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Id: biblio-1133779
Autor: Melo, Carina Mucciolo; Prado, Henrique Pereira; Attie, Gabriela Araújo; Ruiz, Daniel Lacaz; Girão, Manoel João Batista Castello; Pinhal, Maria Aparecida da Silva.
Título: In silico investigation of heparanase-correlated genes in breast cancer subtypes / Investigação in silico dos genes correlacionados à heparanase nos diferentes subtipos de câncer de mama
Fonte: Einstein (Säo Paulo);18:eAO5447, 2020. graf.
Idioma: en.
Projeto: Fundação de Amparo à Pesquisa do Estado de São Paulo; . Conselho Nacional de Desenvolvimento Científico e Tecnológico.
Resumo: ABSTRACT Objective To investigate the possible genes that may be related to the mechanisms that modulate heparanase-1. Methods The analysis was conducted at Universidade Federal de São Paulo, on the data provided by: The Cancer Genome Atlas, University of California Santa Cruz Genome Browser, Kyoto Encyclopedia of Genes and Genomes Pathway Database, Database for Annotation, Visualization and Integrated Discovery Bioinformatics Database and the softwares cBioPortal and Ingenuity Pathway Analysis. Results Using messenger RNA expression pattern of different molecular subtypes of breast cancer, we proposed that heparinase-1 was co-related with its progression. In addition, genes that were analyzed presented co-expression with heparanase-1. The results that showed that heparanase-1 co-expressed with phosphoinositide 3-kinase adapter protein 1, sialic acid-binding immunoglobulin-like lectin 7, and leukocyte-associated immunoglobulin-like receptor 1 are directed related with immune system evasion during breast cancer progression. Furthermore, cathepsin L was co-expressed with heparanase-1 and transformed inactive heparanase-1 form into active heparanase-1, triggering extracellular matrix remodeling, which contributes to enhanced tumor-host interaction of the tumor. Conclusion The signaling pathway analysis using bioinformatics tools gives supporting evidence of possible mechanisms related to breast cancer development. Evasion genes of the immune system co-expressed with heparanase-1, a enzyme related with tumor progression.

RESUMO Objetivo Investigar os genes que podem estar relacionados aos mecanismos que modulam a heparanase-1. Métodos A análise foi realizada na Universidade Federal de São Paulo, utilizando dados fornecidos por: The Cancer Genome Atlas, University of California Santa Cruz Genome Browser, Kyoto Encyclopedia of Genes and Genomes Pathway Database, Database for Annotation, Visualization and Integrated Discovery Bioinformatics Database e os softwares cBioPortal e Ingenuity Pathway Analysis. Resultados Usando o perfil de expressão de RNA mensageiro de diferentes subtipos moleculares de câncer de mama, propusemos que a heparanase-1 esteve correlacionada com a progressão tumoral. Além disso, os genes analisados apresentaram coexpressão com heparanase-1. Os resultados mostraram que a heparanase-1 coexpressa com proteína adaptadora 1 da fosfoinositídeo 3-quinase, lectina 7 tipo Ig de ligação ao ácido siálico e receptor 1 do tipo imunoglobulina associado a leucócitos, estes genes estão diretamente relacionados à evasão do sistema imune durante a progressão do câncer de mama. Além disso, a catepsina L foi coexpressa com a heparanase-1 e transformou a forma inativa da heparanase-1 em heparanase-1 ativa, desencadeando o remodelamento da matriz extracelular, o que contribuiu para a interação do tumor com o ambiente tumoral. Conclusão A análise utilizando bioinformática fornece evidências de possíveis mecanismos relacionados ao desenvolvimento do câncer de mama. Genes de evasão do sistema imune foram coexpressos com a heparanase-1, uma enzima relacionada à progressão tumoral.
Descritores: Neoplasias da Mama/genética
Glucuronidase/genética
-Simulação por Computador
Limites: Humanos
Responsável: BR1.1 - BIREME


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Id: biblio-827746
Autor: Pinhal, Maria Aparecida Silva; Almeida, Maria Carolina Leal; Costa, Alessandra Scorse; Theodoro, Thérèse Rachell; Serrano, Rodrigo Lorenzetti; Machado Filho, Carlos D'Apparecida Santos.
Título: Expression of heparanase in basal cell carcinoma and squamous cell carcinoma
Fonte: An. bras. dermatol;91(5):595-600, Sept.-Oct. 2016. graf.
Idioma: en.
Resumo: Abstract: Background: Heparanase is an enzyme that cleaves heparan sulfate chains. Oligosaccharides generated by heparanase induce tumor progression. Basal cell carcinoma and squamous cell carcinoma comprise types of nonmelanoma skin cancer. Objectives: Evaluate the glycosaminoglycans profile and expression of heparanase in two human cell lines established in culture, immortalized skin keratinocyte (HaCaT) and squamous cell carcinoma (A431) and also investigate the expression of heparanase in basal cell carcinoma, squamous cell carcinoma and eyelid skin of individuals not affected by the disease (control). Methods: Glycosaminoglycans were quantified by electrophoresis and indirect ELISA method. The heparanase expression was analyzed by quantitative RT-PCR (qRTPCR). Results: The A431 strain showed significant increase in the sulfated glycosaminoglycans, increased heparanase expression and decreased hyaluronic acid, comparing to the HaCaT lineage. The mRNA expression of heparanase was significantly higher in Basal cell carcinoma and squamous cell carcinoma compared with control skin samples. It was also observed increased heparanase expression in squamous cell carcinoma compared to the Basal cell carcinoma. Conclusion: The glycosaminoglycans profile, as well as heparanase expression are different between HaCaT and A431 cell lines. The increased expression of heparanase in Basal cell carcinoma and squamous cell carcinoma suggests that this enzyme could be a marker for the diagnosis of such types of non-melanoma cancers, and may be useful as a target molecule for future alternative treatment.
Descritores: Neoplasias Cutâneas/enzimologia
Carcinoma Basocelular/enzimologia
Carcinoma de Células Escamosas/enzimologia
Glucuronidase/metabolismo
Glicosaminoglicanos/metabolismo
-RNA Mensageiro/metabolismo
Queratinócitos/metabolismo
Pálpebras/enzimologia
Reação em Cadeia da Polimerase em Tempo Real/métodos
Glucuronidase/genética
Glicosaminoglicanos/análise
Ácido Hialurônico/análise
Ácido Hialurônico/metabolismo
Limites: Humanos
Responsável: BR1.1 - BIREME


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Id: biblio-1009000
Autor: Liu, Chunming; Yang, Bowen; Ming, Yuetong; Liu, Jianfeng; Cheng, Yunqing.
Título: Construction of an RNAi vector for knockdown of GM-ACS genes in the cotyledonary nodes of soybean
Fonte: Electron. j. biotechnol;26:40-45, Mar. 2017. ilus, graf.
Idioma: en.
Projeto: National Natural Science Foundation of China; . Special Foundation for Young Scientists of Jilin Province, China.
Resumo: Background: Ethylene plays an important role in the regulation of floral organ development in soybean, and 1-aminocyclopropane-1-carboxylate synthase (ACS) is a rate-limiting enzyme for ethylene biosynthesis. However, whether ACS also regulates floral organ differentiation in soybean remains unknown. To address this, we constructed an RNAi vector to inhibit ACS expression in cotyledonary nodes. Linear DNA cassettes of RNAi-ACS obtained by PCR were used to transform soybean cotyledonary nodes. Results: In total, 131 of 139 transiently transformed plants acquired herbicide resistance and displayed GUS activities in the new buds. In comparison to untransformed seedling controls, a greater number of flower buds were differentiated at the cotyledonary node; GM-ACS1 mRNA expression levels and ethylene emission in the transformed buds were reduced. Conclusion: These results indicate that the cotyledonary node transient transformation system may be suitable for stable transformation and that the inhibition of ACS expression may be an effective strategy for promoting floral organ differentiation in soybean.
Descritores: Soja/enzimologia
Soja/genética
Interferência de RNA
Liases/metabolismo
-Soja/crescimento & desenvolvimento
Transformação Genética
Expressão Gênica
Diferenciação Celular
Reação em Cadeia da Polimerase
Regulação da Expressão Gênica de Plantas
Etilenos/biossíntese
Resistência a Herbicidas
Vetores Genéticos
Glucuronidase
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-1008414
Autor: Liu, Juhua; Gao, Pengzhao; Sun, Xiuxiu; Zhang, Jing; Sun, Peiguang; Wang, Jiashui; Jia, Caihong; Zhang, Jianbin; Hu, Wei; Xu, Biyu; Jin, Zhiqiang.
Título: Efficient regeneration and genetic transformation platform applicable to five Musa varieties
Fonte: Electron. j. biotechnol;25:33-38, ene. 2017. tab, ilus.
Idioma: en.
Projeto: Modern Agro-industry Technology Research System; . National Nonprofit Institute Research Grant of Institute of Tropical Bioscience and Biotechnology CATAS-ITBB.
Resumo: Background: Banana (Musa spp.) is an important staple food, economic crop, and nutritional fruit worldwide. Conventional breeding has been seriously hampered by their long generation time, polyploidy, and sterility of most cultivated varieties. Establishment of an efficient regeneration and transformation system for banana is critical to its genetic improvement and functional genomics. Results: In this study, a vigorous and repeatable transformation system for banana using direct organogenesis was developed. The greatest number of shoots per explant for all five Musa varieties was obtained using Murashige and Skoog medium supplemented with 8.9 µM benzylaminopurine and 9.1 µM thidiazuron. One immature male flower could regenerate 380­456, 310­372, 200­240, 130­156, and 100­130 well-developed shoots in only 240­270 d for Gongjiao, Red banana, Rose banana, Baxi, and Xinglongnaijiao, respectively. Longitudinal sections of buds were transformed through particle bombardment combined with Agrobacterium-mediated transformation using a promoterless ß-glucuronidase (GUS) reporter gene; the highest transformation efficiency was 9.81% in regenerated Gongjiao plantlets in an optimized selection medium. Transgenic plants were confirmed by a histochemical assay of GUS, polymerase chain reaction, and Southern blot. Conclusions: Our robust transformation platform successfully generated hundreds of transgenic plants. Such a platform will facilitate molecular breeding and functional genomics of banana.
Descritores: Musa/crescimento & desenvolvimento
Musa/genética
-Regeneração
Transformação Genética
Imuno-Histoquímica
Southern Blotting
Reação em Cadeia da Polimerase
Plantas Geneticamente Modificadas
Agrobacterium tumefaciens/fisiologia
Musa/microbiologia
Organogênese Vegetal
Glucuronidase
Responsável: CL1.1 - Biblioteca Central


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Id: biblio-840021
Autor: Paula, Roberta S; Souza, Vinícius C; Machado-Silva, Wilcelly; Almeida, Bruno Ratier S; Daros, Andersen C; Gomes, Lucy; Ferreira, Aparecido P; Brito, Ciro J; Córdova, Cláudio; Moraes, Clayton F; Nóbrega, Otávio T.
Título: Serum Klotho (but not haplotypes) associate with the post-myocardial infarction status of older adults
Fonte: Clinics;71(12):725-732, Dec. 2016. tab, graf.
Idioma: en.
Projeto: CNPq; . FAPDF.
Resumo: OBJECTIVES: The number of deaths from vascular diseases is incredibly high worldwide, and reliable markers for major events are still needed. The current cross-sectional study investigated the association of Klotho haplotypes and Klotho serum levels with classic risk factors and a clinical history of vascular events. METHODS: Clinical, anthropometric, biochemical and nutritional assessments were conducted with 168 older adults, complemented by genotyping (rs9536314 and rs9527025) and the detection of serum Klotho (ELISA). RESULTS: Klotho levels and haplotypes did not associate with most classic risk factors for vascular events, including markers such as C-reactive protein and homocysteine. A positive association was only found between Klotho levels and the previous occurrence of a myocardial infarction by both correlational (p=0.006) and variance analyses (p<0.001), and these associations were independent of the context. CONCLUSION: Our results suggest that serum Klotho is higher in individuals with a clinical history of myocardial infarction but not with a history of coronary artery disease or stroke. None of the Klotho haplotypes were associated with the variables investigated herein.
Descritores: Glucuronidase/genética
Glucuronidase/sangue
Infarto do Miocárdio/sangue
-Valores de Referência
Doença da Artéria Coronariana/genética
Doença da Artéria Coronariana/sangue
Haplótipos
Ingestão de Energia
Proteína C-Reativa/análise
Ensaio de Imunoadsorção Enzimática
Biomarcadores/sangue
Avaliação Nutricional
Fatores Sexuais
Antropometria
Estudos Transversais
Fatores de Risco
Análise de Variância
Fatores Etários
Estatísticas não Paramétricas
Acidente Vascular Cerebral/genética
Acidente Vascular Cerebral/sangue
Técnicas de Genotipagem
Homocisteína/sangue
Infarto do Miocárdio/genética
Limites: Humanos
Masculino
Feminino
Pessoa de Meia-Idade
Idoso
Idoso de 80 Anos ou mais
Responsável: BR1.1 - BIREME


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Id: biblio-914354
Autor: Silva, André Luís Lopes da; Oliveira, Yohana de; Procopiuk, Marcia; Mudry, Clarissa de Souza; Brondani, Gilvano Ebling; Costa, Jefferson da Luz; Scheidt, Gessiel Newton.
Título: Expressão transiente do gene uidA em explantes foliares de Eucalyptus saligna Sm. transformado via Agrobacterium tumefaciens / Transient expression of uidA gene in leaf explants of Eucalyptus saligna Sm. transformed via Agrobacterium tumefaciens
Fonte: Biosci. j. (Online);29(1):1-7, jan./feb. 2013. ilus, tab.
Idioma: pt.
Resumo: O objetivo desse trabalho foi avaliar a influência do pré-cultivo de explantes foliares e do meio de cultura na ressuspensão de Agrobacterium tumefaciens para infecção dos explantes. Os meios MS/2 (50% da concentração de sais) e MS N/2 (50% da concentração de NH4NO3 e KNO3) + PGR (1,0µM de TDZ (thidiazuron) + 0,1 µM de ANA (ácido naftalenoacético)) foram testados na ressuspensão da bactéria para infecção dos explantes. O pré-cultivo consistiu da manutenção dos explantes em meio de cultura para formação de calos (MS N/2 + PGR) durante um dia, sendo o tratamento sem pré-cultivo consistituído dos explantes após a excissão dos mesmos. Os explantes foram mantidos no escuro a 25 ± 2ºC mediante a utilização de plástico preto. O delineamento usado foi o inteiramente casualisado com 20 explantes. Os experimentos foram repetidos duas vezes. O meio MS/2 promoveu resultados superiores (22,4%) comparado ao meio MS N/2 + PGR (14,5%) para a percentagem de área com expressão do gene uidA. Aos 7 dias de cultivo em meio seletivo, a percentagem de área expressando o gene uidA foi 1,6 no MS/2 e 0% para o MS N/2 + PGR. O pré-cultivo produziu resultados superiores aos encontrados sem pré-cultivo, atingindo 31,4% de expressão transiente e no tratamento sem pré-cultivo 2,1%. Após 7 dias de cultivo em meio seletivo, a percentagem de área de expressão dos explantes do tratamento com pré-cultivo permaneceu 4,8% e 0% para o tratamento sem pré-cultivo. Os resultados indicam que o précultivo e ressuspensão da bactéria em meio MS/2 aumentaram a eficiência da expressão transiente do gene uidA em explantes foliares de E. saligna.

The aim of this research was to evaluate the effect of the pre-culture of leaf explants and the effect of the culture medium for the Agrobacterium tumefaciens resuspension to the explant infection. The media, MS/2 (half strength) and MS N/2 (10.3 mM NH4NO3 and 9.4 mM KNO3) + PGR (1.0 µM TDZ (thidiazuron) and 0.1 µM NAA (1-Naphthaleneacetic acid)) were tested for the bacteria resuspension. The pre-culture consisted of the maintenance of the explants on culture medium for callus formation (MS N/2+PGR) during one day and the treatment without pre-culture consisted of the use of the explants after the excision of the same ones. At the end of the co-culture, the MS/2 promoted results superiors to the MS N/2+PGR, and the area percentage that presented expression of the gene uidA was of 22.4% compared at 14.5%. To the 7 days of culture on a medium with kanamycin, the area percentage expressing the gene uidA was 1.6 in MS/2 and 0% for the MS N/2+PGR. At the end of the co-culture, the pre-culture produced results superiors to the found in the treatment without pre-culture, reaching 31.4% of expression and in the treatment without pre-culture 2.1%. After 7 days of culture on a medium with kanamycin, the area percentage of explant expression of the treatment with pre-culture stayed 4.8% and 0% for the treatment without pre-culture. The results indicate that the pre-culture and the bacteria resuspension in MS/2 increase the efficiency of the transient expression of the gene uidA in leaf explants of E. saligna.
Descritores: Transformação Genética
Agrobacterium tumefaciens
Eucalyptus
Genes
Glucuronidase
Responsável: BR396.4


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Id: lil-733139
Autor: Rodrigues, Ana Paula dos Santos; Silveira, Erika Aparecida da.
Título: Correlação e associação de renda e escolaridade com condições de saúde e nutrição em obesos graves / Correlation and association of income and educational level with health and nutritional conditions among the morbidly obese
Fonte: Ciênc. saúde coletiva;20(1):165-174, 01/2015. tab.
Idioma: pt.
Resumo: O objetivo deste artigo é investigar relações entre renda e escolaridade com condições de saúde e nutrição em obesos graves. Estudo transversal ambulatorial com 79 pacientes de primeira consulta, com Índice de Massa Corporal (IMC) ≥ 35 kg/m2 e idade ≥ 20 anos. Coletaram-se dados: sociodemográficos, antropométricos, estilo de vida, exames bioquímicos e consumo alimentar. O IMC médio foi 48,3 ± 6,9 kg/m2. Observou-se correlação negativa significante de escolaridade com variáveis peso (r = -0,234) e IMC (r = -0,364) e de renda familiar per capita com consumo diário de vegetal A (r = -0,263). Após análise multivariada maior renda familiar per capita se associou à ausência de cardiopatia (RP: 0,51, IC95%: 0,32-0,81), maior consumo diário de vegetal A (RP: 1,79, IC95%: 1,16-2,75) e doces (RP: 3,12, IC95%: 1,21-8,04). Em obesos graves a maior renda familiar per capita se associou à ausência de cardiopatia e maior consumo de vegetais folhosos e doces. Já a escolaridade não se manteve associada às condições de saúde e nutrição.

This article seeks to investigate the relationship between income and educational level and health and nutritional conditions among the morbidly obese. A cross-sectional study was conducted with 79 patients at first appointment, with Body Mass Index (BMI) ≥ 35 kg/m2 and age ≥ 20 years. The following data was collected: demographic, socioeconomic, anthropometric, lifestyle, biochemical and food intake data. Average BMI was 48.3 ± 6.9 kg/m2. There was a significant negative correlation between education level and the variables of weight (r = -0.234) and BMI (r = -0.364) and per capita family income with daily consumption of leafy vegetables (r = -0.263). After multivariate analysis, higher per capita family income was associated with the absence of heart disease (PR: 0.51, CI95%: 0.32-0.81), higher daily consumption of leafy vegetables (PR: 1.79, CI95%: 1.16-2.75) and candy (PR: 3.12, CI95%: 1.21-8.04). In the morbidly obese, per capita household income was associated with absence of heart disease and higher consumption of leafy vegetables and candy. On the other hand, education level was not associated with health and nutrition conditions.
Descritores: Arabidopsis/enzimologia
Arabidopsis/genética
Regulação da Expressão Gênica de Plantas
Ácidos Indolacéticos/metabolismo
Fosfolipases A/metabolismo
-/farmacologia
ABDOMEN,ABDOMINAL NEOPLASMS,ABELSON MURINE LEUKEMIA VIRUS,CONGENITAL ABNORMALITIES-EICOSATETRAYNOIC ACID/farmacologia
Proteínas de Arabidopsis/genética
Proteínas de Arabidopsis/metabolismo
Proteínas de Ligação a DNA/genética
Proteínas de Ligação a DNA/metabolismo
Inibidores Enzimáticos/farmacologia
Glucuronidase/metabolismo
Luciferases/metabolismo
Proteínas Nucleares/genética
Proteínas Nucleares/metabolismo
Fosfolipases A/antagonistas & inibidores
Processamento de Proteína Pós-Traducional/efeitos dos fármacos
RNA Mensageiro/genética
RNA Mensageiro/metabolismo
Plântula/efeitos dos fármacos
Plântula/metabolismo
Fatores de Tempo
/farmacologia
TEMEFOS,ABBREVIATIONS AS TOPIC-DICHLOROPHENOXYACETIC ACID/farmacologia
Tipo de Publ: Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Responsável: BR1.1 - BIREME


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Id: lil-733007
Autor: Melo, Carina Mucciolo; Origassa, Clarice Silvia Taemi; Theodoro, Thérèse Rachell; Matos, Leandro Luongo; Miranda, Thaís Aguilar; Accardo, Camila Melo; Bouças, Rodrigo Ippolito; Suarez, Eloah Rabello; Pares, Madalena Maria Nunes Silva; Waisberg, Daniel Reis; Toloi, Giovanna Canato; Nader, Helena Bonciani; Waisberg, Jaques; Pinhal, Maria Aparecida Silva.
Título: Analysis of heparanase isoforms and cathepsin B in the plasma of patients with gastrointestinal carcinomas: analytical cross-sectional study / Análises das isoformas de heparanase e da catepsina B em plasma de pacientes com carcinomas gastrointestinais: estudo transversal analítico
Fonte: Säo Paulo med. j;133(1):28-35, Jan-Fev/2015. tab, graf.
Idioma: en.
Projeto: Fundação de Amparo à Pesquisa do Estado de São Paulo; . Fundação de Amparo à Pesquisa do Estado de São Paulo.
Resumo: CONTEXT AND OBJECTIVE: Heparanase-1 degrades heparan sulfate and has been correlated with tumor progression. Although the isoform heparanase-2 has no catalytic activity, it seems to be important for modulating heparanase-1 activity. Cathepsin B is a proteinase involved in tumor metastasis. The aim of this study was to analyze heparanase isoform expression and cathepsin B activity in plasma samples from patients with gastrointestinal carcinomas, compared with healthy individuals (control group). DESIGN AND SETTING: This was an analytical cross-sectional study. Peripheral blood samples were collected at a Brazilian public hospital, from 21 patients with histopathological diagnoses of gastrointestinal carcinomas and from 43 healthy individuals. The analyses were performed in two Brazilian medical schools. METHODS: Heparanase isoforms were identified and quantified in plasma samples by means of Western blot. The enzymatic activities of heparanase-1 and cathepsin B were also measured. RESULTS: The results demonstrated that the expression of both heparanase isoforms was significantly greater in plasma samples from gastrointestinal carcinoma patients, compared with the control group. Logistic regression analysis showed that increased heparanase-1 and heparanase-2 expression was exclusively dependent on the ...

CONTEXTO E OBJETIVO: A heparanase-1 degrada heparam sulfato e está relacionada à progressão de tumor. Apesar de a isoforma heparanase-2 não possuir atividade catalítica, parece ser importante para modular a atividade da heparanase-1. A catepsina B é uma proteinase envolvida na metástase de tumores. O objetivo deste estudo foi analisar a expressão das isoformas de heparanase e atividade da catepsina B em amostras de plasma de pacientes com carcinomas gastrointestinais, comparando-se com indivíduos saudáveis (grupo controle). TIPO DE ESTUDO E LOCAL: Este é um estudo transversal analítico. Foram coletadas amostras de sangue periférico, em hospital público brasileiro, de 21 pacientes com diagnóstico histopatológico de carcinoma gastrointestinal e 43 indivíduos saudáveis. As análises foram realizadas em duas faculdades de medicina brasileiras. MÉTODOS: As isoformas da heparanase foram identificadas e quantificadas em amostras de plasma por Western blot. As atividades enzimáticas de heparanase-1 e catepsina B foram também mensuradas. RESULTADOS: Os resultados demonstraram que as expressões das isoformas de heparanase foram significativamente maiores nas amostras de plasma de pacientes com carcinoma gastrointestinal em comparação com ...
Descritores: Biomarcadores Tumorais/sangue
Carcinoma/enzimologia
Catepsina B/sangue
Neoplasias Gastrointestinais/enzimologia
Glucuronidase/sangue
-Western Blotting/métodos
Estudos de Casos e Controles
Estudos Transversais
Técnicas Imunoenzimáticas
Isoenzimas/sangue
Limites: Idoso
Idoso de 80 Anos ou mais
Feminino
Humanos
Masculino
Pessoa de Meia-Idade
Responsável: BR1.1 - BIREME


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Id: lil-730968
Autor: Díaz T, Paula; Bernal G, Adriana; López C, Camilo.
Título: Transient GUS gene expression in cassava (Manihot esculenta Crantz) using Agrobacterium tumefaciens leaf infiltration / Expresión transitoria del gen GUS en yuca (Manihot esculenta Crantz) utilizando infiltración con Agrobacterium tumefaciens
Fonte: Rev. MVZ Córdoba;19(3):4338-4349, Sept.-Dec. 2014. ilus, tab.
Idioma: en.
Resumo: Objective. Assess transient gene expression of GUS in cassava (Manihot esculenta Crantz) leaves using Agrobacterium tumefaciens infiltration. Materials and methods. A. tumefaciens strains GV3101 and AGL1 containing pCAMBIA1305.2 were used to evaluate transient gene expression of β-glucuronidase (GUS). A. tumefaciens infiltration (agroinfiltration) was made using both leaves from in vitro and 1 month old greenhouse plants. Leaves were incubated in X-GLUC buffer, stained and photographed to detect GUS activity. Results. Agroinfiltration assays showed GUS transient expression in leaves of cassava varieties widely cultivated in the north coast and eastern savannah, MCOL2215 (Venezuelan) and CM6438-14 (Vergara), respectively. A. tumefaciens agressive strain AGL1 showed high efficiency inducing GUS expression in cassava leaves. Conclusions. We recommend using A. tumefaciens agressive strain AGL1 for agroinfiltration to assess transient expression in cassava leaves.

Objetivo. Evaluar la expresión transitoria del gen GUS en hojas de yuca (Manihot esculenta Crantz) por medio de infiltración con Agrobacterium tumefaciens. Materiales y métodos. Se utilizaron las cepas GV3101 y AGL1 de A. tumefaciens conteniendo el plásmido pCAMBIA1305.2, para evaluar la expresión transitoria del gen GUS. La infiltración de A. tumefaciens (agroinfiltración) se realizó tanto en hojas de plantas "in Vitro" como de plantas adultas de 1 mes. Las hojas se incubaron en tampón X-GLUC, se destiñeron y se fotografiaron para detectar la actividad de la enzima β-glucuronidasa (GUS). Resultados. Los ensayos de agroinfiltración en hoja muestraron la expresión transitoria del gen GUS en variedades cultivadas en la costa norte y en los llanos orientales, MCOL2215 (Venezolana) y CM6438-14 (Vergara) respectivamente, tanto en plantas "in Vitro" como en plantas adultas. La cepa hipervirulenta de A. tumefaciens AGL1 mostró una mayor eficiencia para la expresión transitoria en hojas de yuca. Conclusiones. Se recomienda utilizar la cepa AGL1 para evaluar la expresión transitoria de genes de interés por agroinfiltración en hojas de yuca.
Descritores: Agrobacterium tumefaciens
-Glucuronidase
Manihot
Responsável: CO332 - Facultad de Medicina


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Aguiar, J.A.K.
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Id: lil-709442
Autor: Peres, G.B.; Juliano, M.A.; Aguiar, J.A.K.; Michelacci, Y.M..
Título: Streptozotocin-induced diabetes mellitus affects lysosomal enzymes in rat liver
Fonte: Braz. j. med. biol. res = Rev. bras. pesqui. méd. biol;47(6):452-460, 06/2014. tab, graf.
Idioma: en.
Projeto: FAPESP; . CNPq.
Resumo: It has been previously shown that dextran sulfate administered to diabetic rats accumulates in the liver and kidney, and this could be due to a malfunction of the lysosomal digestive pathway. The aim of the present study was to evaluate the expression and activities of lysosomal enzymes that act upon proteins and sulfated polysaccharides in the livers of diabetic rats. Diabetes mellitus was induced by streptozotocin in 26 male Wistar rats (12 weeks old), while 26 age-matched controls received only vehicle. The livers were removed on either the 10th or the 30th day of the disease, weighed, and used to evaluate the activity, expression, and localization of lysosomal enzymes. A 50-60% decrease in the specific activities of cysteine proteases, especially cathepsin B, was observed in streptozotocin-induced diabetes mellitus. Expression (mRNA) of cathepsins B and L was also decreased on the 10th, but not on the 30th day. Sulfatase decreased 30% on the 30th day, while glycosidases did not vary (or presented a transitory and slight decrease). There were no apparent changes in liver morphology, and immunohistochemistry revealed the presence of cathepsin B in hepatocyte granules. The decrease in sulfatase could be responsible for the dextran sulfate build-up in the diabetic liver, since the action of sulfatase precedes glycosidases in the digestive pathway of sulfated polysaccharides. Our findings suggest that the decreased activities of cathepsins resulted from decreased expression of their genes, and not from general lysosomal failure, because the levels of glycosidases were normal in the diabetic liver.
Descritores: Catepsina B/metabolismo
Diabetes Mellitus Experimental/enzimologia
Fígado/enzimologia
Lisossomos/enzimologia
-Albuminas/análise
Western Blotting
Glicemia/efeitos dos fármacos
Catepsina L/metabolismo
Creatinina/urina
Cisteína Proteases/metabolismo
Sulfato de Dextrana/farmacologia
Diabetes Mellitus Experimental/induzido quimicamente
Expressão Gênica/efeitos dos fármacos
Glucuronidase/metabolismo
Hexosaminidases/metabolismo
Imuno-Histoquímica
Rim/metabolismo
Ratos Wistar
Reação em Cadeia da Polimerase em Tempo Real
RNA
Sulfatases/metabolismo
Limites: Animais
Masculino
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME



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