilic proteins depending on its amino acid composition.
Descritores: Serina Endopeptidases/genética
Proteínas Periplásmicas/genética
Chromohalobacter/enzimologia
Proteólise
Proteínas de Choque Térmico/genética
-Proteínas Recombinantes
Serina Endopeptidases/metabolismo
Caseínas
Cromatografia em Gel
Dicroísmo Circular
Clonagem Molecular
Proteínas Periplásmicas/metabolismo
Eletroforese em Gel de Poliacrilamida
Escherichia coli
Salinidade
Chromohalobacter/genética
Proteínas de Choque Térmico/metabolismo
Peso Molecular
Responsável: CL1.1 - Biblioteca Central


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Texto completo SciELO Brasil
Texto completo
Id: lil-755800
Autor: McIntosh, Douglas; Bezerra, Rodrigo Alves; Luz, Hermes Ribeiro; Faccini, João Luiz Horacio; Gaiotto, Fernanda Amato; Giné, Gastón Andrés Fernandez; Albuquerque, George Rego.
Título: Detection of Rickettsia bellii and Rickettsia amblyommii in Amblyomma longirostre (Acari: Ixodidae) from Bahia state, Northeast Brazil
Fonte: Braz. j. microbiol;46(3):879-883, July-Sept. 2015. tab.
Idioma: en.
Resumo:

Studies investigating rickettsial infections in ticks parasitizing wild animals in the Northeast region of Brazil have been confined to the detection of Rickettsia amblyommii in immature stages of Amblyomma longirostre collected from birds in the state of Bahia, and in immatures and females of Amblyomma auriculariumcollected from the striped hog-nosed skunk (Conepatus semistriatus) and armadillos (Euphractus sexcinctus) in the state of Pernambuco. The current study extends the distribution of R. amblyommii (strain Aranha), which was detected in A. longirostre collected from the thin-spined porcupine Chaetomys subspinosus and the hairy dwarf porcupine Coendou insidiosus. In addition, we report the first detection of Rickettsia bellii in adults of A. longirostre collected from C. insidiosus in the state of Bahia.

.
Descritores: Ixodidae/microbiologia
Infecções por Rickettsia/microbiologia
Rickettsia/genética
Rickettsia/isolamento & purificação
Infestações por Carrapato/microbiologia
-Animais Selvagens
Tatus
Sequência de Bases
Aves
Brasil
Proteínas da Membrana Bacteriana Externa/genética
Proteínas de Bactérias/genética
Citrato (si)-Sintase/genética
DNA Bacteriano
Proteínas de Choque Térmico/genética
Mephitidae
Dados de Sequência Molecular
Tipagem Molecular
Porcos-Espinhos
Proteínas Periplásmicas/genética
Análise de Sequência de DNA
Serina Endopeptidases/genética
Limites: Animais
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Id: lil-753178
Autor: Maggi, Silviane; Faulhaber, Gustavo Adolpho Moreira.
Título: Lonomia obliqua Walker (Lepidoptera: Saturniidae): hemostasis implications / Lonomia obliqua Walker (Lepidoptera: Saturniidae): implicações em hemostasia.
Fonte: Rev. Assoc. Med. Bras. (1992);61(3):263-268, May-Jun/2015.
Idioma: en.
Resumo: Summary In southern Brazil, since 1989, several cases of accidents produced by unwilling contact with the body of poisonous caterpillars of the moth species Lonomia obliqua Walker, 1855 (Lepidoptera: Saturniidae), were described. L. obliqua caterpillars have gregarious behavior and feed on leaves of host trees during the night, staying grouped in the trunk during the day, which favors the occurrence of accidents with the species. This caterpillar has the body covered with bristles that on contact with the skin of individuals, breaks and release their contents, inoculating the venom into the victim. The basic constitution of the venom is protein and its components produce physiological changes in the victim, which include disturbances in hemostasis. Hemorrhagic syndrome associated with consumption coagulopathy, intravascular hemolysis and acute renal failure are some of the possible clinical manifestations related to poisoning by L. obliqua. Specific laboratory tests for diagnosis of poisoning have not been described previously. The diagnosis of poisoning is made based on the patient's medical history, clinical manifestations, erythrocyte levels, and, primarily, parameters that evaluate blood coagulation. Treatment is performed with the use of supportive care and the administration of specific hyperimmune antivenom. Poisoning can be serious and even fatal.

Na região Sul do Brasil, a partir de 1989, foram descritos inúmeros casos de contato acidental com o corpo de lagartas venenosas da mariposa da espécie Lonomia obliqua Walker, 1855 (Lepidoptera: Saturniidae). As lagartas de L. obliqua possuem comportamento gregário e alimentam- -se de folhas das árvores hospedeiras durante a noite, permanecendo agrupadas no tronco durante o dia, o que favorece a ocorrência de acidentes. Essa lagarta possui o corpo recoberto por cerdas urticantes, que, ao contato com a pele dos indivíduos, se rompem e liberam seu conteúdo, inoculando o veneno na vítima. A constituição básica do veneno é proteica e seus componentes produzem modificações fisiológicas no acidentado, que incluem distúrbios na hemostasia. Síndrome hemorrágica associada a coagulopatia de consumo, hemólise intravascular e insuficiência renal aguda são algumas das manifestações clínicas possíveis relacionadas ao envenenamento por L. obliqua. Ainda não foram descritas avaliações laboratoriais específicas para diagnóstico do envenenamento, que é feito com base na anamnese do paciente, nas manifestações clínicas, nos índices hematimétricos e, principalmente, nos parâmetros que avaliam a coagulação sanguínea. A terapêutica é feita com a utilização de medidas de suporte e com a administração de um soro heterólogo hiperimune específico. Os envenenamentos podem ser graves e até mesmo fatais.
Descritores: Venenos de Artrópodes/envenenamento
Hemostasia
Lepidópteros
-Venenos de Artrópodes/metabolismo
Brasil
Coagulação Sanguínea/efeitos dos fármacos
Coagulação Intravascular Disseminada/etiologia
Hemorragia/induzido quimicamente
Mariposas
Serina Endopeptidases/metabolismo
Limites: Animais
Humanos
Tipo de Publ: Revisão
Responsável: BR1.1 - BIREME


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Id: lil-746873
Autor: Matilde, Leonardo Saboia Vahia.
Título: Análise da expressão protéica e de peptidases em formas pré-imaginais e imagos de Aedes aegypti e Aedes albopictus / Analysis of protein expression and peptidases in pre-imaginal forms and adults of aedes aegypti and aedes albopictus.
Fonte: Rio de Janeiro; s.n; 2013. xiv,140 p. ilus, tab, graf, mapas.
Idioma: pt.
Tese: Apresentada a Instituto Oswaldo Cruz para obtenção do grau de Doutor.
Resumo: Os mosquitos Aedes albopictus (Skuse, 1894) e Aedes aegypti (Linnaeus, 1762) apresentam significativa importância epidemiológica dentro da família Culicidae por serem os principais vetores da febre amarela e dos quatro sorotipos do vírus da dengue. As peptidases ou proteases são enzimas que clivam ligações peptídicas. No sistema digestivo de invertebrados as peptidases, especificamente aquelas da classe das serino-peptidases, são abundantes e participam de vários processos fisiológicos, desempenhando um papel principal na hidrólise de proteínas destinadas à nutrição. Estas enzimas podem apresentar características espécie-específicas e essas diferenças são respostas adaptativas a diferentes estilos de vida, ambientes e habilidade de sobrevivência entre as espécies. Neste trabalho descreve-se a caracterização das atividades proteolíticas detectadas em extratos totais das formas pré-imaginais de A. aegypti e A. albopictus por enzimografia em uma dimensão. A estabilidade da expressão de peptidases nessas espécies foi avaliada pela comparação do perfil proteolítico de formas larvais obtidas de ovos de insetos recém coletados no ambiente com ovos de insetos mantidos em colônia por longo período. Foram também comparados os perfis proteolíticos das pupas com seus respectivos imagos, bem como a estabilidade térmica das peptidases detectadas. Observaram-se complexos perfis de serino-peptidases do tipo tripsina em ambas as espécies vetoras.

Contudo esses perfis apresentam diferenças espécie-específicas e também diferenças entre os distintos estágios evolutivos dentro de uma mesma espécie. Também, neste trabalho, descreve-se a caracterização das atividades proteolíticas detectadas no intestino médio de fêmeas de A. albopictus alimentadas com açúcar bem como o primeiro mapa proteô mico e a identificação de peptidases por eletroforese bi-dimensional e espectrometria de massas neste órgão. São discutidas aqui as estratégias usadas para analisar as peptidases expressas nesse tecido. As proteínas expressas no intestino médio de A. albopictus foram identificadas por similaridade com as sequencias de genoma de A. aegypti e distintas ferramentas bioinformáticas foram usadas para obter informação funcional de muitas dessas sequencias que estão pobremente anotadas. Foram identificadas 59 proteínas entre v Resumo as quais três serino-peptidases, e dessas, duas do tipo tripsina e uma quimiotripsina. Os resultados obtidos nos permitiram atribuir de maneira confiável um local de expressão para os genes de tripsina, tripsina alfa e quimotripsina. Em outras palavras, podemos afirmar que os genes acima mencionados se expressam no intestino médio de fêmeas adultas de A. albopictus alimentadas com açúcar. Este achado representa um pequeno, mas importante passo, para a atribuição funcional, ao nível de proteína, de genes codificantes para serino-peptidases do tipo tripsina e quimiotripsina no gênero Aedes.

The mosquitoes Aedes albopictus (Skuse, 1894) and Aedes aegypti (Linnaeus, 1762) havesignificant epidemiological importance within the family Culicidae because they are themain vectors of the yellow fever virus and the four serotypes of Dengue virus. Peptidasesor proteases are enzymes that hydrolyze peptide bonds. In the digestive system ofinvertebrates, the peptidases, specifically those from the class of serine peptidases, areabundant and participate in various physiological processes, playing a major role in thehydrolysis of proteins destined to nutrition. These enzymes may have species-specificcharacteristics, and such differences reflect adaptive responses to different lifestyles andenvironments. In this work we describe the characterization, using zymography, of theproteolytic activities detected in total extracts of larval forms of A. aegypti and A.albopictus. The stability of expression of peptidases in these species was evaluated bycomparing the proteolytic profile of larval forms obtained from eggs of newly collectedinsects to that from eggs of insect kept in colony for a long period. We also compared theproteolytic profiles of pupae as well as the thermal stability of the peptidases detected.Complex profiles of trypsin-like serine peptidases were observed in both species.

However, these profiles differ between species and also differences between the differentevolutionary stages were detected within the same species. Also, in this work, we describethe characterization of the proteolytic activities detected in the midgut of A. albopictusfemales fed on sugar as well as the first proteome map and proteomic identification ofpeptidases in this organ, using two-dimensional electrophoresis and mass spectrometry.The strategies used to analyze the proteases expressed in this tissue are discussed here. Theproteins expressed in the midgut of A. albopictus were identified by similarity with thegenome sequences of A. aegypti and various bioinformatic tools were used to obtainfunctional information for many of these sequences that are poorly annotated. Weidentified 59 proteins including three serine peptidases, and among these, two types oftrypsin-like and one chymotrypsin. The results obtained here allowed us to reliably assignviiiAbstracta localization for the expression of trypsin, trypsin-alpha and chymotrypsin genes. In otherwords, we can say that the above mentioned genes are expressed in the midgut of adultfemales of A. albopictus fed with sugar. This finding represents a small but important stepfor the functional assignment, at the protein level, of the genes coding for trypsin-like andchymotrypsin serine peptidases of the Aedes genus.
Descritores: Aedes/classificação
Peptídeo Hidrolases
Proteômica
Serina Endopeptidases
-Dengue
Febre Amarela
Responsável: BR15.1 - Biblioteca de Ciências Biomédicas
BR15.1



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