Hypothesis Holiday sudden cardiac death: food and alcohol inhibition of SULT1A enzymes as a precipitant

Science.gov (United States)

Eagle, Ken

2012-01-01

Sudden cardiac death is a significant health issue, causing millions of deaths worldwide annually. Studies have found that the likelihood of such death is higher in winter. Further studies identified that the highest likelihood occurs on Christmas Day and New Years Day, but not the interim period. Thanksgiving, Independence Day and the Islamic holiday Eid Al-Fitr also show significant increases in the rate of cardiac events or death. A number of mechanisms have been proposed, but none have satisfactorily explained the evidence. This article reviews the data supporting the existence of a holiday cardiac death phenomenon, the involvement of catecholamines and the normal modes of human catecholamine deactivation. Further evidence is reviewed that supports a hypothesized mechanism whereby critical SULT1A catecholamine deactivation enzymes can in some patients be inhibited by naturally-occurring phenols and polyphenols in foods and alcohols. If deactivation is inhibited by holiday consumption excesses, holiday stress or excitement could lead to a buildup of catecholamines that can cause fatal arrhythmias. Awareness of this mechanism could reduce deaths, both through doctor/patient education leading to a moderation in consumption and through the potential identification of patients with a predisposition to SULT1A inhibition. This hypothesis also raises parallels between sudden cardiac death in adults and Sudden Infant Death Syndrome (SIDS). The possible involvement of SULT1A inhibition in SIDS is discussed. Copyright © 2012 John Wiley & Sons, Ltd. PMID:22678655

Perspectiva en el desarrollo de las enzimas industriales a partir de la inteligencia tecnológica

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Óscar Fernando Castellanos Domínguez

2006-05-01

Full Text Available El desarrollo de tecnologías emergentes implica el reto permanente de utilizar nuevas herramientas de gestión tecnológica. La ingeniería de enzimas y su respectiva tecnología, en el contexto de la biotecnología, es una de las áreas del conocimiento sobre las cuales reiterativamente se plantean grandes expectativas como posible generador de ventaja competitiva en economías emergentes como la colombiana. Sin embargo, varias décadas de investigación en este campo aún no arrojan resultados importantes en la producción de enzimas y su aplicación en procesos industriales. El presente artículo se propone aplicar los componentes de un sistema de inteligencia tecnológica para el desarrollo de la tecnología de enzimas. Inicialmente se hace énfasis en los procesos de cienciometría retomados en ejercicios de mapeo de patentes, vigilancia tecnológica y evaluación de proyectos colombianos en la red ScienTI de Colciencias, realizados para las enzimas utilizadas, tanto en la industria de alimentos en la obtención de hidrolizados, como en la industria de textiles. Se documenta un ejemplo de estudio Delphi en el contexto de la prospectiva de las enzimas industriales en la biotecnología en general. Posteriormente, se evidencia el aprendizaje que se puede generar a partir del análisis de un líder del sector, a través de un benchmarking organizacional. Finalmente, retomando las bases del sistema de inteligencia tecnológica se discute su aporte a la generación de estrategias de productividad y competitividad de la aplicación de las enzimas industriales.

Actividad y expresión de enzimas convertidoras de angiotensina en neoplasias renales

OpenAIRE

Varela González, Pedro

2015-01-01

146 p. En el presente trabajo de tesis doctoral se realizó un análisis de 3 enzimas convertidoras de angiotensina (ACE, ACE2 y APA) en diferentes histotipos de neoplasias renales. Su objetivo fue conocer la implicación de estas enzimas en la carcinogénesis renal, en el desarrollo de los diferentes histotipos y en el comportamiento agresivo de estos tumores.Mediante diferentes técnicas (PCR, inmunohistoquímica, espectrofluorimetría, espectrofotometría) se analizó la expresión génica y proté...

Combined effects of NQO1 Pro187Ser or SULT1A1 Arg213His polymorphism and smoking on bladder cancer risk: Two meta-analyses.

Science.gov (United States)

Wang, Xiao-Chun; Wang, Jian; Tao, Hui-Hui; Zhang, Chao; Xu, Li-Fa

2017-07-14

Objectives: Cigarette smoking is the major risk factor of bladder cancer via exposure to chemical carcinogens. Nicotinamide adenine dinucleotide phosphate (NADP+): quinine oxidoreductase 1 (NQO1) and sulfotransferase 1A1 (SULT1A1) have been reported to involve in the metabolism of polycyclic aromatic hydrocarbons (PAHs) and aromatic amines. Therefore, the risk of bladder cancer (BC) may be influenced by polymorphisms in the genes that modulate metabolic detoxification in particular by interacting with cigarette smoking. Considering the limited power by the individual studies with a relatively small sample size, especially when analyzing the combined effect of polymorphisms in NQO1 and SULT1A1 genes and smoking, these 2 meta-analyses have aimed to clarify the combined effects of them on BC risk by integrating related studies. Two meta-analyses included 1341 cases and 1346 controls concerning NQO1 Pro187Ser and smoking, and 1921 cases and 1882 controls on SULT1A1 Arg213His and smoking were performed. Odds ratios (OR) and 95% confidence intervals (CI) were used for assessing the strength of the association. The result has demonstrated that smokers with NQO1 Pro/Ser or Ser/Ser genotypes have a prominent association with the risk of BC as compared with non-smokers with NQO1 Pro/Pro genotype, with OR equal to 3.71 (95% CI: 2.87-4.78, pheterogeneity = 0.376). Besides, smokers carrying SULT1A1 Arg/Arg genotypes were observed to confer 2.38 fold increased risk of BC (95% CI: 1.44-3.93, pheterogeneity = 0.001) when compared with non-smokers with SULT1A1 Arg/Arg or His/His genotypes. These findings have suggested that the NQO1 Pro187Ser or SULT1A1 Arg213His polymorphism combination with smoking significantly confer susceptibility to BC. Int J Occup Med Environ Health 2017;30(5):791-802. This work is available in Open Access model and licensed under a CC BY-NC 3.0 PL license.

Combined effects of NQO1 Pro187Ser or SULT1A1 Arg213His polymorphism and smoking on bladder cancer risk: Two meta-analyses

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Xiao-Chun Wang

2017-10-01

Full Text Available Objectives: Objectives: Cigarette smoking is the major risk factor of bladder cancer via exposure to chemical carcinogens. Nicotinamide adenine dinucleotide phosphate (NADP+: quinine oxidoreductase 1 (NQO1 and sulfotransferase 1A1 (SULT1A1 have been reported to involve in the metabolism of polycyclic aromatic hydrocarbons (PAHs and aromatic amines. Therefore, the risk of bladder cancer (BC may be influenced by polymorphisms in the genes that modulate metabolic detoxification in particular by interacting with cigarette smoking. Considering the limited power by the individual studies with a relatively small sample size, especially when analyzing the combined effect of polymorphisms in NQO1 and SULT1A1 genes and smoking, these 2 meta-analyses have aimed to clarify the combined effects of them on BC risk by integrating related studies. Material and Methods: Two meta-analyses included 1341 cases and 1346 controls concerning NQO1 Pro187Ser and smoking, and 1921 cases and 1882 controls on SULT1A1 Arg213His and smoking were performed. Odds ratios (OR and 95% confidence intervals (CI were used for assessing the strength of the association. Results: The result has demonstrated that smokers with NQO1 Pro/Ser or Ser/Ser genotypes have a prominent association with the risk of BC as compared with non-smokers with NQO1 Pro/Pro genotype, with OR equal to 3.71 (95% CI: 2.87–4.78, pheterogeneity = 0.376. Besides, smokers carrying SULT1A1 Arg/Arg genotypes were observed to confer 2.38 fold increased risk of BC (95% CI: 1.44–3.93, pheterogeneity = 0.001 when compared with non-smokers with SULT1A1 Arg/Arg or His/His genotypes. Conclusions: These findings have suggested that the NQO1 Pro187Ser or SULT1A1 Arg213His polymorphism combination with smoking significantly confer susceptibility to BC. Int J Occup Med Environ Health 2017;30(5:791–802

Impacto do polimorfismo genetico da enzima conversora da angiotensina no remodelamento cardiaco

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Felipe Neves de Albuquerque

2014-01-01

Full Text Available Fundamento: O papel dos polimorfismos genéticos da enzima de conversão da angiotensina na insuficiência cardíaca, como preditor de desfechos ecocardiográficos, ainda não está estabelecido. é necessário identificar o perfil local para observar o impacto desses genótipos na população brasileira, sendo inédito o estudo da insuficiência cardíaca de etiologia exclusivamente não isquêmica em seguimento mais longo que 5 anos. Objetivo: Determinar a distribuição das variantes do polimorfismo genético da enzima de conversão da angiotensina e sua relação com a evolução ecocardiográfica de pacientes com insuficiência cardíaca de etiologia não isquêmica. Métodos: Análise secundária de prontuários de 111 pacientes e identificação das variantes do polimorfismo genético da enzima de conversão da angiotensina, classificadas como DD (Deleção/Deleção, DI (Deleção/Inserção ou II (Inserção/Inserção. Resultados: As médias da coorte foram: seguimento de 64,9 meses, idade de 59,5 anos, 60,4% eram homens, 51,4% eram brancos, 98,2% faziam uso de betabloqueadores e 89,2% de inibidores da enzima de conversão da angiotensina ou de bloqueador do receptor da angiotensina. A distribuição do polimorfismo genético da enzima de conversão da angiotensina foi: 51,4% de DD; 44,1% de DI; e 4,5% de II. Não se observou nenhuma diferença das características clínicas ou de tratamento entre os grupos. O diâmetro sistólico do ventrículo esquerdo final foi a única variável ecocardiográfica isolada significativamente diferente entre os polimorfismos genéticos da enzima de conversão da angiotensina: 59,2 ± 1,8 para DD versus 52,3 ± 1,9 para DI versus 59,2 ± 5,2 para II (p = 0,029. No seguimento ecocardiográfico, todas as variáveis (diferença entre a fração de ejeção do ventrículo esquerdo da última e da primeira consulta; diferença entre o diâmetro sistólico do ventrículo esquerdo da última e da primeira

Enzimas fibrolíticas exógenas en la digestibilidad in vitro de cinco ecotipos de Brachiaria

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Juan H. Avellaneda-Cevallos

2007-01-01

Full Text Available En zi mas fi bro lí ti cas exó ge nas en la di ges ti bi li dad in vi tro de cin co eco ti pos de Bra chia ria. Este experimento se llevó a cabo en la Estación Experimental del Colegio de Postgraduados, en Montecillo, Estado de México, México, durante el 25 abril y el 27 noviembre de 2002. Se usó la primera fase de la técnica de Tilley y Terry (3, 6, 12, 24, 48, 72 h incubación con el objetivo de medir el efecto de un compuesto enzimático fibrolítico exógeno (enzima; Fibrozyme®; 0 y 1,5 g enzima/kg MS en la digestibilidad in vitro (DIV de MS, FDN y FDA de henos de cinco ecotipos de Brachiaria [brizantha var. Toledo (BT; ruziziensis x decumbens CIAT 46024 (RD; decumbens var. Señal (DS; ruziziensis x brizantha CIAT 36061 cv. Mulato (RBM; brizantha var. Insurgente (BI]. La DIVMS a las 48 y 72 h, para 0 y 1,5 g enzima, fue mayor (p<0,05 para los ecotipos BT y BI, respecto a RD, DS y RBM. La DIVFDN a las 48 y 72 h, para 0 y 1,5 g enzima, fue mayor (p<0,05 para BT y BI, respecto a RD, DS y RBM. La enzima aumentó (p<0,05 la DIVFDN sólo para el ecotipo RD a las 72 h; además, incrementó la DIVFDA para BT a 12 h; BT y DS a 24 h; BT, RD y BI a 48 h. Por tanto, las enzimas fibrolíticas aumentaron la digestibilidad in vitro de la pared celular de cinco ecotipos de Brachiaria.

Enzimas extracelulares da família Botryosphaeriaceae

OpenAIRE

Pinheiro, Ana Rita Almeida

2015-01-01

As espécies da família Botryosphaeriaceae são morfologicamente diversas e descritas como endofíticas, patogénias e saprófitas. Estas são normalmente encontradas numa grande diversidade de hospedeiros. Os fungos patogénicos para plantas Macrophomina phaseolina, Neofusicoccum parvum e Diplodia corticola secretam uma variedade de enzimas extracelulares, tais como proteases e glicosil hidrolases, algumas das quais envolvidas na interação hospedeiro-patogénio. A fim de elucidar a correlação ent...

INDUCCIÓN DE DOS ENZIMAS PECTOLÍTICAS EN EL MODELO Fusarium oxysporum f. sp. dianthi - CLAVEL

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Liliana Goméz García

2008-03-01

Full Text Available Se estudió por ensayos in vitro la posible participación de las enzimas endopoligalacturonasa (PG (EC.3.2.1.15 y pectato liasa (PL (EC.4.2.2.2, consideradas factores de virulencia en el proceso de infección del clavel por el hongo Fusarium oxysporum f. sp. dianthi (FOD.Los resultados muestran la inducción de la expresión de la enzima PG en presencia de los inductores artificiales, ácido poligalacturónico (APG y pectina, y un nivel de expresión muy bajo en cultivos con pared celular (PC de clavel de variedades resistente y susceptible. La enzima PL no presentó expresión en presencia de inductores artificiales (APG y pectina, mientras que en cultivos inducidos con pared celular de raíz presentó un alto nivel de expresión.

Produção de enzimas lignoceluloliticas e proteinas unicelulares por chrysonilia sitophila TFB 27441

OpenAIRE

Jaime Patricio Rodrigues Gutierrez

1987-01-01

Resumo: Estudou-se as enzimas celulolíticas e lignolíticas produzidas pelo microorganismo Chrysonlila sitophila TFB-27441 que foi isolado do trato intestinal de um inseto xilófago. A produção de celulases foi estudada utilizando como fonte de carbono no meio de cultivo gllcose, a-celuIose e a-celulose pré-tratada fotoquimicamente. As enzimas estudadas foram b-D-glicosidase, endo-1,4-b-D-glicanase e a atividade cetulolítica total foi estimada utilizando papel de filtro como substrato. A ativid...

Targeting Sulfotransferase (SULT) 2B1b as a regulator of Cholesterol Metabolism in Prostate Cancer

Science.gov (United States)

2016-10-01

associated with de novo androgen synthesis will be addressed based on the hypothesis that SULT2B1b promotes PCa proliferation by impacting the...evaluation of sulfonation activity on other sterols using in vitro assays. Seven thousand (7,000) compounds were screened after computational...stim- ulation, and previous studies have demonstrated that cholesterol canbeused as a precursor for androgen synthesis (6, 26). Thus, the impact of

Electrodo biológico con la enzima hidrogenasa, procedimiento de obtención y sus aplicaciones

OpenAIRE

Fernández López, Víctor Manuel; López de Lacey, Antonio; Rüdiger Ortiz, Olaf

2005-01-01

Electrodo biológico con la enzima hidrogenasa, procedimiento de obtención y sus aplicaciones. En la presente invención se protegen electrodos biológicos modificados con enzimas hidrogenasas (ánodos) mediante los cuales es posible obtener energía eléctrica del hidrógeno en una configuración típica de pilas de combustible; asimismo, con estos electrodos modificados con hidrogenasa (cátodos) es posible producir hidrógeno a partir de agua en una configuración típica de cél...

The associations between two vital GSTs genetic polymorphisms and lung cancer risk in the Chinese population: evidence from 71 studies.

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Kui Liu

Full Text Available BACKGROUND: The genetic polymorphisms of glutathione S-transferase (GSTs have been suspected to be related to the development of lung cancer while the current results are conflicting, especially in the Chinese population. METHODS: Data on genetic polymorphisms of glutathione S-transferase Mu 1 (GSTM1 from 68 studies, glutathione S-transferase theta 1 (GSTT1 from 17 studies and GSTM1-GSTT1 from 8 studies in the Chinese population were reanalyzed on their association with lung cancer risk. Odds ratios (OR were pooled using forest plots. 9 subgroups were all or partly performed in the subgroup analyses. The Galbraith plot was used to identify the heterogeneous records. Potential publication biases were detected by Begg's and Egger's tests. RESULTS: 71 eligible studies were identified after screening of 1608 articles. The increased association between two vital GSTs genetic polymorphisms and lung cancer risk was detected by random-effects model based on a comparable heterogeneity. Subgroup analysis showed a significant relationship between squamous carcinoma (SC, adenocarcinoma (AC or small cell lung carcinoma (SCLC and GSTM1 null genotype, as well as SC or AC and GSTT1 null genotype. Additionally, smokers with GSTM1 null genotype had a higher lung cancer risk than non-smokers. Our cumulative meta-analysis demonstrated a stable and reliable result of the relationship between GSTM1 null genotype and lung cancer risk. After the possible heterogeneous articles were omitted, the adjusted risk of GSTs and lung cancer susceptibility increased (fixed-effects model: ORGSTM1 = 1.23, 95% CI: 1.19 to 1.27, P<0.001; ORGSTT1 = 1.18, 95% CI: 1.10 to 1.26, P<0.001; ORGSTM1-GSTT1 = 1.33, 95% CI: 1.10 to 1.61, P = 0.004. CONCLUSIONS: An increased risk of lung cancer with GSTM1 and GSTT1 null genotype, especially with dual null genotype, was found in the Chinese population. In addition, special histopathological classification of lung cancers and a

INDUCCIÓN DE DOS ENZIMAS PECTOLÍTICAS EN EL MODELO Fusarium oxysporum f. sp. dianthi - CLAVEL

OpenAIRE

Gómez García, Liliana; Martínez, Sixta Tulia

2005-01-01

Se estudió por ensayos in vitro la posible participación de las enzimas endopoligalacturonasa (PG) (EC.3.2.1.15) y pectato liasa (PL) (EC.4.2.2.2), consideradas factores de virulencia en el proceso de infección del clavel por el hongo Fusarium oxysporum f. sp. dianthi (FOD). Los resultados muestran la inducción de la expresión de la enzima PG en presencia de los inductores artificiales, ácido poligalacturónico (APG) y pectina, y un nivel de expresión muy bajo en cultivos con pared celular (PC...

Combined effects of GSTO1 and SULT1A1 polymorphisms and cigarette smoking on urothelial carcinoma risk in a Taiwanese population

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Min-Che Tung

2014-09-01

Conclusion: The present study provided epidemiological evidence for a significantly increased risk of UCB in ever smokers with the Ala/Ala genotype of the GSTO1 gene and the Arg/Arg genotype of the SULT1A1 gene.

ASS and SULT2A1 are Novel and Sensitive Biomarkers of Acute Hepatic Injury-A Comparative Study in Animal Models.

Science.gov (United States)

Prima, Victor; Cao, Mengde; Svetlov, Stanislav I

2013-01-10

Liver and kidney damage associated with polytrauma, endotoxic shock/sepsis, and organ transplantation, are among the leading causes of the multiple organ failure. Development of novel sensitive biomarkers that detect early stages of liver and kidney injury is vital for the effective diagnostics and treatment of these life-threatening conditions. Previously, we identified several hepatic proteins, including Argininosuccinate Synthase (ASS) and sulfotransferases which were degraded in the liver and rapidly released into circulation during Ischemia/Reperfusion (I/R) injury. Here we compared sensitivity and specificity of the newly developed sandwich ELISA assays for ASS and the sulfotransferase isoform SULT2A1 with the standard clinical liver and kidney tests Alanine Aminotransferase (ALT) and Aspartate Transaminase (AST) in various pre-clinical models of acute injury. Our data suggest that ASS and SULT2A1 have superior characteristics for liver and kidney health assessment in endotoxemia, Ischemia/Reperfusion (I/R), chemical and drug-induced liver injury and may be of high potential value for clinical applications.

Empleo de polielectrolitos inteligentes para desarrollar matrices insolubles con capacidad bioseparativa de enzimas de importancia biotecnológica

OpenAIRE

Woitovich Valetti, Nadia

2016-01-01

Con los avances de la ciencia moderna, principalmente en el campo de la biotecnología, la obtención de enzimas para uso industrial ha creado la necesidad de desarrollar nuevas metodologías para la purificación de las mismas. Los métodos de escalado clásicos para la purificación de enzimas emplean precipitación con sales o solventes. Dichas técnicas producen gran cantidad de residuos que muchas veces son difíciles de reciclar, incrementando los costos del proceso. Los nuevos protocolos deben r...

Targeting Sulfotransferase (SULT) 2B1b as a Regulator of Cholesterol Metabolism in Prostate Cancer

Science.gov (United States)

2015-10-01

next  subtask.       Subtask 2: Production of infective Lentivirus using co-transfection of HEK293 (pCa cells LNCaP, PC3, DU145, and VCaP...PCa lines. We determined that the level of shRNA expression from a stably intergrated transgene gene delivered by lentivirus was too low...effects  on  SULT2B1b  enzyme  activity  of   cholesterol  sulfate   production  in  vitro.     We  developed  a

Enzimas pécticas de Aspergillus kawachii: aislamiento, purificación y caracterización de pectinesterasa

OpenAIRE

Vita, Carolina Elena

2013-01-01

Estudiar la producción de enzimas pectolíticas (tanto novedosas como previamente reportadas) por Aspergillus kawachii, caracterizarlas y compararlas con sus equivalentes de otros orígenes. Considerando la capacidad de A. kawachii de producir enzimas activas en condiciones de alta acidez y al escaso conocimiento de su sistema pectolítico, se inició este trabajo de investigación para contribuir al estudio de las pectinasas de este microorganismo. Este trabajo incluyó el screening de actividades...

BIOENSAIO PARA DETERMINAÇÃO DA RESISTÊNCIA DE PLANTAS DANINHAS AOS HERBICIDAS INIBIDORES DA ENZIMA ALS

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CHRISTOFFOLETI PEDRO JACOB

2001-01-01

Full Text Available Com o uso intensivo de herbicidas inibidores da acetolactato sintase (ALS em áreas cultivadas com soja, no município de São Gabriel d'Oeste (MS, Brasil, selecionou-se um biótipo resistente da planta daninha Bidens pilosa. A confirmação bioquímica desta resistência, no entanto, é muito difícil de ser feita em campo. Assim, realizou-se um bioensaio com o objetivo de testar a sensibilidade da ALS desse biótipo aos herbicidas inibidores dessa enzima, por meio da aplicação simultânea de herbicidas inibidores da ALS e da KARI (cetoácido reductoisomerase. Para a inibição desta última enzima, utilizou-se o teste CPCA (ácido 1,1-ciclopropanodicarboxílico. Por ele, confirmou-se que a resistência do biótipo de Bidens pilosa é decorrente da insensibilidade da ALS aos herbicidas, que visam inibi-la; pois, quanto maior a quantidade de acetoína, maior é a atividade da enzima e mais intensa a coloração desenvolvida, proporcionando maiores valores nas leituras de absorbância.

Produção de enzimas extracelulares por fungos associados à decomposição materiais vegetais em riachos

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Jéssica Barros Aguiar Silva

2015-12-01

Full Text Available As celulases e xilanases fúngicas, derivadas de sua habilidade degradativa e excretadas, em geral, para o meio, despertam interesse biotecnológico por terem um largo espectro de aplicação. Objetivou-se verificar a produção das enzimas celulase e xilanase por fungos filamentosos isolados de detritos foliares alóctone em um córrego no Cerrado. Os testes foram realizados com 100 cepas utilizando meio de cultura sintético contendo, como única fonte de carbono, carboximetilcelulose e xilana, para celulase e xilanase, respectivamente. Foram realizados inóculos no centro das placas e armazenadas por 4 dias em BOD a 28°C e submetido a choque térmico em estufa a 50°C por 16 horas. Para melhor visualização do halo de hidrólise as placas foram coradas com solução de vermelho congo e lavadas com solução de NaCl. Dos fungos testados 69% apresentaram resultado positivo para celulase dos quais 23% foram consideramos com potencial para aplicação em biotecnologia. Não houve resultados positivos para a produção da enzima xilanase. Os fungos testados, associados ao processo de decomposição de matéria orgânica vegetal, apresentaram produção da enzima celulase, mas não da enzima xilanase.

Determinação da energia metabolizável do farelo residual do milho com e sem enzima em dietas para frangos de corte

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C.G. Valadares

2016-06-01

Full Text Available RESUMO O objetivo deste trabalho foi avaliar o valor nutricional e determinar a energia metabolizável do farelo residual de milho (FRM sem e com o uso da enzima alfa- amilase. Foi realizado um experimento de metabolismo com 180 pintos machos Cobb com 14 dias, distribuídos em um delineamento inteiramente ao acaso, com seis tratamentos, cinco repetições e seis aves por parcela. As dietas experimentais foram: T1: ração referência (RR, T2: 60% T1 + 40% de FRM, T3: RR + enzima, T4: 60% T1 + 40% de FRM com adição de enzima, T5: RR com substituição de 100% do milho pelo FRM e T6: RR com substituição de 100% do milho pelo FRM com adição de enzima. A composição química do FRM foi: 88,33% de matéria seca (MS, 10,23% de proteína bruta (PB, 15,44% de extrato etéreo (EE, 4,33% de cinzas (CZ e 4555kcal/kg de energia bruta (EB. Os valores dos coeficientes de metabolizabilidade aparente para o FRM sem e com adição de enzima foram, respectivamente, de 73,37% e 76,33% para MS (p=0,0136, 70,44% e 70,39% para PB (p=0,9595 e de 74,79% e 76,77% para EB (p=0,0128. Os valores da energia metabolizável aparente (EMA e da EMA corrigida para retenção de nitrogênio (EMAn para o FRM (na base natural foram de 3322±19 e 3241±18kcal/kg e de 3334±16 e 3261±17kcal/kg, respectivamente, sem e com adição de enzima. A adição da enzima não teve efeito estatístico significativo sobre os valores de EMA e EMAn, entretanto houve efeito positivo no coeficiente de metabolização da energia.

A regeneração hepática e os inibidores da enzima conversora da angiotensina

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Fernando Silva Ramalho

2000-01-01

Full Text Available No presente estudo, o autor aborda conceitos fundamentais acerca do fenômeno regenerativo hepático, seguido de alguns aspectos importantes relacionados ao sistema calicreína-cininas. A associação entre os inibidores da enzima conversora da angiotensina, a bradicinina e a regeneração hepática é discutida à luz de estudos recentes, que revelam um papel estimulante das cininas sobre a proliferação hepatocelular. Os inibidores da enzima conversora da angiotensina, extremamente utilizados no tratamento clínico da hipertensão arterial sistêmica, da insuficiência cardíaca congestiva e da nefropatia diabética, também são capazes de favorecer a regeneração hepática.

Alteração de enzimas hepáticas em trabalhadores de refinaria de petróleo

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Carvalho Fernando Martins

2006-01-01

Full Text Available OBJETIVO: A exposição ocupacional típica de uma refinaria de petróleo pode alterar a função hepática de seus trabalhadores. Assim, o objetivo do estudo foi identificar fatores de risco de alterações em enzimas hepáticas em trabalhadores de uma refinaria de petróleo. MÉTODOS: Os trabalhadores de uma refinaria de petróleo, localizada em São Francisco do Conde, Estado da Bahia, eram submetidos a exames periódicos anuais de 1982 a1998. O estudo caso-controle investigou todos os 150 casos de indivíduos com alteração simultânea de gama-glutamil transferase e de alanino amino transferase, de pelo menos 10% acima do valor de referência. Como controles, foram selecionados 150 indivíduos sem quaisquer alterações de enzimas hepáticas ou de bilirrubinas, desde a sua admissão. Foram calculadas as razões de chance e respectivos intervalos de confiança de 95% a partir de modelos de regressão logística. RESULTADOS: Em todos os setores de produção, o risco de alteração de enzimas hepáticas foi significantemente mais elevado do que no setor administrativo (RC=5,7; IC 95%: 1,7-18,4, estando controlados os efeitos do álcool, obesidade e antecedentes médicos de hepatite. No período 1992-1994 foram registrados 89 casos, 88 deles provieram dos diversos setores da produção. CONCLUSÕES: A exposição ocupacional desempenha papel importante na determinação de alterações de enzimas hepáticas em trabalhadores do refino de petróleo, além dos fatores de risco eminentemente biológicos e/ou comportamentais como obesidade e o consumo de álcool.

Correlação entre a incapacidade funcional, idade e enzimas séricas nas doenças neuromusculares

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Lineu Cesar Werneck

1995-03-01

Full Text Available Foram estudados 806 casos de diversas doenças neuromusculares, a fim de verificar se existe correlação entre o grau de incapacidade funcional aferida pela escala de Vignos e Archibald (V&A e enzimas séricas (creatinoquinase, desidrogenase lática, aldolase, transaminase glutâmica oxalacética e pirúvica. Foram utilizados testes para a análise do coeficiente de correlação simples (Pearson e múltiplo. Foi encontrada correlação positiva (piora progressiva da incapacidade da V&A com a idade em algumas doenças, como a distrofia muscular de Duchenne, distrofia fascio-escapulo-umeral, distrofia miotônica, miopatias com defeitos enzimáticos da cadeia respiratória e esclerose lateral amiotrófica. Por outro lado, foi detectada correlação negativa (melhora progressiva dos sintomas na miopatia do multicore, miopatia benigna da infância com predomínio de fibras do tipo 1, deficiência de carnitina e dermatomiosite. A V&A mostrou maiores correlações (p< 0,05 entre as diversas enzimas sérícas quando estudadas isoladamente na distrofia muscular de Duchenne, distrofia óculo-crânio-somática, polimiositcs e periarterite nodosa. Quando as enzimas foram analisadas em conjunto, através de teste de correlação múltipla, verificou-se pequena correlação entre elas e a V&A. Esta reduzida interrelação sugere que a utilização de diversas enzimas na análise longitudinal das doenças neuromusculares é limitada, não tendo aplicação pratica, embora sejam muito importantes no diagnóstico.

Produção e avaliação de enzimas fibrolíticas exógenas na ensilagem de milho

OpenAIRE

Lara, Erika Christina [UNESP

2013-01-01

O presente trabalho avaliou a produção de enzimas fibroliticas exógenas obtidas a partir da fermentação fúngica e sua aplicação em diferentes doses na ensilagem de milho. Para isso, o projeto foi dividido em dois experimentos, sendo o primeiro caracterizado pela produção de enzimas e sua estabilidade em condições ruminais. Cinco amostras de diferentes locais foram coletadas e 45 fungos foram isolados e avaliados quanto a produção de xilanases e celulases. O micro-organismo selecionado como o ...

Enzimas proteolíticas relacionadas con la enfermedad periodontal inflamatoria

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Bárbara E. García Triana

1998-08-01

Full Text Available La enfermedad periodontal inflamatoria ocasiona la destrucción de los tejidos que protegen y soportan al diente; es por eso de gran importancia el papel que pueden desempeñar las enzimas que sean capaces de degradar la matriz del tejido conectivo, como las enzimas proteolíticas. Existen evidencias de que las metaloproteinasas de la matriz, las proteasas leucocitarias y las bacterianas, pueden participar en la etiopatogenia de esta enfermedad. Su acción es regulada en los tejidos, por la presencia de inhibidores específicos, de manera que un desbalance proteasas-inhibidores a favor de los primeros, conduciría a la destrucción de las proteínas de la matriz del tejido conectivo. A su vez, en la actividad proteolítica influyen diferentes factores, que de manera global, inducen un fenotipo degradativo o formativo, y que por lo tanto, podrían estar involucrados en la etiopatogenia de la enfermedad periodontal inflamatoria.The periodontal disease brings about the destruction of the protective and supporting tissues of the teeth, therefore, the role of enzymes capable of degrading connective tissue matrix is of great importance. There are evidences of the possible involvement of the matrix metalloproteinases, leukocyte proteases and bacterial proteases in the pathogenesis of such disease. Their action is controlled by specific inhibitors in the tissues. This means that any protease-inhibitor imbalance favouring the presence of proteases may lead to the destruction of matrix connective tissue. In turn, proteolytic activity is influenced by different factors that globally induce a degradative or formative phenotype, and thus, may be involved in the pathogenesis of the periodontal disease.

Produção in vitro de enzimas extracelulares por fungos e sua relação com os sintomas descritos em planta hospedeira

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Alice Elias

2015-12-01

Full Text Available RESUMOOs fungos fitopatogênicos habitantes de solo causam perdas econômicas em muitas culturas e são difíceis de serem controlados. Esses fungos podem ser agrupados pelos sintomas comuns que causam nas plantas, bem como pelas enzimas extracelulares que podem produzir. O objetivo do trabalho foi verificar a produção in vitro de enzimas extracelulares por fungos de solo e tentar relacionar essas enzimas com os sintomas que cada fungo causa em planta hospedeira. O ensaio foi delineado em esquema inteiramente casualizado, com dois fatores, sete fungos (Cylindrocladium spathiphylli, Rhizoctonia solani, Sclerotium rolfsii, Ceratocystis fimbriata, Sclerotinia sclerotiorum, Fusarium oxysporum f. sp. dianthi e Verticillium dahliae mais testemunha e seis enzimas (amilase, carboximetilcelulase, lipase, lacase, catalase e gelatinase com 10 repetições. Catalase e gelatinase foram mensuradas por escala de notas, enquanto que as demais pelo cálculo da área da coroa circular. O ensaio foi repetido e a análise foi realizada com os dados de dois ensaios. Os fungos que causam podridão na raiz ou no colo da planta apresentaram maior produção de lacase, enquanto os que causam obstrução, fendas ou até a destruição do sistema vascular demonstraram a prevalência da lipase.

OBTENÇÃO E UTILIZAÇÃO DA ENZIMA POLIFENOLOXIDASE EXTRAÍDA DE POLPA DE PINHA (Annona squamosa L.) MADURA NO MELHORAMENTO DO SABOR DO CACAU (Theobroma cacao L.)

OpenAIRE

LIMA,ELIZA DOROTEA POZZOBON DE ALBUQUERQUE; PASTORE,GLÁUCIA MARIA; BARBERY,SANDRA DRINA FERNANDEZ; GARCIA,NELSON HORÁCIO PEZOA; BRITO,EDY SOUSA DE; LIMA,CARLOS ALBERTO DE ALBUQUERQUE

2001-01-01

O presente trabalho teve como objetivo estudar a obtenção e a utilização da enzima polifenoloxidase (PPO) extraída de polpa de pinha madura na redução do teor de compostos polifenólicos com a finalidade de diminuir a adstringência e o amargor das amêndoas de cacau processadas na forma de "nibs". A PPO foi extraída com tampão fosfato de potássio 0,025M (pH 7,5), adicionando sulfato de amônio para a precipitação da enzima. O material em pó obtido foi denominado de enzima parcialmente purificada...

Associação do polimorfismo do gene da enzima conversora da angiotensina com dados ecocardiográficos em jovens normotensos filhos de hipertensos

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Franken Roberto Alexandre

2004-01-01

Full Text Available OBJETIVOS: Os autores objetivaram no presente estudo avaliar o polimorfismo da enzima conversora da angiotensina com dados do ecocardiograma de jovens estudantes de Medicina, filhos de hipertensos, comparados com jovens filhos de normotensos. MÉTODOS: Foram estudados 80 jovens normotensos divididos em dois grupos: 40 filhos normotensos de pais hipertensos e 40 filhos normotensos de pais hipertensos. Critérios de exclusão foram hipertensão arterial, fumo, obesidade, uso de contraceptivos orais. Uso crônico de medicamentos e presença de qualquer doença. Os alunos foram incluídos entre 1994 e 1996. Cinqüenta alunos foram submetidos a ecocardiograma transtoráxico. A análise estatística foi feita através do teste T de Student. A avaliação do polimorfismo do gene da enzima conversora da angiotensina foi feita nos 80 alunos conforme segue: 1 5 ml de sangue em tubo contendo EDTA, 2 extração do DNA, 3 medida da concentração do DNA por eletroforese, 4 reação em cadeia de polimerase com ''primer'' do gene da enzima conversora da angiotensina, 5 análise do polimorfismo do gene da enzima conversora da angiotensina através da eletroforese e 6 análise estatística através do teste do Qui-quadrado. RESULTADOS: O grupo de estudantes filhos de hipertensos mostraram maior espessura do septo interventricular (7,82mm+/-0,69 contra 7,38 +/- 0,8, p 7,82mm; DD 32%, DI 24%, II 20% contra septo 7,38mm: DD28%, DI12%, II 12%, contra septo 131,52g: DD 20,69% DI 13,79%, II 6,9% contra massa 117,11g: DD 30,43%, DI 8,7%, II 8,7% contra massa < 117,11g: DD 13,04%, DI 21,74%, II 17,39% (p=0,17 CONCLUSÃO: Encontramos diferenças entre a espessura do septo interventricular de estudantes normotensos filhos de hipertensos e filhos de normotensos. Por outro lado, não encontramos diferenças entre os grupos considerando o polimorfismo do gene da enzima de conversão da angiotensina, assim como qualquer relação do gene da enzima de conversão da

Docking molecular de derivados de 2-fenilindano-1,3-dionas inibidores da enzima HMG-CoA

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R. Q. Pordeus

2014-11-01

Full Text Available As doenças cardiovasculares constituem uma das principais causas de mortes em todo o mundo. Estudos mostram que a enzima HMG-CoA é considerada uma precursora da via metabólica hipolipidêmica no soro sanguíneo. Na busca por uma nova classe de compostos aptos a inibir esta enzima e consequentemente reduzir os níveis de colesterol, as 2-fenilindano-1,3-dionas apresentam resultados promissores. Uma das maneiras de avaliar o poder farmacológico destes compostos e predizer análogos ainda mais potentes consiste na avaliação da interação entre fármaco (2-fenilindano-1,3-diona e enzima (HMG-CoA, em que se utiliza da técnica de modelagem molecular docking. Neste estudo, o procedimento computacional para obtenção dos resultados de docking foi feito através do software AutoDock 1.5.6. Para avaliar a interação no sítio ativo da HMG-CoA, utilizamos, dentre a série de congêneres, o composto 2-(2-clorofenilindano-1,3-diona. De acordo com os resultados obtidos, foi identificada uma interação hidrofílica importante, do tipo ligação de hidrogênio C=O∙∙∙H–N, a qual apresenta uma distância de 1.62 Å entre os grupos carbonila do anel diona e o aminoácido metionina da HMG-CoA. Outra ligação de hidrogênio p∙∙∙H–N com distância de 3.10 Å formada entre o anel aromático do grupo indano-1,3-diona e o aminoácido glicina também foi identificada.

Enzimas de restricción de bacterias nativas de Nicaragua

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lan Roustan-Espinosa

2000-02-01

Full Text Available Los avances de la ingeniería genética y la biología molecular han propiciado la utilización de bacterias en la industria biotecnológica. En este trabajo se presenta la identificación y caracterización de enzimas de restricción presentes en bacterias recolectadas en medios acuosos de Nicaragua. Se encontró actividad de restricción en el 25% del total de bacterias analizadas. Se abordan los procesos de purificación de extractos de proteínas de bacterias con actividades de Sau961 y Pvull. Este trabajo es un esfuerzo dirigido a la implementación de técnicas modernas de biotecnología en Nicaragua.

Selective reduction in the expression of UGTs and SULTs, a novel mechanism by which piperine enhances the bioavailability of curcumin in rat.

Science.gov (United States)

Zeng, Xiaohui; Cai, Dake; Zeng, Qiaohuang; Chen, Zhao; Zhong, Guoping; Zhuo, Juncheng; Gan, Haining; Huang, Xuejun; Zhao, Ziming; Yao, Nan; Huang, Dane; Zhang, Chengzhe; Sun, Dongmei; Chen, Yuxing

2017-01-01

Curcumin (CUR) is known to exert numerous health-promoting effects in pharmacological studies, but its low bioavailability hinders the development of curcumin as a feasible therapeutic agent. Piperine (PIP) has been reported to enhance the bioavailability of curcumin, but the underlying mechanism remains poorly understood. In an attempt to find the mechanism by which piperine enhances the bioavailability of curcumin, the dosage ratio (CUR: PIP) and pre-treatment with piperine were hypothesized as key factors for improving the bioavailability in this combination. Therefore, combining curcumin with piperine at various dose ratios (1:1 to 100:1) and pre-dosing with piperine (0.5-8 h prior to curcumin) were designed to investigate their contributions to the pharmacokinetic parameters of curcumin in rats and their effects on the expression of UGT and SULT isoforms. It was shown that the C max and AUC 0-t of curcumin were slightly increased by 1.29 and 1.67 fold at a ratio of 20:1, while curcumin exposure was enhanced significantly in all the piperine pre-treated rats (0.5-8 h), peaking at 6 h (a 6.09-fold and 5.97-fold increase in C max and AUC 0-t , p bioavailability of curcumin through the reversible and selective inhibition of UGTs and SULTs. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

ESTUDOS TEÓRICOS E DE MODELAGEM MOLECULAR IN SILICO APLICADOS À INTERAÇÃO ENTRE A ENZIMA DELTA-AMINOLEVULINATO DESIDRATASE E DISSELENETOS DE DIARILA

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Rogério de Aquino Saraiva

2013-01-01

A enzima δ-aminolevulinato desidratase (δ-ALA-D) é uma metaloproteína essencial em vários processos biológicos, uma vez que é responsável por catalisar a formação de porfobilinogênio (PBG), um precursor dos tetrapirrólicos (heme, clorofila). Esta enzima é sensível a metais pesados e outros pró-oxidantes e, dessa forma, tem sido classicamente usada como um marcador na intoxicação por chumbo. Estudos in vitro e in vivo têm demonstrado que o organocalcogênio disseleneto de difenila [(P...

Evaluación de la actividad de la enzima citocromo P-450 1A2 en una población colombiana

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Fanny Cuesta González

2000-02-01

Full Text Available

Introducción: La enzima citocromo P450 1A2 (CYP1A2, presente en tejidos hepáticos humanos, participa en el metabolismo de fármacos tales como Clozapina, Imipramina, Teofilina y Cafeína. Estudios realizados en diferentes poblaciones han mostrado una distribución bimodal y a veces trimodal en la actividad de esta enzima. Este comportamiento ha sido relacionado con variaciones tanto del gen codificante como de las regiones reguladoras, aunque hasta el momento los resultados no son concluyentes. Además del metabolismo de fármacos, la CYP1A2 interviene en la activación de sustancias procarcinogénicas tales como N-nitrosaminas, aminas heterocíclicas y aromáticas; es conocida la correlación entre una actividad incrementada de esta enzima (metabolizadores rápidos con una mayor exposición al riesgo de cáncer.

Debido a la baja incidencia de efectos secundarios y la facilidad en la medición de los metabolitos en orina, la Cafeína es utilizada como fármaco prueba para la determinación de la actividad enzimática CYP1A2, calculando la relación entre las concentraciones molares de sus metabolitos (Actividad CYP1A2 = AFMU+MU+MX/17DMU.

En el presente estudio se evaluará la actividad de la enzima CYP1A2 en una muestra de individuos sanos, con el fin de describir su comportamiento y compararlo con el encontrado en otras poblaciones, las cuales tienen una influencia genética y ambiental diferente a la de la población colombiana.

Metodología: Una vez recolectadas las muestras de orina, se separarán de ellas los

Biological monitoring and the influence of genetic polymorphism of As3MT and GSTs on distribution of urinary arsenic species in occupational exposure workers.

Science.gov (United States)

Janasik, Beata; Reszka, Edyta; Stanislawska, Magdalena; Wieczorek, Edyta; Fendler, Wojciech; Wasowicz, Wojciech

2015-08-01

To examine the differences in urinary arsenic metabolism patterns in men affected by occupational exposure, we performed a study on 149 participants—workers of a copper mill and 52 healthy controls without occupational exposure. To elucidate the role of genetic factors in arsenic (As) metabolism, we studied the associations of six polymorphisms: As3MT Met287Thr (T>C) in exon 9; As3MT A>G in 5'UTR; As3MT C>G in intron 6; As3MT T>G in intron 1; GSTP1 Ile105Val and GSTO2 T>C. Air samples were collected using individual samplers during work shift. Urine samples were analyzed for total arsenic and arsenic chemical forms (As(III); As(V), MMA, DMA, AsB) using HPLC-ICP-MS. A specific polymerase chain reaction was done for the amplification of exons and flanking regions of As3MT and GSTs. The geometric mean arsenic concentrations in the air were 27.6 ± 4.9 µg/m(3). A significant correlation (p iAs +MMA and iAs. As3MT (rs3740400) GG homozygotes showed significantly (p iAs (21.8 ± 2.0) in urine than GC+CC heterozygotes (16.0 ± 2.1). A strong association between the gene variants and As species in urine was observed for GSTO2 (rs156697) polymorphism. The findings of the study point out that the concentration of iAs or the sum of iAs + MMA in urine can be a reliable biological indicator of occupational exposure to arsenic. This study demonstrates that As3MT and/or GSTs genotype may influence As metabolism. Nevertheless, further studies investigating genetic polymorphism in occupational conditions are required.

EFEITO DE ENZIMAS FIBROLÍTICAS SOBRE A COMPOSIÇÃO BROMATOLÓGICA DE SILAGENS DE CAPINS TROPICAIS

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Cirano José Ulhoa

2006-10-01

Full Text Available Este trabalho foi desenvolvido para determinar o efeito de níveis de enzimas fibrolíticas sobre a composição bromatológica e a degradabilidade ruminal da MS e da FDN das silagens de capim-braquiária e de capim-tanzânia. O experimento consistiu de quatro níveis enzimáticos (0, 5, 10e 20 mg de enzimas por kg de matéria natural e dois períodos de armazenamento (45 e 120 dias e três repetições. Após o armazenamento, os silos experimentais foram abertos para avaliação dos teores da MS, da PB, da FDN, do N-NH3/NT e do pH. Utilizou-se a técnica do saco de náilon,com três bovinos canulados no rúmen, horários de incubação de 6, 24 e 96 horas e amostras do período de 45 dias de ensilagem. Verificou-se que as soluções enzimáticas não alteraram o teor de MS, PB, N-NH3/NT e pH das silagens de capins. Para o capim-braquiária, a solução enzimática alterou o teor da FDN, mas foi dependente do nível de enzimas.Verificou-se redução da FDN para o nível de 20 mg (67,99% em relação ao tratamento-testemunha (70,28%. Não houve diferença entre os períodos de armazenamento nem interação entre período de armazenamento e nível enzimático. Para os parâmetros de degradação ruminal da MS e da FDN das silagens de capins não foram observadas diferenças entre os tratamentos. PALAVRAS-CHAVE: Aditivos, celulases, degradabilidade, hemicelulases, parede celular

Expresión diferencial de los genes de algunas enzimas lignocelulolíticas en biopelículas de Aspergillus niger

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Gretty K. Villena

2008-12-01

Full Text Available Se realizó una evaluación génica preliminar a nivel transcripcional de biopelículas de Aspergillus niger ATCC 10864 desarrolladas sobre poliéster respecto a algunas enzimas lignocelulolíticas. El análisis de expresión de genes de enzimas lignocelulolíticas y genes reguladores mediante RT-PCR mostró que los genes eng1, eglC, exo y eglA, eglB y xynB son diferencialmente expresados ya sea temporalmente o mediante más de un transcripto en comparación con cultivos sumergidos. Asimismo, los genes reguladores xlnR y creA mostraron patrones temporales de expresión distintos en ambos sistemas. Los resultados obtenidos aportan la evidencia molecular inicial de expresión diferencial de genes en biopelículas así como patrones de regulación diferencial muy probablemente ligada a la adhesión celular.

INDUCCIÓN DE LA ACTIVIDAD DE LA ENZIMA FENILALANINA AMONIO LIASA EN CLAVEL (Dianthus caryophyllus L POR ELICITORES DEL HONGO Fusarium oxysporum f. sp. Dianthi raza 2

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Harold Ardila

2008-04-01

Full Text Available Con el fin de evaluar el comportamiento a nivel del tallo de la enzima fenilalanina amonio liasa (PAL, por su nombre en inglés phenylalanine ammonia liase, durante la interacción clavel-Fusarium oxysporum f. sp. dianthi raza 2, se seleccionaron las condiciones para su extracción y cuantificación de la actividad. Para la extracción a partir de tallos y raíces se seleccionó un tratamiento previo del material vegetal con acetona y posterior extracción con buffer borato pH 8,8 con EDTA 2mMy -mercaptoetanol 18 mM. Para su cuantificación a nivel del tallo se debe realizar un ensayo discontinuo por 10 min, a 37 oC, pH 8,0 y a una concentración de sustrato de 35 mM. Adicionalmente se muestra mediante un ensayo in vivo el efecto que tiene, como inductor de esta enzima, la aplicación de un extracto crudo del patógeno. Los resultados observados indican que esta enzima se induce significativamente en tallos de claveles de la variedad tolerante “Kiss” durante el tratamiento por aspersión con el extracto crudo del patógeno, mientras que dicha inducción fue inexistente para la infección directamente con el patógeno. La inducción en esta variedad indica que en este extracto del patógeno se presentan elicitores potenciales para la inducción de esta enzima y por ende de la ruta fenilpropanoide.

USO DE NANOPARTÍCULAS DO MESOCARPO DO BABAÇU COMO PLATAFORMA PARA ANCORAGEM DE ENZIMAS NO DESENVOLVIMENTO DE BIOSSENSORES: UM MAPEAMENTO TECNOLÓGICO

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Ana Siqueira do Nascimento Marreiro Teixeira

2018-03-01

Full Text Available A imobilização de enzimas na construção de biossensores torna-se atraente pela capacidade enzimática de catalisar com grande eficiência, reações biológicas com alta sensibilidade e seletividade. A utilização de materiais nanoestruturados, tais como nanopartículas poliméricas, surge como alternativa para aumentar a eficiência da imobilização de enzimas. Dessa forma, surge o interesse na síntese de nanopartículas do mesocarpo do babaçu (Orbignya phalerata Mart, por se tratar de um biopolímero natural, atóxico, facilmente encontrado na região nordeste do País. Diante do relatado, o presente trabalho propõe um estudo prospectivo a respeito do desenvolvimento de um biossensor contendo a enzima polifenol oxidase como monocamada imobilizada, pela técnica de automontagem, sobre a superfície de nanoparticulas poliméricas geradas a partir do mesocarpo de babaçu para detecção de sulfitos em produtos alimentícios, visto que tais compostos são causadores de reações alérgicas em indivíduos que apresentam sensibilidade a este conservante. Para o mapeamento científico e tecnológico, foram realizadas buscas nas bases de periódicos Web of Science, Scopus e Scielo, e nos bancos de patentes INPI, Espacenet e USPTO, para todas patentes depositadas, e artigos publicados no período de 2005 a agosto 2016. A partir dos artigos e patentes analisados, constatou-se que os estudos relacionados ao mesocarpo de babaçu são escassos, sendo que o desenvolvimento de um biossensor para detecção de sulfito a base de nanopartículas poliméricas deste biopolímero como plataforma para a imobilização da enzima polifenol oxidase para pode ser bastante inovador, visto que não foram encontrados relatos na literatura com abordagem parecida.

Síntesis de las enzimas de piruvato, oxidorreductasa de ferredoxina y deshidrogenasa E de alcohol durante el desenquistamiento (excystacion de Giardia intestinalis

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Carlos Alberto Niño

2010-03-01

Conclusiones. Nuestros resultados muestran de forma concluyente que las enzimas PFOR y ADHE son traducidas en el proceso de desenquistamiento (excystacion y esto demuestra que existe un proceso activo de síntesis de proteínas durante él.

La enzima quinasa dependiente de 5´-adenosil monofosfato (ampk): perspectivas terapéuticas

OpenAIRE

Sánchez Junco, Diana

2016-01-01

En este trabajo nos centraremos en el estudio de la enzima quinasa dependiente de 5´-adenosil monofosfato (AMPK). La AMPK actúa como un sensor de energía y nutrientes y regula el balance energético. Se activa con el aumento de la relación AMP/ATP dentro de la célula y está implicada en numerosos procesos metabólicos, destacando la captación de glucosa y de ácidos grasos por la célula. La AMPK está formada por tres subunidades: α, β y γ, cada una de ellas con varias isoformas. Cabe destacar...

Caracterización bioquímica de la enzima bifuncional dihidrofolato reductasa-timidilato sintasa de Leishmania (Viannia y su evaluación como blanco molecular

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Edison Osorio

2013-09-01

Full Text Available Introducción. La dihidrofolato reductasa (DHFR se ha utilizado como blanco molecular en tratamientos antibacterianos, anticancerígenos y antipalúdicos. También, actúa como blanco molecular en Leishmania; sin embargo, no existen reportes de la enzima bifuncional en especies de Leishmania (Viannia. Materiales y métodos. Se ha aislado y expresado en Escherichia coli el gen que codifica para la enzima bifuncional DHFR y la timidilato-sintasa (TS de Leishmania braziliensis. La enzima recombinante se purificó y caracterizó, y se evaluó el efecto inhibitorio de algunos antifolatos, así como de cuatro alcaloides aporfínicos. Resultados. El gen se compone de aproximadamente 1.560 pb y codifica un péptido de 58 kDa que contiene los dominios DHFR y TS ligados en una sola cadena polipeptídica. La enzima recombinante DHFR-TS, utilizando el dihidrofolato (H2F como sustrato, presentó valores de Km y Vmax de 55,35± 4,02 (media ± el error estándar de la media y de 0,02 ± 5,34 x 10-4, respectivamente. La enzimar DHFR-TS de L. braziliensis presentó valores de Ki para los antifolatos en el rango de micras. El metotrexato fue el inhibidor más potente de la actividad enzimática (Ki=22,0 mM en comparación del trimetoprim (Ki=33 mM y la pirimetamina (Ki=68 mM. Estos valores de Ki son significativamente más bajos en comparación con los obtenidos para los alcaloides aporfínicos. Conclusión. Los resultados muestran el efecto inhibitorio de los antifolatos sobre la actividad enzimática, lo cual indica que la rDHFR-TS de L. braziliensis podría ser un modelo para estudiar moléculas antiprotozoarias potenciales. doi: http://dx.doi.org/10.7705/biomedica.v33i3.1434

Relación de la homocisteína y las enzimas involucradas en su metabolismo con enfermedad cardiovascular y otras patologías.

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Natalia Arias Morales

2016-06-01

Full Text Available Introducción: La homocisteína (HCY en la actualidad tiende a considerarse como un posible marcador temprano del desarrollo de diversas enfermedades, en especial de tipo cardiovascular. Los niveles aumentados de HCY (o hiperhomocisteinemia, hHCY también se han asociado con enfermedades neurológicas, neonatales, neoplásicas, metabólicas, entre otras. La hHCY puede ser consecuencia de diferentes factores como el estado nutricional, la edad, el género y por alteraciones genéticas de las enzimas que participan en su metabolismo. La identificación precoz de los polimorfismos genéticos permitiría establecer un esquema de prevención y tratamiento temprano para evitar la evolución de las patologías previamente enunciadas. Objetivo: Realizar una revisión documental descriptiva sobre la relación de la HCY y enzimas involucradas en su metabolismo con enfermedad cardiovascular (ECV y otras patologías. Materiales y métodos: La presente investigación fue de tipo análisis documental. La muestra de estudio correspondió a literatura científica incluida en bases de datos como Springer, PubMed, Scielo, Wiley y material bibliográfico documental relacionada con la homocisteína, enzimas involucradas en su metabolismo, así como su posible asociación con ECV y otras patologías, incluyéndose 140 fuentes bibliográficas. Para el análisis de la información recolectada se utilizó estadística básica descriptiva. Resultados: Del total de fuentes consultadas, predominaron artículos científicos en inglés con fecha de publicación entre 2005 y 2015. De los documentos revisados se evidenció que los países con mayor número de publicaciones fueron Estados Unidos (19%, España (9%, India (7%, China (7%, Brasil (7%, México (5%, Egipto (4%, Colombia (4%, entre otros. En mayor cantidad se encontraron artículos basados en la genética de la HCY, seguidos de causas de hHCY, patologías asociadas, metabolismo, bioquímica, diagnóstico y reportes

EFECTO DE LA GLUCOSA Y NITRATO DE AMONIO SOBRE LAS ENZIMAS LIGNINOLÍTICAS PRODUCIDAS POR Trametes versicolor INMOVILIZADO EN ESPUMA Y LA DECOLORACIÓN DE UN EFLUENTE PAPELERO EN UN BIORREACTOR DE LECHO FLUIDIZADO

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M. Martínez-Salgado

2005-12-01

Full Text Available En Colombia se han realizado varios estudios para el tratamiento de aguas residuales de la industria papelera a escala de laboratorio, con el hongo Trametes versicolor inmovilizado en espuma de poliuretano; este hongo ha sido ampliamente estudiado por la producción de enzimas ligninolíticas y su potencial uso en la decoloración de efluentes. En este trabajo, se estudió el efecto de una fuente de carbono (glucosa y una de nitrógeno (nitrato de amonio en un biorreactor de lecho fluidizado, con adición al medio de cultivo de 0.02% (v/v de Tween 80, 12 ppm de MnSO4 y 1 mM de CuSO4, desarrollando un diseño factorial 22 con dos diferentes concentraciones de los nutrientes, siendo éstos las variables independientes y las variables dependientes fueron la actividad enzimática, las unidades de color y el consumo de sustrato los cuales fueron determinados por métodos colorimétricos, además de la biomasa y pH. El uso del biorreactor de lecho fluidizado con entrada de aire constante, arrojó datos importantes sobre la decoloracióny la producción de enzimas en el agua residual no estéril. Los máximos valores de la enzima Mnperoxidasa fueron hallados en las concentraciones altas y bajas de glucosa y nitrato de amonio con valores de 9.257U y 6.502U; para la enzima Lacasa, al adicionar separadamente las dos variables se obtuvo valores de 8.618U y 8.341U. Las enzimas fueron relacionadas con la disminución de las unidades de color hasta 481.06UC, junto con el aumento de la biomasa del hongo Trametes versicolor hasta 3.365 g junto con flora nativa del agua residual. Los resultados encontrados, determinaron que las variables de consumo de sustrato y biomasa, tienen una gran importancia debido a que el aumento de la biomasa genera un aumento en la actividad de la enzima MnP y en la reducción de las unidades de color.

Desempenho de Frangos de Corte Alimentados com Dietas à Base de Milho e Farelo de Soja, com ou sem Adição de Enzimas Performance of Broilers Fed Corn Soybean Meal Based Diets, with or without Inclusion of Enzymes

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Geferson Fischer

2002-01-01

Full Text Available O experimento foi conduzido com o objetivo de avaliar o efeito da inclusão de um composto multi-enzimático à base de proteases, amilases e celulases (Vegpro, adicionado na proporção de 1 kg para cada 200 kg de farelo de soja, na dieta de frangos de corte. Foram utilizados 2240 pintos, fêmeas, da marca comercial Ross, submetidos a oito tratamentos, com sete repetições. O delineamento experimental utilizado foi em blocos ao acaso, com parcelas divididas. Os tratamentos foram os seguintes: T1 (controle positivo -- ração normal, sem enzima, T2 (controle negativo -- ração com níveis energético, protéico e aminoacídico do farelo de soja, superestimados em 5%, sem enzima, T3 (ração normal, sem enzima até 28 dias, com enzima de 29 a 35 dias, T4 (ração normal, sem enzima até 21 dias, com enzima de 22 a 35 dias, T5 (ração normal, com enzima, T6 (até os 28 dias, ração normal com enzima, e ração superestimada com enzima, dos 29 aos 35 dias, T7 (ração normal, com enzima até os 21 dias e ração superestimada, com enzima, do 22º ao 35º dia e T8 (ração superestimada com enzima. Na primeira fase experimental, à exceção do T2, não houve diferença significativa entre os tratamentos para peso corporal médio, ganho médio de peso e consumo de ração, pois não foi observado efeito da inclusão de enzima. A conversão alimentar diferiu estatisticamente a 5%. Na quarta semana experimental, novamente o T2 foi o único a diferir a 5% para peso corporal médio e ganho médio de peso. Para consumo de ração e conversão alimentar, os tratamentos não diferiram significativamente. Na última semana, foram notadas as maiores diferenças entre os tratamentos. Verificou-se que a inclusão do complexo multi-enzimático não proporcionou ganhos ao desempenho de frangos de corte.This study was run to evaluate the effect of inclusion of an enzymatic cocktail, containing proteases, amilases and cellulases (Vegpro in broiler diets. The

Efeito tóxico dos praguicidas maneb e paraquat sobre a atividade da enzima antioxidante catalase em ratos

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M. D. Arbo

2009-01-01

Full Text Available

Os radicais livres estão envolvidos em um grande número de enfermidades do ser humano. O cérebro tem níveis baixos de enzimas antioxidantes e um conteúdo lípidico elevado, tornando-se muito susceptível ao ataque de espécies reativas de oxigênio. Neste trabalho avaliou- se a lipoperoxidação em hipocampo e a atividade da enzima catalase em estriado e hipocampo de ratos tratados com o fungicida maneb (30 mg/kg e o herbicida paraquat (10 mg/kg. Não houve alteração na lipoperoxidação nem na atividade enzimática no hipocampo dos animais tratados com ambos os praguicidas, porém foi observada uma inibição da catalase no estriado dos ratos tratados com maneb e com paraquat. Com estes resultados pode-se sugerir, de forma preliminar, uma ação tóxica maior sobre centros dopaminérgicos. Estudos sobre a toxicidade destes compostos são essenciais na compreensão do papel destes praguicidas e dos radicais livres na etiologia das doenças. Palavras-chave: catalase; paraquat; maneb; estriado; hipocampo; radicais livres.

Las lipasas: enzimas con potencial para el desarrollo de biocatalizadores inmovilizados por adsorción interfacial

OpenAIRE

Jorge González-Bacerio; Jairo Rodríguez Hernández; Alberto del Monte Martínez

2010-01-01

Las lipasas son enzimas con propiedades funcionales muy interesantes que permiten su utilización práctica en diversos campos de las industrias agroquímica, farmacéutica, de detergentes y alimentaria, así como en química fina. Entre las aplicaciones más importantes de estas moléculas se encuentran: la resolución de mezclas racémicas, la obtención de compuestos ópticamente puros y la bioconversión de principios activos. En este trabajo se presenta una amplia revisión del tema, que abarca desde ...

Estudio de la expresión de enzimas del metabolismo de aminoácidos en lactococcus lactis

OpenAIRE

García Cayuela, Tomás

2011-01-01

Los aminoácidos son fundamentales para la supervivencia y el desarrollo de bacterias. Son las principales fuentes de nitrógeno y están implicados en la producción de energía, el control del pH intracelular y la regeneración de cofactores. Además, son los precursores de una larga variedad de compuestos volátiles en Lactococcus lactis y, por ello, diversas enzimas son consideradas clave para su formación, como aminotransferasas, deshidrogenasas, liasas y decarboxilasas, entre otras. Estas enzim...

Actividad antiinflamatoria de la enzima superóxido dismutasa (SOD), purificada de la especie Allium sativum (ajo)

OpenAIRE

Soberón, Mercedes; Suárez, Silvia; Arnao, Inés; Guija, Emilio; Troncoso, Luzmila; Rojas, Luis; Saldaña, Ítalo; Cordero, Adriana

2013-01-01

Objetivos: Demostrar la actividad antiinflamatoria de la enzima superóxido dismutasa (SOD) purificada de Allium sativum (ajo). Diseño: Experimental. Institución: Centro de Investigación de Bioquímica y Nutrición, Facultad de Medicina, UNMSM Material biológico: Ratas y Allium sativum. Principales medidas de resultados: Muestra de ajo triturada con tampón fosfato 50 mM pH 7,4, incubada 40 minutos y filtrada; fue sometida a extracción con cloroformo:etanol 1:1 (v/v). Se descartó la fase acuosa. ...

Physiological quality and amylase enzyme expression in maize seeds Qualidade fisiológica e expressão das enzimas amilases em sementes de milho

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Gustavo Evangelista Oliveira

2013-02-01

Full Text Available The physiological quality of maize seeds is affected by the genotype. Thus, the study of expression of genes associated with this characteristic is important in the genotype selection process in breeding programs. The aim of this research was to study the expression of amylase enzymes associated with physiological quality of maize seeds with different genotypes and seed sizes. We further sought to assess the expression of these enzymes in dry and soaked seeds The experiment was conducted in the experimental area and the Central Seed Laboratory of the Department of Agriculture of the Universidade Federal de Lavras. Seeds of four maize inbred lines were used, classified in two sizes. The physiological quality of the seeds was evaluated by means of germination, seedling emergence, seedling emergence speed index and accelerated aging test. Expression of the alpha amylase enzyme was evaluated by the electrophoresis technique and expression of the alpha amylase B73, alpha amylase (LOC542522 and beta amylase 5 (amyb5 genes was studied by the qRT-PCR technique in dry and soaked seeds of the inbred lines. There is differentiated expression of amylase enzymes in maize seeds of inbred lines with different levels of physiological quality. higher expression of amylase enzymes is observed in soaked maize seeds. The expression of transcripts is higher in smaller as wellas in soaked maize seeds of inbred lines.A qualidade fisiológica de sementes de milho é influenciada pelo genótipo. Assim, o estudo da expressão de genes associados a essa característica é importante no processo de seleção de genótipos em programas de melhoramento. O objetivo neste trabalho foi estudar a expressão das enzimas amilases associadas à qualidade fisiológica de sementes de milho, de diferentes genótipos e tamanhos de sementes. Objetivou-se ainda avaliar a expressão dessas enzimas em sementes secas e embebidas. O experimento foi conduzido em área experimental e no

EVALUACIÓN DE ENZIMAS PARA LA HIDRÓLISIS DE RESIDUOS (HOJAS Y COGOLLOS DE LA COSECHA CAÑA DE AZÚCAR

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JAIRO G. SALCEDO M.

2011-01-01

Full Text Available El presente trabajo tuvo como objetivo la evaluación de la hidrólisis enzimática de residuos de la cosecha de la caña de azúcar (hojas y cogollos a partir de la preparación de cuatro sustratos con diferentes pretratamientos, entre los que se encuentran: delignificación organosolvente, delignificación enzimática, delignificación con hidróxido de sodio y clorito de sodio y un sustrato base obtenido directamente de la cosecha. Además, se elaboraron cinco enzimas a partir de mezclas de actividades de enzimas comerciales, que fueron evaluadas en los diferentes sustratos. Se introduce un criterio para evaluar la eficiencia de las enzimas a partir de curvas de progreso, porcentaje de sacarificación e índice de sacarificación, a este índice se le denomino "índice global de hidrólisis". Los resultados muestran, que para la hidrólisis con el preparado enzimático E5, que contiene las siguientes actividades enzimáticas: hemicelulusas de 52.75 unidades globales/ml, celulasas total de 27.53 FPU/ml, endoglucanasas de 1782,1 CMC/ml, exoglucanasa de 0.377 UI/ml, betaglucosidasa de 550 pNPG U/ml, xilanasa de 28.23 UI/ml, galactosidasa de 7.1 UI/ml, manasa de 2.76 UI/ml y ranmanasa de 14.63 UI/ml y el sustrato S3, con un contenido de celulosa de 80.53%, hemicelulosa de 13.35, lignina 1.36 y un número de Kappa de 5.85 presenta el mayor índice global de Nhidrólisis que corresponde a un valor de 117.16. El estudio permite concluir que el preparado enzimático E5 presenta un gran potencial para la hidrólisis de los residuos de la caña de azúcar tratados en este proyecto.

Baseline frequency of chromosomal aberrations and sister chromatid exchanges in peripheral blood lymphocytes of healthy individuals living in Turin (North-Western Italy): assessment of the effects of age, sex and GSTs gene polymorphisms on the levels of genomic damage.

Science.gov (United States)

Santovito, Alfredo; Cervella, Piero; Delpero, Massimiliano

2016-05-01

The increased exposure to environmental pollutants has led to the awareness of the necessity for constant monitoring of human populations, especially those living in urban areas. This study evaluated the background levels of genomic damage in a sample of healthy subjects living in the urban area of Turin (Italy). The association between DNA damage with age, sex and GSTs polymorphisms was assessed. One hundred and one individuals were randomly sampled. Sister Chromatid Exchanges (SCEs) and Chromosomal Aberrations (CAs) assays, as well as genotyping of GSTT1 and GSTM1 genes, were performed. Mean values of SCEs and CAs were 5.137 ± 0.166 and 0.018 ± 0.002, respectively. Results showed age and gender associated with higher frequencies of these two cytogenetic markers. The eldest subjects (51-65 years) showed significantly higher levels of genomic damage than younger individuals. GSTs polymorphisms did not appear to significantly influence the frequencies of either markers. The CAs background frequency observed in this study is one of the highest reported among European populations. Turin is one of the most polluted cities in Europe in terms of air fine PM10 and ozone and the clastogenic potential of these pollutants may explain the high frequencies of chromosomal rearrangements reported here.

Cinética do P nos tecidos de suínos em crescimento alimentados com dietas formuladas de acordo com o conceito de proteína ideal e suplementadas com enzima fitase

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J.A. Moreira

2014-10-01

Full Text Available O presente estudo foi desenvolvido para avaliar os efeitos da enzima fitase sobre a cinética do P nos tecidos de suínos, alimentados com dietas formuladas de acordo com o conceito de proteína ideal e suplementadas com enzima fitase. Foram utilizados 20 suínos machos castrados, distribuídos em um delineamento em blocos ao acaso, com cinco tratamentos e quatro repetições. Os animais foram alojados em gaiolas metabólicas durante 17 dias, sendo 10 dias para adaptação e sete dias para coletas de fezes e urina. Amostras de sangue foram coletadas por cinco dias. No primeiro dia da fase experimental, cada animal recebeu, por via endovenosa, uma solução radioativa com 7,4MBq de 32P. No final do período experimental, os animais foram sacrificados, e amostras de tecidos do músculo (lombo, coração, fígado, rins e ossos foram coletadas. A enzima fitase interferiu na cinética do 32P, levando a uma menor incorporação nos tecidos dos ossos.

Hydrolysis of lactose by β-galactosidase from Kluyveromyces fragilis: characterization of the enzyme Hidrólise da lactose pela β-galactosidase de Kluyveromyces fragilis: caracterização da enzima

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Graciette Matioli

2002-03-01

Full Text Available The β-galactosidase enzyme from Kluyveromyces fragilis was characterized in the soluble form using lactose 5% w/v found in skimmed powdered milk as substrate. Enzyme diluted 50 times hydrolyzed the lactose in batch reactor of 50 mL capacity. Enzyme activity and its activation energy were determined as a function of temperature and pH. Temperature ranged from 20 to 55ºC and pH from 5.5 to 8.0. Activation energy was 9.50 kcal/mol. The energy of deactivation was 33.74 kcal/mol. Although the enzyme presented a high specific activity at 45ºC and pH 6.5 (3.312 U/mg protein, values indicate that the best use of the enzymatic activity occur at 40ºC or below, with half-life higher than 12 hours. The activation energy increased proportionally to pH increase. Therefore, the activation energy depends on pH and varies according to the origin of the enzyme.A enzima β-galactosidase de Kluyveromyces fragilis foi caracterizada na forma solúvel, utilizando como substrato, lactose 5% p/v presente no leite em pó desengordurado. A enzima, diluída 50 vezes, hidrolisou a lactose em reator batelada de 50 ml de capacidade. A atividade da enzima e sua energia de ativação foram determinadas em função da temperatura e pH. A faixa de temperatura analisada foi de 20 a 55ºC e de pH de 5,5 a 8,0. A energia de ativação foi de 9,50 kcal/mol. A energia de desativação foi de 33,74 kcal/mol. Embora a enzima tenha apresentado uma atividade específica alta a 45ºC e pH 6,5 (3,312 U/mg proteína, os valores obtidos indicam que o melhor aproveitamento da atividade enzimática se dá a 40ºC ou abaixo, com um tempo de meia-vida superior a 12 horas. A energia de ativação aumentou proporcionalmente com o aumento de pH. Portanto, a energia de ativação depende diretamente do pH da solução e varia com a origem da enzima.

Purificación y caracterización parcial de la enzima xilanasa a partir del preparado comercial Novoban 240

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Dalila Paz-Lago; Marta Hernández

2000-01-01

Novoban 240 es un preparado enzimático comercial de la Novo Industri con actividad amilasa, que se obtiene por fermentación sumergida de Bacillus amilolichuefaciens y es utilizado en la industria de elaboración de vinos. Este preparado de uso industrial posee actividad xilanasa, enzima estrechamente relacionada con la respuesta hipersensible de los tejidos vegetales frente al ataque de microorganismos patógenos. En el presente trabajo se establece una metodología de purificación parcial de es...

Metodologia de seleção de cepas para produção etodologia da ciclodextrina glicosiltransferase e para purificação da enzima = Strains selection methodology for cyclodextrin glycosyltransferase production and enzyme purification

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Glauciane de Lara Costa

2007-01-01

Full Text Available As ciclodextrinas (CDs são maltooligossacarídeos, produzidas a partir do amido, pela enzima ciclodextrina glicosiltransferase (CGTase. Esta pesquisa teve por objetivo estabelecer metodologias de seleção de cepas para produção de CGTase e para purificação da enzima. Os microrganismos foram selecionados a partir de 53 análises de solos de cultura de amido, em placas contendo meio de cultivo específico, para seleção de cepas produtoras de CGTase. As enzimas foram obtidas com cultivo destes microrganismos em meio líquido. As atividades enzimáticas das CGTases foram determinadas pelos métodos espectrofotométricos e precipitação com tricloroetileno. A cepa isolada do solo de aveia foi a que apresentou maior atividade [0,1864 mmol de b-CD (min mL-1]. Esta cepa foi utilizada para a produção da enzima em escala laboratorial e purificação em cromatografia deafinidade bioespecífica. A cepa selecionada nesta pesquisa abre novas perspectivas para produção de enzima e CDs em escala industrial.Cyclodextrins (CDs are maltooligosaccharides produced from starch by cyclodextrin glycosyltransferase (CGTase enzyme. This researchaimed at establishing method for strains selection for CGTase production and enzyme purification. The microorganisms were selected from 53 analyses of starch cultures soils on plates containing specific culture medium for strains selection that produce CGTase. Theenzymes were obtained by culturing these microorganisms in liquid medium. The enzyme activity was determined with photospectrometric methods and precipitation with trichloroethylene. The strain isolated from oat soil was the one that showed the highest activity [0.1864 mmol of b-CD (min mL-1]. This strain was used for enzyme production in laboratory scale and purification by biospecific affinity chromatography. The strain selected in this research opens new perspectives for enzymes production and CDs in industrial scale.

Enzimas exógenas em dietas de frangos de corte: desempenho Exogenous enzymes in broilers fed diets: performance

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Nei André Arruda Barbosa

2012-08-01

Full Text Available O experimento foi conduzido para avaliar a eficiência de enzimas exógenas em dietas sobre o desempenho de frangos de corte. Um total de 1440 pintos de corte machos (Cobb 500® foram distribuídos em um delineamento inteiramente casualizado em esquema fatorial 2x2 (duas dietas com diferentes densidades nutricionais; com e sem a adição enzimática com 8 repetições de 45 aves em cada unidade experimental. As dietas com diferentes densidades nutricionais foram: controle positivo (níveis normais e um controle negativo (com redução de 4,3 e 4,5% de energia metabolizável; 16,7 e 17,7% de cálcio e 35 e 42,7% de fósforo, nas fases inicial e de crescimento, respectivamente. A suplementação enzimática consistiu da combinação das enzimas fitase (100g t-1 e amilase, xilanase e protease (500g t-1. Foi avaliado o desempenho das aves nas fases de 1 a 21 e 1 a 42 dias de idade. Na fase total de criação, as aves alimentadas com a dieta controle negativo com adição enzimática tiveram consumo de ração (5,97%, peso médio (8,47%, ganho de peso (8,64% e conversão alimentar (2,92% melhores (P0,05 ao grupo alimentado com a dieta controle positivo com ou sem enzima. A adição de enzimas exógenas em dieta de frangos de corte com redução de energia metabolizável, cálcio e fósforo, proporciona um consumo de ração, peso vivo médio e ganho de peso similar a uma dieta com níveis nutricionais adequados.The experiment was conducted to evaluate the effect of enzyme efficiency in diets with and without nutrient reduction on the performance of broilers. Thousand and hundred forty male broiler chicks (Cobb 500® were distributed in a completely randomized design with 4 treatments in a factorial 2x2 (two diets with different nutrient densities, with and without enzyme with 8 replications of 45 birds. The diets were: positive control (normal and a negative control (with a reduction of 4.3% and 4.5% of metabolizable energy, 16.7% and 17.7% of

ALTERACIONES EN LA PRODUCCIÓN mRNA DE ENZIMAS INTESTINALES DE CERDOS DURANTE VARIOS PERÍODOS POSDESTETE ALTERAÇÕES NA PRODUÇÃO DE mRNA DE ENZIMAS INTESTINAIS DOS SUINOS DURANTE DIVERSOS PERÍODOS APOS O DESMAME ALTERATIONS IN THE PRODUCTION mRNA OF INTESTINAL ENZYMES IN PIGS DURING SEVERAL POSTWEANING PERIODS

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CAROLINA MONTOYA R.

2012-12-01

Full Text Available El objetivo de este estudio fue determinar la expresión génica de enzimas a nivel de enterocito de cerdos durante el período posdestete. El trabajo de campo se realizó con 16 cerdos destetados a los 21 días de edad en el Centro San Pablo, perteneciente a la Universidad Nacional de Colombia. Los animales fueron alimentados durante 10 días con una dieta basal que tuvo como componentes leche y algunos de sus derivados, y que además cumplía con todos los mínimos nutricionales. Los cerdos se sacrificaron escalonadamente los días 1, 5, 7 y 10 posdestete. Se realizó extracción completa del intestino delgado, el cual fue dividido en tres secciones (duodeno, yeyuno e íleon. Se evaluó la expresión génica de las enzimas digestivas por RT-PCR. El diseño estadístico empleado fue completamente al azar. Para las enzimas intestinales se presentaron disminuciones significativas, donde el día cinco posdestete presentó los menores valores(PO objetivo desta pesquisa foi determinar a expressão dos genes das aminopeptidases e dissacaridasesaonível do enterocito de suinos durante períodos apos o desmame. A experiência foi conduzida no centro de produção de San Pablo da Universidade Nacional de Colômbia (Medellín com 16 suinos desmamados aos 21 dias de vida. Os animais foram alimentados por 10 diascomuma dieta básica comleite e alguns de seus derivados, e aquele igualmente cumpriu todos os requisitos mínimos nutricionaís. Os suinos foram eutanasiados nos dias 1, 5, 7 y 10 pós-desmame. A extração completa do intestino pequeno foi realizada, que foi dividido emtrêsseções (duodeno, jejum e íleo. A expressão dos genes das enzimas digestivas por RT-PCR foi avaliada. O delineamento estatístico empregado foi completamente ao acaso. Para as enzimas intestinais as diminuições significativas foram apresentadas, onde no quinto diaapos o desmame apresentou os valores mais baixos (P0.01. O desmame precoce altera a expressão molecular das

Efeito da peletização e adição de enzimas e vitaminas sobre o desempenho e aproveitamento da energia e nutrientes em frangos de corte de 1 a 21 dias de idade

OpenAIRE

Leite, José Laureano Barbosa; Rodrigues, Paulo Borges; Fialho, Elias Tadeu; Freitas, Rilke Tadeu Fonseca de; Nagata, Adriano Kaneo; Cantarelli, Vinícius de Souza

2008-01-01

Conduziu-se este experimento para avaliar o efeito da peletização e adição de enzimas e vitaminas sobre o desempenho e aproveitamento da energia e nutrientes, em frangos de corte de 1 a 21 dias de idade. Foram utilizados 350 pintos de corte, COBB, machos, com peso inicial médio de 47g ± 2,5g, em um delineamento inteiramente casualizado, com os tratamentos em arranjos fatorial 3 x 2 (três dietas, com e sem enzimas) sendo que as dietas consistiram de uma ração farelada e duas rações peletizadas...

Digestibilidad in situ de dietas con harina de nopal deshidratado conteniendo un preparado de enzimas fibrolíticas exógenas In situ digestibility in dehydrated ground prickly pear diets containing a fybrolitic enzymes product

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Marco Medina Romo

2006-07-01

Full Text Available Se evaluó el efecto de un preparado de enzimas fibrolíticas exógenas (celulasas y xilanasas en la degradabilidad in situ de la materia seca (DisMS, fibra detergente neutro (DFDNr y fibra detergente ácido residual (DFDAr, en dietas altas o bajas en harina de nopal deshidratado. Se aplicaron concentraciones de 0, 1, 2 y 3 g de enzima por kilogramo de materia seca al inicio y 24 horas antes de la degradación in situ. Se determinó la concentración de ácidos grasos volátiles totales y de nitrógeno amoniacal a las 0, 3, 6, 9, 12 y 24 horas después de aplicarse la enzima. No se observaron efectos en DisMS, DFDNr y DFDAr; la aplicación al inicio de la degradación in situ mostró valores más altos que a 24 horas para DisMS y DFDNr, pero fue menor para DFDAr. No se observaron diferencias en las interacciones entre niveles de enzima, tipo de dieta y tiempo de pretratamiento. La aplicación de 1 y 3 g de enzima, en la dieta con bajo contenido de harina de nopal, tuvo efectos en el incremento de los ácidos grasos volátiles totales; para el nitrógeno amoniacal, los mejores resultados ocurrieron con 0 y 1 g de enzima.It was evaluated the effect of a fybrolitic enzyme product (cellulases and xylanases on in situ digestibility of dry matter (DisMS, residual neutral detergent fiber (DFDNr and acid detergent fiber (DFDAr, in dehydrated ground prickly pear diets with a low or high level. Enzyme concentrations of 0, 1, 2, and 3 g kg-1 of dry matter applied at the beginning (0 hour and 24 hours before starting in situ digestibility were used. Total volatile fatty acids and ammonia nitrogen were determined at: 0, 3, 6, 9, 12, and 24 hours after the enzyme application. There were no effects on DisMS, DFDNr, and DFDAr. Initial application of enzyme concentrations (0 hour was higher than 24 hours for DisMS and DFDNr but lower for DFDAr. No differences were observed in interactions among enzyme level, diet and application time. Application of 1 and 3 g of

Dietas à base de milho e farelo de soja suplementadas com enzimas na alimentação de frangos de corte Corn and soybean meal based diets supplemented with enzymes in feed of broiler chicken

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Delma Maria Torres

2003-02-01

Full Text Available Com o objetivo de verificar o efeito da adição de enzimas digestivas exógenas em dietas à base de milho e farelo de soja sobre o desempenho de frangos de corte, conduziu-se este experimento. Foram utilizados 819 pintos Hubbard em DIC e esquema fatorial 3 x 2 x 2 + 1 (enzimas, proteínas, níveis de energia e um tratamento testemunha com 3 repetições de 21 aves por parcela. Os níveis de enzimas foram 0,5; 1,0 e 1,5 g/kg de dieta, e os de proteína foram normais (21,18, 19,95 e 19,43% PB e reduzidos (20,54, 19,35 e 18,46% PB; os de energia também foram normais (3000, 3100 e 3200 kcal/kg de EM e reduzidos (2910, 3007 e 3040 kcal/kg de EM, respectivamente, para as fases inicial, de crescimento e final da criação, e para o testemunha, a ração continha níveis normais de EM e PB, sem enzimas. Os fatores avaliados foram ganho de peso, consumo de ração, conversão alimentar e fator europeu de produção. Pelos resultados, verificou-se manutenção do desempenho zootécnico das aves em razão da aplicação de enzimas, constatado pela não significância desses fatores aos 42 dias de idade e pela melhora na conversão alimentar, redução do consumo de ração e maior fator europeu de produção aos 21 dias de idade. O estudo da interação dos níveis de enzima x proteína mostrou que a adição de 0,5 g/kg de enzima na dieta com nível de proteína reduzido resultou em maior ganho de peso das aves. Conclui-se que é viável o uso de enzimas exógenas adicionadas em rações para frangos de corte.The objective of the experiment was verify the effect of the addition of exogen digestive enzymes in corn and soybean meal based diets on performance of broiler chickens. Were used 819 Hubbard chickens in completely randomized design, and 3 x 2 x 2 + 1 check factorial (enzyme, protein and energy levels and one treatment check with 3 replications, of the 21 birds by replicate. Enzyme levels were 0.5, 1.0 and 1.5 g/kg of diet, energy levels were

Caracterización de las enzimas esterasas implicadas en la interacción polen-pistilo en el olivo (Olea Europaea L.)

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Rejón García, Juan David

2013-01-01

El éxito de la fertilización en las Angiospermas depende en gran medida de la capacidad del polen para germinar y emitir un tubo polínico, el cuál transporta los núcleos espermáticos a través del pistilo hasta el saco embrionario. En este contexto, los objetivos de este proyecto fueron: 1) determinar la existencia de actividad esterolítica en el polen de olivo, 2) identificar y clasificar desde el punto de vista funcional las distintas enzimas esterasas, 3) determinar las funciones del exudad...

Crescimento foliar e atividades das enzimas antioxidativas em plântulas de girassol suplementadas com percolado de aterro sanitário e submetidas a estresse hídrico

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Francisco Holanda Nunes Junior

2017-01-01

Full Text Available Em regiões áridas ou semiáridas, a escassez hídrica tem sido um dos principais fatores limitantes da produção agrícola. Diante disso, o presente trabalho objetivou analisar os efeitos da aplicação de percolado de aterro sanitário no crescimento foliar (número de folhas e área foliar e as atividades das enzimas antioxidativas: superóxido dismutase (SOD, catalase (CAT, peroxidase do ascorbato (APX e peroxidase do guaiacol (GPX em folhas e raízes de plântulas de girassol submetidas às condições de estresse hídrico. O delineamento experimental foi o inteiramente casualizado, disposto em arranjo fatorial dois (irrigadas ou não irrigadas x quatro (areia; areia + adubo orgânico 100 kg N ha-1; areia + percolado de aterro sanitário 100 kg N ha-1; areia + percolado de aterro sanitário 150 kg N ha-1, com cinco repetições. Em relação aos parâmetros foliares, o tratamento 100 kg N ha-1 suplementado com percolado de aterro sanitário obteve os melhores incrementos no número de folhas e área foliar. Além disso, verificaram-se aumentos nas atividades das enzimas antioxidativas em folhas e raízes de plântulas de girassol suplementadas com percolado de aterro sanitário, sendo provável que as reduções dos efeitos deletérios do estresse hídrico nas variáveis foliares dos tratamentos suplementados com percolado de aterro sanitário tenham ocorrido em virtude das maiores atividades das enzimas antioxidativas, especialmente as da CAT nas folhas e GPX nas raízes.

Enzimas amilolíticas de mandioquinha-salsa (Arracacia xanthorrhiza Bancroft. Amylolytic enzymes from peruvian carrot (Arracaia xanthorrhiza Bancroft.

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Tatiana da Costa Raposo Pires

2002-12-01

Full Text Available O objetivo do presente trabalho foi de caracterizar as enzimas amilolíticas de mandioquinha-salsa (Arracacia xanthorrhiza Bancroft.. Foram realizados ensaios mediante a determinação da atividade enzimática, variando-se as condições do meio, tais como temperatura, pH e concentração de cátions. Em gel de eletroforese foram detectadas três bandas protéicas com intensa atividade hidrolítica. As enzimas apresentaram pH ótimo de atividade em torno de 6,0 e mostraram-se mais sensíveis a valores de pH alcalino quando pré-incubadas a 50oC. A temperatura ótima de ativação enzimática foi de 50oC, enquanto aos 70oC, a atividade amilásica foi reduzida em 80%. Em temperaturas altas temperaturas (60 e 70oC, a inativação enzimática ocorreu após 60 e 25min de incubação, respectivamente. Ensaios de estabilidade térmica mostraram que as amilases mantiveram alta atividade após 25h mediante a incubação a 20 e 30oC. Os valores de Km, Vmax e energia de ativação foram de 0,41mg/mL, 1,11mg/mL/min e 7,53kcal/mol, respectivamente. Constatou-se que a presença de Ca+2 ou Mg+2 provoca aumento na atividade amilásica, enquanto íons de Cu+2 causam uma diminuição. Apesar da discreta atividade amilásica apresentada, as enzimas demonstraram ter alta resistência e estabilidade térmicas.This investigation was undertaken to characterize amylases from Peruvian carrot (Arracacia xanthorrhiza Bancroft.. Activity assays were performed in different changing pHs, temperatures, time and cation concentrations. PAGE zymograms and the amylase activity essays were detected by starch-iodine complex. The results showed that Peruvian carrot has three amylase components on the crude extract. The enzymes had optimum pH at 6.0. Amylases were more sensitive to basic pH when the pre-incubation occured at 50oC. The best activation temperature occurred at 50oC, whereas at 70oC the amylase activity was reduced by 80%. Assays of thermal stability at 20 and 30o

Perfil bioquímico de algumas enzimas no plasma sangüíneo de potras da raça Brasileiro de Hipismo (BH criadas em Colina, Estado de São Paulo

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Fernando José Benesi

2009-08-01

Full Text Available Com o propósito de avaliar o perfil bioquímico de algumas enzimas, em potras sadias da raça BH (Brasileiro de Hipismo, utilizaram-se 380 amostras de plasma sangüíneo colhidas de 19 animais desde o nascimento até 24 meses de vida. Na análise dos resultados evidenciou-se que os valores médios das enzimas FA (fosfatase alcalina e CK (creatina quinase foram máximos entre o nascimento e 24 horas de vida (FA-1995.50 UI/ ; CK-189.13UI/L, enquanto que para a LD (lactato desidrogenase e GGT (gama glutamiltransferase as maiores magnitudes ocorreram, respectivamente, entre 3 e 4 dias (LD-479.11UI/L e aos 10 dias de idade (GGT-18.70UI/L. As FA, CK, LD e GGT, mostraram diminuições dos valores médios, respectivamente, até 6 meses (FA-323.50UI/L, 20 dias (CK-51UI/ L, 19 meses (LD-214.00UI/L e 4 meses (GGT-11.40UI/L estabilizando-se a seguir, com pequenas variações. A atividade da AST (aspartato aminotransferase que foi mínima logo após o nascimento (AST- 43.38UI/L, aumentou até os sete dias de vida (AST-110.89UI/L, e a seguir diminuiu progressivamente, com pequenas oscilações, até o final do estudo. Todas as enzimas avaliadas sofreram variações influenciadas pelo fator etário, particularmente no período inicial de vida dos animais estudados.

Estudos de parâmetros que influenciam na produção da enzima CGTase de Bacillus firmus, cepa nº 37 Study of parameters that influence the production of the enzyme CGTase from Bacillus firmus, strain no. 37

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Gisella Maria Zanin

2000-05-01

Full Text Available As ciclodextrinas (CDs são oligossacarídeos cíclicos formados por resíduos de glucopiranose unidos por ligação α-1,4. As mais comuns são α-, β- e γ-CD contendo 6, 7 e 8 resíduos de glucopiranose, respectivamente. Elas são produzidas a partir do amido pela ação da enzima ciclodextrina glicosiltransferase (CGTase. Freqüentemente, β-CD é produzida em maior quantidade. Um estudo da otimização da produção da CGTase de Bacillus firmus cepa nº 37 (β-CGTase foi realizado. A produção da enzima ocorreu durante a fase de crescimento exponencial do microrganismo e a máxima atividade foi observada com três dias de cultivo a 37ºC. O melhor rendimento na produção da enzima foi obtido quando da utilização de pré-inóculo com absorbância entre 0,5 e 1,0 (660 nm. O uso do substrato maltodextrina para produção da enzima proporcionou uma atividade enzimática ao redor de 31% menor que o substrato amido solúvel. Portanto, o substrato maltodextrina não é adequado para melhorar a produção da enzima estudada.The cyclodextrins (CDs are cyclic oligosaccharides formed by residues of glucopyranose linked by α-1,4. The most common are the α-, β- and γ-CD that present 6, 7 and 8 units of glucopyranose, respectively. They are produced from starch by the action of the enzyme cyclodextrin glycosyltransferase (CGTase. Frequently, β-CD is produced in larger amount. A study of optimization of the production of the CGTase from Bacillus firmus, strain no. 37 (β-CGTase was performed. The production of the enzyme occurred during the phase of exponential growth of the microorganism and the maximum activity was observed within three days of cultivation at 37ºC. The best production of the enzyme was obtained with inoculum of optical density between 0.5 and 1.0 (660 nm. The use of the maltodextrin for production of the enzyme provided an enzymatic activity at 31% lower than the substrate, soluble starch. Therefore, the substrate

Development of enzyme-linked immunosorbent assay (ELISA) for glutathione S-transferase (GST-S) protein in the intertidal copepod Tigriopus japonicus and its application for environmental monitoring.

Science.gov (United States)

Rhee, Jae-Sung; Kim, Bo-Mi; Jeong, Chang-Bum; Leung, Kenneth Mei Yee; Park, Gyung Soo; Lee, Jae-Seong

2013-11-01

To utilize the GST-S protein as a useful biomarker for environmental contamination, we developed a polyclonal antibody-based enzyme-linked immunosorbent assay (ELISA) in the intertidal copepod Tigriopus japonicus. Two polyclonal antibodies, TJ-GST-S1 and TJ-GST-S2, were raised against two TJ-GST-S synthetic peptides. Also a recombinant TJ-GST-S protein was purified as a standard for ELISA development. Each polyclonal antibody was tested by Western blot analysis and indirect ELISA. Of two polyclonal antibodies, TJ-GST-S2 ELISA was further employed due to its wide range of detection and the limit of specificity compared to those of TJ-GST-S1 ELISA system. After exposure to 4 metals (Ag, As, Cd, and Cu) to T. japonicus, the amount of TJ-GST-S protein was significantly elevated in a concentration-dependent manner. Also, TJ-GST-S protein was upregulated at relative high concentrations of B[α]P, PCB, and TBT. In this paper, we suggest that T. japonicas ELISA for TJ-GST-S2 is useful as a potential indicator system for marine contaminants. Copyright © 2013 Elsevier Ltd. All rights reserved.

Polimorfismo inserción/deleción del gen de la enzima convertidora de angiontensina y enfermedad coronaria en la población de Montería, Córdoba

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Manolo I Jaramillo

2013-09-01

Full Text Available Antecedentes y objetivo: el polimorfismo inserción/deleción del gen de la enzima convertidora de angiotensina, ha sido identificado como un potente factor de riesgo de enfermedad coronaria. Para la población de Montería se desconocen las frecuencias con las que se expresan los alelos de este gen y el carácter de su interacción con condiciones de riesgo cardiovascular. El objetivo de este trabajo fue determinar, para dicha población, la asociación de este polimorfismo y el riesgo de sufrir enfermedad coronaria. Método: se llevó a cabo un estudio retrospectivo con 70 casos y 70 controles; como casos se consideraron pacientes con padecimientos coronarios confirmados por electrocardiograma, remitidos a la Organización Cardiodiagnóstico de Córdoba, y como controles individuos voluntarios sin antecedentes cardiovasculares y sin relación filial. El ADN requerido se extrajo a partir de sangre periférica. La caracterización del polimorfismo se hizo mediante reacción en cadena de la polimerasa. Resultados: la distribución de genotipos de la enzima convertidora de angiotensina en pacientes casos no fue significativamente diferente a la estimada en pacientes controles (X2=3.687, p=0,1583. El genotipo más frecuente en la población fue ID (40,72%. En el grupo casos, el genotipo II fue más frecuente que el genotipo DD comparado con el grupo control (p<0,05. El modelo de regresión logística múltiple ajustado, indicó no significancia del genotipo DD como factor de riesgo coronario (razón de disparidad = 0,51 IC95% = 0,25 – 1,06. Conclusión: el polimorfismo I/D del gen de la enzima convertidora de angiotensina no mostró ser un factor de riesgo significativo para enfermedad coronaria en la población de Montería.

Hidrólisis del aceite de coco (Cocos nucífera L mediante enzimas estereoespecíficas y sin especificidad posicional.

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Rodríguez, R.

1997-02-01

saponification, is also discussed.

El aceite de coco, por su composición, aporta a la dieta principalmente ácidos grasos de cadena corta y media, los que desde el punto de vista nutricional tienen ventajas, debido a su rápida utilización metabólica que permite brindar energía, principalmente a nivel hepático. Es importante contar con procedimientos que permitan obtener estos ácidos grasos en forma libre a partir del aceite de coco, ya que de esta manera pueden ser utilizados con diferentes fines, ya sea nutricionales, farmacológicos o tecnológicos. En el presente trabajo se estudió la hidrólisis del aceite de coco producida por dos tipos de lipasas; una lipasa sin especificidad posicional obtenida de Candida cylindracea, y otra lipasa sn-1',3' estereoespecífica, obtenida de Mucor miehei, en forma libre e inmovilizada (Lipozyme IM-20. La lipasa de Candida cylindracea hidroliza un 85%-90% de los triacilgliceroles del aceite a las 47-50 horas de acción. La composición del hidrolizado es muy similar a la del aceite, y en los monoacilgliceroles remanentes la composición de ácidos grasos es poco definida, aunque predominan los de cadena corta (C6 y C8 y media (C10-C14. La enzima de Mucor miehei permite alcanzar sólo un 65% de hidrólisis, que se obtiene a las 30 horas en el caso de la enzima libre y a las 10 horas en el caso de la enzima inmovilizada (Lipozyme IM-20. La composición de ácidos grasos del hidrolizado es muy similar en ambos casos, destacando la presencia de ácidos grasos en el rango C8-C14, aunque con una composición muy diferente a la del aceite original. La composición de ácidos grasos del monoacilglicerol remanente muestra una marcada predominancia de ácido láurico (C12:0 en el producto de ambas formas de la enzima, lo cual indicaría que este ácido graso ocupa mayoritariamente la posición sn-2' de los triacilgliceroles del aceite de coco. Se discute la utilidad de las lipasas para obtener fracciones

Metodologia de seleção de cepas para produção etodologia da ciclodextrina glicosiltransferase e para purificação da enzima - DOI: 10.4025/actascihealthsci.v29i1.133 Strains selection methodology for cyclodextrin glycosyltransferase production and enzyme purification - DOI: 10.4025/actascihealthsci.v29i1.133

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Graciette Matioli

2007-12-01

Full Text Available As ciclodextrinas (CDs são maltooligossacarídeos, produzidas a partir do amido, pela enzima ciclodextrina glicosiltransferase (CGTase. Esta pesquisa teve por objetivo estabelecer metodologias de seleção de cepas para produção de CGTase e para purificação da enzima. Os microrganismos foram selecionados a partir de 53 análises de solos de cultura de amido, em placas contendo meio de cultivo específico, para seleção de cepas produtoras de CGTase. As enzimas foram obtidas com cultivo destes microrganismos em meio líquido. As atividades enzimáticas das CGTases foram determinadas pelos métodos espectrofotométricos e precipitação com tricloroetileno. A cepa isolada do solo de aveia foi a que apresentou maior atividade [0,1864 mol de -CD (min mL-1]. Esta cepa foi utilizada para a produção da enzima em escala laboratorial e purificação em cromatografia de afinidade bioespecífica. A cepa selecionada nesta pesquisa abre novas perspectivas para produção de enzima e CDs em escala industrial. Palavras-chave: ciclodextrina, glicosiltransferase, CGTase.Cyclodextrins (CDs are maltooligosaccharides produced from starch by cyclodextrin glycosyltransferase (CGTase enzyme. This research aimed at establishing method for strains selection for CGTase production and enzyme purification. The microorganisms were selected from 53 analyses of starch cultures soils on plates containing specific culture medium for strains selection that produce CGTase. The enzymes were obtained by culturing these microorganisms in liquid medium. The enzyme activity was determined with photospectrometric methods and precipitation with trichloroethylene. The strain isolated from oat soil was the one that showed the highest activity [0.1864 µmol of ß-CD (min mL-1]. This strain was used for enzyme production in laboratory scale and purification by biospecific affinity chromatography. The strain selected in this research opens new perspectives for enzymes

Sucrose metabolizing enzymes in cell suspension cultures of Bauhinia forficata, Curcuma zedoaria and Phaseolus vulgaris Enzimas do metabolismo da sacarose em cultura celular de Bauhinia forficata, Curcuma zedoaria e Phaseolus vulgaris

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Marcia Ometto de Mello

2001-09-01

Full Text Available The objective of this work was to study the activity of sucrose metabolizing enzymes in extracts of cell suspension cultures of Bauhinia forficata Link, Curcuma zedoaria Roscoe and Phaseolus vulgaris L. Invertase pathway was identified in the three studied species. Sucrose synthase pathway was also responsible for sucrose metabolism in Curcuma zedoaria and Phaseolus vulgaris cells. Activity values higher than 300 nmol min-1 mg-1 of protein were found for acid and neutral invertases, UDPglucose pyrophosphorylase and phosphoglucomutase in the cell extract of the three plant species. Sucrose synthase showed low activity in Bauhinia forficata cells. As sucrose concentration in the culture medium decreased, sucrose synthase activity increased in C. zedoaria and P. vulgaris cells. The glycolytic enzymes activity gradually reduced at the end of the culture period, when carbohydrate was limited.O objetivo deste trabalho foi estudar as enzimas do metabolismo da sacarose em culturas de célula em suspensão de Bauhinia forficata Link, Curcuma zedoaria Roscoe e Phaseolus vulgaris L. A via da invertase foi identificada nas três espécies estudadas. A via da sacarose sintase também foi responsável pelo metabolismo da sacarose em células de Curcuma zedoaria e Phaseolus vulgaris. Foram encontradas atividades maiores que 300 nmol min-1 mg-1 de proteína das enzimas invertase ácida e alcalina, UDPglicose pirofosforilase e fosfoglicomutase no extrato celular das três espécies de plantas. A sacarose sintase mostrou atividade baixa nas células de Bauhinia forficata. À medida que a concentração de sacarose no meio de cultura diminuiu, a atividade da sacarose sintase aumentou em células de Curcuma zedoaria e Phaseolus vulgaris. Ao final do período de cultura, quando os carboidratos se tornaram limitantes, as atividades das enzimas glicolíticas reduziram-se gradualmente.

Mice plasma fibrinogen consumption by thrombin-like enzyme present in rattlesnake venom from the North-East region of Argentina Consumo de fibrinógeno plasmático en ratones por acción de enzima con actividad trombínica presente en el veneno de cascabel del nordeste argentino

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Silvana L. Maruñak

2004-12-01

Full Text Available Due to variability of venom components from the same species of snakes that inhabit different regions, particular properties of the venom of Crotalus durissus terrificus that inhabits the North-East of Argentina were studied. Gyroxin, a thrombin-like enzyme, was isolated from this venom by gel filtration and affinity chromatography, it was found to be homogeneous according to SDS-PAGE, with a molecular weight of 33 kDa. "Gyroxin syndrome" in mice was tested and it showed changes in the animal behavior, confirming that the isolated thrombin-like enzyme is gyroxin. Effects of this enzyme and the crude venom on mice plasmatic fibrinogen levels were determined. The mice plasma fibrinogen decreased rapidly until incoagulability during the first hour after thrombin-Iike enzyme injection, then reaching its normal level 10 hours after injection; whereas crude venom resulted in a 60% decrease of the mice plasma fibrinogen, reaching its normal level after the same period of time. After 1 hour of gyroxin inoculation, intravascular coagulation was observed in histological cuttings of lung, cardiac muscle and liver. The isolated enzyme showed strong hydrolyzing activity on fibrinogen and fibrin in vitro, whereas the crude venom exhibited weak hydrolyzing activity on both substrates. It is probable that this very low activity is due to the low percentage of the enzyme in the crude venom. Decreasing of plasmatic fibrinogen levels may be due to either the coagulant or hydrolyzing actions of the enzyme.Teniendo en cuenta la variabilidad de los componentes del veneno de serpientes de una misma especie que habitan regiones diferentes, se decidió estudiar las propiedades particulares del veneno de Crotalus durissus terrificus que habita el nordeste de Argentina, Giroxina, una enzima con actividad trombínica, fue aislada del veneno por cromatografía de filtración por gel y de afinidad; se comprobó su homogeneidad y se determinó su peso molecular, 33 kDa, por

Molecular cloning, expression, and characterization of mouse amine N-sulfotransferases

International Nuclear Information System (INIS)

Takahashi, Saki; Sakakibara, Yoichi; Mishiro, Emi; Kouriki, Haruna; Nobe, Rika; Kurogi, Katsuhisa; Yasuda, Shin; Liu, M.-C.; Suiko, Masahito

2008-01-01

By searching the GenBank database, we recently identified a novel mouse cytosolic sulfotransferase (SULT) cDNA (IMAGE Clone ID 679629) and a novel mouse SULT gene (LOC 215895). Sequence analysis revealed that both mouse SULTs belong to the cytosolic SULT3 gene family. The recombinant form of these two newly identified SULTs, designated SULT3A1 and SULT3A2, were expressed using the pGEX-4T-1 glutathione S-transferase fusion system and purified from transformed BL21 Escherichia coli cells. Both purified SULT3A1 and SULT3A2 exhibited strong amine N-sulfonating activities toward 1-naphthylamine among a variety of endogenous and xenobiotic compounds tested as substrates. Kinetic constants of the sulfation of 1-naphthylamine and 1-naphthol by these two enzymes were determined. Collectively, these results imply that these two amine-sulfonating SULT3s may play essential roles in the metabolism and detoxification of aromatic amine compounds in the body

Glutathione transferases are structural and functional outliers in the thioredoxin fold.

Science.gov (United States)

Atkinson, Holly J; Babbitt, Patricia C

2009-11-24

Glutathione transferases (GSTs) are ubiquitous scavengers of toxic compounds that fall, structurally and functionally, within the thioredoxin fold suprafamily. The fundamental catalytic capability of GSTs is catalysis of the nucleophilic addition or substitution of glutathione at electrophilic centers in a wide range of small electrophilic compounds. While specific GSTs have been studied in detail, little else is known about the structural and functional relationships between different groupings of GSTs. Through a global analysis of sequence and structural similarity, it was determined that variation in the binding of glutathione between the two major subgroups of cytosolic (soluble) GSTs results in a different mode of glutathione activation. Additionally, the convergent features of glutathione binding between cytosolic GSTs and mitochondrial GST kappa are described. The identification of these structural and functional themes helps to illuminate some of the fundamental contributions of the thioredoxin fold to catalysis in the GSTs and clarify how the thioredoxin fold can be modified to enable new functions.

Avaliação da celulase e pectinase como enzimas complementares, no processo de hidrólise-sacarificação do farelo de mandioca para produção de etanol Evaluation of the cellulase and pectinase by complementary enzymes in the process of hydrolysis-saccharification of cassava fibrous waste for alcohol production

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Magali LEONEL

1999-01-01

Full Text Available Neste trabalho objetivou-se avaliar o uso de enzimas complementares no processo enzimático de hidrólise e sacarificação para a produção de etanol a partir do resíduo fibroso das fecularias. Os resultados obtidos demonstraram que 63,42% do amido foram hidrolisados no tratamento em que não se utilizaram enzimas complementares. No tratamento com as duas enzimas complementares foram hidrolisados 89,55%, no tratamento com celulase 65,42% e no tratamento com pectinase 88,73%. A prensagem do resíduo após o processo de hidrólise e sacarificação mostrou-se eficiente, ficando 10,43% do total de açúcares obtidos retidos no resíduo fibroso final. Portanto, o tratamento em que se utilizou a pectinase como enzima complementar na hidrólise foi o melhor. A celulase não apresentou efeito significativo no rendimento do processo.This work it was proposed to evaluate the use of complementary enzymes (cellulase and pectinase in the enzymatic process of hydrolysis-saccharification of the cassava fibrous waste for alcohol production. The results indicated that 63,42% of starch was hydrolyzed in the treatment without complementary enzymes, 89,55% in the treatment with the enzymes, 65,42% with the cellulase by complementary and 88,73% in the pectinase treatment. The pressing was efficacious for sugar recuperation and 10% of total sugar was retaining in the final fibrous residue. The pectinase was the better complementary enzyme enhance the yield.

SERUM ACTIVITIES OF ASPARTATE AMINOTRANSFERASE, CREATINE KINASE AND LACTATE DEHYDROGENASE IN HORSES WITH COLIC ATIVIDADE SÉRICA DAS ENZIMAS ASPARTATO AMINOTRANSFERASE, CREATINA QUINASE E LACTATO DESIDROGENASE EM EQÜINOS COM CÓLICA

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Aureo Evangelista Santana

2008-12-01

Full Text Available Seventy equines distributed in two experimental groups were used, G1 (20 healthy equines, and G2 (50 equines with colic. Blood samples were obtained by jugular vein puncture in ten different moments. The variables aspartate aminotransferase (AST, creatine kinase (CK, and lactate dehydrogenase (LDH were determined by spectrophotometric assay using specific reagents. The average values presented by the animals of the G2 for variables CK, AST, and LDH were higher (P<0.05 than the values presented by the animals of the G1 in all the evaluation moments. The results showed for G2 animals suggest the existence of acute muscle injury. The muscle injuries in equines with colic were attributed to the tissue hypoperfusion, and the muscular damage.

KEY WORDS: Acute abdomen, horses, muscles enzyme. De setenta eqüinos, distribuídos em dois grupos experimentais – G1 (vinte eqüinos hígidos e G2 (cinqüenta eqüinos com cólica –, colheram-se amostras de sangue em dez diferentes momentos, mediante punção da jugular, para a determinação da atividade sérica das enzimas aspartato aminotransferase (AST, creatina quinase (CK e lactato desidrogenase (LDH. Os valores médios apresentados pelos animais do G2, para as variáveis CK, AST e LDH, foram superiores (P<0,05 aos valores médios apresentados pelos animais do G1 em todos os momentos de avaliação. Os resultados apresentados pelos animais com cólica (G2 sugerem a existência de lesão muscular aguda, porém com tendência a cura, e foram atribuídos a hipoperfusão tecidual e a traumas musculares. A análise seriada das enzimas CK, AST e LDH auxilia tanto no diagnóstico de lesões musculares em eqüinos com cólica como no acompanhamento da evolução do processo de cura.

PALAVRAS-CHAVES: Abdômen agudo, cavalos, enzimas musculares.

Atividade β-D-N-Acetilglucosaminidásica de enzimas imobilizadas extraídas da Artemia franciscana e possíveis aplicações biotecnológicas

OpenAIRE

Santos, Pablo de Castro

2008-01-01

A β-D-N-acetilglucosaminidase, extraída e parcialmente isolada do crustáceo Artemia franciscana através de precipitação com sulfato de amônio e cromatografia em gel filtração Bio Gel A 1.5m foi imobilizada em Dacron ferromagnético rendendo um derivado insolúvel ativo contendo 5,0 unid/mg de proteína e retendo 10,35% da atividade da enzima solúvel. A β-D-N-acetilglucosaminidase-Dacron ferromagnético foi facilmente removida do meio reacional com o auxílio de um campo magnético e pô...

Influencia de la alimentación sobre el ritmo circadiano de las enzimas digestivas en el cultivo de juveniles de la langosta de agua dulce Cherax quadricarinatus (Parastacidae

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Hernán J Sacristán

2013-09-01

Full Text Available Se analiza en juveniles en etapa de engorde (7 g de Cherax quadricarinatus, el ritmo circadiano de los niveles de proteínas totales, la actividad de las proteasas, lipasas y amilasas, y su posible modificación en función del momento de alimentación matutino (8 h y vespertino (17 h. En general, la actividad de las enzimas digestivas de los juveniles de C. quadricarinatus no mostró un patrón circadiano de secreción, a excepción de las lipasas, donde se observó un incremento de actividad hacia la tarde-noche y disminución hacia las primeras horas de la mañana. Los niveles de proteína total, la actividad de proteinasas y amilasas registradas en la glándula digestiva, permanecieron prácticamente inalterados en todos los experimentos. A pesar de ello, se observó tendencia a que los niveles de actividad de proteinasas disminuyan luego del pulso de alimentación, recuperando los niveles anteriores 3 h después, independientemente del horario de alimentación. En cambio la actividad de amilasa no mostró ninguna tendencia por efecto del horario de alimentación. El estudio de los patrones de secreción de las enzimas digestivas y sus posibles modificaciones, podría ser utilizado como una herramienta para establecer los momentos del día más propicios para la alimentación de los juveniles en cultivo.

Palmito de pupunha (Bactris gasipaes Kunth. composição mineral e cinética de enzimas oxidativas Heart of palm (Bactris gasipaes Kunth.: mineral composition and kinetics of oxidative enzymes

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Nathália Ottoboni Galdino

2008-09-01

Full Text Available A análise da presença de enzimas oxidativas como a peroxidase (POD e a polifenoloxidase (PPO e o controle da atividade destas enzimas são importantes na preservação e no processamento de alimentos. Este trabalho teve por objetivo determinar a atividade enzimática da polifenoloxidase (PPO e da peroxidase (POD do palmito de pupunha, bem como avaliar o comportamento destas enzimas frente ao tratamento térmico e assim calcular a cinética de inativação térmica das mesmas para suas porções termorresistente e termolábil. Para a extração de peroxidase (POD e polifenoloxidase (PPO de palmito, utilizou-se solução tampão fosfato de sódio 100 mM com diferentes pHs (5,5; 6,0; 6,5 e 7,0. O melhor pH de extração da POD foi 5,5 e da PPO, 6,5. Estes extratos foram tratados em diferentes temperaturas (65, 70, 75 e 80 °C por períodos de 1 a 10 minutos. A POD e a PPO sofreram um decréscimo de 70 e 80%, respectivamente, em relação às suas atividades iniciais. As energias de ativação, nas temperaturas estudadas, para a porção termolábil e termorresistente da peroxidase foram 154,0 e 153,0 kJ.mol-1, respectivamente, enquanto que para a polifenoloxidase foram 26,3 e 27,0 kJ.mol-1, respectivamente. Resultados apresentaram valores que estão dentro da faixa de energia de ativação reportada para o processo de inativação térmica de enzimas.Analysis of oxidative enzymes such as peroxidase (POD and polyphenoloxidase (PPO and the control of the activity of these enzymes are important in food preservation and also in food processing. The aim of this work was to determine polyphenoloxidase (PPO and peroxidase (POD enzymatic activity in heart of palm, as well as to evaluate enzyme behavior during thermal treatment, determining the kinetics of thermal inactivation of the heat resistant and heat labile portions. For the extraction of peroxidase (POD and polyphenoloxidase (PPO from the heart of palm solution, 100 mM sodium phosphate buffer with

Efeito da peletização e adição de enzimas e vitaminas sobre o desempenho e aproveitamento da energia e nutrientes em frangos de corte de 1 a 21 dias de idade Effect of pelleting and addition of enzymes and vitamins on the performance and advantage of energy and nutrients in broiler chickens from 1 to 21 days old

OpenAIRE

José Laureano Barbosa Leite; Paulo Borges Rodrigues; Elias Tadeu Fialho; Rilke Tadeu Fonseca de Freitas; Adriano Kaneo Nagata; Vinícius de Souza Cantarelli

2008-01-01

Conduziu-se este experimento para avaliar o efeito da peletização e adição de enzimas e vitaminas sobre o desempenho e aproveitamento da energia e nutrientes, em frangos de corte de 1 a 21 dias de idade. Foram utilizados 350 pintos de corte, COBB, machos, com peso inicial médio de 47g ± 2,5g, em um delineamento inteiramente casualizado, com os tratamentos em arranjos fatorial 3 x 2 (três dietas, com e sem enzimas) sendo que as dietas consistiram de uma ração farelada e duas rações peletizadas...

Characterization of glutathione transferases involved in the pathogenicity of Alternaria brassicicola.

Science.gov (United States)

Calmes, Benoit; Morel-Rouhier, Mélanie; Bataillé-Simoneau, Nelly; Gelhaye, Eric; Guillemette, Thomas; Simoneau, Philippe

2015-06-18

Glutathione transferases (GSTs) represent an extended family of multifunctional proteins involved in detoxification processes and tolerance to oxidative stress. We thus anticipated that some GSTs could play an essential role in the protection of fungal necrotrophs against plant-derived toxic metabolites and reactive oxygen species that accumulate at the host-pathogen interface during infection. Mining the genome of the necrotrophic Brassica pathogen Alternaria brassicicola for glutathione transferase revealed 23 sequences, 17 of which could be clustered into the main classes previously defined for fungal GSTs and six were 'orphans'. Five isothiocyanate-inducible GSTs from five different classes were more thoroughly investigated. Analysis of their catalytic properties revealed that two GSTs, belonging to the GSTFuA and GTT1 classes, exhibited GSH transferase activity with isothiocyanates (ITC) and peroxidase activity with cumene hydroperoxide, respectively. Mutant deficient for these two GSTs were however neither more susceptible to ITC nor less aggressive than the wild-type parental strain. By contrast mutants deficient for two other GSTs, belonging to the Ure2pB and GSTO classes, were distinguished by their hyper-susceptibility to ITC and low aggressiveness against Brassica oleracea. In particular AbGSTO1 could participate in cell tolerance to ITC due to its glutathione-dependent thioltransferase activity. The fifth ITC-inducible GST belonged to the MAPEG class and although it was not possible to produce the soluble active form of this protein in a bacterial expression system, the corresponding deficient mutant failed to develop normal symptoms on host plant tissues. Among the five ITC-inducible GSTs analyzed in this study, three were found essential for full aggressiveness of A. brassicicola on host plant. This, to our knowledge is the first evidence that GSTs might be essential virulence factors for fungal necrotrophs.

Glutathione transferase supergene family in tomato: Salt stress-regulated expression of representative genes from distinct GST classes in plants primed with salicylic acid.

Science.gov (United States)

Csiszár, Jolán; Horváth, Edit; Váry, Zsolt; Gallé, Ágnes; Bela, Krisztina; Brunner, Szilvia; Tari, Irma

2014-05-01

A family tree of the multifunctional proteins, glutathione transferases (GSTs, EC 2.5.1.18) was created in Solanum lycopersicum based on homology to known Arabidopsis GSTs. The involvement of selected SlGSTs was studied in salt stress response of tomato primed with salicylic acid (SA) or in un-primed plants by real-time qPCR. Selected tau GSTs (SlGSTU23, SlGSTU26) were up-regulated in the leaves, while GSTs from lambda, theta, dehydroascorbate reductase and zeta classes (SlGSTL3, SlGSTT2, SlDHAR5, SlGSTZ2) in the root tissues under salt stress. Priming with SA exhibited a concentration dependency; SA mitigated the salt stress injury and caused characteristic changes in the expression pattern of SlGSTs only at 10(-4) M concentration. SlGSTF4 displayed a significant up-regulation in the leaves, while the abundance of SlGSTL3, SlGSTT2 and SlGSTZ2 transcripts were enhanced in the roots of plants primed with high SA concentration. Unexpectedly, under high salinity the SlDHAR2 expression decreased in primed roots as compared to the salt-stressed plants, however, the up-regulation of SlDHAR5 isoenzyme contributed to the maintenance of DHAR activity in roots primed with high SA. The members of lambda, theta and zeta class GSTs have a specific role in salt stress acclimation of tomato, while SlGSTU26 and SlGSTF4, the enzymes with high glutathione conjugating activity, characterize a successful priming in both roots and leaves. In contrast to low concentration, high SA concentration induced those GSTs in primed roots, which were up-regulated under salt stress. Our data indicate that induction of GSTs provide a flexible tool in maintaining redox homeostasis during unfavourable conditions. Copyright © 2014 Elsevier Masson SAS. All rights reserved.

Optimação de biocatalisadores: desenvolvimento de estratégias para modulação de propriedades de enzimas por técnicas físicas e químicas

OpenAIRE

Santos, José Cleiton Sousa dos

2015-01-01

Tesis doctoral inédita cotutelada por la Universidad Federal do Ceara (Benfica-Fortaleza), Centro de Tecnología, Departamento de Engenharía Química y por la Universidad Autónoma de Madrid, Facultad de Ciencias, Departamento de Biología Molecular. Fecha de lectura: 27-02-2015 Este trabalho de tese apresenta suportes ativados com divilsulfona (DVS) como úteis para a estabilização de enzimas por ligação covalente multipontual, os suportes são muito estáveis, e reagem com diferentes grup...

Modeling and Stability Assessment of Single-Phase Grid Synchronization Techniques

DEFF Research Database (Denmark)

Golestan, Saeed; Guerrero, Josep M.; Vasquez, Juan

2018-01-01

(GSTs) is of vital importance. This task is most often based on obtaining a linear time-invariant (LTI) model for the GST and applying standard stability tests to it. Another option is modeling and dynamics/stability assessment of GSTs in the linear time-periodic (LTP) framework, which has received...... a very little attention. In this letter, the procedure of deriving the LTP model for single-phase GSTs is first demonstrated. The accuracy of the LTP model in predicting the GST dynamic behavior and stability is then evaluated and compared with that of the LTI one. Two well-known single-phase GSTs, i...

Production of hydrolytic enzymes by the plant pathogenic fungus Myrothecium verrucaria in submerged cultures Produção de enzimas hidrolíticas pelo fungo fitopatogênico Myrothecium verrucaria em culturas submersas

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Fabiana Guillen Moreira

2005-03-01

Full Text Available The capability of the plant pathogenic fungus Myrothecium verrucaria to produce extracellular hydrolytic enzymes in submerged cultures was studied using several substrates. The fungus was able to produce different depolymerases and glycosidases, being xylanase, pectinase and protease the most important. Lipase was found in cultures developed in the presence of olive oil, while protease activity was detected in all cultures. Xylanase and pectinase were optimally active at pH 4.5-5.5, while protease was active in a large range of pH 3.5 to 11.0. All three enzymes were maximally active at 40ºC and they were stable for several hours at temperature up to 50ºC.A capacidade do fungo fitopatogênico Myrothecium verrucaria produzir enzimas hidrolíticas extracelulares em culturas submersas foi estudada utilizando diversos substratos. O fungo foi capaz de produzir diferentes depolimerases e glicosidases, sendo xilanases, pectinases e proteases as mais importantes. Atividade lipase foi encontrada nos filtrados das culturas desenvolvidas na presença de óleo de oliva, enquanto atividade proteolítica foi detectada em todas as culturas. Xilanase e pectinase foram otimamente ativas em pH 4,5 a 5,5, enquanto protease foi ativa em ampla faixa de pH (3,5 a 11,0. As três enzimas foram otimamente ativas 40ºC e estáveis por várias horas a temperaturas até 50ºC.

Topical estradiol does not interfere with the expression of the metalloproteinase-1 enzyme in photo exposed skin cells Estradiol tópico não interfere na expressão da enzima metaloproteinase-1 em células da pele fotoexposta

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Luciana Neder

2012-02-01

Full Text Available BACKGROUND: In postmenopausal women there is a rapid destruction of dermal collagen, resulting in accelerated skin ageing, which is manifested by cutaneous atrophy, increased number and depth of wrinkles and sagging. This accelerated catabolism of the collagen is due to estrogen deficiency and increased synthesis of the metalloproteinase-1 enzyme, which degrades the dermal collagen. OBJECTIVES: To assess whether the use of topical estradiol 0.05% cream on photo exposed skin can inhibit the expression of the metalloproteinase-1 enzyme on the dermis and subsequently the rapid loss of collagen in women after menopause. METHODS: We included 40 postmenopausal women without hormone replacement therapy. Information about lifestyle, lipid profile, blood glucose level, thyroid hormones, mammography, Pap smear and transvaginal ultrasound were obtained to rule out associated diseases. Skin biopsy of the right preauricular region was performed before and after treatment with topical estradiol 0.05% for 30 days. The biopsy specimens were subjected to immunohistochemistry to identify the expression of the metalloproteinase-1 enzyme. RESULTS: There was no statistically significant difference on the expression of the metalloproteinase-1 enzyme in keratinocytes, fibroblasts and endothelial cells before and after treatment with topical estradiol for 30 days. CONCLUSION: Treatment with estradiol 0.05% cream, in photo exposed skin for 30 days, does not inhibit the production of metalloproteinase-1.FUNDAMENTOS: Na pós-menopausa, ocorre rápida destruição do colágeno dérmico, com consequente envelhecimento acelerado da pele, que se expressa com atrofia cutânea, aumento do número e da profundidade das rugas e flacidez. Esse catabolismo acelerado do colágeno ocorre por deficiência estrogênica e aumento na síntese da enzima metaloproteinase-1, que degrada o colágeno dérmico. OBJETIVOS: Avaliar se o uso de estradiol tópico a 0,05% em creme na pele

Purification and properties of the glutathione S-transferases from the anoxia-tolerant turtle, Trachemys scripta elegans.

Science.gov (United States)

Willmore, William G; Storey, Kenneth B

2005-07-01

Glutathione S-transferases (GSTs) play critical roles in detoxification, response to oxidative stress, regeneration of S-thiolated proteins, and catalysis of reactions in nondetoxification metabolic pathways. Liver GSTs were purified from the anoxia-tolerant turtle, Trachemys scripta elegans. Purification separated a homodimeric (subunit relative molecular mass =34 kDa) and a heterodimeric (subunit relative molecular mass = 32.6 and 36.8 kDa) form of GST. The enzymes were purified 23-69-fold and 156-174-fold for homodimeric and heterodimeric GSTs, respectively. Kinetic data gathered using a variety of substrates and inhibitors suggested that both homodimeric and heterodimeric GSTs were of the alpha class although they showed significant differences in substrate affinities and responses to inhibitors. For example, homodimeric GST showed activity with known alpha class substrates, cumene hydroperoxide and p-nitrobenzylchloride, whereas heterodimeric GST showed no activity with cumene hydroperoxide. The specific activity of liver GSTs with chlorodinitrobenzene (CDNB) as the substrate was reduced by 2.6- and 8.7-fold for homodimeric and heterodimeric GSTs isolated from liver of anoxic turtles as compared with aerobic controls, suggesting an anoxia-responsive stable modification of the protein that may alter its function during natural anaerobiosis.

Versatile Gene-Specific Sequence Tags for Arabidopsis Functional Genomics: Transcript Profiling and Reverse Genetics Applications

Science.gov (United States)

Hilson, Pierre; Allemeersch, Joke; Altmann, Thomas; Aubourg, Sébastien; Avon, Alexandra; Beynon, Jim; Bhalerao, Rishikesh P.; Bitton, Frédérique; Caboche, Michel; Cannoot, Bernard; Chardakov, Vasil; Cognet-Holliger, Cécile; Colot, Vincent; Crowe, Mark; Darimont, Caroline; Durinck, Steffen; Eickhoff, Holger; de Longevialle, Andéol Falcon; Farmer, Edward E.; Grant, Murray; Kuiper, Martin T.R.; Lehrach, Hans; Léon, Céline; Leyva, Antonio; Lundeberg, Joakim; Lurin, Claire; Moreau, Yves; Nietfeld, Wilfried; Paz-Ares, Javier; Reymond, Philippe; Rouzé, Pierre; Sandberg, Goran; Segura, Maria Dolores; Serizet, Carine; Tabrett, Alexandra; Taconnat, Ludivine; Thareau, Vincent; Van Hummelen, Paul; Vercruysse, Steven; Vuylsteke, Marnik; Weingartner, Magdalena; Weisbeek, Peter J.; Wirta, Valtteri; Wittink, Floyd R.A.; Zabeau, Marc; Small, Ian

2004-01-01

Microarray transcript profiling and RNA interference are two new technologies crucial for large-scale gene function studies in multicellular eukaryotes. Both rely on sequence-specific hybridization between complementary nucleic acid strands, inciting us to create a collection of gene-specific sequence tags (GSTs) representing at least 21,500 Arabidopsis genes and which are compatible with both approaches. The GSTs were carefully selected to ensure that each of them shared no significant similarity with any other region in the Arabidopsis genome. They were synthesized by PCR amplification from genomic DNA. Spotted microarrays fabricated from the GSTs show good dynamic range, specificity, and sensitivity in transcript profiling experiments. The GSTs have also been transferred to bacterial plasmid vectors via recombinational cloning protocols. These cloned GSTs constitute the ideal starting point for a variety of functional approaches, including reverse genetics. We have subcloned GSTs on a large scale into vectors designed for gene silencing in plant cells. We show that in planta expression of GST hairpin RNA results in the expected phenotypes in silenced Arabidopsis lines. These versatile GST resources provide novel and powerful tools for functional genomics. PMID:15489341

“extracción, purificación parcial y secado de la enzima bromelina obtenida a partir del corazón de la piña (ananas comosus)”.

OpenAIRE

Quinde Fuentes, Carmina Stefania; Sánchez Lluguín, Nancy Viviana

2013-01-01

El presente artículo tuvo como objetivo la purificación parcial de la enzima bromelina mediante el sistema bifásico acuoso (SBA), evaluando el efecto del pH y las concentraciones de los componentes PEG 8000 y sales fosfato del sistema. La bromelina fue extraída del corazón de la piña de la variedad Milagreña mediante solución buffer a 4°c con EDTA, cisteína y fosfato disódico anhídrido. El mejor resultado de purificación obtenido en el sistema bifásico acuoso fue el que consistió de 16% PEG, ...

Phytase enzyme in diets containing defatted rice bran for growing swine Enzima fitase dietas com farelo de arroz desengordurado para suínos em crescimento

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José Aparecido Moreira

2003-12-01

Full Text Available Organic phosphorus is poorly utilized by monogastric animals because they lack phytase, the enzyme that cleaves the ortho-phosphate groups from the phytate molecule. Diets fed to pigs are supplemented with inorganic P, and this can increase environmental pollution and diet costs. Sixty mixed sex, half-breed pigs, were used to evaluate the effect of increasing dietary levels of phytase (253, 759, 1265 and 1748 PU kg-1 feed on animal performance as compared to a control without phytase but supplemented with dicalcium phosphate. Enzyme levels did not affect daily feed intake, food conversion, average daily weight gain, plasma P and Ca, calcium and phosphorus in bone ash, and the calcium/phosphorus ratio in the plasma and bones. A quadratic relationship between phytase levels and the percentages of P and Ca in bone ash was observed, reaching a maximum at the 880 and 879 PU levels, respectively. Animals fed diets containing phytase presented low plasma P values when compared to the control, but no effects were observed for the regression analysis. Using 759 PU phytase in rations containing corn, soybean bran and defatted rice bran for growing pigs can eliminate the use of traditional sources of P.Os animais monogástricos não aproveitam eficientemente o P orgânico das dietas, pois não sintetizam a enzima fitase, sendo necessária a suplementação das rações com P inorgânico, podendo elevar o custo das dietas e a poluição ambiental. Foram utilizados 60 leitões mestiços (machos castrados e fêmeas para avaliar a eficácia dos níveis dietéticos crescentes da enzima fitase (253, 759, 1265 e 1748 UF kg-1 de ração sobre os parâmetros de desempenho e comparar com o tratamento testemunha que diferia dos demais por não conter fitase e por conter fosfato bicálcico. Os níveis da enzima fitase não afetaram o consumo diário de ração, conversão alimentar, ganho diário de peso, P e Ca no plasma, cinzas no osso e relação cálcio e f

Bioactivation of food genotoxicants 5-hydroxymethylfurfural and furfuryl alcohol by sulfotransferases from human, mouse and rat: a comparative study.

Science.gov (United States)

Sachse, Benjamin; Meinl, Walter; Sommer, Yasmin; Glatt, Hansruedi; Seidel, Albrecht; Monien, Bernhard H

2016-01-01

5-Hydroxymethylfurfural (HMF) and furfuryl alcohol (FFA) are moderately potent rodent carcinogens that are present in thermally processed foodstuffs. The carcinogenic effects were hypothesized to originate from sulfotransferase (SULT)-mediated bioactivation yielding DNA-reactive and mutagenic sulfate esters, a confirmed metabolic pathway of HMF and FFA in mice. It is known that orthologous SULT forms substantially differ in substrate specificity and tissue distribution. This could influence HMF- and FFA-induced carcinogenic effects. Here, we studied HMF and FFA sulfoconjugation by 30 individual SULT forms of humans, mice and rats. The catalytic efficiencies (k cat/K M) of HMF sulfoconjugation of human SULT1A1 (13.7 s(-1) M(-1)), mouse Sult1a1 (15.8 s(-1) M(-1)) and 1d1 (4.8 s(-1) M(-1)) and rat Sult1a1 (5.3 s(-1) M(-1)) were considerably higher than those of all other SULT forms investigated (≤0.73 s(-1 )M(-1)). FFA sulfoconjugation was monitored using adenosine as a nucleophilic scavenger for the reactive 2-sulfoxymethylfuran (t 1/2 = 20 s at 37 °C). The resulting adduct N (6)-((furan-2-yl)methyl)-adenosine (N (6)-MF-A) was quantified by isotope-dilution UPLC-MS/MS. The rates of N (6)-MF-A formation showed that hSULT1A1 and its orthologues in mice and rats were also the most important contributors to FFA sulfoconjugation in each of the species. Taken together, the catalytic capacity of hSULT1A1 is comparable to that of mSult1a1 in mice, the species in which carcinogenic effects of HMF and FFA were detected. This is of primary concern due to the expression of hSULT1A1 in many different tissues.

MANNAN OLIGOSACCHARIDES AND ENZYMES IN CORN AND SOYBEAN MEAL BASED-DIETS FOR BROILERS MANANOLIGOSSACARÍDEOS E ENZIMAS EM DIETAS À BASE DE MILHO E FARELO DE SOJA PARA AVES

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Vera Maria Barbosa de Moraes

2007-09-01

Full Text Available

Mannan oligosaccharides, as an alternative to an-tibiotics, show promising results related to bird perfor-mance, immunity stimulus and improvement of intestinal mucosa. Enzyme inclusion in diets containing mannan oli-gosaccharides could improve these results. Poultry diets consist of corn and soybean meal, and approximately 30% of the phosphorus (P in these ingredients is bound to phytic acid. Phytase is capable of release not only the P but also other nutrients that can be better used by the birds while corn is relatively free of viscosous nonstarch poly-saccharides (NSP, soybean has about 20% of NSP, whose digestibility is almost null. NSP`s insoluble component of corn and soybean encapsulate nutrients and are responsive to exogenous enzymes. Negative environmental and nutri-tional consequences of dietary phytic acid are reduced by phytase inclusion in monogastric diets. Additionally, the enzymatic complex supplementation improves the nutri-tional value of feedstuffs through higher digestibility in birds. Finally, the use of exogenous enzymes allows re-duction in feed costs.

KEY-WORDS: Animal nutrition, enzymatic supplementation, prebiotic.

Os mananoligossacarídeos, como alternativa aos antibióticos, têm apresentado resultados promissores no que se refere tanto ao desempenho das aves quanto ao estímulo da imunidade e à melhora da mucosa intestinal. A inclusão de enzimas nas dietas contendo mananoligossacarídeos poderia melhorar ainda mais esses resultados. As dietas para aves são compostas de milho e farelo de soja, principalmente, e cerca de 30% do fósforo (P desses in-gredientes estão ligados ao ácido fítico. A fitase libera não só o P, mas também outros nutrientes para que sejam me-lhor utilizados pelas aves. O milho é relativamente isento de polissacarídeos não-amiláceos (PNAs viscosos e a soja apresenta cerca de 20% de PNAs, com digestibilidade praticamente nula. Os componentes insolúveis dos PNAs do

On the sulfation of O-desmethyltramadol by human cytosolic sulfotransferases.

Science.gov (United States)

Rasool, Mohammed I; Bairam, Ahsan F; Kurogi, Katsuhisa; Liu, Ming-Cheh

2017-10-01

Previous studies have demonstrated that sulfate conjugation is involved in the metabolism of the active metabolite of tramadol, O-desmethyltramadol (O-DMT). The current study aimed to systematically identify the human cytosolic sulfotransferases (SULTs) that are capable of mediating the sulfation of O-DMT. The sulfation of O-DMT under metabolic conditions was demonstrated using HepG2 hepatoma cells and Caco-2 human colon carcinoma cells. O-DMT-sulfating activity of thirteen known human SULTs and four human organ specimens was examined using an established sulfotransferase assay. pH-Dependency and kinetic parameters were also analyzed using, respectively, buffers at different pHs and varying O-DMT concentrations in the assays. Of the thirteen human SULTs tested, only SULT1A3 and SULT1C4 were found to display O-DMT-sulfating activity, with different pH-dependency profiles. Kinetic analysis revealed that SULT1C4 was 60 times more catalytically efficient in mediating the sulfation of O-DMT than SULT1A3 at respective optimal pH. Of the four human organ specimens tested, the cytosol prepared from the small intestine showed much higher O-DMT-sulfating activity than cytosols prepared from liver, lung, and kidney. Both cultured HepG2 and Caco-2 cells were shown to be capable of sulfating O-DMT and releasing sulfated O-DMT into cultured media. SULT1A3 and SULT1C4 were the major SULTs responsible for the sulfation of O-DMT. Collectively, the results obtained provided a molecular basis underlying the sulfation of O-DMT and contributed to a better understanding about the pharmacokinetics and pharmacodynamics of tramadol in humans. Copyright © 2017 Institute of Pharmacology, Polish Academy of Sciences. Published by Elsevier Urban & Partner Sp. z o.o. All rights reserved.

The stereoselective sulfate conjugation of 4'-methoxyfenoterol stereoisomers by sulfotransferase enzymes.

Science.gov (United States)

Iyer, Lalitha V; Ramamoorthy, Anuradha; Rutkowska, Ewelina; Furimsky, Anna M; Tang, Liang; Catz, Paul; Green, Carol E; Jozwiak, Krzysztof; Wainer, Irving W

2012-10-01

The presystemic sulfate conjugation of the stereoisomers of 4'-methoxyfenoterol, (R,R')-MF, (S,S')-MF, (R,S')-MF, and (S,R')-MF, was investigated using commercially available human intestinal S9 fractions, a mixture of sulfotransferase (SULT) enzymes. The results indicate that the sulfation was stereospecific and that an S-configuration at the β-OH carbon of the MF molecule enhanced the maximal formation rates with (S,R')-MF  (S,S')-MF  (R,S')-MF ≈ (R,R')-MF, and competition studies demonstrated that (S,R')-MF is an effective inhibitor of (R,R')-MF sulfation (IC(50) = 60 μM). In addition, the results from a cDNA-expressed human SULT isoform screen indicated that SULT1A1, SULT1A3, and SULT1E1 can mediate the sulfation of all four MF stereoisomers. Previously published molecular models of SULT1A3 and SULT1A1 were used in docking simulations of the MF stereoisomers using Molegro Virtual Docker. The models of the MF-SULT1A3 and MF-SULT1A1 complexes indicate that each of the two chiral centers of MF molecule plays a role in the observed relative stabilities. The observed stereoselectivity is the result of multiple hydrogen bonding interactions and induced conformational changes within the substrate-enzyme complex. In conclusion, the results suggest that a formulation developed from a mixture of (R,R')-MF and (S,R')-MF may increase the oral bioavailability of (R,R')-MF. Copyright © 2012 Wiley Periodicals, Inc.

Identification, characterization and expression profiles of Chironomus riparius glutathione S-transferase (GST) genes in response to cadmium and silver nanoparticles exposure

Energy Technology Data Exchange (ETDEWEB)

Nair, Prakash M. Gopalakrishnan [School of Environmental Engineering and Graduate School of Energy and Environmental System Engineering, University of Seoul, 90 Jeonnong-dong, Dongdaemun-gu, Seoul 130-743 (Korea, Republic of); Choi, Jinhee, E-mail: jinhchoi@uos.ac.kr [School of Environmental Engineering and Graduate School of Energy and Environmental System Engineering, University of Seoul, 90 Jeonnong-dong, Dongdaemun-gu, Seoul 130-743 (Korea, Republic of)

2011-02-15

In this study, we report the identification and characterization of 13 cytosolic GST genes in Chironomus riparius from Expressed Sequence Tags (ESTs) database generated using pyrosequencing. Comparative and phylogenetic analyses were undertaken with Drosophila melanogaster and Anopheles gambiae GSTs and 3 Delta, 4 Sigma, 1 each in Omega, Epsilon, Theta, Zeta and 2 unclassified classes of GSTs were identified and characterized. The relative mRNA expression levels of all of the C. riparius GSTs (CrGSTs) genes under different developmental stages were varied with low expression in the larval stage. The antioxidant role of CrGSTs was studied by exposing fourth instar larvae to a known oxidative stress inducer Paraquat and the relative mRNA expression to different concentrations of cadmium (Cd) and silver nanoparticles (AgNPs) for various time intervals were also studied. All the CrGSTs showed up- or down regulation to varying levels based upon the concentration, and duration of exposure. The highest mRNA expression was noticed in Delta3, Sigma4 and Epsilon1 GST class in all treatments. These results show the role of CrGST genes in defense against oxidative stress and its potential as a biomarker to Cd and AgNPs exposure.

RELACION DE LA EXPRESION DEL RECEPTOR DE INSULINA CON LA EXPRESION DE LAS ENZIMAS GLICOGENICAS (GS1 Y GS2) Y GLUCONEOGENICAS EN LAS CELULAS DE TUBULO PROXIMAL DE RATAS NORMALES Y DIABETICAS

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GATICA GUTIERREZ, RODRIGO ORLANDO; GATICA GUTIERREZ, RODRIGO ORLANDO

2012-01-01

En el presente estudio se determinó los efectos sobre la expresión y localización del receptor de insulina (InsR), de glicógeno sin tasa (GS) y de las enzimas glicolíticas y gluconeogénicas en diabetes de largo plazo. Insulina es esencial para el manejo energético del cuerpo, sin embargo, su rol a nivel renal es poco conocido. Su unión al receptor de insulina provoca una cascada de señalización intracelular, que regula múltiples procesos biológicos, tales corno el metabolismo lipídico y pr...

Influencia de especies forestales sobre la actividad de las enzimas fosfatasa ácida y proteasas en un suelo de bosque Influence of tree species on the activity of acid phosphatase and protease in a forest soil

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Rl Defrieri

2008-12-01

Full Text Available Los distintos tipos de cubiertas vegetales y especialmente las especies arbóreas dominantes en los sistemas forestales, afectan las propiedades químicas y bioquímicas de los suelos. Dado que la concentración de algunos nutrientes en la hojarasca, entre ellos N y P, es considerada crítica para la descomposición del material vegetal, el objetivo de este trabajo ha sido estudiar a dos profundidades, la influencia del aporte de biomasa vegetal proveniente de especies forestales dominantes, sobre la actividad de las enzimas fosfatasa ácida y proteasas en un suelo de un monte del Parque Chaqueño Argentino. Se extrajeron muestras de suelo, ubicadas debajo de árboles de cuatro especies del monte, a dos profundidades (0-10 y 10-20 cm, en verano y en invierno sobre las que se determinaron las actividades enzimáticas mencionadas y algunas propiedades edáficas. Para todas las especies y en las dos épocas de muestreo se comprobó que los valores de las actividades de las dos enzimas y los parámetros edáficos analizados resultaron significativamente menores en las muestras a la profundidad de 10-20 cm. La influencia de las especies forestales sobre la actividad de las enzimas estudiadas se puso de manifiesto en el muestreo superficial en donde es mayor la influencia de la incorporación de los residuos orgánicos y en verano. En estas condiciones los valores de las actividades enzimáticas medidas debajo de cada especie oscilaron entre 1.600 y 900 μg p-nitrofenol g-1 h-1 para fosfatasa ácida y entre 850 y 450 mg tirosina g-1h-1 para proteasa. Para dos de las especies estudiadas se pudo vincular la magnitud de su aporte vegetal, su tasa de descomposición y los valores de N y P en hojas senescentes con la magnitud de la liberación de ambas enzimas al suelo según la especie. No se encontró asociación de la influencia de las especies con algunos parámetros edáficos analizados. Por lo tanto la actividad de las enzimas medidas en este trabajo

Identification and characterisation of multiple glutathione S-transferase genes from the diamondback moth, Plutella xylostella.

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Chen, Xi'en; Zhang, Ya-lin

2015-04-01

The diamondback moth (DBM), Plutella xylostella, is one of the most harmful insect pests on crucifer crops worldwide. In this study, 19 cDNAs encoding glutathione S-transferases (GSTs) were identified from the genomic and transcriptomic database for DBM (KONAGAbase) and further characterized. Phylogenetic analysis showed that the 19 GSTs were classified into six different cytosolic classes, including four in delta, six in epsilon, three in omega, two in sigma, one in theta and one in zeta. Two GSTs were unclassified. RT-PCR analysis revealed that most GST genes were expressed in all developmental stages, with higher expression in the larval stages. Six DBM GSTs were expressed at the highest levels in the midgut tissue. Twelve purified recombinant GSTs showed varied enzymatic properties towards 1-chloro-2,4-dinitrobenzene and glutathione, whereas rPxGSTo2, rPxGSTz1 and rPxGSTu2 had no activity. Real-time quantitative PCR revealed that expression levels of the 19 DBM GST genes were varied and changed after exposure to acephate, indoxacarb, beta-cypermethrin and spinosad. PxGSTd3 was significantly overexpressed, while PxGSTe3 and PxGSTs2 were significantly downregulated by all four insecticide exposures. The changes in DBM GST gene expression levels exposed to different insecticides indicate that they may play individual roles in tolerance to insecticides and xenobiotics. © 2014 Society of Chemical Industry.

Characterization and expression profiling of glutathione S-transferases in the diamondback moth, Plutella xylostella (L.).

Science.gov (United States)

You, Yanchun; Xie, Miao; Ren, Nana; Cheng, Xuemin; Li, Jianyu; Ma, Xiaoli; Zou, Minming; Vasseur, Liette; Gurr, Geoff M; You, Minsheng

2015-03-05

Glutathione S-transferases (GSTs) are multifunctional detoxification enzymes that play important roles in insects. The completion of several insect genome projects has enabled the identification and characterization of GST genes over recent years. This study presents a genome-wide investigation of the diamondback moth (DBM), Plutella xylostella, a species in which the GSTs are of special importance because this pest is highly resistant to many insecticides. A total of 22 putative cytosolic GSTs were identified from a published P. xylostella genome and grouped into 6 subclasses (with two unclassified). Delta, Epsilon and Omega GSTs were numerically superior with 5 genes for each of the subclasses. The resulting phylogenetic tree showed that the P. xylostella GSTs were all clustered into Lepidoptera-specific branches. Intron sites and phases as well as GSH binding sites were strongly conserved within each of the subclasses in the GSTs of P. xylostella. Transcriptome-, RNA-seq- and qRT-PCR-based analyses showed that the GST genes were developmental stage- and strain-specifically expressed. Most of the highly expressed genes in insecticide resistant strains were also predominantly expressed in the Malpighian tubules, midgut or epidermis. To date, this is the most comprehensive study on genome-wide identification, characterization and expression profiling of the GST family in P. xylostella. The diversified features and expression patterns of the GSTs are inferred to be associated with the capacity of this species to develop resistance to a wide range of pesticides and biological toxins. Our findings provide a base for functional research on specific GST genes, a better understanding of the evolution of insecticide resistance, and strategies for more sustainable management of the pest.

Molecular cloning and expression of five glutathione S-transferase (GST) genes from Banana (Musa acuminata L. AAA group, cv. Cavendish).

Science.gov (United States)

Wang, Zhuo; Huang, Suzhen; Jia, Caihong; Liu, Juhua; Zhang, Jianbin; Xu, Biyu; Jin, Zhiqiang

2013-09-01

Three tau class MaGSTs responded to abiotic stress, MaGSTF1 and MaGSTL1 responded to signaling molecules, they may play an important role in the growth of banana plantlet. Glutathione S-transferases (GST) are multifunctional detoxification enzymes that participate in a variety of cellular processes, including stress responses. In this study, we report the molecular characteristics of five GST genes (MaGSTU1, MaGSTU2, MaGSTU3, MaGSTF1 and MaGSTL1) cloned from banana (Musa acuminate L. AAA group, cv. Cavendish) using a RACE-PCR-based strategy. The predicted molecular masses of these GSTs range from 23.4 to 27.7 kDa and their pIs are acidic. At the amino acid level, they share high sequence similarity with GSTs in the banana DH-Pahang (AA group) genome. Phylogenetic analysis showed that the deduced amino acid sequences of MaGSTs also have high similarity to GSTs of other plant species. Expression analysis by semi-quantitative RT-PCR revealed that these genes are differentially expressed in various tissues. In addition, their expression is regulated by various stress conditions, including exposure to signaling molecules, cold, salinity, drought and Fusarium oxysporum f specialis(f. Sp) cubense Tropical Race 4 (Foc TR4) infection. The expression of the tau class MaGSTs (MaGSTU1, MaGSTU2 and MaGSTU3) mainly responded to cold, salinity and drought while MaGSTF1 and MaGSTL1 expressions were upregulated by signaling molecules. Our findings suggest that MaGSTs play a key role in both development and abiotic stress responses.

Enzyme expression in indica and japonica rice cultivars under saline stress=Expressão de enzimas em cultivares de arroz indica e japonica sob estresse salino

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Cristina Rodrigues Mendes

2012-10-01

indicas amplamente utilizadas no RS demonstram maior suscetibilidade à salinidade quando comparadas às japonicas em outros aspectos fisiológicos. O objetivo da pesquisa foi analisar a expressão de enzimas envolvidas na respiração de cultivares de arroz, indica e japonica, submetidas à salinidade. Foram utilizadas cultivares de Oryza sativa L. spp. japonica S. Kato (BRS Bojuru, IAS 12-9 Formosa e Goyakuman e de Oryza sativa L. spp. indica S. Kato (BRS-7 Taim, BRS Querência e BRS Atalanta. As plântulas foram transferidas para bacias de 15 L, contendo solução nutritiva de Hoagland meia força acrescida de 0, 25, 50, 75 e 100 mM de NaCl. A coleta foi aos 14, 28 e 42 dias. Os tecidos vegetais foram macerados e colocados em tubos eppendorf com solução extratora de Scandálios. A eletroforese foi realizada em géis de poliacrilamida 7% em cubas eletroforéticas verticais. As bandas foram reveladas para os sistemas enzimáticos esterase, álcool desidrogenase, fosfoglico isomerase, malato desidrogenase, enzima málica e alfa amilase. A expressão das enzimas foi maior na subespécie japonica, com bandas mais intensas conforme o aumento da salinidade. Conclui-se que tais sistemas enzimáticos estejam envolvidos em mecanismos de tolerância ao estresse salino nas cultivares de O. sativa spp. japonica.

USO DE ENZIMAS EN LA CRÍA Y ENGORDE DE POLLOS BROILERS EN ÉPOCA LLUVIOSA EN LAS LOCALIDADES DE QUEVEDO, SALCEDO Y SANTO DOMINGO DE LOS COLORADOS

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Ricardo Luna Murillo

2010-12-01

Full Text Available Con la finalidad de evaluar el uso de enzimas comerciales Ronozyme, SSF, VegPro e Hyboteck (Yucca schidigera en la cría y engorde de pollos broilers en época lluviosa se estableció un ensayo en tres provincias: Los Ríos (Quevedo, Santo Domingo de los Tsáchilas (Santo Domingo de los Colorados y Cotopaxi (Salcedo, utilizando un total de 1,440 aves de línea Hubbard (480 por localidad. Se empleó un Diseño Completamente al Azar, con cuatro tratamientos (enzimas y seis repeticiones con 20 pollos como unidad experimental. Las variables evaluadas fueron: consumo de alimento (g, ganancia de peso (g, conversión alimenticia, rendimiento a la canal (%, relación Costo/Beneficio y análisis bromatológicos de las canales. Se observó que los mayores consumo de alimento en las tres localidades se presentó con Hyboteck, (5,181.21; 6,713.42 y 5,680.37 g; la mejor ganancia de peso se observó con VegPro en Quevedo (2,543.34 g, SSF Santo Domingo de los Colorados (3,497.83 g e Hyboteck en Salcedo (2,286.08 g, la conversión más eficiente se reportó con Ronozyme en Quevedo (1.76, SSF en Santo Domingo de los Colorados (1.88, y VegPro en Salcedo (2.55. La mayor relación costo / beneficio se presentó con SSF en las localidades de Santo Domingo de los Colorados (0.10 y Salcedo (0.21 mientras en Quevedo la mejor fue Ronozyme (0.26. El mejor valor proteico de las canales de pollos se reportaron en las localidades de Quevedo y Salcedo con Ronozyme (26.05 y 22.80% respectivamente, y para la localidad de Santo Domingo de los Colorados fue con SSF (25.35% de proteína.

Efeitos genotóxicos e alterações de enzimas hepáticas em trabalhadores do refino de petróleo Genotoxic effects and hepatic enzymes alterations among petroleum refinery workers

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Rozana Oliveira Gonçalves

2005-10-01

Full Text Available Um estudo de casos e controles, aninhado num estudo de coorte, investigou a associação entre efeitos genotóxicos e alteração de enzimas hepáticas em trabalhadores de uma refinaria de petróleo do Nordeste. Foram examinados todos os dez novos casos de alterações de enzimas hepáticas - gama-glutamil transferase (GGT e alanina aminotransferase (ALT - ocorridos em 2002. Dez trabalhadores sem alterações de GGT ou ALT foram selecionados como controles. Os efeitos do fumo, sexo, idade e consumo de café foram controlados. O efeito genotóxico foi avaliado pela técnica de trocas entre cromátides irmãs (TCI e alterações cromossômicas (AC estruturais. As médias de TCI por célula (3,92 ± 1,04 versus 4,25 ± 1,47 e de ACE (8,85 ± 3,4 versus 9,1 ± 3,7 não diferiram de forma significante entre casos e controles respectivamente.A case-control study, nested in a cohort study, investigated the association between genotoxic effects and hepatic enzymes alterations among workers in a petroleum refinery, Northeast Brazil. Ten cases of hepatic enzymes alterations - gamma-glutamyltransferase (GGT and Alanine aminotransferase (ALT - representing all incident cases occurring in the refinery during 2002, were examined. Ten workers without GGT and ALT alterations were selected as controls. The effects of smoking, sex, age and coffee consumption were controlled. The genotoxic effects were evaluated by the sister chromatid exchange (SCE and by the chromosomal aberrations (CA techniques. Mean SCE per cell (3.92 ± 1.04 versus 4.25 ± 1.47 and CA per cell (8.85 ± 3.4 versus 9.1 ± 3.7 did not differ significantly between cases and controls respectively.

Procedimiento para la extracción de gluten contenido en alimentos, procesados y no procesados por calor, compatible con un ensayo de inmunoabsorción ligado a enzimas, composición y kits que comprenden dicha composición

OpenAIRE

López Villar, Elena; Llorente Gómez, María de las Mercedes; Méndez Cormán, Enrique

2001-01-01

La presente invención se refiera a, en general, el análisis de alimentos para enfermos celiacos, y, especialmente, se refiere a un procedimiento para la extracción de gluten de alimentos, compatible con un ensayo de inmunoabsorción ligado a enzimas (ELISA), a composiciones adecuadas para la puesta en práctica de dicho procedimiento, a kits que comprenden dichas composiciones y a un procedimiento para la cuantificación por ELISA del gluten presente en los alimentos.

Producción y purificación parcial de enzimas hidrolíticas de Aspergillus ficuum en fermentación sólida sobre residuos agroindustriales

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Marcia Costa

2010-07-01

Full Text Available Título en inglés: Production and partial purification of Aspergillus ficuum hydrolytic enzymes in solid state fermentation of agroindustrial residues Resumen En el presente trabajo se describe la producción de las enzimas fitasa, celulasa, xilanasa y proteasa con Aspergillus ficuum cepa DSM 932 mediante fermentación en estado sólido (SSF usando torta de canola y pomaza de cranberry como sustratos. Como medida indirecta de la producción de las enzimas se usó en cada caso la actividad enzimática. la torta de canola resultó ser un mejor sustrato para fitasa, celulasa y xilanasa, en tanto que la pomaza de cranberry resultó ser un sustrato potencial para proteasa. Mediante ultrafiltración escalonada fue posible purificar parcialmente los extractos enzimáticos de fitasa, celulasas y xilanasas, obtenidos a partir de torta de canola. La fitasa resultó tener un tamaño >100 kDa, en tanto que las celulasas y xilanasas presentan actividad en los retenidos de 10, 30 y 50 kDa, lo que indicaría que las isoenzimas de ambos complejos tienen pesos moleculares que oscilan entre 10 y 100 kDa. Palabras clave: fermentación sólida; torta de canola; Aspergillus ficuum; fitasa; xilanasa. Abstract In this paper, describes the production of the enzymes phytase, cellulase, xylanase and protease by Aspergillus ficuum DSM 932 strain, in solid state fermentation (SSF using canola cake and cranberry pomace as substrates. The enzyme activity was used in each case as an indirect measure of the enzymes production. Canola meal turned out to be a better substrate for phytase, cellulase and xylanase, while cranberry pomace was found to be a potential substrate for protease. Various ultrafiltration operations were carried out, decreasing the cut off membranes out in order to purify partially extracts of enzymes phytase, cellulase and xylanase, obtained from canola meal. Phytase was found to have a size >100 kDa, whereas cellulase and xylanase activity present in the

Bioprospección de enzimas de restricción en bacterias de suelos y ambientes volcánicos de Nicaragua

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Julio A, Gómez Rodríguez

2008-04-01

Full Text Available LA BIOPROSPECCIÓN ES LA BÚSQUEDA SISTEMÁTICA DE USOS SOSTENIBLES y con fines comerciales de los elementos genéticos y bioquímicos de la biodiversidad. En Nicaragua realizamos bioprospección de enzimas de restricción (ER de interés biotecnológico desde el año 1999. A través de esta investigación buscamos nuevas ER en bacterias de suelos en ambientes aún inexplorados, como zonas volcánicas y agrícolas del país. Aquí reportamos la identificación de isoesquizómeros de 14 ER conocidas: BamHI, EcoRV, MboI, MaeI, HaeIII, EarI, BsoBI, BsaBI, Acl I, BsaWI, Nci I, BstNI, BssSI y HpaII, las 8 últimas termoestables. La información de las ER encontradas en Nicaragua fue registrada en REBASE (Restriction Enzyme Database. Resaltamos la importancia del estudio de la biodiversidad microbianade los ambientes volcánicos nicaragüenses, ya que favorece la investigación de la evolución de la vida, los extremos donde podemos encontrarla y sus aplicaciones a la biotecnología.

The role of glutathione transferases in renal cell carcinoma

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Ćorić Vesna

2016-01-01

Full Text Available Mounting evidence suggest that members of the subfamily of cytosolic glutathione S-transferases (GSTs possess roles far beyond the classical glutathione-dependent enzymatic conjugation of electrophilic metabolites and xenobiotics. Namely, monomeric forms of certain GSTs are capable of forming protein: protein interactions with protein kinases and regulate cell apoptotic pathways. Due to this dual functionality of cytosolic GSTs, they might be implicated in both the development and the progression of renal cell carcinoma (RCC. Prominent genetic heterogeneity, resulting from the gene deletions, as well as from SNPs in the coding and non-coding regions of GST genes, might affect GST isoenzyme profiles in renal parenchyma and therefore serve as a valuable indicator for predicting the risk of cancer development. Namely, GSTs are involved in the biotransformation of several compounds recognized as risk factors for RCC. The most potent carcinogen of polycyclic aromatic hydrocarbon diol epoxides, present in cigarette smoke, is of benzo(apyrene (BPDE, detoxified by GSTs. So far, the relationship between GST genotype and BPDE-DNA adduct formation, in determining the risk for RCC, has not been evaluated in patients with RCC. Although the association between certain individual and combined GST genotypes and RCC risk has been debated in a the literature, the data on the prognostic value of GST polymorphism in patients with RCC are scarce, probably due to the fact that the molecular mechanism supporting the role of GSTs in RCC progression has not been clarified as yet.

GST Theta null genotype is associated with an increased risk for ulcerative colitis: a case-control study and meta-analysis of GST Mu and GST Theta polymorphisms in inflammatory bowel disease

NARCIS (Netherlands)

Broekman, M.M.T.J.; Bos, C.; Morsche, R.H.M. te; Hoentjen, F.; Roelofs, H.M.J.; Peters, W.H.M.; Wanten, G.J.A.; Jong, D.J. de

2014-01-01

Glutathione S-transferases (GSTs) are important in the detoxification of many compounds, including reactive oxygen species. Polymorphisms in GSTs resulting in a decreased enzyme activity might enhance the risk for inflammatory bowel disease by eliciting a state of oxidative stress. Previous

UDP-glucuronosyltransferase and sulfotransferase polymorphisms, sex hormone concentrations, and tumor receptor status in breast cancer patients

International Nuclear Information System (INIS)

Sparks, Rachel; Yuan, Xiaopu; Lin, Ming Gang; McVarish, Lynda; Aiello, Erin J; McTiernan, Anne; Ulrich, Cornelia M; Bigler, Jeannette; Tworoger, Shelley S; Yasui, Yutaka; Rajan, Kumar B; Porter, Peggy; Stanczyk, Frank Z; Ballard-Barbash, Rachel

2004-01-01

UDP-glucuronosyltransferase (UGT) and sulfotransferase (SULT) enzymes are involved in removing sex hormones from circulation. Polymorphic variation in five UGT and SULT genes – UGT1A1 ((TA) 6 /(TA) 7 ), UGT2B4 (Asp 458 Glu), UGT2B7 (His 268 Tyr), UGT2B15 (Asp 85 Tyr), and SULT1A1 (Arg 213 His) – may be associated with circulating sex hormone concentrations, or the risk of an estrogen receptor-negative (ER - ) or progesterone receptor-negative (PR - ) tumor. Logistic regression analysis was used to estimate the odds ratios of an ER - or PR - tumor associated with polymorphisms in the genes listed above for 163 breast cancer patients from a population-based cohort study of women in western Washington. Adjusted geometric mean estradiol, estrone, and testosterone concentrations were calculated within each UGT and SULT genotype for a subpopulation of postmenopausal breast cancer patients not on hormone therapy 2–3 years after diagnosis (n = 89). The variant allele of UGT1A1 was associated with reduced risk of an ER - tumor (P for trend = 0.03), and variants of UGT2B15 and SULT1A1 were associated with non-statistically significant risk reductions. There was some indication that plasma estradiol and testosterone concentrations varied by UGT2B15 and SULT1A1 genotypes; women with the UGT2B15 Asp/Tyr and Tyr/Tyr genotypes had higher concentrations of estradiol than women with the Asp/Asp genotype (P = 0.004). Compared with women with the SULT1A1 Arg/Arg and Arg/His genotypes, women with the His/His genotype had elevated concentrations of testosterone (P = 0.003). The risk of ER - breast cancer tumors may vary by UGT or SULT genotype. Further, plasma estradiol and testosterone concentrations in breast cancer patients may differ depending on some UGT and SULT genotypes

DETERMINAÇÃO DOS VALORES MÉDIOS DAS ENZIMAS AST, DHL, gGT E FAS NO SORO DE EQUINOS SADIOS EM SANTA MARIA, RS

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Sônia Terezinha dos Anjos Lopes

1993-12-01

Full Text Available Foram usados 50 eqüinos sadios provenientes do Batalhão de Polícia Montada da Brigada Militar em Santa Maria, RS, sendo 43 machos e 7 fêmeas com idade variadas a partir de 3 anos. Foram colhidos 10ml de sangue da jugular para determinação dos valores da atividade sérica das enzimas aspartato-aminotransferase (AST, desidrogenase lática (DHL, gama-glutamiltransferase (gGT e fosfatase alcalina sérica (FAS. Os resultados encontrados para AST foi de 101 - 190U/I com média de 130UI; DHL foi de 100 - 421 U/l com média de 182U/I; gGT foi de 2 - 27U/I com média de 6.5U/I e FAS foi de 103 - 335U/I com média de 190U/I. A partir de outubro/1992 estes valores passaram a ser referência no laboratório de Patologia Clínica do Hospital de Clinicas Veterinárias da Universidade Federal de Santa Maria.

Molecular evolution and the role of oxidative stress in the expansion and functional diversification of cytosolic glutathione transferases

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Vasconcelos Vítor

2010-09-01

Full Text Available Abstract Background Cytosolic glutathione transferases (cGST are a large group of ubiquitous enzymes involved in detoxification and are well known for their undesired side effects during chemotherapy. In this work we have performed thorough phylogenetic analyses to understand the various aspects of the evolution and functional diversification of cGSTs. Furthermore, we assessed plausible correlations between gene duplication and substrate specificity of gene paralogs in humans and selected species, notably in mammalian enzymes and their natural substrates. Results We present a molecular phylogeny of cytosolic GSTs that shows that several classes of cGSTs are more ubiquitous and thus have an older ancestry than previously thought. Furthermore, we found that positive selection is implicated in the diversification of cGSTs. The number of duplicate genes per class is generally higher for groups of enzymes that metabolize products of oxidative damage. Conclusions 1 Protection against oxidative stress seems to be the major driver of positive selection in mammalian cGSTs, explaining the overall expansion pattern of this subfamily; 2 Given the functional redundancy of GSTs that metabolize xenobiotic chemicals, we would expect the loss of gene duplicates, but by contrast we observed a gene expansion of this family, which likely has been favored by: i the diversification of endogenous substrates; ii differential tissue expression; and iii increased specificity for a particular molecule; 3 The increased availability of sequence data from diversified taxa is likely to continue to improve our understanding of the early origin of the different cGST classes.

EFFECT OF FIBROLYTIC ENZYMES ON CHEMICAL COMPOSITION OF MAIZE SILAGE EFEITO DE ENZIMAS FIBROLÍTICAS SOBRE A COMPOSIÇÃO QUÍMICA DASILAGEM DE MILHO

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José Mauro da Silva Diogo

2006-12-01

Full Text Available This experiment was carried out to determine theeffect of enzymatic solution on bromatologic compositionand in situ degradability for neutral detergent fiber (NDFof maize silage. The treatments were four enzymatic levels,two fermentation periods and three replications. The levelsadopted were 0, 5, 10 and 20 mg of enzyme per kg of naturalmatter (MN of grass. Experimental silos, in triplicate, wereopened 45 and 120 days after ensiling, and DM, NDF, crudeprotein (CP, N-NH3 and pH were analyzed. In the study ofin situ degradability of DM and NDF, silage of 45 days wascollected. Nylon bag technique was used with three canulatedbovine and three incubation times (6, 24 and 96 hours. Theexperimental period was of the 36 days. The solution didn’tchange the content of DM, pH and N-NH3 values of maize silage. There was no difference on the CP amongfermentation periods of, but there was interaction betweenperiod of storage and enzymatic level. The solution increasedfinal CP of maize silage after 45 days of ensiling and thiseffect was higher to 20 mg (10.14% compared the control(8.77%. In case of NDF, maize silage treated with 10 mglevel showed lower fiber content (47.25% versus untreatedcontrol (49.96%, suggesting an improvement in the nutritivevalue. There was no difference among period of storageneither interaction period of storage and enzymatic level.In situ rumen degradation showed no difference among thetreatments on NDF degradation of maize silage. KEY WORDS: additives, cellulases, degradability, hemicellulases, cell wall. Objetivou-se avaliar o efeito de níveis de enzimasfibrolíticas sobre a composição bromatológica e adegradabilidade ruminal da fibra em detergente neutro(FDN da silagem de milho. O experimento consistiu dequatro níveis enzimáticos, dois períodos de armazenamentoe três repetições. Aplicaram-se, por aspersão, na ensilagem,os níveis enzimáticos de 0, 5, 10 e 20 mg de enzimas por kgde matéria natural (MN. Após 45 e

Prevalencia del déficit de vitamina B12 en mayores de 60 años hospitalizados. Estudio del polimorfismo C677T de la enzima 5-10 MTHFR en pacientes con déficit de vitamina B12

OpenAIRE

Alonso Ortiz, María Belén

2014-01-01

Programa de doctorado: Avances en Medicina Interna. [ES] El déficit de Vitamina B12 es frecuente en los pacientes mayores. Sin embargo, son escasos los trabajos realizados en el ámbito hospitalario. Este déficit se asocia frecuentemente con elevación de la homocisteína plasmática, conocido factor de riesgo cardiovascular. Además la mutación C677T del gen de la enzima MTHFR se relaciona con hiperhomocistenemia, que puede coexistir con déficit de Vitamina B12, cuya frecuencia y consecuencias...

Quantum Mechanical/Molecular Mechanical Free Energy Simulations of the Glutathione S-Transferase (M1-1) Reaction with Phenanthrene 9,10-Oxide

NARCIS (Netherlands)

Ridder, L.; Rietjens, I.M.C.M.; Vervoort, J.J.M.; Mulholland, A.J.

2002-01-01

Glutathione S-transferases (GSTs) play an important role in the detoxification of xenobiotics in mammals. They catalyze the conjugation of glutathione to a wide range of electrophilic compounds. Phenanthrene 9,10-oxide is a model substrate for GSTs, representing an important group of epoxide

Stress-inducible GmGSTU4 shapes transgenic tobacco plants metabolome towards increased salinity tolerance

NARCIS (Netherlands)

Kissoudis, Christos; Kalloniati, Chrissanthi; Flemetakis, Emmanouil; Madesis, Panagiotis; Labrou, Nikolaos E.; Tsaftaris, Athanasios; Nianiou-Obeidat, Irini

2015-01-01

The involvement of glutathione transferases (GSTs) in plant’s tolerance to abiotic stresses has been extensively studied; however, the metabolic changes occurring in the plants with altered GSTs expression have not been studied in detail. We have previously demonstrated that GmGSTU4

Purification of a membrane-bound trypsin-like enzyme from the gut of the velvetbean caterpillar (Anticarsia gemmatalis Hübner =Purificação de uma enzima “tipo tripsina” não-solúvel do intestino da lagarta da soja (Anticarsia gemmatalis Hübner

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Marcelo Matos Santoro

2012-07-01

Full Text Available Disruption of protein digestion in insects by specific endoprotease inhibitors is being regarded as an alternative to conventional insecticides for pest control. To optimize the effectiveness of this strategy, the understanding of the endoprotease diversity of the target insect is crucial. In this sense, a membrane-bound trypsin-like enzyme from the gut of Anticarsia gemmatalis fifth-instar larvae was purified. Non-soluble fraction of the gut extract was solubilized with 3-[(3-cholamidopropyldimethylammonio]-1-propanesulfonate (CHAPS and subjected to a p-aminobenzamidine affinity chromatography followed by anion-exchange chromatography. The yield of the purified enzyme was 11% with a purification factor of 143 and a final specific activity of 18.6 µM min.-1 mg-1 protein using N-α-benzoyl-L- Arg-p-nitroanilide (L-BApNA as substrate. The purified sample showed a single band with proteolytic activity active and apparent molecular mass of 25 kDa on SDS-PAGE. Molecular mass determined by MALDI-TOF mass spectrometry was 28,632 ± 26 Da. Although the low recovery and the difficulties in purifying large enzyme amounts limited its further characterization, the results contribute for the understanding of the proteases present on A. gemmatalis gut, which are potential targets for natural or specifically designed protease inhibitors.Comprometer a digestão de proteínas dos insetos pelo uso de inibidores específicos de endoproteases tem sido amplamente estudado como um método de controle de pragas alternativo ao uso dos inseticidas convencionais. No processo de otimização desta estratégia, o conhecimento da diversidade das endoproteases do inseto alvo torna-se crucial. Neste sentido, uma enzima “tipo-tripsina” ligada à membrana obtida do intestino de larvas do 5° instar de A. gemmatalis foi purificada. A fração insolúvel do extrato do intestino foi solubilizada com 3-[(3-cholamidopropyldimethylammonio]-1-propanesulfonate (CHAPS e submetida �

In silico genome-wide identification and characterization of the glutathione S-transferase gene family in Vigna radiata.

Science.gov (United States)

Vaish, Swati; Awasthi, Praveen; Tiwari, Siddharth; Tiwari, Shailesh Kumar; Gupta, Divya; Basantani, Mahesh Kumar

2018-05-01

Plant glutathione S-transferases (GSTs) are integral to normal plant metabolism and biotic and abiotic stress tolerance. The GST gene family has been characterized in diverse plant species using molecular biology and bioinformatics approaches. In the current study, in silico analysis identified 44 GSTs in Vigna radiata. Of the total 44 GSTs identified, chromosomal locations of 31 GSTs were confirmed. The pI value of GST proteins ranged from 5.10 to 9.40. The predicted molecular weights ranged from 13.12 to 50 kDa. Subcellular localization analysis revealed that all GSTs were predominantly localized in the cytoplasm. The active site amino acids were confirmed to be serine in tau, phi, theta, zeta, and TCHQD; cysteine in lambda, DHAR, and omega; and tyrosine in EF1G. The gene architecture conformed to the two-exon/one-intron and three-exon/two-intron organization in the case of tau and phi classes, respectively. MEME analysis identified 10 significantly conserved motifs with the width of 8-50 amino acids. The motifs identified were either specific to a specific GST class or were shared by multiple GST classes. The results of the current study will be of potential importance in the characterization of the GST gene family in V. radiata, an economically important leguminous crop.

Human cytosolic glutathione-S-transferases: quantitative analysis of expression, comparative analysis of structures and inhibition strategies of isozymes involved in drug resistance.

Science.gov (United States)

Mohana, Krishnamoorthy; Achary, Anant

2017-08-01

Glutathione-S-transferase (GST) inhibition is a strategy to overcome drug resistance. Several isoforms of human GSTs are present and they are expressed in almost all the organs. Specific expression levels of GSTs in various organs are collected from the human transcriptome data and analysis of the organ-specific expression of GST isoforms is carried out. The variations in the level of expressions of GST isoforms are statistically significant. The GST expression differs in diseased conditions as reported by many investigators and some of the isoforms of GSTs are disease markers or drug targets. Structure analysis of various isoforms is carried out and literature mining has been performed to identify the differences in the active sites of the GSTs. The xenobiotic binding H site is classified into H1, H2, and H3 and the differences in the amino acid composition, the hydrophobicity and other structural features of H site of GSTs are discussed. The existing inhibition strategies are compared. The advent of rational drug design, mechanism-based inhibition strategies, availability of high-throughput screening, target specific, and selective inhibition of GST isoforms involved in drug resistance could be achieved for the reversal of drug resistance and aid in the treatment of diseases.

Inhibition of rat, mouse, and human glutathione S-transferase by eugenol and its oxidation products

NARCIS (Netherlands)

Rompelberg, C.J.M.; Ploemen, J.H.T.M.; Jespersen, S.; Greef, J. van der; Verhagen, H.; Bladeren, P.J. van

1996-01-01

The irreversible and reversible inhibition of glutathione S-transferases (GSTs) by eugenol was studied in rat, mouse and man. Using liver cytosol of human, rat and mouse, species differences were found in the rate of irreversible inhibition of GSTs by eugenol in the presence of the enzyme

Disponibilidade de fósforo estimada por três métodos químicos e pela atividade de duas enzimas em solos que receberam incorporação de materiais orgânicos

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R. S. Berton

1997-12-01

Full Text Available A disponibilidade de fósforo em amostras dos solos Aquatibia (solo aluvial e Hoda (podzólico vermelho-amarelo eutrófico, incubados com quantidades crescentes de materiais orgânicos (palha de cevada, parte aérea de feijão-caupi, esterco de curral e lodo de esgoto, foi estimada por três métodos químicos e pela atividade das enzimas fosfatase ácida e fosfodiesterase, no Departamento de Ciências do Solo e Ambientais da Universidade da Califórnia/Riverside, de janeiro a novembro de 1985. Em estudo em casa de vegetação, a produção de matéria seca e a quantidade de P absorvida pela parte aérea de plantas de milho cultivadas nos solos que receberam os mesmos materiais orgânicos foram determinadas. A disponibilidade de P medida pela extração com água correlacionou-se, significativamente, apenas quando o esterco de curral e o lodo de esgoto foram adicionados aos solos Aquatibia e Hoda, respectivamente. A resina de troca aniônica e o Mehlich 1 foram os métodos mais indicados para determinar a disponibilidade de P para o milho quando os solos receberam quantidades crescentes dos materiais orgânicos. As altas quantidades de P extraídas pela resina de troca aniônica do solo com alta capacidade de adsorção de P indicaram que esse método foi o que melhor considerou os fatores intensidade e capacidade do solo. A atividade da enzima fosfodiesterase do solo revelou ser um bom índice para a determinação do P disponível para o milho, quando os solos receberam a adição do feijão-caupi e do esterco de curral.

Efeito de enzimas de maceração na textura do palmito (Euterpe edulis Mart Influence of cellulase, pectinase and hemicellulase on the texture of hearts of palm (Euterpe edulis Mart.

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Regina Kitagawa GRIZOTTO

1997-12-01

Full Text Available Com a finalidade de ampliar o aproveitamento da palmeira produtora do palmito estudou-se a influência da poligalacturonase e de enzimas maceradoras na textura das partes semi-rígidas do vegetal não-comestíveis, incubando-se preparados comerciais de celulase, hemicelulase e poligalacturonase com o palmito preparado na forma de pequenos toletes (1-3 cm de comprimento e em porções de 2cm do raquis do vegetal. Embora os tratamentos com hemicelulase e mistura de hemicelulase e poligalacturonase tenham promovido ligeiro amaciamento do palmito, os resultados mostraram ,de modo geral, acréscimo na textura do palmito cortado em porções de 3,0 cm e em fatias de 1,0 cm indicando solubilização intensa das regiões suscetíveis a hidrólise com a permanência das regiões duras mais ricas em lignina. Como nos outros tecidos do palmito, no raquis fibroso, não foi comprovada estatiscamente a ação das enzimas na textura do vegetal.With the aim of amplifying the utilization of the palm tree the influence of poligalacturonase and macerating enzymes on the texture of the hard tips of hearts of palm was studied, incubating commercial preparations of cellulase , hemicellulase and poligalacturonase with the hearts of palm, prepared as small stalks of 1-3 cm length and with 2 cm portions of the rachis of the plant. Although the treatments containing hemicellulase and a mixture of hemicellulase and poligalacturonase promoted a slight softening of the hearts of palm, the results generally showed a hardening of the texture of the hearts of palm prepared as stalks of 1 to 3 cm indicating great solubilization of the parts susceptible to hydrolysis, the hard lignin rich parts remaining intact. As in the other tissue of the plant, in the rachis of the fibrous hearts of palm, the action of the enzymes was not statiscally proven.

Geospatial Technology in Geography Education

NARCIS (Netherlands)

Muniz Solari, Osvaldo; Demirci, A.; van der Schee, J.A.

2015-01-01

The book is presented as an important starting point for new research in Geography Education (GE) related to the use and application of geospatial technologies (GSTs). For this purpose, the selection of topics was based on central ideas to GE in its relationship with GSTs. The process of geospatial

Overlapping protective roles for glutathione transferase gene family members in chemical and oxidative stress response in Agrobacterium tumefaciens.

Science.gov (United States)

Skopelitou, Katholiki; Muleta, Abdi W; Pavli, Ourania; Skaracis, Georgios N; Flemetakis, Emmanouil; Papageorgiou, Anastassios C; Labrou, Nikolaos E

2012-03-01

In the present work, we describe the characterisation of the glutathione transferase (GST) gene family from Agrobacterium tumefaciens C58. A genome survey revealed the presence of eight GST-like proteins in A. tumefaciens (AtuGSTs). Comparison by multiple sequence alignment generated a dendrogram revealing the phylogenetic relationships of AtuGSTs-like proteins. The beta and theta classes identified in other bacterial species are represented by five members in A. tumefaciens C58. In addition, there are three "orphan" sequences that do not fit into any previously recognised GST classes. The eight GST-like genes were cloned, expressed in Escherichia coli and their substrate specificity was determined towards 17 different substrates. The results showed that AtuGSTs catalyse a broad range of reactions, with different members of the family exhibiting quite varied substrate specificity. The 3D structures of AtuGSTs were predicted using molecular modelling. The use of comparative sequence and structural analysis of the AtuGST isoenzymes allowed us to identify local sequence and structural characteristics between different GST isoenzymes and classes. Gene expression profiling was conducted under normal culture conditions as well as under abiotic stress conditions (addition of xenobiotics, osmotic stress and cold and heat shock) to induce and monitor early stress-response mechanisms. The results reveal the constitutive expression of GSTs in A. tumefaciens and a modulation of GST activity after treatments, indicating that AtuGSTs presumably participate in a wide range of functions, many of which are important in counteracting stress conditions. These functions may be relevant to maintaining cellular homeostasis as well as in the direct detoxification of toxic compounds.

Screening of filamentous fungi for production of enzymes of biotechnological interest Seleção de fungos filamentosos para a produção de enzimas de interesse biotecnológico

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Luis Henrique S. Guimarães

2006-12-01

Full Text Available Many enzymes produced by fungi have relevant biotechnological applications in several industrial areas. The purpose of this study was to collect and isolate filamentous fungi from soil and humus, plants and sugar cane bagasse of different regions of the São Paulo state. Forty isolates were examined for their ability to produce xylanase, glucose-oxidase, alkaline phosphatase, acid phosphatase, phytase, pectinase and amylase. Among these, twenty three isolates exhibited enzymatic potential. The xylanases produced by two of these isolates (Aspergillus caespitosus and A. phoenicis showed good potential for pulp bleaching. Among seventeen isolates, at least three produced high levels of glucose-oxidase, being Rhizopus stolonifer and A. versicolor the best producer strains. A. caespitosus, Mucor rouxii, and nine others still not identified were the best producers of phosphatases in submerged fermentation. Pectinase was best produced by IF II and C-8 belong R. stolonifer. Significant levels of amylase were produced by Paecilomyces variotii and A. phoenicis. A remarkable enzyme producer was Rhizopus microsporus var. rhizopodiformis that produced high levels of amylase, alkaline and acid phosphatases, and pectinase. Some morphological structures of this fungus were illustrated using light microscopy (LM and scanning electron microscopy (SEM. This study contributes to catalogue soil fungi isolated in the state of São Paulo, and provides additional information to support future research about the industrial potential of these microorganisms that may produce enzymes and, eventually, also secondary metabolites with anti-microbial or anti-parasitic activities.Muitas enzimas produzidas por fungos têm relevantes aplicações em diferentes áreas industriais. O objetivo desse trabalho foi coletar e isolar fungos filamentosos do solo e humus, plantas e bagaço de cana de açúcar de diferentes regiões do Estado de São Paulo. Quarenta isolados foram

Estrogen sulfotransferases in breast and endometrial cancers.

Science.gov (United States)

Pasqualini, Jorge Raul

2009-02-01

Estrogen sulfotransferase is significantly more active in the normal breast cell (e.g., Human 7) than in the cancer cell (e.g., MCF-7). The data suggest that in breast cancer sulfoconjugated activity is carried out by another enzyme, the SULT1A, which acts at high concentration of the substrates. In breast cancer cells sulfotransferase (SULT) activity can be stimulated by various progestins: medrogestone, promegestone, and nomegestrol acetate, as well as by tibolone and its metabolites. SULT activities can also be controlled by other substances including phytoestrogens, celecoxib, flavonoids (e.g., quercetin, resveratrol), and isoflavones. SULT expression was localized in breast cancer cells, which can be stimulated by promegestone and correlated with the increase of the enzyme activity. The estrogen sulfotransferase (SULT1E1), which acts at nanomolar concentration of estradiol, can inactivate most of this hormone present in the normal breast; however, in the breast cancer cells, the sulfotransferase denoted as SULT1A1 is mainly present, and this acts at micromolar concentrations of E(2). A correlation was postulated among breast cancer cell proliferation, the effect of various progestins, and sulfotransferase stimulation. In conclusion, it is suggested that factors involved in the stimulation of the estrogen sulfotransferases could provide new possibilities for the treatment of patients with hormone-dependent breast and endometrial cancers.

Alterations in muscular enzymes of horses competing long-distance endurance rides under tropical climate Alterações em enzimas musculares de cavalos em competições de enduro de longa distância em clima tropical

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A.R. Teixeira-Neto

2008-06-01

Full Text Available This study tried to monitor the alterations of muscle enzymes activity - creatinokinase (CK, lactate dehydrogenase (LDH, and aspartate aminotransferase (AST - in a group of horses that participated of 70 and 100km distance rides in five competitions of an annual endurance championship under tropical climate. Pre ride levels (U/l were 245.13±9.84 for CK, 496.61±14.76 for LDH, and 328.95±8.65 for AST. When compared to these levels, the results revealed a significant decrease in all enzymes activities in the first moment of the rides. Peak levels, significantly different, were reached, immediately after rides by CK (413.591±50.75; 24 hours post rides by LDH (628.61±33.30; and 48 hours after rides by AST (389.89±16.96. Monitoring of recovery period revealed different behavior among enzymes activities with CK values returning to pre ride values (279.61±23.05 24 hours post rides, while LDH and AST values returned to pre ride values (505.25±33.78 and 359.35±24.90, respectively 72 hours post rides. These data revealed different alterations in concentration of muscular enzymes in endurance horses directly related to the duration of the effort.Estudaram-se as alterações de atividade das enzimas musculares creatino quinase (CK, lactato desidrogenase (LDH e aspartato aminotransferase (AST em um grupo de cavalos que utilizados em provas de enduro de 70 e 100km de distância, em cinco competições. Os valores (U/l basais (antes da largada foram 245,13±9,84 para CK, 496,61±14,76 para LDH e 328,95±8,65 para AST. Todas as atividades das enzimas decresceram no primeiro momento das provas (~30km. Valores de pico, significativamente diferentes, foram alcançados para CK (413,59±50,75 imediatamente após 70km de distância; 24 horas após para LDH (628,61±33,30; e 48 horas após as provas para AST (389,89±16,96. A monitoração do período de recuperação revelou diferente comportamento entre as concentrações enzimáticas com CK retornando aos

Nueve casos del sindrome HELLP (hemolisis, enzimas hepaticas Elevadas y plaquetopenia

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M. F. Capellino

2003-10-01

Full Text Available El síndrome HELLP (hemólisis, enzimas hepáticas elevadas y plaquetopenia descripto por Weinstein en 1982, se asocia a alta morbi-mortalidad materna y perinatal. Se evaluaron retrospectivamente las pacientes que presentaron síndrome HELLP entre marzo 1998 y marzo 2001 en el Hospital Privado de Córdoba. Se identificaron nueve pacientes con Síndrome HELLP (incidencia 2.3‰. La edad media de las pacientes fue 24.5 años (15-36 con una edad gestacional media de 34.5 semanas (29-40. Cinco pacientes fueron nulíparas. El síntoma principal fue epigastralgia (77.7%. El parto fue por cesárea en siete pacientes y tres tuvieron síndrome HELLP post-parto. Tres pacientes presentaron hipertensión de difícil manejo, una eclampsia y otra coagulación intravascular diseminada, insuficiencia renal aguda y muerte. Tres requirieron transfusión de derivados sanguíneos y el promedio de internación fue 4.4 días. Seis (75% neonatos fueron pretérmino. El peso medio de nacimiento fue 2030 gramos (736-3200. Cuatro recién nacidos tuvieron un score de Apgar menor de 7 al minuto y todos mayor de 7 a los cinco minutos. Tres neonatos presentaron trastornos alimentarios, uno hipoglucemia y otro enfermedad de membrana hialina, ductus arterioso permeable, sepsis y plaquetopenia. Debido a la morbi-mortalidad materna y perinatal el síndrome HELLP requiere atención en un centro de alta complejidad mediante un equipo multidisciplinario.HELLP syndrome (Hemolysis, Elevated Liver Enzymes and Low Platelets was described by Weinstein in 1982. It has a high maternal and perinatal morbi-mortality rate. We undertook this study to evaluate perinatal outcome in patients with HELLP syndrome. Patients with HELLP syndrome were identified in a retrospective study between March 1998 and March 2001 at the Hospital Privado de Córdoba. Maternal and neonatal variables were analized. Nine patients with HELLP syndrome were identified (incidence 2.3‰. Mean maternal age was 24.5 (15

OBTENÇÃO E UTILIZAÇÃO DA ENZIMA POLIFENOLOXIDASE EXTRAÍDA DE POLPA DE PINHA (Annona squamosa L. MADURA NO MELHORAMENTO DO SABOR DO CACAU (Theobroma cacao L. OBTAINING AND USE OF POLYPHENOLOXIDASE ENZYME EXTRACTED FROM RIPE CUSTARD APPLE (Annona squamosa L. PULP ON THE COCOA (Theobroma cacao L. NIBS IN TASTE IMPROVEMENT

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ELIZA DOROTEA POZZOBON DE ALBUQUERQUE LIMA

2001-12-01

Full Text Available O presente trabalho teve como objetivo estudar a obtenção e a utilização da enzima polifenoloxidase (PPO extraída de polpa de pinha madura na redução do teor de compostos polifenólicos com a finalidade de diminuir a adstringência e o amargor das amêndoas de cacau processadas na forma de "nibs". A PPO foi extraída com tampão fosfato de potássio 0,025M (pH 7,5, adicionando sulfato de amônio para a precipitação da enzima. O material em pó obtido foi denominado de enzima parcialmente purificada, sendo que a análise de atividade enzimática foi realizada, utilizando-se de catecol como substrato. As características bioquímicas apresentadas foram pH de estabilidade de 6,0 a 6,5 e temperatura de estabilidade de 10 a 30°C. Os "nibs" foram autoclavados (121°C por 15 minutos e não autoclavados de amêndoas cruas insuficientemente fermentadas e secas, da mesma origem, sendo embebidas em 25 mL de uma solução da enzima contendo 200 unidades/min/mL, durante 30; 60; 90; 210 e 360 minutos, a 23°C e pH 6,0. Os "nibs" foram homogeneizados com a solução de enzima a cada 15 minutos, secos, moídos e desengordurados. Após o tratamento enzimático durante 210 minutos realizado nos "nibs" de cacau desengordurado não autoclavados foi possível observar diminuição de 15% nas concentrações de fenóis totais, 15% de taninos, 10% de flavan-3-ois e 18% de antocianidinas. Os "nibs" de cacau desengordurado autoclavados apresentaram diminuição de 25% nas concentrações de fenóis totais, 26% de taninos, 23% de flavan-3-ois e 51% de antocianidinas.The present work had as aim to study the obtaining and the uses of polyphenoloxidase enzyme (PPO extracted from ripe custard apple pulp on the reduction of polyphenolic compounds with decrease adstringency and bitterness of cocoa nuts processed at nibs form. The PPO was extracted with 0.025M potassium phosphate buffer (pH 7.5, adding ammonium sulfate to the enzyme precipitation. The powdered

Humanizing π-class glutathione S-transferase regulation in a mouse model alters liver toxicity in response to acetaminophen overdose.

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Matthew P Vaughn

Full Text Available Glutathione S-transferases (GSTs metabolize drugs and xenobiotics. Yet despite high protein sequence homology, expression of π-class GSTs, the most abundant of the enzymes, varies significantly between species. In mouse liver, hepatocytes exhibit high mGstp expression, while in human liver, hepatocytes contain little or no hGSTP1 mRNA or hGSTP1 protein. π-class GSTs are known to be critical determinants of liver responses to drugs and toxins: when treated with high doses of acetaminophen, mGstp1/2+/+ mice suffer marked liver damage, while mGstp1/2-/- mice escape liver injury.To more faithfully model the contribution of π-class GSTs to human liver toxicology, we introduced hGSTP1, with its exons, introns, and flanking sequences, into the germline of mice carrying disrupted mGstp genes. In the resultant hGSTP1+mGstp1/2-/- strain, π-class GSTs were regulated differently than in wild-type mice. In the liver, enzyme expression was restricted to bile duct cells, Kupffer cells, macrophages, and endothelial cells, reminiscent of human liver, while in the prostate, enzyme production was limited to basal epithelial cells, reminiscent of human prostate. The human patterns of hGSTP1 transgene regulation were accompanied by human patterns of DNA methylation, with bisulfite genomic sequencing revealing establishment of an unmethylated CpG island sequence encompassing the gene promoter. Unlike wild-type or mGstp1/2-/- mice, when hGSTP1+mGstp1/2-/- mice were overdosed with acetaminophen, liver tissues showed limited centrilobular necrosis, suggesting that π-class GSTs may be critical determinants of toxin-induced hepatocyte injury even when not expressed by hepatocytes.By recapitulating human π-class GST expression, hGSTP1+mGstp1/2-/- mice may better model human drug and xenobiotic toxicology.

Influence of carbon source, pH, and temperature on the polygalacturonase activity of Kluyveromyces marxianus CCMB 322 Influência da fonte de carbono, pH e temperatura na atividade da enzima poligalacturonase de Kluyveromyces marxianus CCMB 322

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Rodrigo de Queiroz Oliveira

2012-09-01

Full Text Available Microbial pectinolytic enzymes are known to play a commercially important role in a number of industrial processes. Two kinds of yeast can be discerned regarding the production of enzymes. One group includes those which can produce enzymes in the absence of an inducer, and the other group comprises the yeasts that produce enzymes in the presence of an inducer. The objective of this study was to investigate the influence of pectic substances, glucose, pH, and temperature on the polygalacturonase activity by Kluyveromyces marxianus CCMB 322. The yeast was grown in a fermentation broth containing different concentrations of glucose and pectic substances. The polygalacturonase activity was determined by the DNS method, and the pH and temperature were optimized using a central composite experimental design. The polygalacturonase secreted by K. marxianus CCMB 322 was partially constitutive showing optimum pH and temperature of 7.36 and 70 °C, respectively, and maintained approximately 93% of its original activity for 50 minutes at 50 °C. Thermal stability of the polygalacturonase enzyme was studied at different temperatures (50, 60, 70, and 80 °C and different incubation times (0, 10, 20, 30, 40, and 50 minutes. This study showed that glucose can influence the regulation of the synthesis of polygalacturonase.Enzimas pectinolíticas de origem microbiana são conhecidas por desempenhar um papel importante comercialmente em uma série de processos industriais. Dois tipos de levedura podem ser distinguidos para a produção dessas enzimas. Um grupo inclui aqueles que têm capacidade de produzi-las na ausência de um indutor e outro grupo compreende as leveduras que as produzem na presença de um indutor. O objetivo deste estudo foi investigar a influência de substâncias pécticas, de glicose, do pH e da temperatura na atividade da poligalacturonase de Kluyveromyces marxianus CCMB 322. O cultivo foi em caldo de fermentação contendo diferentes

Phylogenomic approaches to common problems encountered in the analysis of low copy repeats: The sulfotransferase 1A gene family example

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Benner Steven A

2005-03-01

Full Text Available Abstract Background Blocks of duplicated genomic DNA sequence longer than 1000 base pairs are known as low copy repeats (LCRs. Identified by their sequence similarity, LCRs are abundant in the human genome, and are interesting because they may represent recent adaptive events, or potential future adaptive opportunities within the human lineage. Sequence analysis tools are needed, however, to decide whether these interpretations are likely, whether a particular set of LCRs represents nearly neutral drift creating junk DNA, or whether the appearance of LCRs reflects assembly error. Here we investigate an LCR family containing the sulfotransferase (SULT 1A genes involved in drug metabolism, cancer, hormone regulation, and neurotransmitter biology as a first step for defining the problems that those tools must manage. Results Sequence analysis here identified a fourth sulfotransferase gene, which may be transcriptionally active, located on human chromosome 16. Four regions of genomic sequence containing the four human SULT1A paralogs defined a new LCR family. The stem hominoid SULT1A progenitor locus was identified by comparative genomics involving complete human and rodent genomes, and a draft chimpanzee genome. SULT1A expansion in hominoid genomes was followed by positive selection acting on specific protein sites. This episode of adaptive evolution appears to be responsible for the dopamine sulfonation function of some SULT enzymes. Each of the conclusions that this bioinformatic analysis generated using data that has uncertain reliability (such as that from the chimpanzee genome sequencing project has been confirmed experimentally or by a "finished" chromosome 16 assembly, both of which were published after the submission of this manuscript. Conclusion SULT1A genes expanded from one to four copies in hominoids during intra-chromosomal LCR duplications, including (apparently one after the divergence of chimpanzees and humans. Thus, LCRs may

In Silico Mechanistic Profiling to Probe Small Molecule Binding to Sulfotransferases

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Martiny, Virginie Y.; Carbonell, Pablo; Lagorce, David; Villoutreix, Bruno O.; Moroy, Gautier; Miteva, Maria A.

2013-01-01

Drug metabolizing enzymes play a key role in the metabolism, elimination and detoxification of xenobiotics, drugs and endogenous molecules. While their principal role is to detoxify organisms by modifying compounds, such as pollutants or drugs, for a rapid excretion, in some cases they render their substrates more toxic thereby inducing severe side effects and adverse drug reactions, or their inhibition can lead to drug–drug interactions. We focus on sulfotransferases (SULTs), a family of phase II metabolizing enzymes, acting on a large number of drugs and hormones and showing important structural flexibility. Here we report a novel in silico structure-based approach to probe ligand binding to SULTs. We explored the flexibility of SULTs by molecular dynamics (MD) simulations in order to identify the most suitable multiple receptor conformations for ligand binding prediction. Then, we employed structure-based docking-scoring approach to predict ligand binding and finally we combined the predicted interaction energies by using a QSAR methodology. The results showed that our protocol successfully prioritizes potent binders for the studied here SULT1 isoforms, and give new insights on specific molecular mechanisms for diverse ligands’ binding related to their binding sites plasticity. Our best QSAR models, introducing predicted protein-ligand interaction energy by using docking, showed accuracy of 67.28%, 78.00% and 75.46%, for the isoforms SULT1A1, SULT1A3 and SULT1E1, respectively. To the best of our knowledge our protocol is the first in silico structure-based approach consisting of a protein-ligand interaction analysis at atomic level that considers both ligand and enzyme flexibility, along with a QSAR approach, to identify small molecules that can interact with II phase dug metabolizing enzymes. PMID:24039991

In silico mechanistic profiling to probe small molecule binding to sulfotransferases.

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Virginie Y Martiny

Full Text Available Drug metabolizing enzymes play a key role in the metabolism, elimination and detoxification of xenobiotics, drugs and endogenous molecules. While their principal role is to detoxify organisms by modifying compounds, such as pollutants or drugs, for a rapid excretion, in some cases they render their substrates more toxic thereby inducing severe side effects and adverse drug reactions, or their inhibition can lead to drug-drug interactions. We focus on sulfotransferases (SULTs, a family of phase II metabolizing enzymes, acting on a large number of drugs and hormones and showing important structural flexibility. Here we report a novel in silico structure-based approach to probe ligand binding to SULTs. We explored the flexibility of SULTs by molecular dynamics (MD simulations in order to identify the most suitable multiple receptor conformations for ligand binding prediction. Then, we employed structure-based docking-scoring approach to predict ligand binding and finally we combined the predicted interaction energies by using a QSAR methodology. The results showed that our protocol successfully prioritizes potent binders for the studied here SULT1 isoforms, and give new insights on specific molecular mechanisms for diverse ligands' binding related to their binding sites plasticity. Our best QSAR models, introducing predicted protein-ligand interaction energy by using docking, showed accuracy of 67.28%, 78.00% and 75.46%, for the isoforms SULT1A1, SULT1A3 and SULT1E1, respectively. To the best of our knowledge our protocol is the first in silico structure-based approach consisting of a protein-ligand interaction analysis at atomic level that considers both ligand and enzyme flexibility, along with a QSAR approach, to identify small molecules that can interact with II phase dug metabolizing enzymes.

Sex differences in apolipoprotein A1 and nevirapine-induced toxicity

OpenAIRE

Aline Marinho; Clara Dias; Alexandra Antunes; Umbelina Caixas; Teresa Branco; Matilde Marques; Emília Monteiro; Sofia Pereira

2014-01-01

Nevirapine (NVP) is associated with severe liver and skin toxicity through sulfotransferase (SULT) bioactivation of the phase I metabolite 12-hydroxy-NVP [1–3]. The female sex, a well-known risk factor for NVP-induced toxicity, is associated with higher SULT expression [4] and lower plasma levels of 12-hydroxy-NVP [3]. Interestingly, apolipoprotein A1 (ApoA1) increases SULT2B1 activity and ApoA1 synthesis is increased by NVP [5, 6]. Herein, we explore the effect of ApoA1 levels on NVP metabol...

Glutathione S-Transferases: Role in Combating Abiotic Stresses Including Arsenic Detoxification in Plants

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Smita Kumar

2018-06-01

Full Text Available Arsenic (As, naturally occurring metalloid and a potential hazardous material, is found in low concentrations in the environment and emerges from natural sources and anthropogenic activities. The presence of As in ground water, which is used for irrigation, is a matter of great concern since it affects crop productivity and contaminates food chain. In plants, As alters various metabolic pathways in cells including the interaction of substrates/enzymes with the sulfhydryl groups of proteins and the replacement of phosphate in ATP for energy. In addition, As stimulates the generation of free radicals and reactive oxygen species (ROS, resulting in oxidative stress. Glutathione S-transferases (GSTs quench reactive molecules with the addition of glutathione (GSH and protect the cell from oxidative damage. GSTs are a multigene family of isozymes, known to catalyze the conjugation of GSH to miscellany of electrophilic and hydrophobic substrates. GSTs have been reported to be associated with plant developmental processes and are responsive to multitude of stressors. In past, several studies suggested involvement of plant GST gene family in As response due to the requirement of sulfur and GSH in the detoxification of this toxic metalloid. This review provides updated information about the role of GSTs in abiotic and biotic stresses with an emphasis on As uptake, metabolism, and detoxification in plants. Further, the genetic manipulations that helped in enhancing the understanding of the function of GSTs in abiotic stress response and heavy metal detoxification has been reviewed.

ACTIVIDAD DE ENZIMAS ANTIOXIDANTES EN TEJIDOS EMBRIOGÉNICOS Y NO EMBRIOGÉNICOS DE CAÑA DE AZÚCAR

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Marina Medeiros de Araújo Silva

2014-05-01

Full Text Available Este trabajo tuvo el objetivo de inducir la embriogénesis directa en segmentos de hojas jóvenes no-expandidas en la variedad de caña de azúcar RB872552 e investigar la correlación de este evento morfogénico con el estrés oxidativo. Se utilizaron dos protocolos de inducción de embriogénesis somática, previamente descritos para caña de azúcar, con distintas suplementaciones en el medio de cultivo y en las condiciones de incubación. Para la conversión de embriones en plantas se utilizó el medio MS sin fitorreguladores. Se realizaron además análisis histológicas y de actividad enzimática in tejidos embriogénicos y no embriogénicos. La formación de embriones somáticos por la vía directa ocurrió en 81% de los explantes, con la combinación de los reguladores de crecimiento 2,4-D (ácido 2,4-diclorofenoxiacético y BAP (6-bencilaminopurina e incubación bajo un fotoperiodo de 16 horas. Respecto a las enzimas antioxidantes, se observó un incremento en la actividad de peroxidase y en el contenido de proteínas solubles en los tejidos embriogénicos, mientras que la polifenoloxidase y la catalase presentaron más bajos valores de actividad en estos tejidos, en comparación con los no embriogénicos. Se podría inferir que el estrés oxidativo juega un papel importante en la inducción de la embriogénesis somática en caña de azúcar.

Efeito de enzimas hidrolíticas no comportamento reológico do óleo de palma cru

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FREITAS Suely P.

1998-01-01

Full Text Available No processamento tradicional de extração de óleo de palma, uma quantidade substancial do produto é perdida na fibra - durante a etapa de prensagem do fruto - e na borra - durante a etapa de clarificação do óleo cru. A viscosidade deste constitui a principal dificuldade de separação do óleo dos demais componentes da mistura. A eficiência de recuperação do óleo é usualmente melhorada por adição de água, no tanque de clarificação. Neste trabalho avaliou-se os efeitos da diluição e do tratamento enzimático no comportamento reológico do óleo de palma cru. Estes dados são importantes para estimar a velocidade de separação do óleo nos tanques de clarificação e/ou nos decanters das usinas comerciais. O óleo cru utilizado neste estudo foi recolhido em uma unidade comercial sendo composto de 65% de água, 30% de óleo e 5% de sólidos. A hidrólise enzimática foi conduzida usando como agentes hidrolisantes uma enzima comercial (viscozyme da Novo Nordisk e um preparado enzimático produzido no CTAA. Pode-se constatar que quanto mais diluída a amostra maior é a redução na viscosidade após a hidrólise enzimática. Para uma taxa de deformação fixa em 6s-1, o tratamento enzimático do óleo cru a 60°C reduziu a viscosidade do mesmo em cerca de 29% enquanto a adição de 20% de água em cerca de 35%. A combinação da diluição com 20% de água seguida de tratamento enzimático reduziu a viscosidade do óleo cru em cerca de 75%.

Modulation of xenobiotic metabolising enzymes by anticarcinogens-focus on glutathione S-transferases and their role as targets of dietary chemoprevention in colorectal carcinogenesis

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Pool-Zobel, Beatrice [Department of Nutritional Toxicology, Institute for Nutrition, Friedrich Schiller University Jena, 07743 Jena (Germany)]. E-mail: b8pobe@uni-jena.de; Veeriah, Selvaraju [Department of Nutritional Toxicology, Institute for Nutrition, Friedrich Schiller University Jena, 07743 Jena (Germany); Boehmer, Frank-D. [Institute of Molecular Cell Biology, University Hospital, Friedrich Schiller University Jena, 07743 Jena (Germany)

2005-12-11

There is evidence that consumption of certain dietary ingredients may favourably modulate biotransformation of carcinogens. Associated with this is the hypothesis that the risk for developing colorectal cancer could be reduced, since its incidence is related to diet. Two main groups of biotransformation enzymes metabolize carcinogens, namely Phase I enzymes, which convert hydrophobic compounds to more water-soluble moieties, and Phase II enzymes (e.g. glutathione S-transferases [GST]), which primarily catalyze conjugation reactions. The conjugation of electrophilic Phase I intermediates with glutathione, for instance, frequently results in detoxification. Several possible colon carcinogens may serve as substrates for GST isoenzymes that can have marked substrate specificity. The conjugated products could be less toxic/genotoxic if GSTs are induced, thereby reducing exposure. Thus, numerous studies have shown that the induction of GSTs by antioxidants enables experimental animals to tolerate exposure to carcinogens. One important mechanism of GST induction involves an antioxidant-responsive response element (ARE) and the transcription factor nuclear factor E2-related factor 2 (Nrf2), which is bound to the Kelch-like ECH associated protein 1 (Keap1) in the cytoplasm. Antioxidants may disrupt the Keap-Nrf2 complex, allowing Nrf2 to translocate to the nucleus and mediate expression of Phase II genes via interaction with the ARE. GSTs are also induced by butyrate, a product of gut flora-derived fermentation of plant foods, which may act via different mechanisms, e.g. by increasing histone acetylation. GSTs are expressed with high inter-individual variability in human colonocytes, which points to large differences in cellular susceptibility to xenobiotics. Enhancing expression of GSTs in human colon tissue could therefore contribute to reducing cancer risks. However, it has not been demonstrated in humans that this mechanism is associated with cancer prevention. In the

Modulation of xenobiotic metabolising enzymes by anticarcinogens-focus on glutathione S-transferases and their role as targets of dietary chemoprevention in colorectal carcinogenesis

International Nuclear Information System (INIS)

Pool-Zobel, Beatrice; Veeriah, Selvaraju; Boehmer, Frank-D.

2005-01-01

There is evidence that consumption of certain dietary ingredients may favourably modulate biotransformation of carcinogens. Associated with this is the hypothesis that the risk for developing colorectal cancer could be reduced, since its incidence is related to diet. Two main groups of biotransformation enzymes metabolize carcinogens, namely Phase I enzymes, which convert hydrophobic compounds to more water-soluble moieties, and Phase II enzymes (e.g. glutathione S-transferases [GST]), which primarily catalyze conjugation reactions. The conjugation of electrophilic Phase I intermediates with glutathione, for instance, frequently results in detoxification. Several possible colon carcinogens may serve as substrates for GST isoenzymes that can have marked substrate specificity. The conjugated products could be less toxic/genotoxic if GSTs are induced, thereby reducing exposure. Thus, numerous studies have shown that the induction of GSTs by antioxidants enables experimental animals to tolerate exposure to carcinogens. One important mechanism of GST induction involves an antioxidant-responsive response element (ARE) and the transcription factor nuclear factor E2-related factor 2 (Nrf2), which is bound to the Kelch-like ECH associated protein 1 (Keap1) in the cytoplasm. Antioxidants may disrupt the Keap-Nrf2 complex, allowing Nrf2 to translocate to the nucleus and mediate expression of Phase II genes via interaction with the ARE. GSTs are also induced by butyrate, a product of gut flora-derived fermentation of plant foods, which may act via different mechanisms, e.g. by increasing histone acetylation. GSTs are expressed with high inter-individual variability in human colonocytes, which points to large differences in cellular susceptibility to xenobiotics. Enhancing expression of GSTs in human colon tissue could therefore contribute to reducing cancer risks. However, it has not been demonstrated in humans that this mechanism is associated with cancer prevention. In the

Molecular characterization of novel sulfotransferases from the tick, Ixodes scapularis

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King Roberta S

2011-06-01

Full Text Available Abstract Background Ixodes scapularis, commonly known as the blacklegged or deer tick, is the main vector of Lyme disease in the United States. Recent progress in transcriptome research has uncovered hundreds of different proteins expressed in the salivary glands of hard ticks, the majority of which have no known function, and include many novel protein families. We recently identified transcripts coding for two putative cytosolic sulfotransferases in these ticks which recognized phenolic monoamines as their substrates. In this current study, we characterize the genetic expression of these two cytosolic sulfotransferases throughout the tick life cycle as well as the enzymatic properties of the corresponding recombinant proteins. Interestingly, the resultant recombinant proteins showed sulfotransferase activity against both neurotransmitters dopamine and octopamine. Results The two sulfotransferase genes were coded as Ixosc SULT 1 & 2 and corresponding proteins were referred as Ixosc Sult 1 and 2. Using gene-specific primers, the sulfotransferase transcripts were detected throughout the blacklegged tick life cycle, including eggs, larvae, nymphs, adult salivary glands and adult midgut. Notably, the mRNA and protein levels were altered upon feeding during both the larval and nymphal life stages. Quantitative PCR results confirm that Ixosc SULT1 was statistically increased upon blood feeding while Ixosc SULT 2 was decreased. This altered expression led us to further characterize the function of these proteins in the Ixodid tick. The sulfotransferase genes were cloned and expressed in a bacterial expression system, and purified recombinant proteins Ixosc Sult 1(R and 2(R showed sulfotransferase activity against neurotransmitters dopamine and octopamine as well as the common sulfotransferase substrate p-nitrophenol. Thus, dopamine- or octopamine-sulfonation may be involved in altering the biological signal for salivary secretion in I. scapularis

Metabolic Phase I (CYPs) and Phase II (GSTs) Gene Polymorphisms and Their Interaction with Environmental Factors in Nasopharyngeal Cancer from the Ethnic Population of Northeast India.

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Singh, Seram Anil; Ghosh, Sankar Kumar

2017-09-26

Multiple genetic and environmental factors and their interaction are believed to contribute in the pathogenesis of Nasopharyngeal Cancer (NPC). We investigate the role of Metabolic Phase I (CYPs) and Phase II (GSTs) gene polymorphisms, gene-gene and gene-environmental interaction in modulating the susceptibility to NPC in Northeast India. To determine the association of metabolic gene polymorphisms and environmental habits, 123 cases and 189 controls blood/swab samples were used for PCR and confirmed by Sanger sequencing. Analysis for GSTM1 and GSTT1 gene polymorphism was done by multiplex PCR. The T3801C in the 3'- flanking region of CYP1A1 gene was detected by PCR-RFLP method. The Logistic regression analysis was used to estimate odds ratios (OR) and 95% confidence intervals (95% CI). The GSTM1 null genotype alone (OR = 2.76) was significantly associated with NPC risk (P < 0.0001). The combinations of GSTM1 null and GSTT1 null genotypes also higher, 3.77 fold (P < 0.0001), risk of NPC, while GSTM1 null genotype along with CYP1A1 T3801C TC + CC genotype had 3.22 (P = 0.001) fold risk. The most remarkable risk was seen among individual carrying GSTM1 null, GSTT1 null genotypes and CYP1A1 T3801C TC + CC genotypes (OR = 5.71, P = 0.001). Further; analyses demonstrate an enhanced risk of NPC in smoked meat (OR = 5.56, P < 0.0001) and fermented fish consumers (OR = 5.73, P < 0.0001) carrying GSTM1 null genotype. An elevated risk of NPC was noted in smokers (OR = 12.67, P < 0.0001) and chewers (OR = 5.68, P < 0.0001) with GSTM1 null genotype. However, smokers had the highest risk of NPC among individuals carrying GSTT1 null genotype (OR = 4.46, P = 0.001) or CYP1A1 T3801C TC + CC genotype (OR = 7.13, P < 0.0001). The association of null genotypes and mutations of metabolic neutralizing genes along with the environmental habits (tobacco smokers and chewers, smoke meat, fermented fishes) can be used as a possible biomarker for

High accuracy in silico sulfotransferase models.

Science.gov (United States)

Cook, Ian; Wang, Ting; Falany, Charles N; Leyh, Thomas S

2013-11-29

Predicting enzymatic behavior in silico is an integral part of our efforts to understand biology. Hundreds of millions of compounds lie in targeted in silico libraries waiting for their metabolic potential to be discovered. In silico "enzymes" capable of accurately determining whether compounds can inhibit or react is often the missing piece in this endeavor. This problem has now been solved for the cytosolic sulfotransferases (SULTs). SULTs regulate the bioactivities of thousands of compounds--endogenous metabolites, drugs and other xenobiotics--by transferring the sulfuryl moiety (SO3) from 3'-phosphoadenosine 5'-phosphosulfate to the hydroxyls and primary amines of these acceptors. SULT1A1 and 2A1 catalyze the majority of sulfation that occurs during human Phase II metabolism. Here, recent insights into the structure and dynamics of SULT binding and reactivity are incorporated into in silico models of 1A1 and 2A1 that are used to identify substrates and inhibitors in a structurally diverse set of 1,455 high value compounds: the FDA-approved small molecule drugs. The SULT1A1 models predict 76 substrates. Of these, 53 were known substrates. Of the remaining 23, 21 were tested, and all were sulfated. The SULT2A1 models predict 22 substrates, 14 of which are known substrates. Of the remaining 8, 4 were tested, and all are substrates. The models proved to be 100% accurate in identifying substrates and made no false predictions at Kd thresholds of 100 μM. In total, 23 "new" drug substrates were identified, and new linkages to drug inhibitors are predicted. It now appears to be possible to accurately predict Phase II sulfonation in silico.

Purification, molecular cloning and heterologous expression of a glutathione S-transferase involved in insecticide resistance from the rice brown planthopper, Nilaparvata lugens.

OpenAIRE

Vontas, John G; Small, Graham J; Nikou, Dimitra C; Ranson, Hilary; Hemingway, Janet

2002-01-01

A novel glutathione S-transferase (GST)-based pyrethroid resistance mechanism was recently identified in Nilaparvata lugens [Vontas, Small and Hemingway (2001) Biochem. J. 357, 65-72]. To determine the nature of GSTs involved in conferring this resistance, the GSTs from resistant and susceptible strains of N. lugens were partially purified by anion exchange and affinity chromatography. The majority of peroxidase activity, previously correlated with resistance, was confined to the fraction tha...

Phylogenetic characterization of Clonorchis sinensis proteins homologous to the sigma-class glutathione transferase and their differential expression profiles.

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Bae, Young-An; Kim, Jeong-Geun; Kong, Yoon

2016-01-01

Glutathione transferase (GST) is one of the major antioxidant proteins with diverse supplemental activities including peroxidase, isomerase, and thiol transferase. GSTs are classified into multiple classes on the basis of their primary structures and substrate/inhibitor specificity. However, the evolutionary routes and physiological environments specific to each of the closely related bioactive enzymes remain elusive. The sigma-like GSTs exhibit amino acid conservation patterns similar to the prostaglandin D synthases (PGDSs). In this study, we analyzed the phylogenetic position of the GSTs of the biocarcinogenic liver fluke, Clonorchis sinensis. We also observed induction profile of the GSTs in association with the parasite's maturation and in response to exogenous oxidative stresses, with special attention to sigma-class GSTs and PGDSs. The C. sinensis genome encoded 12 GST protein species, which were separately assigned to cytosolic (two omega-, one zeta-, two mu-, and five sigma-class), mitochondrial (one kappa-class), and microsomal (one membrane-associated proteins in eicosanoid and glutathione metabolism-like protein) GST families. Multiple sigma GST (or PGDS) orthologs were also detected in Opisthorchis viverrini. Other trematode species possessed only a single sigma-like GST gene. A phylogenetic analysis demonstrated that one of the sigma GST lineages duplicated in the common ancestor of trematodes were specifically expanded in the opisthorchiids, but deleted in other trematodes. The induction profiles of these sigma GST genes along with the development and aging of C. sinensis, and against various exogenous chemical stimuli strongly suggest that the paralogous sigma GST genes might be undergone specialized evolution to cope with the diverse hostile biochemical environments within the mammalian hepatobiliary ductal system. Copyright © 2016 Elsevier B.V. All rights reserved.

Identification and expression profiles of nine glutathione S-transferase genes from the important rice phloem sap-sucker and virus vector Laodelphax striatellus (Fallén) (Hemiptera: Delphacidae).

Science.gov (United States)

Zhou, Wen-Wu; Li, Xi-Wang; Quan, Yin-Hua; Cheng, Jiaan; Zhang, Chuan-Xi; Gurr, Geoff; Zhu, Zeng-Rong

2012-09-01

Glutathione S-transferases (GSTs) have received considerable attention in insects for their roles in insecticide resistance. Laodelphax striatellus (Fallén) is a serious rice pest. L. striatellus outbreaks occur frequently throughout eastern Asia. A key problem in controlling this pest is its rapid adaptation to numerous insecticides. In this research, nine cDNAs encoding GSTs in L. striatellus were cloned and characterised. The cloned GSTs of L. striatellus belonged to six cytosolic classes and a microsomal subgroup. Exposure to sublethal concentrations of each of the six insecticides, DDT, chlorpyrifos, fipronil, imidacloprid, buprofezin and beta-cypermethrin, quickly induced (6 h) up-expression of LsGSTe1. The expression of LsGSTs2 was increased by chlorpyrifos, fipronil and beta-cypermethrin. Furthermore, exposure of L. striatellus to fipronil, imidacloprid, buprofezin and beta-cypermethrin increased the expression of the LsGSTm gene after 24 or 48 h. This work is the first identification of GST genes from different GST groups in Auchenorrhyncha species and their induction characteristics with insecticide types and time. The elevated expression of GST genes induced by insecticides might be related to the enhanced tolerance of this insect to insecticides and xenobiotics. Copyright © 2012 Society of Chemical Industry.

Proteomic Profiling of Cytosolic Glutathione Transferases from Three Bivalve Species: Corbicula fluminea, Mytilus galloprovincialis and Anodonta cygnea

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José Carlos Martins

2014-01-01

Full Text Available Suspension-feeding bivalves are considered efficient toxin vectors with a relative insensitivity to toxicants compared to other aquatic organisms. This fact highlights the potential role of detoxification enzymes, such as glutathione transferases (GSTs, in this bivalve resistance. Nevertheless, the GST system has not been extensively described in these organisms. In the present study, cytosolic GSTs isoforms (cGST were surveyed in three bivalves with different habitats and life strategies: Corbicula fluminea, Anodonta cygnea and Mytilus galloprovincialis. GSTs were purified by glutathione-agarose affinity chromatography, and the collection of expressed cGST classes of each bivalve were identified using a proteomic approach. All the purified extracts were also characterized kinetically. Results reveal variations in cGST subunits collection (diversity and properties between the three tested bivalves. Using proteomics, four pi-class and two sigma-class GST subunits were identified in M. galloprovincialis. C. fluminea also yielded four pi-class and one sigma-class GST subunits. For A. cygnea, two mu-class and one pi-class GST subunits were identified, these being the first record of GSTs from these freshwater mussels. The affinity purified extracts also show differences regarding enzymatic behavior among species. The variations found in cGST collection and kinetics might justify diverse selective advantages for each bivalve organism.

Comparative structural analysis of a novel glutathioneS-transferase (ATU5508) from Agrobacterium tumefaciens at 2.0 A resolution.

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Kosloff, Mickey; Han, Gye Won; Krishna, S Sri; Schwarzenbacher, Robert; Fasnacht, Marc; Elsliger, Marc-André; Abdubek, Polat; Agarwalla, Sanjay; Ambing, Eileen; Astakhova, Tamara; Axelrod, Herbert L; Canaves, Jaume M; Carlton, Dennis; Chiu, Hsiu-Ju; Clayton, Thomas; DiDonato, Michael; Duan, Lian; Feuerhelm, Julie; Grittini, Carina; Grzechnik, Slawomir K; Hale, Joanna; Hampton, Eric; Haugen, Justin; Jaroszewski, Lukasz; Jin, Kevin K; Johnson, Hope; Klock, Heath E; Knuth, Mark W; Koesema, Eric; Kreusch, Andreas; Kuhn, Peter; Levin, Inna; McMullan, Daniel; Miller, Mitchell D; Morse, Andrew T; Moy, Kin; Nigoghossian, Edward; Okach, Linda; Oommachen, Silvya; Page, Rebecca; Paulsen, Jessica; Quijano, Kevin; Reyes, Ron; Rife, Christopher L; Sims, Eric; Spraggon, Glen; Sridhar, Vandana; Stevens, Raymond C; van den Bedem, Henry; Velasquez, Jeff; White, Aprilfawn; Wolf, Guenter; Xu, Qingping; Hodgson, Keith O; Wooley, John; Deacon, Ashley M; Godzik, Adam; Lesley, Scott A; Wilson, Ian A

2006-11-15

Glutathione S-transferases (GSTs) comprise a diverse superfamily of enzymes found in organisms from all kingdoms of life. GSTs are involved in diverse processes, notably small-molecule biosynthesis or detoxification, and are frequently also used in protein engineering studies or as biotechnology tools. Here, we report the high-resolution X-ray structure of Atu5508 from the pathogenic soil bacterium Agrobacterium tumefaciens (atGST1). Through use of comparative sequence and structural analysis of the GST superfamily, we identified local sequence and structural signatures, which allowed us to distinguish between different GST classes. This approach enables GST classification based on structure, without requiring additional biochemical or immunological data. Consequently, analysis of the atGST1 crystal structure suggests a new GST class, distinct from previously characterized GSTs, which would make it an attractive target for further biochemical studies. (c) 2006 Wiley-Liss, Inc.

Resistance to ACCase inhibitors in Eleusine indica from Brazil involves a target site mutation Resistência aos inibidores de ACCase em Eleusine indica do Brasil envolve uma mutação na enzima alvo

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M.D. Osuna

2012-09-01

Full Text Available Eleusine indica (goosegrass is a diploid grass weed which has developed resistance to ACCase inhibitors during the last ten years due to the intensive and frequent use of sethoxydim to control grass weeds in soybean crops in Brazil. Plant dose-response assays confirmed the resistant behaviour of one biotype obtaining high resistance factor values: 143 (fenoxaprop, 126 (haloxyfop, 84 (sethoxydim to 58 (fluazifop. ACCase in vitro assays indicated a target site resistance as the main cause of reduced susceptibility to ACCase inhibitors. PCR-generated fragments of the ACCase CT domain of the resistant and sensitive reference biotype were sequenced and compared. A point mutation was detected within the triplet of aspartate at the amino acid position 2078 (referred to EMBL accession no. AJ310767 and resulted in the triplet of glycine. These results constitute the first report on a target site mutation for a Brazilian herbicide resistant grass weed.Eleusine indica (ELEIN é uma espécie monocotiledônea, diploide. No Brasil, ela desenvolveu resistência aos inibidores da ACCase durante os últimos dez anos, devido ao uso intensivo e frequente desses graminicidas para controlar plantas daninhas em lavouras de soja. Experimentos de dose-resposta realizados com a planta confirmaram a resistência de um biótipo. Houve elevada tolerância aos herbicidas, com fatores de resistência da ordem de 143 (fenoxaprop, 126 (haloxyfop, 84 (sethoxydim e 58 (fluazifop. Ensaios com a enzima ACCase in vitro indicaram a insensibilidade desta como a principal causa de suscetibilidade reduzida a esses herbicidas. Fragmentos de PCR gerados do domínio CT da enzima ACCase dos biótipos resistente e sensível de referência foram sequenciados e comparados. Foi detectada uma mutação dentro do tripleto de asparagina na posição do aminoácido 2078 (referente ao acesso número AJ310767 no EMBL, que resultou no tripleto de glicina. Esses resultados constituem o primeiro caso

Matriz extracelular e enzimas degradatórias na hematopoese e doenças onco-hematológicas Extracellular matrix in hematopoiesis and hematologic malignancies

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Juliana L. Dreyfuss

2008-10-01

Full Text Available A matriz extracelular (MEC é uma rede complexa composta por quatro grandes classes de macromoléculas: colágenos, proteoglicanos (PGs, glicosaminoglicanos (GAGs e glicoproteínas adesivas. As interações entre as células e a MEC são cruciais para determinar os padrões de comportamento celular, tais como crescimento, morte, diferenciação e motilidade. A hematopoese é o sistema responsável pela produção das células sangüíneas. O controle da proliferação e diferenciação destas células é feito através da interação das células com o microambiente da medula óssea (matriz extracelular. A adesão de progenitores hematopoéticos a moléculas da MEC e a ativação das integrinas são modulados por uma variedade de citocinas e fatores de crescimento, e esta modulação parece ser o mecanismo de regulação que influencia a proliferação de células-tronco e progenitores hematopoéticos, migração transendotelial ou transestromal e homing. Tanto no processo de migração, homing e invasão tumoral, as células seguem os seguintes passos: 1 - Degradação da MEC por enzimas secretadas pelas células: metaloproteinases, colagenases, plasmina, catepsinas, glicosidases e heparanases; 2 - Locomoção das células na região da MEC previamente degradada pelas enzimas; 3 - Adesão das células via receptores específicos da superfície celular, que geralmente interagem com componentes da MEC. Nas doenças onco-hematológicas, a interação das células neoplásicas com a matriz extracelular também influencia na agressividade e prognóstico da doença.The extracellular matrix (ECM is a complex structure composed of collagens, proteoglycans, glycosaminoglycans and adhesive glycoproteins. Interactions between the cells and the ECM are crucial to determine cell behavior, such as growth, death, differentiation and motility. Hematopoiesis is the system responsible for the production of blood cells. The control of proliferation and

Inhibidores de la enzima de conversión de la angiotensina en el tratamiento de la hipertensión arterial

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Alfredo Vázquez Vigoa

1998-12-01

Full Text Available Se presenta una revisión actualizada de la función de los inhibidores de la enzima de conversión de angiotensina (IECA en el tratamiento del paciente hipertenso. Se explica el mecanismo de acción de dichos fármacos, basado en el bloqueo de la generación de angiotensina II y en el incremento de péptidos vasodilatadores (bradiquinina, efectos que logran tanto a nivel del sistema renina-angiotensina circulante, como sobre el sistema renina-angiotensina hístico. Se resalta su utilidad en el paciente diabético hipertenso, donde aumenta la sensibilidad a la insulina y evita la aparición de nefropatía diabética. Se discute el impacto de los IECA sobre la regresión de la hipertrofia ventricular izquierda y su valor en la cardiopatía isquémica. Se concluye que los IECA constituyen una opción eficaz en el paciente hipertenso con normalización de las cifras de tensión arterial en el 60 % de los casos.An updated review of the function of angiotensin-converting enzyme inhibitors (ACEI in the treatment of hypertensive patient is set forth. The mechanism of the action of such drugs, based on the blocking of II angiotension generation and the increase of vasodilatador peptides (bradichinin, and their effects on both the circulating renin- -angiotensin system and the hystic renin-angiotensin system are explained. Their usefulness for treating hypertensive diabetic patient is underline since they invease sensitivity to insuline and prevent diabetic nephropathy. The impact of ACEI on regression of left ventricular hypertrophy and their importance in ischemic heart disease are discussed. It is concluded that ACEI are an effective alternative for hypertensive patient treatment because they help to normalize blood presure in 60 % of cases.

Technological application of an extracellular cell lytic enzyme in xanthan gum clarification Aplicação tecnológica de uma enzima celulolítica para clarificação de goma xantana

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Suresh Shastry

2005-03-01

Full Text Available An extracellular cell lytic enzyme from Pseudomonas sp. was active on heat killed cells of Xanthomonas campestris. The lytic activity caused enzymatic digestion of X.campestris xanthan gum. Digestion was effective for highly viscous native xanthan 2.0% (w/v and 2.5% (w/v commercial Sigma xanthan. Scanning electron microscopy and SDS-PAGE observations confirmed the cell lytic action on X.campestris cells.Uma enzima extracelular celulolítica produzida por Pseudomonas sp. foi ativa sobre células de Xanthomonas campestris mortas pelo calor. A atividade lítica causou a digestão enzimática de goma xantana de X. campestris. A digestão foi eficiente tanto para xantana nativa altamante viscosa (2,0% w/v como para xantana comercial Sigma (2,5% w/v. Observações por microscopia eletrônica de varredura demonstraram a ação celulolítica sobre células de X. campestris.

Effect of resveratrol on 17beta-estradiol sulfation by human hepatic and jejunal S9 and recombinant sulfotransferase 1E1.

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Furimsky, Anna M; Green, Carol E; Sharp, Lewanne E Hunt; Catz, Paul; Adjei, Araba A; Parman, Toufan; Kapetanovic, Izet M; Weinshilboum, Richard M; Iyer, Lalitha V

2008-01-01

The purpose of this study was to investigate the sulfation of resveratrol (3,5,4'-trihydroxystilbene) and its potential to exhibit drug-drug interactions via sulfation. The possible interaction of resveratrol with 17beta-estradiol (E2), a major estrogen hormone and prototypic substrate for sulfate conjugation, was studied. Resveratrol and E2 are both known to undergo sulfate conjugation catalyzed by human sulfotransferases (SULTs). Resveratrol is a phytoestrogen with mixed estrogen agonist/antagonist properties that is being developed as a chemopreventive agent. The sulfate conjugation of E2 and resveratrol were studied individually using S9 fractions from human liver and jejunum as well as recombinant human SULT isoforms. The sulfation of E2 (3-20 nM) was then investigated in the presence of various concentrations (0, 0.5, 1, and 2 microM) of resveratrol using the two S9 preparations as well as recombinant SULT1E1, the major isoform responsible for E2 sulfation. Resveratrol inhibited E2 sulfation with estimated K(i) values of 1.1 microM (liver), 0.6 microM (jejunum), and 2.3 microM (SULT1E1), concentrations that could be pharmacologically relevant. The results suggest that these phytoestrogens can potentially alter the homeostasis of estrogen levels. These findings also imply that resveratrol may inhibit the metabolism of other estrogen analogs or therapeutic agents such as ethinylestradiol or dietary components that are also substrates for SULT1E1.

Glutathione-S-transferase profiles in the emerald ash borer, Agrilus planipennis.

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Rajarapu, Swapna Priya; Mittapalli, Omprakash

2013-05-01

The emerald ash borer, Agrilus planipennis Fairmaire is a recently discovered invasive insect pest of ash, Fraxinus spp. in North America. Glutathione-S-transferases (GST) are a multifunctional superfamily of enzymes which function in conjugating toxic compounds to less toxic and excretable forms. In this study, we report the molecular characterization and expression patterns of different classes of GST genes in different tissues and developmental stages plus their specific activity. Multiple sequence alignment of all six A. planipennis GSTs (ApGST-E1, ApGST-E2, ApGST-E3, ApGST-O1, ApGST-S1 and ApGST-μ1) revealed conserved features of insect GSTs and a phylogenetic analysis grouped the GSTs within the epsilon, sigma, omega and microsomal classes of GSTs. Real time quantitative PCR was used to study field collected samples. In larval tissues high mRNA levels for ApGST-E1, ApGST-E3 and ApGST-O1 were obtained in the midgut and Malpighian tubules. On the other hand, ApGST-E2 and ApGST-S1 showed high mRNA levels in fat body and ApGST-μ1 showed constitutive levels in all the tissues assayed. During development, mRNA levels for ApGST-E2 were observed to be the highest in feeding instars, ApGST-S1 in prepupal instars; while the others showed constitutive patterns in all the developmental stages examined. At the enzyme level, total GST activity was similar in all the tissues and developmental stages assayed. Results obtained suggest that A. planipennis is potentially primed with GST-driven detoxification to metabolize ash allelochemicals. To our knowledge this study represents the first report of GSTs in A. planipennis and also in the family of wood boring beetles. Copyright © 2013 Elsevier Inc. All rights reserved.

Improving the redistribution of the security lessons in healthcare: An evaluation of the Generic Security Template.

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He, Ying; Johnson, Chris

2015-11-01

The recurrence of past security breaches in healthcare showed that lessons had not been effectively learned across different healthcare organisations. Recent studies have identified the need to improve learning from incidents and to share security knowledge to prevent future attacks. Generic Security Templates (GSTs) have been proposed to facilitate this knowledge transfer. The objective of this paper is to evaluate whether potential users in healthcare organisations can exploit the GST technique to share lessons learned from security incidents. We conducted a series of case studies to evaluate GSTs. In particular, we used a GST for a security incident in the US Veterans' Affairs Administration to explore whether security lessons could be applied in a very different Chinese healthcare organisation. The results showed that Chinese security professional accepted the use of GSTs and that cyber security lessons could be transferred to a Chinese healthcare organisation using this approach. The users also identified the weaknesses and strengths of GSTs, providing suggestions for future improvements. Generic Security Templates can be used to redistribute lessons learned from security incidents. Sharing cyber security lessons helps organisations consider their own practices and assess whether applicable security standards address concerns raised in previous breaches in other countries. The experience gained from this study provides the basis for future work in conducting similar studies in other healthcare organisations. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Purification, molecular cloning and heterologous expression of a glutathione S-transferase involved in insecticide resistance from the rice brown planthopper, Nilaparvata lugens.

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Vontas, John G; Small, Graham J; Nikou, Dimitra C; Ranson, Hilary; Hemingway, Janet

2002-03-01

A novel glutathione S-transferase (GST)-based pyrethroid resistance mechanism was recently identified in Nilaparvata lugens [Vontas, Small and Hemingway (2001) Biochem. J. 357, 65-72]. To determine the nature of GSTs involved in conferring this resistance, the GSTs from resistant and susceptible strains of N. lugens were partially purified by anion exchange and affinity chromatography. The majority of peroxidase activity, previously correlated with resistance, was confined to the fraction that bound to the affinity column, which was considerably elevated in the resistant insects. A cDNA clone encoding a GST (nlgst1-1) - the first reported GST sequence from Hemiptera with up to 54% deduced amino-acid identity with other insect class I GSTs - was isolated from a pyrethroid-resistant strain. Northern analysis showed that nlgst1-1 was overexpressed in resistant insects. nlgst1-1 was expressed in Escherichia coli, purified and characterized. The ability of the recombinant protein to bind to the S-hexylglutathione affinity matrix, its substrate specificities and its immunological properties confirmed that this GST was one from the elevated subset of N. lugens GSTs. Peroxidase activity of the recombinant nlgst1-1 indicated that it had a role in resistance, through detoxification of lipid peroxidation products induced by pyrethroids. Southern analysis of genomic DNA from the resistant and susceptible strains indicated that GST-based insecticide resistance may be associated with gene amplification in N. lugens.

Cantharidin Impedes Activity of Glutathione S-Transferase in the Midgut of Helicoverpa armigera Hübner

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Ya Lin Zhang

2013-03-01

Full Text Available Previous investigations have implicated glutathione S-transferases (GSTs as one of the major reasons for insecticide resistance. Therefore, effectiveness of new candidate compounds depends on their ability to inhibit GSTs to prevent metabolic detoxification by insects. Cantharidin, a terpenoid compound of insect origin, has been developed as a bio-pesticide in China, and proves highly toxic to a wide range of insects, especially lepidopteran. In the present study, we test cantharidin as a model compound for its toxicity, effects on the mRNA transcription of a model Helicoverpa armigera glutathione S-transferase gene (HaGST and also for its putative inhibitory effect on the catalytic activity of GSTs, both in vivo and in vitro in Helicoverpa armigera, employing molecular and biochemical methods. Bioassay results showed that cantharidin was highly toxic to H. armigera. Real-time qPCR showed down-regulation of the HaGST at the mRNA transcript ranging from 2.5 to 12.5 folds while biochemical assays showed in vivo inhibition of GSTs in midgut and in vitro inhibition of rHaGST. Binding of cantharidin to HaGST was rationalized by homology and molecular docking simulations using a model GST (1PN9 as a template structure. Molecular docking simulations also confirmed accurate docking of the cantharidin molecule to the active site of HaGST impeding its catalytic activity.

Ambient pH-regulated enzime secretion in endophytic and pathogenic isolates of the fungal genus Colletotrichum Secreção de enzimas mediada pelo pH do ambiente em isolados patogênicos e endofíticos do fungo Colletotrichum

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Walter Maccheroni Jr.

2004-06-01

Full Text Available In fungi a genetic system ensures that enzymes are secreted mainly at ambient pH values corresponding to their optima of activity. Although a great deal of information has been obtained concerning this environmental response, there is a lack of studies involving phytopathogenic, endophytic and entomopathogenic fungi as well as different aspects of fungus-host interactions. This study compares in a plate-clearing assays, the effect of ambient pH in the secretion of amylase, cellulase, lipase, pectinase and protease by endophytic, phytopathogenic, and entomopathogenic isolates belonging to several species of Colletotrichum. All enzymes were secreted in a pH-dependent manner by all isolates. Endophytes and pathogens showed distinct patterns of protease secretion, with optima at alkaline and acid growth conditions, respectively. In liquid medium, a Pi-repressible acid phosphatase of an endophytic isolate responded to ambient pH, having a 14-fold increase in secreted specific activity at acid pH, as compared to alkaline pH. Furthermore, part of a Colletotrichum pacC homologue gene, coding for a transcriptional factor responsible for pH-regulated gene expression, was cloned. Ambient pH seems to be a general factor controlling enzyme secretion in fungus-host interactions through a conserved genetic circuit.Em fungos, um sistema de regulação gênica garante que enzimas sejam secretadas predominantemente em valores de pH do ambiente próximos aos pH ótimos de atividade correspondentes. Embora muita informação tenha sido acumulada sobre essa resposta adaptativa, não existem estudos envolvendo fungos fitopatogênicos, endofíticos e entomopatogênicos, bem como sobre outros aspectos relacionados às interações fungo-hospedeiro. No presente trabalho foi comparado, em meio sólido, o efeito do pH do ambiente na secreção das enzimas amilase, celulase, lipase, pectinase e protease por isolados endofíticos, fitopatogênico e entomopatog

THE ROLES OF DETOXIFYING ENZYMES AND AChE INSENSITIVITY IN METHAMIDOPHOS RESISTANCE DEVELOPMENT AND DECLINE IN NILAPARVATA LUGENS

Institute of Scientific and Technical Information of China (English)

Ze-wenLiu; Zhao-junHan; Ling-chunZhang

2003-01-01

Methamidophos resistance of brown planthopper (Nilaparvata lugens Stal, BPH) was selected in laboratory. After successive selection for 9 generations, the selection was ceased by rearing BPH without contact with any insecticide for 9 generations. In the full course, the successive changes of esterase activity, MFO activity, GSTs activity and AChE insensitivity were analyzed. The results showed that the change of esterase activity was high correlated with that of methamidophos in the full course, which indicated that esterase played very important role both in the resistance development and in the resistance decline. However, the change of AChE insensitivity only significantly correlated with that of resistance in the development stage, and the change of MFO activity or GSTs activity only significantly correlated with that of the resistance in the decline stage, which indicated the changes of AChE insensitivity, MFO activity or GSTs activity only played some roles in different stages of the resistance change.

Radiation Resistance of Enzymes in Foods Irradiated Against Microbial Damage; Radioresistance des Enzymes dans les Denrees Alimentaires Irradiees pour les Proteger Contre les Alterations Dues a des Bacteries; Radiatsionnaya ustojchivost' ehnzimov v pishchevykh produktakh,obluchennykh protiv vrednogo vozdejstviya mikrobov; Radiorresistencia de las Enzimas en los Alimentos Irradiados

Energy Technology Data Exchange (ETDEWEB)

Vas, K. [Joint FAO/IAEA Division of Atomic Energy in Agriculture, International Atomic Energy Agency, Vienna (Austria)

1966-11-15

, on sait que les substances radiomoueificatrices exercent une influence determinante sur la radioinactivation des enzymes, l'auteur estime qu'il faudrait entreprendre des etudes plus poussees sur les radioprotecteurs et les radiosensibilisateurs avec des enzymes fortement purifies. La grande radiosensibilite qui caracterise ordinairement les enzymes tres purs et, par contraste, l'extreme resistance des enzymes dans les tissus et dans leurs extraits non purifies, demontrent la necessite de determiner les raisons de cette difference, ce qui facilitera sans doute la mise au point d'une methode, rationnelle permettant de controler l'activite des enzymes dans le cadre de l'irradiation des aliments. De cette maniere, on pourrait peut-etre mettre au point une 'methode mixte' inedite, laquelle viendrait s'ajouter aux methodes combinees qui ont deja ete elaborees pour le controle de l'action des enzymes; en d'autres termes, on combinerait l'irradiation avec a) le traitement thermique, b) la refrigeration et c) les methodes physiologiques (par exemple, controle de la giycogenolyse). Le memoire fait brievement le point des applications de ces divers procedes. (author) [Spanish] La mayor parte de las enzimas que actuan durante el almacenamiento, la elaboracion y la conservacion de los alimentos son bastante resistentes a las radiaciones ionizantes. Por lo general, esa resistencia en su propio medio suele ser superior en un orden de magnitud a la de los microorganismos que causan la descomposicion o la alteracion de los alimentos. Debido a ello, dosis de irradiacion suficientes para la estabilizacion microbiologica y cuya aplicacion se considera actualmente apenas economica, no impiden la deterioracion causada por las enzimas. Se ha intentado ya muchas veces inactivar por irradiacion las enzimas de los alimentos, pero a juzgar por el escaso exito de la mayor parte de los ensayos efectuados, es necesario estudiar mucho mas a fondo los aspectos generales de la cuestion a fin de

Multiple analytical approaches reveal distinct gene-environment interactions in smokers and non smokers in lung cancer.

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Rakhshan Ihsan

Full Text Available Complex disease such as cancer results from interactions of multiple genetic and environmental factors. Studying these factors singularly cannot explain the underlying pathogenetic mechanism of the disease. Multi-analytical approach, including logistic regression (LR, classification and regression tree (CART and multifactor dimensionality reduction (MDR, was applied in 188 lung cancer cases and 290 controls to explore high order interactions among xenobiotic metabolizing genes and environmental risk factors. Smoking was identified as the predominant risk factor by all three analytical approaches. Individually, CYP1A1*2A polymorphism was significantly associated with increased lung cancer risk (OR = 1.69;95%CI = 1.11-2.59,p = 0.01, whereas EPHX1 Tyr113His and SULT1A1 Arg213His conferred reduced risk (OR = 0.40;95%CI = 0.25-0.65,p<0.001 and OR = 0.51;95%CI = 0.33-0.78,p = 0.002 respectively. In smokers, EPHX1 Tyr113His and SULT1A1 Arg213His polymorphisms reduced the risk of lung cancer, whereas CYP1A1*2A, CYP1A1*2C and GSTP1 Ile105Val imparted increased risk in non-smokers only. While exploring non-linear interactions through CART analysis, smokers carrying the combination of EPHX1 113TC (Tyr/His, SULT1A1 213GG (Arg/Arg or AA (His/His and GSTM1 null genotypes showed the highest risk for lung cancer (OR = 3.73;95%CI = 1.33-10.55,p = 0.006, whereas combined effect of CYP1A1*2A 6235CC or TC, SULT1A1 213GG (Arg/Arg and betel quid chewing showed maximum risk in non-smokers (OR = 2.93;95%CI = 1.15-7.51,p = 0.01. MDR analysis identified two distinct predictor models for the risk of lung cancer in smokers (tobacco chewing, EPHX1 Tyr113His, and SULT1A1 Arg213His and non-smokers (CYP1A1*2A, GSTP1 Ile105Val and SULT1A1 Arg213His with testing balance accuracy (TBA of 0.6436 and 0.6677 respectively. Interaction entropy interpretations of MDR results showed non-additive interactions of tobacco chewing with

A novel method for screening the glutathione transferase inhibitors

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Węgrzyn Grzegorz

2009-03-01

Full Text Available Abstract Background Glutathione transferases (GSTs belong to the family of Phase II detoxification enzymes. GSTs catalyze the conjugation of glutathione to different endogenous and exogenous electrophilic compounds. Over-expression of GSTs was demonstrated in a number of different human cancer cells. It has been found that the resistance to many anticancer chemotherapeutics is directly correlated with the over-expression of GSTs. Therefore, it appears to be important to find new GST inhibitors to prevent the resistance of cells to anticancer drugs. In order to search for glutathione transferase (GST inhibitors, a novel method was designed. Results Our results showed that two fragments of GST, named F1 peptide (GYWKIKGLV and F2 peptide (KWRNKKFELGLEFPNL, can significantly inhibit the GST activity. When these two fragments were compared with several known potent GST inhibitors, the order of inhibition efficiency (measured in reactions with 2,4-dinitrochlorobenzene (CDNB and glutathione as substrates was determined as follows: tannic acid > cibacron blue > F2 peptide > hematin > F1 peptide > ethacrynic acid. Moreover, the F1 peptide appeared to be a noncompetitive inhibitor of the GST-catalyzed reaction, while the F2 peptide was determined as a competitive inhibitor of this reaction. Conclusion It appears that the F2 peptide can be used as a new potent specific GST inhibitor. It is proposed that the novel method, described in this report, might be useful for screening the inhibitors of not only GST but also other enzymes.

Análisis del consumo de inhibidores de la enzima convertidora de angioténsina en el territorio oeste de La Habana, 2005-2009

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José Ramón Cabrera Cepero

2011-12-01

Full Text Available La planificación de los recursos es un problema trascendental en los países en desarrollo y también en Cuba, por lo que hacer el mejor uso de los presupuestos limitados y de las escasas divisas, es de vital importancia. Planificar las cantidades de medicamentos necesarias, para lograr garantizar una disponibilidad adecuada de estos en todos los niveles de asistencia, es una tarea en la cual intervienen un sinnúmero de factores. El objetivo de este trabajo fue demostrar en qué medida la introducción del enalapril tabletas influyó en el consumo del captopril tabletas, mediante el análisis de los patrones de consumo de los medicamentos inhibidores de la enzima convertidora de angiotensina en el territorio oeste de La Habana entre marzo de 2005 y diciembre de 2009. Para ello se realizó un estudio de utilización de medicamentos de consumo, de tipo descriptivo, observacional y retrospectivo. Se calcularon las DHD (dosis por mil habitantes día. Los resultados de este trabajo demuestran cómo en este grupo hay un desplazamiento del consumo hacia el enalapril. Este es un comportamiento lógico por la comodidad de la administración y la menor incidencia de efectos adversos. Sin embargo, el captopril se mantiene en valores entre 20 y 30 DHD x 1 000 habitantes ya que hay un grupo de pacientes que continúan con este tratamiento y es de elección en la crisis hipertensiva.

Transcrição reversa na determinação da expressão do mRNA para a enzima conversora de angiotensina testicular em animais tratados com zinco Assessment of the reverse transcriptase polymerase chain reaction technique in the determination of the mRNA expression for the testicular angiotensin-converting enzyme in zinc treated rats

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Gilberto Simeone Henriques

2005-12-01

Full Text Available OBJETIVO: O objetivo deste trabalho foi otimizar as condições reacionais capazes de ocasionar variabilidade e de introduzir erros sistemáticos na reação em cadeia pela polimerase aplicada à análise da expressão gênica da isoforma testicular da enzima conversora de angiotensina. MÉTODOS: Avaliaram-se a concentração de cDNA, a concentração dos iniciadores, a temperatura de hibridização e o número de ciclos de desnaturação, hibridização e extensão. Para tanto, extraiu-se o RNA total por meio da reação com fenol-clorofórmio e isotiocianato de guanidina de amostras de testículos de ratos Wistar alimentados com uma ração contendo zinco. Em seguida, gerou-se o cDNA por transcrição reversa. Utilizando-se iniciadores específicos, amplificaram-se o gene de interesse (isoforma testicular da enzima conversora de angiotensina e o gene controle Gliceraldeído-3-Fosfato-Desidrogenase. As amostras foram então aplicadas em gel de agarose e submetidas à eletroforese, coradas em brometo de etídio e visualizadas sob luz ultravioleta. RESULTADOS: Demonstrou-se que a melhor condição reacional para a reação em cadeia pela polimerase da isoforma testicular da enzima conversora de angiotensina e do Gliceraldeído-3-Fosfato-Desidrogenase foi: (1 concentração inicial de cDNA de 2µg, (2 concentração de iniciadores de 200nM, (3 temperatura de hibridização entre 57,5ºC e 60,1ºC e (4 33 ciclos. CONCLUSÃO: Com essa otimização pôde-se minimizar as interferências sobre a técnica, contribuindo-se para a obtenção de dados comparativos a respeito da expressão gênica da enzima conversora de angiotensina testicular.OBJETIVE: The aim of the present work was to optimize the reaction conditions capable of generating variability and introducing systematic errors in the chain reaction of the polymerase used to analyze the gene expression for the testicular isoform of the angiotensin-converting enzyme. METHODS:The cDNA concentration

Meta-análise da digestibilidade ileal de aminoácidos e minerais em suínos alimentados com dietas contendo enzimas

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Cheila Roberta Lehnen

2011-04-01

Full Text Available O objetivo deste trabalho foi avaliar, por meio da meta-análise, o efeito da fitase e da xilanase sobre a digestibilidade ileal aparente (DIa de aminoácidos, cálcio e fósforo, em suínos em fase de crescimento. A base de dados consistiu de 21 artigos publicados entre 1998 e 2009, no total de 82 tratamentos e 644 suínos. A meta-análise foi realizada por análise gráfica, de correlação, de variância-covariância. As concentrações de fósforo fítico e as frações fibra em detergente neutro, fibra em detergente ácido e lignina em detergente ácido, nas dietas, apresentaram correlações baixas e negativas com a DIa do cálcio, fósforo e aminoácidos. A adição de fitase às dietas aumentou em 2% a DIa da arginina, em 14% a do cálcio e em 34% a do fósforo. A DIa da arginina, fenilalanina, isoleucina e lisina foi 3,3% superior em suínos alimentados com dietas com xilanase, em relação às dietas sem a enzima. O fósforo fítico e as fibras, nas dietas, reduzem a DIa do cálcio, do fósforo e dos aminoácidos essenciais. O uso de fitase e xilanase, nas dietas, melhora o aproveitamento de cálcio, fósforo e alguns aminoácidos. No entanto, o excesso de cálcio e fósforo nas dietas reduz a ação da fitase sobre a digestibilidade ileal dos nutrientes.

Ethylene evolution and endo-beta-mannanase activity during lettuce seed germination at high temperature Evolução de etileno e atividade da enzima endo-beta-mananase durante a germinação de sementes de alface sob altas temperaturas

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Warley Marcos Nascimento

2004-04-01

Full Text Available High temperatures during lettuce seed imbibition can delay or completely inhibit germination and the endosperm layer appears to restrict the radicle protrusion. The role of endo-beta-mannanase during lettuce seed germination at 35°C and the influence of ethylene in endo-beta-mannanase regulation were investigated. Seeds of 'Dark Green Boston' (DGB and 'Everglades' (EVE were germinated in water, or 10 mmol L-1 of 1-aminocyclopropane-1-carboxylic acid (ACC, or 10 mmol L-1 of aminoethoxyvinylglycine (AVG, or 20 mmol L-1 of silver thiosulphate (STS. Seeds were also primed in polyethylene glycol (PEG, or PEG + ACC, PEG + AVG, or PEG + STS. Untreated seeds germinated 100% at 20°C. At 35°C, EVE seeds germinated 100%, whereas DGB seeds germinated only 33%. Seed priming or adding ACC during incubation increased germination at 35°C. Higher ethylene evolution was detected in EVE than in DGB during germination at 35°C. AVG did not inhibit seed germination of DGB at 35°C, but STS did. Higher endo-beta-mannanase activity was observed in EVE compared with DGB seeds. Providing ACC either during priming or during germination increased endo-beta-mannanase activity, whereas AVG and STS led to decreased or no activity. Ethylene may overcome the inhibitory effect of high temperature in thermosensitive lettuce seeds due to increased endo-beta-mannanase, possibly leading to weakening of the endosperm.Altas temperaturas durante a embebição das sementes de alface podem atrasar ou inibir a germinação e o endosperma parece ser o responsável na restrição da protrusão da radícula. O envolvimento da enzima endo-beta-mananase durante a germinação de sementes de alface a 35°C e a influência do etileno na regulagem desta enzima foram estudados. Sementes das cultivares Dark Green Boston (DGB e Everglades (EVE foram germinadas em água ou em soluções de 10 mmol L-1 de 1-aminociclopropano-1-ácido carboxilico (ACC, 10 mmol L-1 de amino-etoxi-vinil-glicina (AVG

Aparência, compostos fenólicos e enzimas oxidativas em uva 'Itália' sob influência do cálcio e do armazenamento refrigerado

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LIMA MARIA AUXILIADORA COÊLHO DE

2002-01-01

Full Text Available O objetivo deste trabalho foi determinar o efeito da aplicação pré-colheita de cálcio na aparência (secamento do engaço, danos mecânicos e podridões, teor de fenólicos e enzimas oxidativas (polifenoloxidase e peroxidase em uva. Os cachos de uva 'Itália' de um cultivo comercial em Petrolina, Pernambuco, Brasil, foram marcados e imersos por 10 segundos, em soluções de Ca a 0 e 1,5%, na forma de cloreto de cálcio, aos 57 dias após o início da formação dos frutos (quando as bagas começaram a mudar de cor e amolecer. Após a colheita, os frutos foram armazenados a 3,5±0,2°C e 93±6% UR e avaliados aos 0; 14; 28; 42; 56 e 70 dias. Houve um incremento no secamento do engaço, no aparecimento de sintomas de danos mecânicos e de podridões nas bagas com o tempo de armazenamento. A aplicação de cálcio reduziu a atividade de polifenoloxidase e, conseqüentemente, os sintomas de danos mecânicos, resultando numa melhor aparência. A vida útil das uvas foi de aproximadamente 56 dias, quando sintomas de senescência, podridões e o nível dos sintomas de danos mecânicos começaram a aumentar de forma significativa.

Epistasis Analysis for Estrogen Metabolic and Signaling Pathway Genes on Young Ischemic Stroke Patients

Science.gov (United States)

Hsieh, Yi-Chen; Jeng, Jiann-Shing; Lin, Huey-Juan; Hu, Chaur-Jong; Yu, Chia-Chen; Lien, Li-Ming; Peng, Giia-Sheun; Chen, Chin-I; Tang, Sung-Chun; Chi, Nai-Fang; Tseng, Hung-Pin; Chern, Chang-Ming; Hsieh, Fang-I; Bai, Chyi-Huey; Chen, Yi-Rhu; Chiou, Hung-Yi; Jeng, Jiann-Shing; Tang, Sung-Chun; Yeh, Shin-Joe; Tsai, Li-Kai; Kong, Shin; Lien, Li-Ming; Chiu, Hou-Chang; Chen, Wei-Hung; Bai, Chyi-Huey; Huang, Tzu-Hsuan; Chi-Ieong, Lau; Wu, Ya-Ying; Yuan, Rey-Yue; Hu, Chaur-Jong; Sheu, Jau- Jiuan; Yu, Jia-Ming; Ho, Chun-Sum; Chen, Chin-I; Sung, Jia-Ying; Weng, Hsing-Yu; Han, Yu-Hsuan; Huang, Chun-Ping; Chung, Wen-Ting; Ke, Der-Shin; Lin, Huey-Juan; Chang, Chia-Yu; Yeh, Poh-Shiow; Lin, Kao-Chang; Cheng, Tain-Junn; Chou, Chih-Ho; Yang, Chun-Ming; Peng, Giia-Sheun; Lin, Jiann-Chyun; Hsu, Yaw-Don; Denq, Jong-Chyou; Lee, Jiunn-Tay; Hsu, Chang-Hung; Lin, Chun-Chieh; Yen, Che-Hung; Cheng, Chun-An; Sung, Yueh-Feng; Chen, Yuan-Liang; Lien, Ming-Tung; Chou, Chung-Hsing; Liu, Chia-Chen; Yang, Fu-Chi; Wu, Yi-Chung; Tso, An-Chen; Lai, Yu- Hua; Chiang, Chun-I; Tsai, Chia-Kuang; Liu, Meng-Ta; Lin, Ying-Che; Hsu, Yu-Chuan; Chen, Chih-Hung; Sung, Pi-Shan; Chern, Chang-Ming; Hu, Han-Hwa; Wong, Wen-Jang; Luk, Yun-On; Hsu, Li-Chi; Chung, Chih-Ping; Tseng, Hung-Pin; Liu, Chin-Hsiung; Lin, Chun-Liang; Lin, Hung-Chih; Hu, Chaur-Jong

2012-01-01

Background Endogenous estrogens play an important role in the overall cardiocirculatory system. However, there are no studies exploring the hormone metabolism and signaling pathway genes together on ischemic stroke, including sulfotransferase family 1E (SULT1E1), catechol-O-methyl-transferase (COMT), and estrogen receptor α (ESR1). Methods A case-control study was conducted on 305 young ischemic stroke subjects aged ≦ 50 years and 309 age-matched healthy controls. SULT1E1 -64G/A, COMT Val158Met, ESR1 c.454−397 T/C and c.454−351 A/G genes were genotyped and compared between cases and controls to identify single nucleotide polymorphisms associated with ischemic stroke susceptibility. Gene-gene interaction effects were analyzed using entropy-based multifactor dimensionality reduction (MDR), classification and regression tree (CART), and traditional multiple regression models. Results COMT Val158Met polymorphism showed a significant association with susceptibility of young ischemic stroke among females. There was a two-way interaction between SULT1E1 -64G/A and COMT Val158Met in both MDR and CART analysis. The logistic regression model also showed there was a significant interaction effect between SULT1E1 -64G/A and COMT Val158Met on ischemic stroke of the young (P for interaction = 0.0171). We further found that lower estradiol level could increase the risk of young ischemic stroke for those who carry either SULT1E1 or COMT risk genotypes, showing a significant interaction effect (P for interaction = 0.0174). Conclusions Our findings support that a significant epistasis effect exists among estrogen metabolic and signaling pathway genes and gene-environment interactions on young ischemic stroke subjects. PMID:23112845

Genotype of metabolic enzymes and the benefit of tamoxifen in postmenopausal breast cancer patients

International Nuclear Information System (INIS)

Wegman, Pia; Vainikka, Linda; Stål, Olle; Nordenskjöld, Bo; Skoog, Lambert; Rutqvist, Lars-Erik; Wingren, Sten

2005-01-01

Tamoxifen is widely used as endocrine therapy for oestrogen-receptor-positive breast cancer. However, many of these patients experience recurrence despite tamoxifen therapy by incompletely understood mechanisms. In the present report we propose that tamoxifen resistance may be due to differences in activity of metabolic enzymes as a result of genetic polymorphism. Cytochrome P450 2D6 (CYP2D6) and sulfotransferase 1A1 (SULT1A1) are polymorphic and are involved in the metabolism of tamoxifen. The CYP2D6*4 and SULT1A1*2 genotypes result in decreased enzyme activity. We therefore investigated the genotypes of CYP2D6 and SULT1A1 in 226 breast cancer patients participating in a trial of adjuvant tamoxifen treatment in order to validate the benefit from the therapy. The patients were genotyped using PCR followed by cleavage with restriction enzymes. Carriers of the CYP2D6*4 allele demonstrated a decreased risk of recurrence when treated with tamoxifen (relative risk = 0.28, 95% confidence interval = 0.11–0.74, P = 0.0089). A similar pattern was seen among the SULT1A1*1 homozygotes (relative risk = 0.48, 95% confidence interval = 0.21–1.12, P = 0.074). The combination of CYP2D6*4 and/or SULT1A1*1/*1 genotypes comprised 60% of the patients and showed a 62% decreased risk of distant recurrence with tamoxifen (relative risk = 0.38, 95% confidence interval = 0.19–0.74, P = 0.0041). The present study suggests that genotype of metabolic enzymes might be useful as a guide for adjuvant endocrine treatment of postmenopausal breast cancer patients. However, results are in contradiction to prior hypotheses and the present sample size is relatively small. Findings therefore need to be confirmed in a larger cohort

Association of GSTO1 and GSTO2 Polymorphism with Risk of End-Stage Renal Disease Development and Patient Survival

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Cimbaljevic Slavica

2016-09-01

Full Text Available Background: Oxidative stress in patients with end-stage renal disease (ESRD is associated with long-term cardiovascular complications. The cytosolic family of glutathione S-transferases (GSTs is involved in the detoxication of various toxic compounds and antioxidant protection. GST omega class members, GSTO1 and GSTO2 possess, unlike other GSTs, dehydroascorbate reductase and deglutathionylation activities. The aim of this study was to clarify the role of genetic polymorphisms of GSTO1 (rs4925 and GSTO2 (rs156697 as risk determinants for ESRD development, as well as in the survival of these patients.

Producción de endoproteasa ácida extracelular (mrAE) por el hongo fitopatógeno Moniliophthora roreri cultivado en el sistema de fermentación en el estado sólido con bagazo de caña de azúcar como sustrato.

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Soberano Mayo, María Isabel

2012-01-01

Las proteasas son enzimas industriales muy importantes y comprenden alrededor del 60% de las enzimas comerciales en el mundo. Dos terceras partes de estas proteasas son de fuentes microbianas, principalmente de origen fúngico. El presente trabajo tuvo como objetivos estudiar la cinética de producción de enzima endoproteasa ácida extracelular (mrAe) del hongo fitopatógeno Moniliophthora roreri MRO1, cultivado en sistema de fermentación en estado sólido (FES), utilizando como sustrato inductor ...

Enzymatic sulfation of tocopherols and tocopherol metabolites by human cytosolic sulfotransferases.

Science.gov (United States)

Hashiguchi, Takuyu; Kurogi, Katsuhisa; Sakakibara, Yoichi; Yamasaki, Masao; Nishiyama, Kazuo; Yasuda, Shin; Liu, Ming-Cheh; Suiko, Masahito

2011-01-01

Tocopherols are essential micronutrients for mammals widely known as potent lipid-soluble antioxidants that are present in cell membranes. Recent studies have demonstrated that most of the carboxychromanol (CEHC), a tocopherol metabolite, in the plasma exists primarily in sulfate- and glucuronide-conjugated forms. To gain insight into the enzymatic sulfation of tocopherols and their metabolites, a systematic investigation was performed using all 14 known human cytosolic sulfotransferases (SULTs). The results showed that the members of the SULT1 family displayed stronger sulfating activities toward tocopherols and their metabolites. These enzymes showed a substrate preference for γ-tocopherol over α-tocopherol and for γ-CEHC over other CEHCs. Using A549 human lung epithelial cells in a metabolic labeling study, a similar trend in the sulfation of tocopherols and CEHCs was observed. Collectively, the results obtained indicate that SULT-mediated enzymatic sulfation of tocopherols and their metabolites is a significant pathway for regulation of the homeostasis and physiological functions of these important compounds.

Enzimas de função hepática na aflatoxicose aguda experimental em frangos de corte Hepatic enzimes function in experimental acute enzimes aflatoxicosis in broilers

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Adriana Borsa

1998-12-01

Full Text Available Com o objetivo de avaliar a função hepática de aves experimentalmente intoxicadas por aflatoxina, com e sem uso de bentonita sódica, foram utilizados 40 (quarenta frangos de corte, machos, linhagem Ross, divididos em 4 (quatro grupos de 10 (dez. animais, sendo que cada grupo foi submetido a um tratamento: T1 - controle (ração sem aflatoxina ou bentonita, T2 - ração com 5ppm de aflatoxina, T3 - ração com 5ppm de aflatoxina e 0,5% de bentonita sódica e T4 - ração com 0,5% de bentonita sódica. Todos estes tratamentos foram aplicados do 1° ao 42 ° dia de vida das aves. Aos 21, 35 e 42 dias de idade, foram analisados os níveis séricos das enzimas aspartato aminotransferase (AST, alanina aminotransferase (ALT, lactato desidrogenase (LDH e gama glutamiltransferase (GGT, A análise da variância mostrou que houve interação entre os tratamentos e datas de colheita de material, para as seguintes variáveis: AST, LDH e GGT. Para estas, foi aplicado o teste de Tukey, comparando-se as médias de cada tratamento em cada data. Observou-se que as enzimas AST, ALT e GGT não apresentaram diferenças significativas entre tratamentos, porém, nos tratamentos l e 2, a AST apresentou um aumento linear (pThe aim ofthis study is to evaluate ïhe hepatic function of experimentally intoxicated broilers by aflatoxin with and without sodium bentonite. Forty Ross mole broilers, were used divided into 4 groups of 10 birds, and such groups have been submitted to the following treatments: T1- control (feed without aflatoxin or sodium bentonite, T2- feed containing 5ppm of aflatoxin, T3- feed containing 5ppm of aflatoxin and 0.5% of sodium bentonite and T4- feed containing 0.5% of sodium bentonite. Ali these treatments have been appiied from the l st to the 42nd day of lif e. On the days 21, 35 and 42, the serum leveis of the enzimes aspartate aminotransferase (AST, alanino aminotransferase (ALT, lactate dehydrogenase (LDH and gamma glutamiltransferase (GGT

INVESTIGACION, DESARROLLO Y PRODUCCION DE PROTEASAS Y LIPASAS CREIOFILICAS COMERCIALES DE KRILL Y BACTERIAS ANTARTICAS

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ASENJO DE LEUZE, JUAN ALFONSO

2005-01-01

El uso de enzimas a bajas temperaturas posee un gran potencial, fundamentalmente por los menores costos en el uso de energía. Los organismos antárticos son muy interesantes como fuente de enzimas de propiedades únicas, específicamente de proteasas y lipasas que poseen altas actividades a bajas temperaturas (criofílicas), ideales para ser utilizadas como enzimas de detergentes. Esto ha sido demostrado claramente en dos proyectos Fondef anteriores que fueron exitosos. Además no existen hoy en d...

Produção e caracterização de aromas de frutas por microrganismos pectinoliticos utilizando-se residuos agroindustriais.

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Mariana Uenojo

2003-01-01

Resumo: As pectinases foram algumas das primeiras enzimas a serem utilizadas comercialmente nas preparações de vinhos e sucos de frutas na década de 1930 e seu uso tem aumentado consideravelmente, especialmente nas indústrias de alimentos, bebidas e vinhos, têxtil e de papel e celulose. Atualmente essas enzimas correspondem à cerca de 20% do mercado mundial de enzimas e são produzidas naturalmente por plantas, fungos, leveduras e bactérias. Estes microrganismos podem ser inoculados em meios c...

Avaliação da atividade das enzimas pectina metilesterase e β-Galactosidase em mamões cv. Golden armazenados sob diferentes concentrações de oxigênio Activity of pectin methylesterase and β-Galactosidase enzymes in 'Golden' papaya stored under different oxygen concentrations

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Luciana Konda de Azevedo Pinto

2013-03-01

Full Text Available Este trabalho foi realizado com o objetivo de avaliar o efeito de atmosferas controladas contendo diferentes concentrações de oxigênio sobre a atividade das enzimas β-galactosidase e pectina metilesterase, e sobre a cor da casca e a firmeza da polpa de mamões 'Golden'. Os frutos foram mantidos por 36 dias, nas seguintes atmosferas controladas: 1% de O2 e 0,03% CO2 com adsorvedor de etileno, 3% de O2 e 0,03% de CO2 com adsorvedor de etileno, 5% O2 e 0,03% de CO2 com adsorvedor de etileno e atmosfera ambiente sem adsorvedor de etileno. A UR e a temperatura foram mantidas entre 85-95% e a 13º C, respectivamente. Os frutos estocados sob atmosfera de 1% de O2 e 0,03% CO2 apresentaram retardamento nas atividades das enzimas β-galactosidase e pectina metilesterase comparado com os frutos estocados nas outras atmosferas avaliadas. Os frutos armazenados sob atmosfera de 1% de O2 e 0,03% O2 também apresentaram atraso no desenvolvimento da cor da casca e amolecimento da polpa.This study was carried out with the objective to evaluate the effect of controlled atmospheres containing different concentration of oxygen on the activity of β-galactosidase and pectinmethylesterase enzymes, on the skin color and pulp firmness in 'Golden' papaya. The fruits were kept for 36 days under the following controlled atmospheres: 1% of O2 and 0.03% CO2 with ethylene absorber, 3% of O2 and 0.03% of CO2 with ethylene absorber, 5% O2 and 0.03% of CO2 with ethylene absorber and environmental atmosphere without ethylene absorber. Relative humidity was set at the range 85-95% and temperature maintained at 13ºC. Fruit stored under atmospheres of 1% of O2 and 0.03% CO2 had a β-galactosidase and pectimethylesterase activities delayed compared with the fruits storage under the other atmospheres evaluated. There was also, a delay in the development of color and pulp softening in fruits stored under atmospheres of 1% of O2 and 0.03% O2.

Glutathione S-Transferase (GST Gene Diversity in the Crustacean Calanus finmarchicus--Contributors to Cellular Detoxification.

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Vittoria Roncalli

Full Text Available Detoxification is a fundamental cellular stress defense mechanism, which allows an organism to survive or even thrive in the presence of environmental toxins and/or pollutants. The glutathione S-transferase (GST superfamily is a set of enzymes involved in the detoxification process. This highly diverse protein superfamily is characterized by multiple gene duplications, with over 40 GST genes reported in some insects. However, less is known about the GST superfamily in marine organisms, including crustaceans. The availability of two de novo transcriptomes for the copepod, Calanus finmarchicus, provided an opportunity for an in depth study of the GST superfamily in a marine crustacean. The transcriptomes were searched for putative GST-encoding transcripts using known GST proteins from three arthropods as queries. The identified transcripts were then translated into proteins, analyzed for structural domains, and annotated using reciprocal BLAST analysis. Mining the two transcriptomes yielded a total of 41 predicted GST proteins belonging to the cytosolic, mitochondrial or microsomal classes. Phylogenetic analysis of the cytosolic GSTs validated their annotation into six different subclasses. The predicted proteins are likely to represent the products of distinct genes, suggesting that the diversity of GSTs in C. finmarchicus exceeds or rivals that described for insects. Analysis of relative gene expression in different developmental stages indicated low levels of GST expression in embryos, and relatively high expression in late copepodites and adult females for several cytosolic GSTs. A diverse diet and complex life history are factors that might be driving the multiplicity of GSTs in C. finmarchicus, as this copepod is commonly exposed to a variety of natural toxins. Hence, diversity in detoxification pathway proteins may well be key to their survival.

Avaliação da produção de lipases por diferentes cepas de microrganismos isolados em efluentes de laticínios por fermentação submersa Evaluation of lipase production using different strains of microorganisms isolated from dairy effluent through submerged fermentation

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Mirela Roveda; Marcelo Hemkemeier; Luciane Maria Colla

2010-01-01

A produção enzimática é um dos campos mais promissores dentro das tecnologias para a síntese de compostos de alto valor agregado, estando em constante crescimento pela grande capacidade dos microrganismos de realizarem transformações químicas. As enzimas produzidas por processos fermentativos têm sido utilizadas para o controle ambiental. Muitas destas enzimas podem ser produzidas a partir de resíduos industriais, diminuindo os custos de produção. As lipases são enzimas que catalisam a hidról...

Avaliação da produção de lipases por diferentes cepas de microrganismos isolados em efluentes de laticínios por fermentação submersa

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Roveda,Mirela; Hemkemeier,Marcelo; Colla,Luciane Maria

2010-01-01

A produção enzimática é um dos campos mais promissores dentro das tecnologias para a síntese de compostos de alto valor agregado, estando em constante crescimento pela grande capacidade dos microrganismos de realizarem transformações químicas. As enzimas produzidas por processos fermentativos têm sido utilizadas para o controle ambiental. Muitas destas enzimas podem ser produzidas a partir de resíduos industriais, diminuindo os custos de produção. As lipases são enzimas que catalisam a hidról...

Inmovilización de lacasa : métodos y potenciales aplicaciones industriales

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Fernández Fernández, María

2013-01-01

La utilización de enzimas en el sector industrial supone un gran avance para muchos procesos antes realizados mediante reacciones químicas. Desde el punto de vista ambiental la utilización de enzimas en lugar de otro tipo de catalizadores para reacciones de tipo industrial mejora notablemente el impacto ambiental que generan este tipo de procesos. El empleo de estos biocatalizadores está limitado por su escasa estabilidad y alto coste, por lo que la inmovilización de enzimas podría superar es...

Desenvolvimento de novas tecnologias para produção de xarope de glicose a partir do amido

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Freitas, Aline Costa de [UNESP

2012-01-01

As amilases são enzimas que catalisam a hidrólise do amido, dentre as quais se destaca a α-amilase, a β-amilase e a glicoamilase. Apresentam grande importância biotecnológica devido sua aplicação em vários processos industriais, principalmente alimentício. Podem ser obtidas de plantas, animais e microorganismos, no entanto, enzimas microbianas encontram maior demanda industrial. Além da importância econômica destas enzimas, novas metodologias têm sido desenvolvidas com resíduos como substrato...

ENZIMAS LIPOLITICAS DE KRILL ANTARTICO: PURIFICACION Y CARACTERIZACION, ENZIMAS ADAPTADAS AL FRIO?.

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BARRIGA GONZALEZ, ANDRES ANTONIO; BARRIGA GONZALEZ, ANDRES ANTONIO

2006-01-01

La temperatura es uno de los factores ambientales más importantes para la vida, siendo los ambientes fríos los que presentan la mayor distribución en la biosfera. Diferentes organismos han desarrollado diversas estrategias de tolerancia y adaptación al fr 160p.

Inhibition of glutathione S-transferases (GSTs) activity from cowpea ...

African Journals Online (AJOL)

STORAGESEVER

2009-10-19

Oct 19, 2009 ... 1Department of Biochemistry, Federal University of Technology, Akure, Nigeria. ... have positive results for alkaloids, saponin, tannins and flavonoids. ... binding of the extract was competitive by the Dixon plot with Ki of 84, 132 ...

Inhibition of glutathione S-transferases (GSTs) activity from cowpea ...

African Journals Online (AJOL)

STORAGESEVER

2009-10-19

Oct 19, 2009 ... Inhibition effect of the plant extracts on the GST was studied by spectrophotometric method. The ... of assuring food security in developing countries like ..... studies on African cat fish (Clarias gariepinus) liver glutathione s-.

Filamentous fungi and agro-industrial residues selection for enzyme production of biotechnological interestSeleção de fungos filamentosos e de resíduos agroindustriais para a produção de enzimas de interesse biotecnológico

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Erivelton César Stroparo

2012-12-01

Full Text Available Many fungal enzymes have relevant applications in different industrial areas. The objective of this work was to select fungi producing hydrolytic enzymes, as well as establish agro-industrial wastes capable of inducing higher production levels. Xylanase, endoglucanase, amylase and poligalacturonase activities were determined by incubating the culture filtrates with their respective substrates. Subsequently, the reducing sugars determination was carried out using 3,5-dinitrosalicylic acid reagent.The protein determination was performed according the modified Bradford method. Among the fungal strains evaluated, Aspergillus niger J4 showed higher levels of xylanase production (8.73 ± 0.34 U/ml and this was greatest when brewer’s spent grain was used as substrate (9.80 ± 0.02 U/ml. Penicillium miczynskii produced the highest levels of endoglucanasic activity (0.13 ± 0.03 U/ml, which, in turn, was favored in the pineapple peel presence (0.18 ± 0.02 U/ml. In relation to amylase, A. niger J26 was selected as the best producer strain (6.10 ± 0.30 U/ml with wheat bran as the best substrate for their production (7.32 ± 0.14 U/ml. Penicillium verruculosum exhibited the highest level of poligalacturonase activity (8.65 ± 0.12 U/ml, especially when grown in orange peel presence (10.32 ± 0.10 U/ml. These wastes use in these enzymes production may not only reduce their production cost, but also substantially reduce the environmental impact caused by the deposition of these wastes on the environment.Muitas enzimas produzidas por fungos têm relevantes aplicações em diferentes áreas industriais. O objetivo deste trabalho foi selecionar fungos filamentosos produtores de enzimas hidrolíticas, bem como estabelecer os resíduos agroindustriais capazes de induzir maiores níveis de produção. As atividades xilanásica, endoglucanásica, amilásica e poligalacturonásica foram determinadas incubandose os filtrados de cultura com seus respectivos

PREDICCIÓN DE LA ESTRUCTURA Y CARACTERÍSTICAS BIOQUÍMICAS DE LA ACETATO TIOQUINASA

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Jairo A. Tovart

2001-12-01

Full Text Available Las actividades de las enzimas implicadas en el metabolismo de los cuerpos cetónicos están presentes tanto en neuronas, como en astrocitos y en oligodendroglía. La utilización de cultivos primarios ha puesto de manifiesto que estos cuerpos cetónicos pueden ser metabolizados en estas células.La· acetato tioquinasa reportada en tejido neural total es una enzima que reduce significativamente el costo energético de la utilización del acetato y por lo tanto se propone como una ruta de elección en metabolismo de esta sustancia.Los datos de predicción para propiedades bioquímicas y estructurales de la enzima muestran una estructura estable, con segmentos conservados entre todas las secuencias existentes y una alta flexibilidad estructural, propia de la mayoría de la enzimas.

Análise sérica das enzimas aspartato aminotransferase, alanina aminotransferase e gama glutamiltranspeptidase de coelhos adultos tratados com extrato bruto de própolis

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J. N. Ribeiro

2009-01-01

Full Text Available

Diversos trabalhos têm atribuído a própolis inúmeras propriedades farmacológicas, dentre elas podemos citar, como exemplo, efeitos antibacteriano, antiviral, antiinflamatório, regenerador do tecido cartilaginoso, inibidor da formação de radicais livres e redutor de níveis sangüíneo de glicose e triacilglicerol. Alguns efeitos colaterais são atribuídos à própolis principalmente em doses elevadas. Muitos efeitos tóxicos da própolis são atribuídos ao álcool etílico presente no extrato.Dentre alguns efeitos tóxicos citados em literatura como realmente da própolis temos a dermatite e o aumento da uréia sangüínea. O presente estudo teve como objetivo investigar se o extrato bruto de própolis ocasiona algum efeito adverso nos níveis séricos de alanina aminotransferase, aspartato aminotransferase e gama – glutamiltranspeptidase de coelhos saudáveis. O experimento teve 30 dias de duração, sendo as dosagens dos constituintes do sangue (alanina aminotransferase, aspartato aminotransferase e gama – glutamiltranspeptidase realizadas a 0, 15 e 30 dias. Os resultados indicaram que, de o extrato bruto de própolis na forma testadea, não ocasionou alteração relevante nos níveis séricos das enzimas marcadoras de metabolismo hepático. Palavras-chave: Própolis, alanina aminotransferase, aspartato aminotransferase, gama glutamiltranpeptidase, toxicologia.

Efeito da protease sobre o coeficiente de metabolizabilidade dos nutrientes em frangos de corte

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C.F.Q. Matias

2015-04-01

Full Text Available Avaliaram-se os efeitos da protease sobre o coeficiente de metabolizabilidade dos nutrientes em dietas contendo farinha de penas (2,0% na fase inicial e 3,0% na fase de crescimento para frangos de corte machos, Cobb(r, de um a 32 dias de idade. Foram alojadas 336 aves em gaiolas metálicas equipadas com bandeja para coleta total de excretas. Foi utilizado delineamento inteiramente ao acaso, em arranjo fatorial 2x2 (com e sem adição da enzima protease 0,05% e duas valorizações da matriz nutricional da enzima, sendo sete repetições por tratamento, de 12 aves cada. Observou-se, na fase inicial, maior CMPB (P≤0,05 para as aves que, independentemente da adição de protease, consumiram ração com valorização da matriz nutricional da enzima. Não houve efeito dos tratamentos sobre o CMMS e o CMEE (P>0,05. Na fase de crescimento, houve interação entre os tratamentos para as variáveis CMPB e CMEE. Os frangos alimentados com dieta valorizada sem adição de enzima (controle negativo apresentaram melhores resultados (P≤0,05 para CMPB e CMEE. Nos tratamentos com enzima, o melhor CMEE (P≤0,05 foi obtido com o tratamento sem valorização da matriz nutricional (over the top. Conclui-se que níveis reduzidos e enzima sem valorização melhoram os coeficientes de metabolizabilidade dos nutrientes na fase adulta de frangos de corte.

Clonación, expresión y caracterización de una nueva esterasa derivada de metagenomas de suelos agrícolas colombianos

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Carolina Villamil

2016-07-01

Full Text Available El presente trabajo tuvo como objetivo la bioprospección de ADN metagenómico derivado de comunidades microbianas asociadas a un agroecosistema de importancia nacional. Este análisis permitió realizar la producción, expresión, purificación y caracterización de una enzima novedosa con actividad esterasa. Esta enzima, denominada LipM, había sido previamente identificada en clones metagenómicos derivados de suelos dedicados al cultivo de papa criolla (Solanum pureja, mediante secuencia de nueva generación y análisis bioinformáticos. La secuencia codificante de la enzima fue clonada en el vector pBADgiii y expresada en E. coli como sistema de expresión, lo que permitió optimizar el proceso de producción recombinante y su posterior purificación. Funcionalmente la enzima presentó una mayor afinidad por sustratos de p-nitrofenil con ácidos grasos de cadena corta (enzima exhibió buena estabilidad en presencia de varios iones metálicos, inhibidores y  0.1% (p/v de SDS. Su alto nivel de estabilidad en presencia de iones metálicos e inhibidores, así como su particular especificidad en cuanto a sustratos, la hacen una enzima óptima para utilización en diferentes aplicaciones biotecnológicas.

Enzibióticos bactericidas mejorados frente a neumococo y otras bacterias

OpenAIRE

García, Pedro; Menéndez, Margarita; García, Ernesto; Díez-Martínez, Roberto; De Paz, Héctor D.; Bustamante, Noemí

2013-01-01

La presente invención se encuadra dentro del campo de la biotecnología. En la presente invención se presenta una secuencia polipeptídica derivada del módulo de unión a pared del enzima lítica del fago Cp7 (Cpl-7), que permite la construcción de nuevas enzimas líticas con actividad bactericida mejorada y amplio espectro. Igualmente, en esta invención se incluyen enzimas quiméricas que contienen dicho módulo de unión a pared mejorado y se dan ejemplos de su a...

Human glutathione transferases catalyzing the bioactivation of anticancer thiopurine prodrugs.

Science.gov (United States)

Eklund, Birgitta I; Gunnarsdottir, Sjofn; Elfarra, Adnan A; Mannervik, Bengt

2007-06-01

cis-6-(2-Acetylvinylthio)purine (cAVTP) and trans-6-(2-acetylvinylthio)guanine (tAVTG) are thiopurine prodrugs provisionally inactivated by an alpha,beta-unsaturated substituent on the sulfur of the parental thiopurines 6-mercaptopurine (6-MP) and 6-thioguanine (6-TG). The active thiopurines are liberated intracellularly by glutathione (GSH) in reactions catalyzed by glutathione transferases (GSTs) (EC 2.5.1.18). Catalytic activities of 13 human GSTs representing seven distinct classes of soluble GSTs have been determined. The bioactivation of cAVTP and tAVTG occurs via a transient addition of GSH to the activated double bond of the S-substituent of the prodrug, followed by elimination of the thiopurine. The first of these consecutive reactions is rate-limiting for thiopurine release, but GST-activation of this first addition is shifting the rate limitation to the subsequent elimination. Highly active GSTs reveal the transient intermediate, which is detectable by UV spectroscopy and HPLC analysis. LC/MS analysis of the reaction products demonstrates that the primary GSH conjugate, 4-glutathionylbuten-2-one, can react with a second GSH molecule to form the 4-(bis-glutathionyl)butan-2-one. GST M1-1 and GST A4-4 were the most efficient enzymes with tAVTG, and GST M1-1 and GST M2-2 had highest activity with cAVTP. The highly efficient GST M1-1 is polymorphic and is absent in approximately half of the human population. GST P1-1, which is overexpressed in many cancer cells, had no detectable activity with cAVTP and only minor activity with tAVTG. Other GST-activated prodrugs have targeted GST P1-1-expressing cancer cells. Tumors expressing high levels of GST M1-1 or GST A4-4 can be predicted to be particularly vulnerable to chemotherapy with cAVTP or tAVTG.

INFLUÊNCIA DE ENZIMAS DE MACERAÇÃO NA PRODUÇÃO DE PUBA INFLUENCE OF MACERATING ENZYMES IN THE PRODUCTION OF "PUBA"

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Tobias José Barreto de MENEZES

1998-10-01

Full Text Available Pubagem é a fermentação natural de raízes de mandioca para produção da puba, um alimento popular do Nordeste do Brasil. Além da fermentação lática predominante, os microrganismos também causam o amolecimento das raízes, importante para a obtenção de um produto de boa qualidade. O objetivo deste trabalho foi detectar a presença de enzimas de maceração e verificar a influência da adição de preparados comerciais de celulase e pectinase na pubagem. Constatou-se que as atividades de celulases, xilanase e poligalacturonase aumentaram durante a fermentação natural. Ao se adicionar preparados comerciais de celulase e pectinase este aumento foi maior do que o esperado pela fermentação natural. A adição desses preparados enzímicos acelerou a fermentação aumentando a acidez e reduzindo o pH mais rapidamente do que o tratamento-testemunha.Retting is the natural fermentation of cassava for the production of puba, a common food of the Northeast of Brazil. In addition to a predominant lactic acid fermentation, the microoganisms also cause the softening of the roots which is important for obtaining a good quality product. The objective of this work was to detect the presence of macerating enzymes and to verify the influence of the addition of a commercial preparation of cellulase and pectinase on the retting of cassava. It was found that the activities of cellulases, xilanase and poligalacturonase increased during natural fermentation. This increase was higher than that expected by natural fermentation when commercial preparations of cellulase and pectinase were added. The additions of these enzymic preparations speeded up the fermentation process increasing the acidity and reducing the pH more rapidly than in the control samples.

Novel enzymatic assay predicts minoxidil response in the treatment of androgenetic alopecia.

Science.gov (United States)

Goren, Andy; Castano, Juan Antonio; McCoy, John; Bermudez, Fernando; Lotti, Torello

2014-01-01

Topical minoxidil is the most common drug used for the treatment of androgenetic alopecia (AGA) in men and women. Although topical minoxidil exhibits a good safety profile, the efficacy in the overall population remains relatively low at 30-40%. To observe significant improvement in hair growth, minoxidil is typically used daily for a period of at least 3-4 months. Due to the significant time commitment and low response rate, a biomarker for predicting patient response prior to therapy would be advantageous. Minoxidil is converted in the scalp to its active form, minoxidil sulfate, by the sulfotransferase enzyme SULT1A1. We hypothesized that SULT1A1 enzyme activity in the hair follicle correlates with minoxidil response for the treatment of AGA. Our preliminary retrospective study of a SULT1A1 activity assay demonstrates 95% sensitivity and 73% specificity in predicting minoxidil treatment response for AGA. A larger prospective study is now under way to further validate this novel assay. © 2013 Wiley Periodicals, Inc.

A glutathione S-transferase gene associated with antioxidant properties isolated from Apis cerana cerana

Science.gov (United States)

Liu, Shuchang; Liu, Feng; Jia, Haihong; Yan, Yan; Wang, Hongfang; Guo, Xingqi; Xu, Baohua

2016-06-01

Glutathione S-transferases (GSTs) are an important family of multifunctional enzymes in aerobic organisms. They play a crucial role in the detoxification of exogenous compounds, especially insecticides, and protection against oxidative stress. Most previous studies of GSTs in insects have largely focused on their role in insecticide resistance. Here, we isolated a theta class GST gene designated AccGSTT1 from Apis cerana cerana and aimed to explore its antioxidant and antibacterial attributes. Analyses of homology and phylogenetic relationships suggested that the predicted amino acid sequence of AccGSTT1 shares a high level of identity with the other hymenopteran GSTs and that it was conserved during evolution. Quantitative real-time PCR showed that AccGSTT1 is most highly expressed in adult stages and that the expression profile of this gene is significantly altered in response to various abiotic stresses. These results were confirmed using western blot analysis. Additionally, a disc diffusion assay showed that a recombinant AccGSTT1 protein may be roughly capable of inhibiting bacterial growth and that it reduces the resistance of Escherichia coli cells to multiple adverse stresses. Taken together, these data indicate that AccGSTT1 may play an important role in antioxidant processes under adverse stress conditions.

Glutathione Transferases Superfamily: Cold-Inducible Expression of Distinct GST Genes in Brassica oleracea

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Harshavardhanan Vijayakumar

2016-07-01

Full Text Available Plants, as sessile organisms, can suffer serious growth and developmental consequences under cold stress conditions. Glutathione transferases (GSTs, EC 2.5.1.18 are ubiquitous and multifunctional conjugating proteins, which play a major role in stress responses by preventing oxidative damage by reactive oxygen species (ROS. Currently, understanding of their function(s during different biochemical and signaling pathways under cold stress condition remain unclear. In this study, using combined computational strategy, we identified 65 Brassica oleracea glutathione transferases (BoGST and characterized them based on evolutionary analysis into 11 classes. Inter-species and intra-species duplication was evident between BoGSTs and Arabidopsis GSTs. Based on localization analyses, we propose possible pathways in which GST genes are involved during cold stress. Further, expression analysis of the predicted putative functions for GST genes were investigated in two cold contrasting genotypes (cold tolerance and susceptible under cold condition, most of these genes were highly expressed at 6 h and 1 h in the cold tolerant (CT and cold susceptible (CS lines, respectively. Overall, BoGSTU19, BoGSTU24, BoGSTF10 are candidate genes highly expressed in B. oleracea. Further investigation of GST superfamily in B. oleracea will aid in understanding complex mechanism underlying cold tolerance in plants.

Allyl isothiocyanate that induces GST and UGT expression confers oxidative stress resistance on C. elegans, as demonstrated by nematode biosensor.

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Koichi Hasegawa

Full Text Available BACKGROUND: Electrophilic xenobiotics and endogenous products from oxidative stresses induce the glutathione S-transferases (GSTs, which form a large family within the phase II enzymes over both animal and plant kingdoms. The GSTs thus induced in turn detoxify these external as well as internal stresses. Because these stresses are often linked to ageing and damage to health, the induction of phase II enzymes without causing adverse effects would be beneficial in slowing down ageing and keeping healthy conditions. METHODOLOGY/PRINCIPAL FINDINGS: We have tested this hypothesis by choosing allyl isothiocyanate (AITC, a functional ingredient in wasabi, as a candidate food ingredient that induces GSTs without causing adverse effects on animals' lives. To monitor the GST induction, we constructed a gst::gfp fusion gene and used it to transform Caenorhabditis elegans for use as a nematode biosensor. With the nematode biosensor, we found that AITC induced GST expression and conferred tolerance on the nematode against various oxidative stresses. We also present evidence that the transcription factor SKN-1 is involved in regulating the GST expression induced by AITC. CONCLUSIONS/SIGNIFICANCE: We show the applicability of the nematode biosensor for discovering and evaluating functional food substances and chemicals that would provide anti-ageing or healthful benefits.

Identification of nevadensin as an important herb-based constituent inhibiting estragole bioactivation and physiology-based biokinetic modeling of its possible in vivo effect

International Nuclear Information System (INIS)

Alhusainy, W.; Paini, A.; Punt, A.; Louisse, J.; Spenkelink, A.; Vervoort, J.; Delatour, T.; Scholz, G.; Schilter, B.; Adams, T.; Bladeren, P.J. van; Rietjens, I.M.C.M.

2010-01-01

Estragole is a natural constituent of several herbs and spices including sweet basil. In rodent bioassays, estragole induces hepatomas, an effect ascribed to estragole bioactivation to 1'-sulfooxyestragole resulting in DNA adduct formation. The present paper identifies nevadensin as a basil constituent able to inhibit DNA adduct formation in rat hepatocytes exposed to the proximate carcinogen 1'-hydroxyestragole and nevadensin. This inhibition occurs at the level of sulfotransferase (SULT)-mediated bioactivation of 1'-hydroxyestragole. The Ki for SULT inhibition by nevadensin was 4 nM in male rat and human liver fractions. Furthermore, nevadensin up to 20 μM did not inhibit 1'-hydroxyestragole detoxification by glucuronidation and oxidation. The inhibition of SULT by nevadensin was incorporated into the recently developed physiologically based biokinetic (PBBK) rat and human models for estragole bioactivation and detoxification. The results predict that co-administration of estragole at a level inducing hepatic tumors in vivo (50 mg/kg bw) with nevadensin at a molar ratio of 0.06, representing the ratio of their occurrence in basil, results in almost 100% inhibition of the ultimate carcinogen 1'-sulfooxyestragole when assuming 100% uptake of nevadensin. Assuming 1% uptake, inhibition would still amount to more than 83%. Altogether these data point at a nevadensin-mediated inhibition of the formation of the ultimate carcinogenic metabolite of estragole, without reducing the capacity to detoxify 1'-hydroxyestragole via glucuronidation or oxidation. These data also point at a potential reduction of the cancer risk when estragole exposure occurs within a food matrix containing SULT inhibitors compared to what is observed upon exposure to pure estragole.

OBTAINING AND USE OF POLYPHENOLOXIDASE ENZYME EXTRACTED FROM RIPE CUSTARD APPLE (Annona squamosa L.) PULP ON THE COCOA (Theobroma cacao L.) NIBS IN TASTE IMPROVEMENT

OpenAIRE

LIMA, ELIZA DOROTEA POZZOBON DE ALBUQUERQUE; PASTORE, GLÁUCIA MARIA; BARBERY, SANDRA DRINA FERNANDEZ; GARCIA, NELSON HORÁCIO PEZOA; BRITO, EDY SOUSA DE; LIMA, CARLOS ALBERTO DE ALBUQUERQUE

2001-01-01

O presente trabalho teve como objetivo estudar a obtenção e a utilização da enzima polifenoloxidase (PPO) extraída de polpa de pinha madura na redução do teor de compostos polifenólicos com a finalidade de diminuir a adstringência e o amargor das amêndoas de cacau processadas na forma de "nibs". A PPO foi extraída com tampão fosfato de potássio 0,025M (pH 7,5), adicionando sulfato de amônio para a precipitação da enzima. O material em pó obtido foi denominado de enzima parcialmente purificada...

Estudo da produção do suco clarificado de cajá (Spondias lutea L.)

OpenAIRE

SILVA,Antonio de Pádua Valença da; MAIA,Geraldo Arraes; OLIVEIRA,Gerardo Sérgio Francelino de; FIGUEIREDO,Raimundo Wilane de; BRASIL,Isabella Montenegro

1999-01-01

Cajás ( Spondias lutea L.), oriundos do município de Maranguape, Ceará, Brasil, foram processados em nível de escala-piloto para obtenção de suco clarificado através do emprego de enzimas pectinolíticas e agentes clarificantes. Aplicaram-se 120ppm de enzima pectinolítica (Pectinex Ultra SP-L) na polpa para obtenção do suco polposo e, em seguida, 500ppm de enzima pectinolítica (Pectinex-AR) no suco extraído, utilizando-se a "Prova do álcool" como indicador da presença de pectina. Para o proces...

Estudo e otimização da produção da dextranasacarase e caracterização da dextrana produzida por Leuconostoc mesenteroides FT045B

OpenAIRE

Vettori, Mary Helen Palmuti Braga [UNESP

2011-01-01

Um estudo comparativo de onze métodos para determinação da atividade da dextranasacarase e de enzimas relacionadas foi realizado. Atualmente existem dois métodos usualmente empregados na determinação da atividade da dextranasacarase. Um método relativamente difundido para a reação da enzima com a sacarose consiste na medição dos valores redutores da D-frutose pelo reagente 3,5-dinitrosalicilato (DNS). Outro método é a reação da enzima com 14Csacarose, pela adição de uma alíquota da reação em ...

PURIFICAÇÃO DA ASCORBATO-OXIDASE POR ELETROELUIÇÃO PARA DETERMINAÇÃO DE VITAMINA C

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Ligia Renata FIRMINO

2009-07-01

Full Text Available

O método de eletroeluição revelou-se eficiente, rápido e simples para purificar a enzima ascorbato-oxidase de abobrinha (Cucurbita pepo medulosa A atividade específica da enzima, após a eletroeluição, foi da ordem de 1.421 U/mg de proteína, obtendo-se nessa etapa um fator de purificação de 152,8 vezes. O conteúdo de vitamina C de alimentos e medicamento foi determinado com a enzima purificada e comparado com o método titulométrico que utiliza o DFI.

Valores energéticos do farelo de arroz integral suplementado com complexos enzimáticos para frangos de corte Energy values of whole rice bran supplemented with enzymes complexes for broilers

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Renato Alberto Giacometti

2003-06-01

Full Text Available Este experimento foi realizado com o objetivo de estudar o efeito de enzimas carboidrases exógenas com atividade xilanase sobre a energia metabolizável aparente (EMA e aparente corrigida (EMAn do farelo de arroz integral (FAI. As enzimas carboidrases com atividade xilanase utilizadas foram as da marca comercial Rovabio TM Excel AP, Allzyme Rice e Ronozyme WX, denominadas, respectivamente, neste trabalho, de enzimas A, B, e C. Foram utilizadas 240 aves de corte da linhagem Coob, com 21 dias de idade, alojadas em gaiolas metálicas com bandejas coletoras de excretas instaladas em uma sala de metabolismo com ambiente controlado. O delineamento foi inteiramente casualizado, constituído por 4 dietas-referência e 4 dietas-teste e 6 repetições, totalizando 48 parcelas com 5 aves cada uma. As dietas-referência e as dietas-teste estudadas foram as seguintes: Dieta-referência à base de milho, farelo de soja, vitaminas e minerais (DR; DR + enzima A; DR + enzima B; DR + enzima C; 70% de DR + 30% de FAI; 70% de DR + 30% de FAI + enzima A; 70% de DR + 30% de FAI + enzima B e 70% de DR + 30% de FAI + enzima C. Utilizou-se o método de coleta total de excretas (Sibbald & Slinger, 1963 durante 5 dias em cada unidade experimental, precedido de 5 dias de adaptação das aves às dietas. As determinações de EMA e EMAn foram realizadas conforme metodologia de Matterson et al. (1965. A EMAn do FAI foi de 2897 kcal/kg de MS. Com o uso das enzimas carboidrases, o melhor valor de EMAn do FAI foi obtido quando utilizou-se a enzima C, sendo de 3083 kcal/kg de MS, com um aumento percentual de 6,4%. A dieta-referência que recebeu a adição da enzima C teve sua EMAn aumentada, o que não ocorreu com a utilização das outras carboidrases.An experiment was conducted to study the effect of exogen carbohydrase enzymes with xylanase activity on the apparent metabolizable energy (AME and apparent corrected metabolizable energy (AMEn of whole rice bran (WRB. The

AISLAMIENTO, CLONACIÓN Y EXPRESIÓN DE UNA ENZIMA LIPOLÍTICA DE Natronococcus sp. TC6, A PARTIR DE UNA LIBRERÍA GENÓMICA

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Alberto López Barrera

2017-01-01

Full Text Available Los procesos biotecnológicos e industriales demandan de biocatalizadores robustos. Para conseguir este atributo, se recurre a la búsqueda de enzimas de microorganismos que naturalmente se han adaptado a ambientes inhóspitos. La generación de bibliotecas genómicas y el análisis genómico son herramientas moleculares poderosas para descubrir biomoléculas con características únicas y sobresalientes. Empleando el genoma fragmentado mecánicamente de un arquea halófila, Natronococcus sp. TC6, se construyó una genoteca de cósmidos, los cuales fueron usados para transformar a la cepa E. coli. En total, fueron generados 2181 clones, de los cuales, se seleccionaron seis con actividad lipolítica, ya que hidrolizaron ésteres de p-nitrofenilo como sustratos, en un medio de reacción conteniendo 2.5 M de NaCl. Las clonas que hidrolizaron ambos sustratos (clonas positivas fueron aisladas y los cósmidos fueron purificados y fragmentados para posteriores análisis enzimáticos. Los fragmentos de DNA de las clonas positivas, fueron subclonados y secuenciados. El análisis bioinformático de las secuencias obtenidas permitió identificar una tioesterasa putativa, la cual fue ligada en un vector de expresión, y éste utilizado para su introducción en células de E. coli BL21 para la producción de la proteína recombinante. La tioesterasa identificada posee 158 aminoácidos con un peso molecular de 15.4 kDa y con una similitud del 99% con el gene de la bacteria Stenotrophomonas maltophilia. La tioesterasa putativa de Natronococcus sp. TC6, presenta tres sitios altamente conservados como en S. maltophilia, posee dos motivos cercanos al extremo N-terminal, GHVNN y YXEXAR. También presenta una variante del pentapéptido involucrado en el sitio activo y característico de la mayoría de las éster hidrolasas, GXSXV.

Efecto de la incorporación de transglutaminasa microbiana en las propiedades sensoriales de hamburguesas de desmenuzado de merluza (Merluccius hubbsi

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Alicia Panuncio

2013-12-01

Full Text Available Se elaboraron hamburguesas de pescado a partir de desmenuzado de merluza (Merluccius hubbsi con la incorporación de transglutaminasa microbiana, con la finalidad de mejorar sus propiedades sensoriales y elaborar un producto pesquero que fomente el consumo de pescado en Uruguay. Se evaluó desde el punto de vista sensorial y de textura instrumental el efecto del agregado de la enzima en la formulación de las hamburguesas. Se elaboraron tres tipos de hamburguesas: sin enzima y con incorporación de 0,5% y 1,0% de enzima transglutaminasa, respectivamente. Para la evaluación sensorial se analizaron las muestras con un panel de seis jueces entrenados y para la determinación de los parámetros de textura instrumental se utilizó un analizador de textura TA.XT2i. Los resultados de ambos estudios indicaron que el agregado de enzima mejoró sustancialmente la apariencia, la textura y el sabor del producto. Se obtuvo un producto pesquero innovador, con características que pueden resultar muy atractivas para el consumidor uruguayo.

Uso de enzimas na alimentação de frangos de corte Enzymes in the broiler diets

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Evani Souza de Oliveira Strada

2005-12-01

Full Text Available Dois experimentos foram realizados para se avaliar o efeito de enzimas microbianas sobre o ganho de peso, o consumo de ração e a conversão alimentar frangos de corte. No primeiro experimento, utilizaram-se 288 pintos de 8 a 21 dias de idade em um delineamento inteiramente casualizado (DIC, em arranjo fatorial 2 x 2, (duas dietas, farelo de soja + sorgo e farelo de soja + milheto, com ou sem complexo multienzimático - CM, com quatro repetições por tratamento, em que cada parcela foi constituída por 18 aves. As dietas foram formuladas para conter níveis idênticos de energia metabolizável (EM, aminoácidos, cálcio e fósforo. No segundo experimento, foram utilizados 384 frangos de corte de 21 a 42 dias de idade em delineamento inteiramente casualizado, com seis tratamentos, quatro repetições e 16 aves por unidade experimental. As dietas experimentais, à base de milho e farelo de soja, foram formuladas para conterem 7 e 9% de EM e 5 e 7% dos aminoácidos metionina (Met, Met + cistina (Cis e lisina (Lis, com a presença ou não de complexo multienzimático. A inclusão do CM, tanto em rações à base de farelo de soja e sorgo como à base de farelo de soja e milheto, não proporcionou ganhos nos desempenhos das aves na fase inicial de crescimento. Os valores de EM e de aminoácidos (Met, Met+Cis e Lis da soja podem ser superestimados em 9 e 7%, respectivamente, quando há adição de CM às rações, sem prejuízo ao desempenho das aves.Two experiments were conducted to evaluate the effects of microbial enzymes on weight gain, feed intake and feed: gain ratio. In the first trial, 288 chicks from 8-21 days old were assigned to a complete randomized design with a 2 x 2 fatorial arrangement: meal (soybean meal + sorghum and soybean meal + millet and with or without multi-enzymatic complex (MC, with four replications of 18 bird per experimental unit. Diets were formulated to contain the same levels of metabolizable energy (ME, amino acids

Association of GSTs polymorphisms with risk of gestational diabetes mellitus.

Science.gov (United States)

Li, Yan; Li, Shaoru; Zhai, Qianqian; Hai, Jie; Wang, Di; Cao, Meng; Zhang, Qinggui

2015-01-01

We conducted a case-control study to investigate the association between GSTM1, GSTT1 and GSTP1 IIe105Val polymorphisms and development of gestational diabetes mellitus in a Chinese population. A total of 320 patients with gestational diabetes mellitus and 358 pregnancy subjects were consecutively collected between January 2013 and December 2014. Genotyping for detection of GSTM1, GSTT1 and GSTP1 IIe105Val was conducted by using PCR-RFLP (polymerase chain reaction-restriction fragment length polymorphisms) method. By Fisher's exact test, we found that the genotype distributions of GSTP1 IIe105Val were in line with the Hardy-Weinberg equilibrium in control subjects (P=0.57). By Chi-square test, we found significant differences in the genotype distributions of GSTM1 (χ(2)=11.49, P=0.001) and GSTT1 (χ(2)=18.50, Pgestational diabetes mellitus when compared with the present genotype, and the adjusted Ors (95% CI) were 1.71 (1.24-2.36) and 2.00 (1.44-2.79), respectively. However, the GSTP1 IIe105Val polymorphism was not associated with an elevated risk of gestational diabetes mellitus. In conclusion, we suggest that the GSTM1 null genotype and GSTT1 null genotype are correlated with an increased risk of gestational diabetes mellitus in a Chinese population.

Glutathione S-transferase expression and isoenzyme composition during cell differentiation of Caco-2 cells

International Nuclear Information System (INIS)

Scharmach, E.; Hessel, S.; Niemann, B.; Lampen, A.

2009-01-01

The human colon adenocarcinoma cell line Caco-2 is frequently used to study human intestinal metabolism and transport of xenobiotica. Previous studies have shown that both Caco-2 cells and human colon cells constitutively express the multigene family of detoxifying enzymes glutathione S-transferases (GSTs), particularly GST alpha and GST pi. GSTs may play a fundamental role in the molecular interplay between phase I, II enzymes and ABC-transporters. The gut fermentation product, butyrate, can modulate the potential for detoxification. The aim of this study was to investigate the basal expression of further cytosolic GSTs in Caco-2 cells during cell differentiation. In addition, a comparison was made with expression levels in MCF-7 and HepG2, two other cell types with barrier functions. Finally, the butyrate-mediated modulation of gene and protein expression was determined by real time PCR and western blot analysis. In Caco-2, gene and protein expression levels of GST alpha increased during cell differentiation. High levels of GSTO1 and GSTP1 were constantly expressed. No expression of GSTM5 and GSTT1 was detected. HepG2 expressed GSTO1 and MCF-7 GSTZ1 most intensively. No expression of GSTA5, GSTM5, or GSTP1 was detected in either cell. Incubation of Caco-2 cells with butyrate (5 mM) significantly induced GSTA1 and GSTM2 in proliferating Caco-2 cells. In differentiated cells, butyrate tended to increase GSTO1 and GSTP1. The results of this study show that a differentiation-dependent expression of GSTs in Caco-2 cells may reflect the in vivo situation and indicate the potential of butyrate to modify intestinal metabolism. GSTA1-A4 have been identified as good markers for cell differentiation. The Caco-2 cell line is a useful model for assessing the potential of food-related substances to modulate the GST expression pattern.

Differential roles for VviGST1, VviGST3 and VviGST4 in proanthocyanidin and anthocyanin transport in Vitis vinífera

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Ricardo Pérez-Díaz

2016-08-01

Full Text Available In plant cells, flavonoids are synthesized in the cytosol and then are transported and accumulated in the vacuole. Glutathione S-transferase-mediated transport has been proposed as a mechanism involved in flavonoid transport however whether binding of flavonoids to GST or their transport is glutathione-dependent is not well understood. Glutathione S-transferases from Vitis vinífera (VviGSTs have been associated with the transport of anthocyanins, however their ability to transport other flavonoids such as proanthocyanidins (PAs has not been established. Following bioinformatics approaches, we analyzed the capability of VviGST1, VviGST3, VviGST4 and Arabidopsis TT19 to bind different flavonoids. Analyses of protein-ligand interactions indicate that these GSTs can bind glutathione and monomers of anthocyanin, PAs and flavonols. A total or partial overlap of the binding sites for glutathione and flavonoids was found in VviGST1, and a similar condition was observed in VviGST3 using anthocyanin and flavonols as ligands, whereas VviGST4 and TT19 have both sites for GSH and flavonoids separated. To validate the bioinformatics predictions, functional complementation assays using the Arabidopsis tt19 mutant were performed. Overexpression of VviGST3 in tt19-1 specifically rescued the dark seed coat phenotype associated to correct PA transport, which correlated with higher binding affinity for PA precursors. VviGST4, originally characterized as an anthocyanin-related GST, complemented both the anthocyanin and PA deposition, resembling the function of TT19. By contrast, VviGST1 only partially rescued the normal seed color. Furthermore the expression pattern of these VviGSTs showed that each of these genes could be associated with the accumulation of different flavonoids in specific tissues during grapevine fruit development. These results provide new insights into GST-mediated PA transport in grapevine and suggest that VviGSTs present different

Differential Roles for VviGST1, VviGST3, and VviGST4 in Proanthocyanidin and Anthocyanin Transport in Vitis vinífera.

Science.gov (United States)

Pérez-Díaz, Ricardo; Madrid-Espinoza, José; Salinas-Cornejo, Josselyn; González-Villanueva, Enrique; Ruiz-Lara, Simón

2016-01-01

In plant cells, flavonoids are synthesized in the cytosol and then are transported and accumulated in the vacuole. Glutathione S-transferase-mediated transport has been proposed as a mechanism involved in flavonoid transport, however, whether binding of flavonoids to glutathione S-transferase (GST) or their transport is glutathione-dependent is not well understood. Glutathione S-transferases from Vitis vinífera (VviGSTs) have been associated with the transport of anthocyanins, however, their ability to transport other flavonoids such as proanthocyanidins (PAs) has not been established. Following bioinformatics approaches, we analyzed the capability of VviGST1, VviGST3, VviGST4, and Arabidopsis TT19 to bind different flavonoids. Analyses of protein-ligand interactions indicate that these GSTs can bind glutathione and monomers of anthocyanin, PAs and flavonols. A total or partial overlap of the binding sites for glutathione and flavonoids was found in VviGST1, and a similar condition was observed in VviGST3 using anthocyanin and flavonols as ligands, whereas VviGST4 and TT19 have both sites for GSH and flavonoids separated. To validate the bioinformatics predictions, functional complementation assays using the Arabidopsis tt19 mutant were performed. Overexpression of VviGST3 in tt19-1 specifically rescued the dark seed coat phenotype associated to correct PA transport, which correlated with higher binding affinity for PA precursors. VviGST4, originally characterized as an anthocyanin-related GST, complemented both the anthocyanin and PA deposition, resembling the function of TT19. By contrast, VviGST1 only partially rescued the normal seed color. Furthermore the expression pattern of these VviGSTs showed that each of these genes could be associated with the accumulation of different flavonoids in specific tissues during grapevine fruit development. These results provide new insights into GST-mediated PA transport in grapevine and suggest that VviGSTs present

Glutathione S-transferase expression and isoenzyme composition during cell differentiation of Caco-2 cells.

Science.gov (United States)

Scharmach, E; Hessel, S; Niemann, B; Lampen, A

2009-11-30

The human colon adenocarcinoma cell line Caco-2 is frequently used to study human intestinal metabolism and transport of xenobiotica. Previous studies have shown that both Caco-2 cells and human colon cells constitutively express the multigene family of detoxifying enzymes glutathione S-transferases (GSTs), particularly GST alpha and GST pi. GSTs may play a fundamental role in the molecular interplay between phase I, II enzymes and ABC-transporters. The gut fermentation product, butyrate, can modulate the potential for detoxification. The aim of this study was to investigate the basal expression of further cytosolic GSTs in Caco-2 cells during cell differentiation. In addition, a comparison was made with expression levels in MCF-7 and HepG2, two other cell types with barrier functions. Finally, the butyrate-mediated modulation of gene and protein expression was determined by real time PCR and western blot analysis. In Caco-2, gene and protein expression levels of GST alpha increased during cell differentiation. High levels of GSTO1 and GSTP1 were constantly expressed. No expression of GSTM5 and GSTT1 was detected. HepG2 expressed GSTO1 and MCF-7 GSTZ1 most intensively. No expression of GSTA5, GSTM5, or GSTP1 was detected in either cell. Incubation of Caco-2 cells with butyrate (5 mM) significantly induced GSTA1 and GSTM2 in proliferating Caco-2 cells. In differentiated cells, butyrate tended to increase GSTO1 and GSTP1. The results of this study show that a differentiation-dependent expression of GSTs in Caco-2 cells may reflect the in vivo situation and indicate the potential of butyrate to modify intestinal metabolism. GSTA1-A4 have been identified as good markers for cell differentiation. The Caco-2 cell line is a useful model for assessing the potential of food-related substances to modulate the GST expression pattern.

Efeito da alta pressão hidrostática na atividade de enzimas da polpa de açaí High hydrostatic pressure effect on enzyme activity of açai pulp

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Ellen Mayra da Silva Menezes

2008-12-01

Full Text Available O Brasil vem conquistando o mercado externo de sucos de frutos tropicais, com destaque para o açaí, muito procurado por ser conhecido como um alimento funcional, devido à concentração de antocianinas, fibras dietéticas e ácidos graxos monoinsaturados. Entretanto, o açaí é altamente perecível, necessitando de intervenções tecnológicas para prolongar sua vida de prateleira. Nesse caso, o uso da Alta Pressão Hidrostática (APH pode ser uma alternativa aos processamentos tradicionais, por sua capacidade de ativar ou inativar enzimas. Peroxidases (POD e polifenoloxidases (PFO são as principais enzimas responsáveis por alterações indesejáveis das características originais de produtos vegetais e, com a sua inativação pela APH, pode-se evitar o escurecimento enzimático e manter as suas propriedades sensoriais. Com o objetivo de avaliar o efeito da APH em POD e PFO de polpa de açaí, adotaram-se as variáveis de pressão, temperatura e tempo. As atividades enzimáticas expressas em percentuais revelaram a POD mais estável, atingindo percentuais próximos a 100% (controle. As menores atividades foram a 90,74%, e quando se aplicaram 300 e 500 MPa a 25 °C por 5 minutos, as atividades aumentaram (112,34 e 132,98%, respectivamente. A atividade da PFO aumentou até 83,03% em relação ao controle, embora apresentasse inativação na maioria dos processos, evidenciando-se atividade a 35 °C/5 minutos de 53,25 e 53,75% a 300 e 500 MPa, respectivamente. Diante das condições experimentais, a APH mostrou-se eficaz na inativação parcial de oxidases na polpa de açaí.Brazil has been conquering tropical fruit juice international market , especially with açaí (native fruit with the highest penetration in the international, considered a functional food for its anthocyanin, dietetic fibe, and monounsaturated fat content. However, açaí is very perishable, so the use of technology is needed in order to ensure long shelf life. In this

Short-term calorie restriction feminizes the mRNA profiles of drug metabolizing enzymes and transporters in livers of mice.

Science.gov (United States)

Fu, Zidong Donna; Klaassen, Curtis D

2014-01-01

Calorie restriction (CR) is one of the most effective anti-aging interventions in mammals. A modern theory suggests that aging results from a decline in detoxification capabilities and thus accumulation of damaged macromolecules. The present study aimed to determine how short-term CR alters mRNA profiles of genes that encode metabolism and detoxification machinery in the liver. Male C57BL/6 mice were fed CR (0, 15, 30, or 40%) diets for one month, followed by mRNA quantification of 98 xenobiotic processing genes (XPGs) in the liver, including 7 uptake transporters, 39 phase-I enzymes, 37 phase-II enzymes, 10 efflux transporters, and 5 transcription factors. In general, 15% CR did not alter mRNAs of most XPGs, whereas 30 and 40% CR altered over half of the XPGs (32 increased and 29 decreased). CR up-regulated some phase-I enzymes (fold increase), such as Cyp4a14 (12), Por (2.3), Nqo1 (1.4), Fmo2 (5.4), and Fmo3 (346), and numerous number of phase-II enzymes, such as Sult1a1 (1.2), Sult1d1 (2.0), Sult1e1 (33), Sult3a1 (2.2), Gsta4 (1.3), Gstm2 (1.3), Gstm3 (1.7), and Mgst3 (2.2). CR feminized the mRNA profiles of 32 XPGs in livers of male mice. For instance, CR decreased the male-predominantly expressed Oatp1a1 (97%) and increased the female-predominantly expressed Oatp1a4 (11). In conclusion, short-term CR alters the mRNA levels of over half of the 98 XPGs quantified in livers of male mice, and over half of these alterations appear to be due to feminization of the liver. Copyright © 2013 Elsevier Inc. All rights reserved.

Efeito da aplicação de enzimas pectinolíticas no rendimento da extração de polpa de cupuaçu

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BASTOS MARIA DO SOCORRO ROCHA

2002-01-01

Full Text Available O Brasil apresenta uma diversidade de frutas regionais, principalmente nas regiões Norte e Nordeste. Dentre estas, destaca-se o cupuaçu, que é uma fruta considerada exótica e de grande potencial comercial nos mercados dos estados do Sudeste do Brasil e dos países europeus. A polpa e a semente são importantes para o desenvolvimento de produtos industriais. A polpa tem sido estudada como matéria-prima para néctar; entretanto, é usada em sorvetes, geléias, purês e polpa enlatada. Atualmente, o processo de extração da polpa ainda é feito de maneira empírica, utilizando-se de acessórios como facas, colheres ou tesouras de uso doméstico, devido à forte aderência da mesma ao caroço, causando baixo rendimento no produto final. Com o objetivo de aumentar o rendimento durante o processo de extração da polpa de cupuaçu, adicionaram-se à polpa bruta dois tipos de preparações enzimáticas de ação pectolítica (Citrozym-L e Rohament PL. As concentrações estabelecidas foram de 100; 300; 500 e 750 ppm. Após todo o processo, obteve-se a polpa, e o rendimento foi comparado com a polpa-controle. O experimento foi realizado nos laboratórios de Tecnologia de Alimentos da Embrapa Agroindústria Tropical. Pelos resultados obtidos, observou-se que o rendimento da polpa com adição de Citrozym-L foi superior a 60% nas concentrações de 300; 500 e 750 ppm, enquanto, com a utilização de Rohament-PL, o rendimento variou de 43 a 57% nas mesmas concentrações. O rendimento da polpa-controle foi de 44 e 41 %, para os experimentos realizados com a primeira e segunda preparação enzimática, respectivamente. Conclui-se que a utilização de enzimas é uma alternativa para melhorar o processo de extração da polpa; adicionando-se acima de 300ppm, obtém-se um rendimento próximo a 60%.

Nonhemocyte sources of selected lysosomal enzymes in Biomphalaria glabrata, B. tenagophila and B. straminea (Mollusca: Pulmonata

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Gary E. Rodrick

1981-09-01

Full Text Available The specific activities of acid phosphatase, alkaline phosphatase, β-glucuronidase, lysozymes, glutamate-oxalacetate transaminase and glutamate-pyruvate transaminate were determined in the head-foot and digestive gland of Brazilian Biomphalaria glabrata (Touros, B. tenagophila (Caçapava and B. straminea (Monsenhor Gil. All six enzymes were detected inthe 3000g supernatant. Both cytoplasmic enzymes, glutamate-oxalacetate and glutamate-pyruvate transaminase exhibited the highest specific activities. In the case of the four hydrolytic enzymes assayed, β-glucuronidase exhibited the highest specific activity while lysozyme showed the lowest activity. All six enzymes are thought to be produced by cells within the head-foot and digestive gland of B. glabrata, B. tenagophila and B. straminea.Foram determinadas, na massa cefalopedal e na glândula digestiva de Biomphalaria glabrata de Touros (Rio Grande do Norte B. tenagophila de Cacapava (Sao Paulo e B. straminea de Monsenhor Gil (Piauí, as atividades específicas das seguintes enzimas: fosfatase acida, fosfatase alcalina, beta-glucuronidase, lisozima, transaminase glutâmico-oxalacetica e transaminase glutâmico-piruvica. As seis enzimas referidas foram detectadas no sobrenadante a 3000g. Ambas as enzimas citoplasmaticas - transaminases glutamico-oxalacetica e glutamico-piruvica - mostraram as atividades específicas mais altas. No caso das quatro enzimas hidrolíticas, a beta-glucuronidase revelou a mais alta atividade específica, enquanto a lisozima revelou a mais baixa. E admitido que todas as seis enzimas sao produzidas por celulas presentes na massa cefalopedal e na glândula digestiva das tres especies de moluscos examinadas.

A View of DANIDAs Policy Note on Food Security through A Policy Coherent Lens

DEFF Research Database (Denmark)

Robertson, Aileen

2014-01-01

-usikkerhed hæmmer udvikling’ understreges det, at hvis ‘underernæring’ og ‘sult’ forveksles, findes disse synergier ikke og sparsomme resourcer spildes. I Danida`s ’Policy Note on Food Security’ nævnes det, at ‘852 million are living in hunger’, altså sult. Det er imidlertid snarere kronisk underernæring end sult...... Greener World for All (NEC)’. Faktisk peger man på, at landbrugets og fødevaresektorens primære rolle er at brødføde befolkningen ved at øge tilgængeligheden og forbruget af sikre, miljørigtige madvarer, der er i overensstemmelse med ernæringsmæssige anbefalinger og til overkommelige priser. Derfor...

Molecular characterization of zeta class glutathione S-transferases ...

Indian Academy of Sciences (India)

In higher plants, studies on GSTs have focussed largely on agricultural plants. There is ... Mediterranean region and have to cope with several environmental stress conditions. ..... Naliwajski M. R. and Skłodowska M 2014 The oxidative stress.

Avaliação de métodos para manutenção e preservação de bactéria esporulada produtora da enzima CGTase = Evaluation of methods for maintenance and preservation of sporulating bacteria producer of CGTase enzyme

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Cristiane Moriwaki

2009-04-01

Full Text Available A conservação de células sem mudanças morfológicas, fisiológicas ou genéticas é uma necessidade da biotecnologia. Bacillus firmus cepa 37 é uma bactéria esporulada produtora da enzima ciclodextrina glicosiltransferase (CGTase, que transforma o amido em ciclodextrinas (CDs. O objetivo deste estudo foi avaliar a manutenção e preservação de B. firmus cepa 37 estocada em meio de cultivo sólido, solo estéril e em glicerol a baixa temperatura (-70ºC. Para avaliação do melhor método de manutenção da bactéria foram utilizados procedimentos de imobilização das células em matrizes inorgânicas. As células imobilizadas foram submetidas ao teste do efeito da biomassa inicial e à microscopia eletrônica de varredura (MEV. O repique não foi um método adequado, pois a cepadiminuiu a produção de CGTase. A estocagem em solo estéril mostrou-se eficaz e a produção da enzima mantida constante. A conservação a baixas temperaturas também foi satisfatória, com contagem de células praticamente a mesma após 360 dias. A imobilização, avaliada por MEV, não mostrou diferença na adsorção das células conservadas pelosdiferentes métodos. O mesmo ocorreu para o teste do efeito da biomassa inicial, que apresentou maior produção de beta-CD quando do uso de 1,5 g de células.The conservation of cells without morphologic, physiologic or genetic changes is a biotechnology necessity. Bacillus firmus strain 37 is a sporulating bacteria that produces the cyclodextrin lycosyltransferase(CGTase enzyme, which transforms starch into cyclodextrins (CDs. This study aimed to evaluate the maintenance and preservation of B. firmus strain 37 stored in a solid medium, sterile soil and in glycerol at low temperature (-70ºC. In order to evaluate the best bacteria maintenance method, cell immobilization procedures were used on inorganic matrices. The immobilized cells were submitted to the initial biomass effect test and scanning electron

Avaliação da concentração de pectinase no processo de hidrólise-sacarificação do farelo de mandioca para obtenção de etanol

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LEONEL M.

2000-01-01

Full Text Available Neste trabalho objetivou-se avaliar a concentração de pectinase (Pectinex Ultra SP-L no processo de hidrólise-sacarificação do farelo de mandioca para produção de etanol. Foram avaliadas quatro concentrações da enzima pectinase com enzima complemetar as amilases e o tratamento com apenas as amilases. Realizou-se a caracterização do hidrolisado e resíduo fibroso resultantes do processo, e os resultados obtidos demonstraram que a concentração mínima de pectinase para um bom rendimento do processo foi 8Kg enzima/t fibras, com. 89,4% do amido hidrolisado. Quanto ao resíduo fibroso, este apresenta potencialidade de aproveitamento com base para produtos dietéticos ricos em fibras.

Pretreatment of cottonseed flakes with proteases and an amylase for higher oil yields

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Hassanein, Minar M.

2007-09-01

Full Text Available The effect of enzymatic pretreatment of cottonseed flakes on oil extractability was studied. The enzymes investigated included bacterial protease (Bp, papain (Pa, savinase (S, termamyl (T, pectinase (Pe and cellulase (C. The variables studied during the enzymatic hydrolysis experiments were: enzyme concentration, moisture: cottonseed flakes ratio, and time of hydrolysis. Enzymatic hydrolysis experiments were first carried out with a single enzyme, then with enzyme mixtures formulated according to the results of single enzyme treatments. Results were evaluated based on the relative increase in oil extractability, and some oil characteristics in comparison with untreated cottonseed flakes (control. Pretreatment with enzyme mixtures resulted in a relative increase in oil extractability that was higher than single enzyme pretreatment and the control. Statistical analysis showed a significant difference (at 5% level between the control and all enzymatically treated oils as well as among different enzymatically treated oils. The relative increase in oil extractability due to pretreatment with enzyme mixtures were in the following order: S: Pe: Bp>S: P>S: C: Pe>S: Bp>S:T>S: C >S: Pa with values 44.9%, 38.9%, 37.1%, 34.9%, 30.1%, 28.9%, respectively. Enzymatic pretreatment of cottonseed flakes resulted in oils with fatty acid composition, acid value, iodine value and peroxide values that were generally comparable to the control.En este trabajo se estudió el efecto del pretratamiento con enzimas sobre la extractabilidad del aceite en hojuelas de semilla de algodón. Las enzimas que se investigaron fueron proteasa bacteriana (Bp, papaína (Pa, savinasa (S, temamil (T, pectinasa (Pe y celulasa (C. Las variables estudiadas durante los experimentos de hidrólisis enzimática fueron: concentración de la enzima, ratio humedad:cantidad de hojuelas y tiempo de hidrólisis. Estos experimentos se realizaron primeramente con una sola enzima y posteriormente con

Isolation and characterization of a cDNA clone coding for a glutathione S-transferase class delta enzyme from the biting midge Culicoides variipennis sonorensis Wirth and Jones.

Science.gov (United States)

Abdallah, M A; Pollenz, R S; Droog, F N; Nunamaker, R A; Tabachnick, W J; Murphy, K E

2000-12-01

Culicoides variipennis sonorensis is the primary vector of bluetongue viruses in North America. Glutathione S-transferases (GSTs) are enzymes that catalyze nucleophilic substitutions, converting reactive lipophilic molecules into soluble conjugates. Increased GST activity is associated with development of insecticide resistance. Described here is the isolation of the first cDNA encoding a C. variipennis GST. The clone consists of 720 translated bases encoding a protein with a M(r) of approximately 24,800 composed of 219 amino acids. The deduced amino acid sequence is similar (64%-74%) to class Delta (previously named Theta) GSTs from the dipteran genera Musca, Drosophila, Lucilia and Anopheles. The cDNA was subcloned into pET-11b, expressed in Epicurian coli BL21 (DE3) and has a specific activity of approximately 28,000 units/mg for the substrate 1-chloro-2,4-dinitrobenzene.

Isolation and purification of glutathione S-transferases from Brachionus plicatilis and B. calyciflorus (Rotifera).

Science.gov (United States)

Bowman, B P; Snell, T W; Cochrane, B J

1990-01-01

1. The enzyme glutathione S-transferase (GST), a critical element in xenobiotic metabolism, was isolated from the marine rotifer Brachionus plicatilis and its freshwater congener B. calyciflorus. 2. In B. plicatilis, GST comprised 4.2% of cytosolic protein and was present as three separate isozymes with mol. wts 30,000, 31,400 and 33,700. Specific activity of crude homogenates was 56 nmol min-1 mg-1 protein, while that of affinity chromatography purified GST was 1850. 3. In B. calyciflorus, GST was present as two isozymes with mol. wts of 26,300 and 28,500, representing 1.0% of cytosolic protein. Crude GST specific activity was 1750 nmol min-1 mg-1 protein and purified was 72,400. 4. Rotifer GSTs are unusual because they are monomers whereas all other animals thus far investigated posses dimeric GSTs.

Influence of supplying bullfrog tadpoles with feed containing 28% crude protein on performance and enzymatic activities Influência do fornecimento de uma ração comercial com 28% de proteína bruta sobre o desempenho e a atividade de enzimas digestivas de girinos de rã-touro

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José Teixeira de Seixas Filho

2010-01-01

Full Text Available The objective of this study was evaluate the influence of feeding bullfrog tadpoles on commercial feed containing 28% crude protein (CP, on their digestive enzyme performance and activities. The experiment lasted 60 days, at the density of one tadpole/L in boxes containing 30 L water. A hundred and twenty tadpoles at the 25 Gosner stage averaging weight and length was 0.046 g and 6.22 mm, respectively, were used. Survival rate, length, final weight, weight gain, feed consumption, apparent food conversion, specific growth rate and activities of chime, amylase, lipase and trypisin were the parameters evaluated, biweekly, in five biometries. Quadratic effect was observed for the length and the weight. There was larger growth of the tadpoles from the 15th to the 45th day (19.82 mm. On the 15th day, the tadpoles presented the largest specific growth rate (16.93%/day, and the largest weight gain (5.460 g, feed intake (14.099 g and the best apparent food conversion (2.46 was from the 45th to the 60th day. The specific activity of amylase was 205 times greater at 60 days when compared to the beginning of the experiment. The results demonstrated that, for the three enzymes studied, the action capacity over the tadpole chime increased significantly after the 30th experimental day. Moreover, they suggested a greater capacity of tadpoles to digest carbohydrates in detriment to proteins, and this fact was accentuated in the initial phase of the exogenous feeding of this amphibian. The commercial feed with 28% CP provide good performance in the bullfrog tadpoles, indicating the juvenile formation within commercial bullfrog farming standards.Objetivou-se com este trabalho avaliar a influência da alimentação de girinos de rã-touro com ração comercial contendo 28% de proteína bruta (PB no desempenho e nas atividades de enzimas digestivas desses animais. Utilizaram-se 120 girinos no estágio 25 de Gosner com 0,046 g e 6,22 mm, respectivamente, mantidos em

PRODUÇÃO DE BIODIESEL POR BIOCATÁLISE UTILIZANDO MÉTODO ALTERNATIVO DE IMOBILIZAÇÃO DA LIPASE EM HIDROGEL

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Fernanda Marder

2008-12-01

Full Text Available A sociedade vem buscando alternativas para reduzir o processo de destruição ambiental instalada no planeta, porém, a partirdo século passado, uma nova visão sobre o uso indiscriminado dos recursos ambientais passou a ser discutido levando a utilizaçãode tecnologias alternativas de produção de combustíveis mais limpos, como o biodiesel, produzido a partir de óleos vegetais. Osprocessos enzimáticos constituem-se numa alternativa de produção, por serem métodos biotecnológicos e menos impactantes aomeio ambiente, apresentando até então, elevado custo de produção. Com o objetivo de diminuir custos e aumentar a eficiênciainvestigou-se a imobilização das enzimas por método alternativo e o emprego destes biocatalisadores na produção de biodiesel. Aimobilização da lipase PS Amano foi realizada utilizando esferas formadas pela hidratação de hidrogéis comerciais em soluçãoaquosa contendo a enzima. Com a enzima imobilizada realizou-se a transesterificação, na qual obteve-se conversões em ésteresmetílicos com rendimento de até 70%, em solvente orgânico, no tempo de 24h, temperatura de 55°C, 100 rpm e 40 esferas dehidrogel com enzima imobilizada. O método de transesterificação enzimática com as enzimas imobilizadas em hidrogéisapresentou-se como uma tecnologia alternativa e promissora para a obtenção de biodiesel.

Effect of cadmium on glutathione S-transferase and metallothionein gene expression in coho salmon liver, gill and olfactory tissues

International Nuclear Information System (INIS)

Espinoza, Herbert M.; Williams, Chase R.; Gallagher, Evan P.

2012-01-01

Highlights: ► Developed qPCR assays to distinguish closely related GST isoforms in salmon. ► Examined the effect of cadmium on GST and metallothionein genes in 3 tissues. ► Modulation of GST varied among isoforms, tissues, and included a loss of expression. ► Metallothionein outperformed, but generally complemented, GSTs as biomarkers. ► Salmon olfactory genes were among the most responsive to cadmium. - Abstract: The glutathione S-transferases (GSTs) are a multifunctional family of phase II enzymes that detoxify a variety of environmental chemicals, reactive intermediates, and secondary products of oxidative damage. GST mRNA expression and catalytic activity have been used as biomarkers of exposure to environmental chemicals. However, factors such as species differences in induction, partial analyses of multiple GST isoforms, and lack of understanding of fish GST gene regulation, have confounded the use of GSTs as markers of pollutant exposure. In the present study, we examined the effect of exposure to cadmium (Cd), a prototypical environmental contaminant and inducer of mammalian GST, on GST mRNA expression in coho salmon (Oncorhynchus kisutch) liver, gill, and olfactory tissues. GST expression data were compared to those for metallothionein (MT), a prototypical biomarker of metal exposure. Data mining of genomic databases led to the development of quantitative real-time PCR (qPCR) assays for salmon GST isoforms encompassing 9 subfamilies, including alpha, mu, pi, theta, omega, kappa, rho, zeta and microsomal GST. In vivo acute (8–48 h) exposures to low (3.7 ppb) and high (347 ppb) levels of Cd relevant to environmental scenarios elicited a variety of transient, albeit minor changes (<2.5-fold) in tissue GST profiles, including some reductions in GST mRNA expression. In general, olfactory GSTs were the earliest to respond to cadmium, whereas, more pronounced effects in olfactory and gill GST expression were observed at 48 h relative to earlier time

Efeitos da farinha de folhas de mandioca sobre a atividade das enzimas AST, ALT, FA e lipídios hepáticos de ratos Wistar Effects of cassava leaves flour on the AST, ALT, ALP enzymes activity and hepatic lipids of Wistar rats

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Daniela Séfora de Melo

2008-12-01

Full Text Available Folhas de mandioca possuem substâncias como ligninas e saponinas que podem apresentar efeito hipolipidêmico. Todavia, um estudo recente relatou aumento no peso do fígado de ratos alimentados com dietas contendo farinha de folhas de mandioca (FFM - Manihot esculenta Crantz cv. Cacao, tornando-se necessário um estudo mais aprofundado dos efeitos desta farinha sobre os parâmetros hepáticos. Para este estudo, um ensaio biológico com 32 ratos machos Wistar foi conduzido por um período de 7 semanas, sendo os tratamentos: dieta controle e dietas contendo 5, 10 e 15% de FFM. As dietas contendo FFM não apresentaram efeitos sobre as atividades das enzimas Aspartato Aminotransferase (AST e Fosfatase Alcalina (FA, mas aumentaram significativamente a atividade da enzima alanina aminotransferase (ALT. O estudo histopatológico revelou vacuolização do citoplasma dos hepatócitos para todos os grupos. No entanto, a freqüência de animais com vacuolização acentuada foi superior nos grupos que receberam dietas com FFM, apresentando também maiores teores de lipídios e colesterol total hepáticos e maior relação peso fígado/peso corporal. Estes resultados indicam que os antinutrientes presentes nas folhas de mandioca, como taninos, cianeto e saponinas, podem ser responsáveis pela redução da função hepática nos animais alimentados com FFM.Cassava leaves contain substances such as lignins and saponins that can present the hypolipidemic effect. However, a recent study has reported an increase in liver weight of rats fed diet containing cassava leaves flour (CLF - Manihot esculenta Crantz cv. Cacao. Thus, a further study of the effect of this flour on the hepatic parameters is necessary. For the development of this study, a biological assay with 32 male Wistar rats was conducted for a period of 7 weeks with the following treatments: control diet and diets containing 5, 10, and 15% of CLF. The diets containing CLF showed no effects on the

Cold induced changes on sugar contents and respiratory enzyme activities in coffee genotypes Alterações dos teores de açúcares e da atividade de enzimas do metabolismo respiratório em genótipos de café submetidos ao frio

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Fábio Luiz Partelli

2010-04-01

Full Text Available The present research aimed to characterize some biochemical responses of Coffea canephora (clones 02 and 153 and C. arabica (Catucaí IPR 102 genotypes subjected to low positive temperatures, helping to elucidate the mechanisms involved in cold tolerance. For that, one year old plants were subjected successively to 1 a temperature decrease (0.5°C a day from 25/20°C to 13/8°C (acclimation period, 2 a three day chilling cycle (3x13/4°C and to 3 a recovery period of 14 days (25/20°C. In Catucaí (less cold sensitive when compared to clone 02 there was an increased activity in the respiratory enzymes malate dehydrogenase and pyruvate kinase. Furthermore, Catucaí showed significant increases along the cold imposition and the higher absolute values after chilling exposure of the soluble sugars (sucrose, glucose, fructose, raffinose, arabinose and mannitol that are frequently involved in osmoregulation and membrane stabilization/protection. The analysis of respiratory enzymes and of soluble sugar balance may give valuable information about the cold acclimation/tolerance mechanisms, contributing to a correct selection and breeding of Coffea sp. genotypes.A pesquisa teve por objetivo caracterizar respostas bioquímicas de genótipos de Coffea canephora (clones 02 and 153 e C. arabica (Catucaí IPR 102 submetidos a baixas temperaturas positivas, ajudando a elucidar os mecanismos envolvidos na tolerância ao frio. Plantas com um ano de idade foram submetidas sucessivamente a 1 decréscimo da temperatura (0,5°C por dia desde 25/20°C até 13/8°C (período de aclimatização, 2 um ciclo de três dias a 13/4°C e 3 14 dias de recuperação (25/20°C. Em Catucaí, genótipo menos sensível ao frio quando comparado com o clone 02, observou-se um aumento das atividades das enzimas malato desidrogenase e piruvate cinase, relacionadas com a respiração. Nesse genótipo, os níveis de açúcares solúveis sacarose, glucose, frutose, rafinose, arabinose e

Insecticide resistance and glutathione S-transferases in mosquitoes ...

African Journals Online (AJOL)

Mosquito glutathione S-transferases (GSTs) have received considerable attention in the last 20 years because of their role in insecticide metabolism producing resistance. Many different compounds, including toxic xenobiotics and reactive products of intracellular processes such as lipid peroxidation, act as GST substrates.

Differential transcription of cytochrome P450s and glutathione S transferases in DDT-susceptible and resistant Drosophila melanogaster strains in response to DDT and oxidative stress

Science.gov (United States)

Metabolic DDT resistance in Drosophila melanogaster has previously been associated with constitutive over-transcription of cytochrome P450s. Increased P450 activity has also been associated with increased oxidative stress. In contrast, over-transcription of glutathione S transferases (GSTs) has been...

Functional characterization of glutathione S-transferases associated with insecticide resistance in Tetranychus urticae

NARCIS (Netherlands)

Pavlidi, N.; Tseliou, V.; Riga, M.; Nauen, R.; Van Leeuwen, T.; Labrou, N.E.; Vontas, J.

2015-01-01

The two-spotted spider mite Tetranychus urticae is one of the most important agricultural pests world-wide. It is extremely polyphagous and develops resistance to acaricides. The overexpression of several glutathione S-transferases (GSTs) has been associated with insecticide resistance. Here, we

Disease: H01321 [KEGG MEDICUS

Lifescience Database Archive (English)

Full Text Available e caused by Streptococcus pneumoniae (pneumococcus). PD is particularly common in younger children and in ol...sult of pneumococcal vaccination in children. However, especially elderly patients with comorbidities remain

The liver transcriptome of suckermouth armoured catfish (Pterygoplichthys anisitsi, Loricariidae): Identification of expansions in defensome gene families

International Nuclear Information System (INIS)

Parente, Thiago E.; Moreira, Daniel A.; Magalhães, Maithê G.P.; Andrade, Paula C.C. de; Furtado, Carolina; Haas, Brian J.; Stegeman, John J.; Hahn, Mark E.

2017-01-01

Pterygoplichthys is a genus of related suckermouth armoured catfishes native to South America, which have invaded tropical and subtropical regions worldwide. Physiological features, including an augmented resistance to organic xenobiotics, may have aided their settlement in foreign habitats. The liver transcriptome of Pterygoplichthys anisitsi was sequenced and used to characterize the diversity of mRNAs potentially involved in the responses to natural and anthropogenic chemicals. In total, 66,642 transcripts were assembled. Among the identified defensome genes, cytochromes P450 (CYP) were the most abundant, followed by sulfotransferases (SULT), nuclear receptors (NR) and ATP binding cassette transporters (ABC). A novel expansion in the CYP2Y subfamily was identified, as well as an independent expansion of the CYP2AAs. Two expansions were also observed among SULT1. Thirty-two transcripts were classified into twelve subfamilies of NR, while 21 encoded ABC transporters. The diversity of defensome transcripts sequenced herein could contribute to this species' resistance to organic xenobiotics. - Highlights: • Resistance of P. anisitsi to organic pollutants (e.g. Diesel oil) is elevated. • > 60 thousand transcripts were assembled at high sequencing depth. • > 20 thousand transcripts were annotated using UniProt and GO databases. • Complete coding sequence for 183 defensome transcripts were identified. • Most abundant defensome families were P450 followed by SULT, NR and ABC transporters.

EFEITOS DO pH, SAL E DESNATURANTES NA HIDRÓLISE IN VITRO DA GLOBULINA PRINCIPAL DE GRÃO-DE-BICO (Cicer arietinum L., var. IAC-Marrocos*

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V. A. NEVES

2009-03-01

Full Text Available

As leguminosas caracterizam-se por apresentar alto teor protéico que na sua grande totalidade são globulinas de reserva,e cujas características moleculares influenciam algumas de suas propriedades com destaque para sua hidrólise frente às enzimas digestivas. As frações 7 e 11S, apesar de certa homologia estrutural, surgem como determinantes das propriedades nutritivas e funcionais das globulinas. Nesse trabalho a globulina principal de grão-de-bico (tipo legumina, 11S foi isolada e purificada, verificando-se o efeito da exposição a diferentes fatores na hidrólise in vitro com as enzimas tripsina e quimotripsina. Comparada à caseína, a globulina principal na forma nativa, e na presença de NaCl 0,3 M, mostrou-se menos susceptível à hidrólise pelas enzimas. A exposição a pHs ácidos extremos revela a retirada desse efeito protetor observado com o sal. As maiores alterações ocorreram pela exposição ao pH 1,0, temperatura de 121°C/15 min e presença de NaCl, no entanto ainda inferiores aos obtidos com a caseína e para ambas enzimas. O tratamento alcalino apesar de aumentar a susceptibilidade às enzimas mostrou valores de hidrólise inferiores à caseína. A exposição a diferentes concentrações dos desnaturantes uréia e hidrocloreto de guanidínio revelou que apenas o segundo teve pequeno efeito sobre a hidrólise pelas enzimas e, mesmo assim, em concentração tão alta como 8M. PALAVRAS-CHAVE: Grão-de-bico; Cicer arietinum L; globulina principal; hidrólise in vitro.

PURIFICACIÓN Y CARACTERIZACIÓN DE alfa-AMILASA DE PENICILLIUM COMMUNE PRODUCIDA MEDIANTE FERMENTACIÓN EN FASE SÓLIDA

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Esperanza Espinel

2010-03-01

Full Text Available Este estudio reporta la purificación y caracterización parcial de una alfa-amilasa producida por Penicillium commune mediante fermentación en fase sólida, empleandoyuca blanca colombiana (Manihot esculenta Crantz como soporte. La enzima fue purificada por precipitación fraccionada con sulfato de amonio, cromatografía de intercambio aniónico (DEAE-Sephadex A-50, cromatografía de filtración por gel (Sephadex G-75 y cromatografía de intercambio catiónico (CM-Sephadex C-50 obteniendo una actividad específica final de 314,82 U/mg, un grado de purificación del orden de 62 y un rendimiento de 9%. La purificación hasta la homogeneidad fue confirmada por SDS-PAGE. El peso molecular estimado fue 35 kDa. La enzima mostró máxima actividad de hidrólisis de almidón soluble con pH 6,0, y estabilidad en un intervalo de pH de 5,0-7,0. La estabilidad térmica de la enzima se presentó en el intervalo de temperatura 0-50 °C y su temperatura óptima fue 70 °C. Los iones Ca2+,Ba2+ y Ag+ aumentaron significativamente la actividad de la enzima, siendo el ión Ca2+ el que tuvo el más alto poder activador. Cu2+ no alteró significativamente la actividad de la enzima, mientras que Li+ y Fe3+ la disminuyeron ligeramente (13%, y Co2+ y Hg2+ la disminuyeron 25% y 40% respectivamente. Los valores de Km y Vmáx fueron calculados usando la linealización de Lineweaver- Burk, con el resultado Km= 0,48 mg/mL y Vmáx = 5,85 micromol glucosa/min. Entre los principales productos de hidrólisis del almidón de yuca se encuentran la maltosa y la glucosa, este resultado proporciona evidencia de que la enzima es capaz de romper los enlaces glicosídicos alfa-1,4 del almidón, comportamiento característico de una alfa-amilasa.

Purificação de três diferentes beta-galactosidades microbianas por partição em sistemas de duas fases aquosas Purification of three different microbial beta-galactosidases by partitioning in aqueous two-phase systems

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Maria Estela SILVA

1997-12-01

Full Text Available Este trabalho tratou da investigação do efeito do peso molecular de polietilenoglicol (PEG sobre a partição de enzimas beta-galactosidases de diferentes origens microbianas: Escherichia coli, Klueveromyces lactis e Aspergillus orizae em sistemas de duas fases aquosas (SDFA.Foi observado que os melhores sistemas para purificação da enzima de E. coli foram os formados por PEG 4000, 6000 e 8000/fosfato, fornecendo os mais elevados fatores de purificação da enzima. As enzimas de Klueveromyces lactis e Aspergillus orizae não foram eficientemente purificadas nestes sistemas sendo insensíveis à alterações do peso molecular do PEG. Portanto, um outro sistema de duas fases aquosas foi desenvolvido contendo um ligante específico, p-aminofenil 1-tio-beta-D-galactopiranosídeo (APGP, acoplado ao PEG para purificar a enzima de Klueveromyces lactis. Uma etapa simples de partição no SDFA formado por 6% APGP-PEG4000 + 12% dextrana T505.000 foi capaz de recuperar 83% da enzima na fase superior do sistema e de aumentar 1,6 vezes o fator de purificação.This work investigated the effect of the molecular weight of polyethyleneglycol (PEG upon the partition coefficient of beta-galactosidases from three different microorganisms: Escherichia coli, Klueveromyces lactis and Aspergillus orizae. It was found that PEG 6,000 and PEG 8,000/phosphate were the best systems for achieving the highest purification factors of E. coli beta-galactosidase. However, the other two yeast beta-galactosidases were not efficiently separated from their contaminants in any of the PEG/salt systems. In order to improve the separation of Klueveromyces lactis beta-galactosidase from the main protein contaminants, the biospecific ligand p-aminophenyl 1-thio-beta-D-galactopyranoside (APGP was attached to activated PEG 4000. The affinity PEG having APGP bound to its backbone was synthesized in two steps. The partitioning of Klueveromyces lactis beta-galactosidase in aqueous two

Estudio de las propiedades de las colinesterasas en epitelios de las vías aéreas superiores humanas y sus alteraciones en el cáncer

OpenAIRE

Castillo González, Ana Cristina

2016-01-01

Objetivos Los objetivos principales de la presente Tesis Doctoral son: 1. Determinar los niveles de actividad Acetilcolinesterasa (AChE) y Butirilcolinesterasa (BChE) en epitelios sanos y tumorales de laringe. 2. Analizar los componentes de formas moleculares de las enzimas en tejidos sanos y patológicos. 3. Identificar y cuantificar los distintos transcritos de AChE y BChE, de la enzima colina-acetil transferasa (ChAT) y de distintas subunidades de los receptores nicotínicos...

Determinación de frecuencia cardiaca, frecuencia respiratoria, lactato deshidrogenasa, creatinkinasa y ácido láctico en caballos durante competencia de salto en la Sabana de Bogotá

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Paula Andrea Guerrero Nieto

2009-06-01

Full Text Available El estudio presenta los primeros resultados de investigación en variables fisiológicas y sus cambios por el ejercicio en equinos en competencia de salto en el país. Se utilizaron 24 ejemplares de las razas Silla Argentina, PSI y mestizo de la Escuela de Equitación del Ejército Nacional de Colombia, que participaron en pruebas de salto (1,10 – 1,20 m en diferentes centros ecuestres de la Sabana de Bogotá. Se determinó frecuencia cardiaca, frecuencia respiratoria y enzimas musculares (creatinquinasa (CK, lactato deshidrogenasa (LDH y el ácido láctico.. Se tomaron tres muestras: reposo, inmediatamente después del ejercicio y a las 6 horas posejercicio. Para la frecuencia cardiaca se utilizó un monitor POLAR S625; la frecuencia respiratoria se determinó mediante fonendoscopio; y las enzimas y el ácido láctico se analizaron en laboratorio. Se utilizó el método estadístico ANAVA para enzimas musculares y acido láctico y estadística descriptiva para frecuencia cardiaca y respiratoria. En los resultados se encontró un comportamiento estadísticamente significativo (p < 0.05 del ácido láctico y la enzima creatinkinasa, a diferencia del comportamiento de la enzima lactato deshidrogenasa, que fue no significativo. Las frecuencias cardiaca y respiratoria se elevaron de manera significativa posejercicio y regresaron a lo normal alrededor de las 6 seis horas. El ácido láctico se incrementó posejercicio y disminuyó a las 6 horas, contrario a lo reportado por otros autores. Se concluye que una vez analizadas estas variables, hay diferencias con los valores obtenidos por otros autores, que pueden deberse a la altitud, al tipo y tiempo de ejercicio realizado. Por tanto es indispensable continuar haciendo estudios en este campo.

INTERACCIÓN DE LA GLUTAMINA SINTETASA (GS Y EL PÉPTIDO β -AMILOIDE COMO UNA ESTRATEGIA DE PURIFICACIÓN

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Sonia Luz Albarracín

2013-08-01

Full Text Available La enfermedad de Alzheimer (EA esla forma de demencia más común en laedad adulta. Se manifiesta con la pérdidaprogresiva de la memoria a medida quelas neuronas en la corteza cerebral y elhipocampo mueren. En todas las formasde EA se evidencia aumento de la expresiónde diferentes proteínas, así comola presencia de agregados insolubles depéptido-β-amiloide (PBA. La glutaminasintetasa (GS es una enzima clave en elmetabolismo del glutamato y en la detoxificaciónde amonio (NH4+. Previamentese ha reportado una posible interacciónGS-PBA que puede estar asociada conEA. En este trabajo se realizó la purificaciónde la enzima cerebral de rata a partirde un extracto sometido a precipitaciónfraccionada con (NH42SO4 del 20-60 %de saturación y posteriormente a travésde cromatografías sucesivas de filtraciónen gel, intercambio iónico y afinidad. Elpeso molecular del complejo fue calculadoen 137 kDa por el orden de eluciónen la columna de filtración. Se identificóinteracción de la enzima con PBA1-40, lográndose la purificación de unasola banda de 45 kDa, correspondiente ala forma monomérica de la GS. En estetrabajo se presenta un nuevo método depurificación de la enzima y se demuestrala interacción de GS con el PBA. Se proponeque esta interacción GS-PBA puedeser uno de los procesos que se presentanen la enfermedad al explicar la reducciónde la actividad de la enzima en pacientes con EA, ya que podría alterar el cicloglutamato-glutamina y generar cambiosen el entorno celular que favorecen excitotoxicidadpor glutamato típica de losprocesos de neurodegeneración.

Polymorphic Human Sulfotransferase 2A1 Mediates the Formation of 25-Hydroxyvitamin D3-3-O-Sulfate, a Major Circulating Vitamin D Metabolite in Humans.

Science.gov (United States)

Wong, Timothy; Wang, Zhican; Chapron, Brian D; Suzuki, Mizuki; Claw, Katrina G; Gao, Chunying; Foti, Robert S; Prasad, Bhagwat; Chapron, Alenka; Calamia, Justina; Chaudhry, Amarjit; Schuetz, Erin G; Horst, Ronald L; Mao, Qingcheng; de Boer, Ian H; Thornton, Timothy A; Thummel, Kenneth E

2018-04-01

Metabolism of 25-hydroxyvitamin D 3 (25OHD 3 ) plays a central role in regulating the biologic effects of vitamin D in the body. Although cytochrome P450-dependent hydroxylation of 25OHD 3 has been extensively investigated, limited information is available on the conjugation of 25OHD 3 In this study, we report that 25OHD 3 is selectively conjugated to 25OHD 3 -3- O -sulfate by human sulfotransferase 2A1 (SULT2A1) and that the liver is a primary site of metabolite formation. At a low (50 nM) concentration of 25OHD 3 , 25OHD 3 -3- O -sulfate was the most abundant metabolite, with an intrinsic clearance approximately 8-fold higher than the next most efficient metabolic route. In addition, 25OHD 3 sulfonation was not inducible by the potent human pregnane X receptor agonist, rifampicin. The 25OHD 3 sulfonation rates in a bank of 258 different human liver cytosols were highly variable but correlated with the rates of dehydroepiandrosterone sulfonation. Further analysis revealed a significant association between a common single nucleotide variant within intron 1 of SULT2A1 (rs296361; minor allele frequency = 15% in whites) and liver cytosolic SULT2A1 content as well as 25OHD 3 -3- O -sulfate formation rate, suggesting that variation in the SULT2A1 gene contributes importantly to interindividual differences in vitamin D homeostasis. Finally, 25OHD 3 -3- O -sulfate exhibited high affinity for the vitamin D binding protein and was detectable in human plasma and bile but not in urine samples. Thus, circulating concentrations of 25OHD 3 -3- O -sulfate appear to be protected from rapid renal elimination, raising the possibility that the sulfate metabolite may serve as a reservoir of 25OHD 3 in vivo, and contribute indirectly to the biologic effects of vitamin D. Copyright © 2018 by The American Society for Pharmacology and Experimental Therapeutics.

Association study of genetic variants in estrogen metabolic pathway genes and colorectal cancer risk and survival.

Science.gov (United States)

Li, Shuwei; Xie, Lisheng; Du, Mulong; Xu, Kaili; Zhu, Lingjun; Chu, Haiyan; Chen, Jinfei; Wang, Meilin; Zhang, Zhengdong; Gu, Dongying

2018-05-16

Although studies have investigated the association of genetic variants and the abnormal expression of estrogen-related genes with colorectal cancer risk, the evidence remains inconsistent. We clarified the relationship of genetic variants in estrogen metabolic pathway genes with colorectal cancer risk and survival. A case-control study was performed to assess the association of single-nucleotide polymorphisms (SNPs) in ten candidate genes with colorectal cancer risk in a Chinese population. A logistic regression model and Cox regression model were used to calculate SNP effects on colorectal cancer susceptibility and survival, respectively. Expression quantitative trait loci (eQTL) analysis was conducted using the Genotype-Tissue Expression (GTEx) project dataset. The sequence kernel association test (SKAT) was used to perform gene-set analysis. Colorectal cancer risk and rs3760806 in SULT2B1 were significantly associated in both genders [male: OR = 1.38 (1.15-1.66); female: OR = 1.38 (1.13-1.68)]. Two SNPs in SULT1E1 were related to progression-free survival (PFS) [rs1238574: HR = 1.24 (1.02-1.50), P = 2.79 × 10 -2 ; rs3822172: HR = 1.30 (1.07-1.57), P = 8.44 × 10 -3 ] and overall survival (OS) [rs1238574: HR = 1.51 (1.16-1.97), P = 2.30 × 10 -3 ; rs3822172: HR = 1.53 (1.67-2.00), P = 2.03 × 10 -3 ]. Moreover, rs3760806 was an eQTL for SULT2B1 in colon samples (transverse: P = 3.6 × 10 -3 ; sigmoid: P = 1.0 × 10 -3 ). SULT2B1 expression was significantly higher in colorectal tumor tissues than in normal tissues in the Cancer Genome Atlas (TCGA) database (P colorectal cancer susceptibility and survival.

Short-term calorie restriction feminizes the mRNA profiles of drug metabolizing enzymes and transporters in livers of mice

International Nuclear Information System (INIS)

Fu, Zidong Donna; Klaassen, Curtis D.

2014-01-01

Calorie restriction (CR) is one of the most effective anti-aging interventions in mammals. A modern theory suggests that aging results from a decline in detoxification capabilities and thus accumulation of damaged macromolecules. The present study aimed to determine how short-term CR alters mRNA profiles of genes that encode metabolism and detoxification machinery in the liver. Male C57BL/6 mice were fed CR (0, 15, 30, or 40%) diets for one month, followed by mRNA quantification of 98 xenobiotic processing genes (XPGs) in the liver, including 7 uptake transporters, 39 phase-I enzymes, 37 phase-II enzymes, 10 efflux transporters, and 5 transcription factors. In general, 15% CR did not alter mRNAs of most XPGs, whereas 30 and 40% CR altered over half of the XPGs (32 increased and 29 decreased). CR up-regulated some phase-I enzymes (fold increase), such as Cyp4a14 (12), Por (2.3), Nqo1 (1.4), Fmo2 (5.4), and Fmo3 (346), and numerous number of phase-II enzymes, such as Sult1a1 (1.2), Sult1d1 (2.0), Sult1e1 (33), Sult3a1 (2.2), Gsta4 (1.3), Gstm2 (1.3), Gstm3 (1.7), and Mgst3 (2.2). CR feminized the mRNA profiles of 32 XPGs in livers of male mice. For instance, CR decreased the male-predominantly expressed Oatp1a1 (97%) and increased the female-predominantly expressed Oatp1a4 (11). In conclusion, short-term CR alters the mRNA levels of over half of the 98 XPGs quantified in livers of male mice, and over half of these alterations appear to be due to feminization of the liver. - Highlights: • Utilized a graded CR model in male mice • The mRNA profiles of xenobiotic processing genes (XPGs) in liver were investigated. • CR up-regulates many phase-II enzymes. • CR tends to feminize the mRNA profiles of XPGs

DYNAMIC OF BIOMECHANIC TECHNOLOGY

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Duško Bjelica

2007-05-01

Full Text Available The mo ving con sists of a set of mo ve ments ma de on right ti me with a ca re fully cho sen in ten sity. Every com po sed mo ving is har mo ni o us but if we ru i ned that har mony in only one se ge ment, if only one mo ve ment was ma de too early or too la te with a gre a ter for ce, it wo uld de stroy the who le pro ce du re and the re sult wo uldn’t be op ti mal.The analysis of every com po sed sport mo ve ment is usu ally do ne in two pha ses: in the fi rst one it sho uld be esta blis hed the le vel of the tec hni cal know led ge, and in the se cond that know led ge sho uld be com pa red to the tec hi que of the sport man in that di sci pli ne. The only in di ca tors of the tec hni que of com plex mo ve ment are the fi rst and the second re sults of the ex ce e ded way and the se i zed an gle from the ac tual and re pre sen ta ti ve po int that are re pre sen ted di gi tally and by ana lo gue. When we com pa re tho se re sults with the re sults of a top spor tsman we will be co me awa re of the de vi a tion from the op ti mal compo si tion of the mo ve ments in to a mo ving and we will esta blish a re la ti on ship bet we en the fa i lu res and the ir con se qu en ces.

Short-term calorie restriction feminizes the mRNA profiles of drug metabolizing enzymes and transporters in livers of mice

Energy Technology Data Exchange (ETDEWEB)

Fu, Zidong Donna [Department of Pharmacology, Toxicology, and Therapeutics, University of Kansas Medical Center, Kansas City, KS 66160 (United States); Klaassen, Curtis D., E-mail: cklaasse@kumc.edu [Department of Internal Medicine, University of Kansas Medical Center, Kansas City, KS 66160 (United States)

2014-01-01

Calorie restriction (CR) is one of the most effective anti-aging interventions in mammals. A modern theory suggests that aging results from a decline in detoxification capabilities and thus accumulation of damaged macromolecules. The present study aimed to determine how short-term CR alters mRNA profiles of genes that encode metabolism and detoxification machinery in the liver. Male C57BL/6 mice were fed CR (0, 15, 30, or 40%) diets for one month, followed by mRNA quantification of 98 xenobiotic processing genes (XPGs) in the liver, including 7 uptake transporters, 39 phase-I enzymes, 37 phase-II enzymes, 10 efflux transporters, and 5 transcription factors. In general, 15% CR did not alter mRNAs of most XPGs, whereas 30 and 40% CR altered over half of the XPGs (32 increased and 29 decreased). CR up-regulated some phase-I enzymes (fold increase), such as Cyp4a14 (12), Por (2.3), Nqo1 (1.4), Fmo2 (5.4), and Fmo3 (346), and numerous number of phase-II enzymes, such as Sult1a1 (1.2), Sult1d1 (2.0), Sult1e1 (33), Sult3a1 (2.2), Gsta4 (1.3), Gstm2 (1.3), Gstm3 (1.7), and Mgst3 (2.2). CR feminized the mRNA profiles of 32 XPGs in livers of male mice. For instance, CR decreased the male-predominantly expressed Oatp1a1 (97%) and increased the female-predominantly expressed Oatp1a4 (11). In conclusion, short-term CR alters the mRNA levels of over half of the 98 XPGs quantified in livers of male mice, and over half of these alterations appear to be due to feminization of the liver. - Highlights: • Utilized a graded CR model in male mice • The mRNA profiles of xenobiotic processing genes (XPGs) in liver were investigated. • CR up-regulates many phase-II enzymes. • CR tends to feminize the mRNA profiles of XPGs.

Polymorphisms in the GST (M1 andT1) gene and their possible ...

African Journals Online (AJOL)

Glutathione S-transferases (GSTs) are enzymes involved in the detoxification of several environmental mutagens, carcinogens and anticancer drugs. GST polymorphisms resulting in decreased enzymatic activity have been associated with several types of solid tumors. We determined the frequencies of the deletion of two ...

Märt Sultsi kavandatud kasiinosaar vajab 300 miljonit krooni laenu / Kertu Kalmus

Index Scriptorium Estoniae

Kalmus, Kertu

2008-01-01

Tallinna linnavolikogu liige Märt Sults palub linnavalitsused toetada ja aktsepteerida SA Kasiinosaar laenu taotlemist Saksamaalt 300 mln. krooni ulatuses. Tema ideekavandi järgi kerkiksid Tallinnas Paljassaare lahte tehissaared: lastesaar, kultuurisaar ja kasiinosaar

Influencia de la transglutaminasa en el rendimiento de la producción de queso dambo uruguayo

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Daniela Escobar

2014-12-01

Full Text Available El queso dambo uruguayo es un queso de mediana humedad de origen danés con formulación modificada. Esta investigación estudió el uso de la enzima transglutaminasa en la producción de queso dambo tipo barra para aumentar el rendimiento del proceso.Se fabricaron quesos aplicando un diseño experimental completamente al azar con parcelas divididas medidas en el tiempo, considerando dos factores (concentración de la enzima y momento de agregado en tres niveles, por triplicado, tomando como referencia el queso sin enzima.Los quesos fueron analizados a lo largo de cuatro meses en peso, pH, contenido de proteínas, humedad y materia grasa. Se calculó el rendimiento relativo al queso referencia, la recuperación de extracto seco y el contenido en proteínas y materia grasa en el queso respecto a la leche. Se analizó el perfil de textura y se determinaron las propiedades sensoriales con un panel de jueces entrenados.Los resultados mostraron que la única condición con diferencia significativa al 5% en el rendimiento relativo fue el momento de agregado de la enzima, resultando como momento óptimo la incorporación de la misma junto con el cuajo, independientemente de la dosis utilizada. El rendimiento logrado fue un 6% superior al queso referencia. 

Digestibilidade aparente dos nutrientes e energia de ração suplementada com enzimas digestivas exógenas para juvenis de tambaqui (Colosssoma macropomum Cuvier, 1818 Apparent digestibility of nutrients and crude energy in diets with addition of exogenous digestive enzymes in tambaqui juveniles ( Colosssoma macropomum Cuvier, 1818

Directory of Open Access Journals (Sweden)

Jorge Antonio Moreira da Silva

2007-01-01

Full Text Available O experimento foi conduzido com o objetivo de avaliar o efeito da adição de um compelxo multienzimático exógeno composto de amilase, protease, lipase e celulase, em rações de juvenis de tambaqui, sobre os coeficientes de digestibilidade aparente (CDa da proteína bruta (PB, extrato etéreo (EE, carboidratos (ENN e energia bruta (EB. O delineamento experimental foi inteiramente casualizado com quatro tratamentos (quatro níveis de inclusão de enzimas, 0,0; 0,05; 0,10; e 0,15 %, três repetições (no tempo e 10 peixes por unidade experimental. Foram utilizados 40 juvenis de tambaqui, com peso médio de 155,0 ± 0,49 g, distribuídos em quatro tanques de alimentação de 500 l, recebendo refeições à vontade das 8 às 12h, a cada hora. Em seguida os animais foram transferidos para coletores de fezes (200 l, onde permaneceram até às 18h, sendo a coleta de dejetos realizada a cada hora. A determinação dos CDa foi realizada pelo método indireto, sendo utilizado como indicador externo 0,5% de óxido de cromo-III (Cr2O3 incorporado à ração. Os resultados demonstraram que a suplementação das dietas com enzimas exógenas para juvenis de tambaqui aumenta a digestibilidade aparente dos nutrientes e energia bruta, no nível de inclusão de 0,05% (PThe experiment evaluated the effect of dietary supplementation a multienzyme complex containing amylase, protease, lipase and cellulase for tambaqui juveniles on the apparent digestibility coefficients (ADCs of crude protein (CP, ether extract (EE, carbohydrates (CHO and crude energy (CE. A completely randomized experimental design was used with four treatments (level of enzymes 0.0; 0.05; 0.10; and 0.15 % and three replicates (over time, with ten fish per experimental unity. Forty fish weighting 155.0 ± 0.49g were distributed in four 500 l feeding tanks and were fed hourly, from 8 to 12 AM. After this period, fish were transferred for fecal collection units until 6 PM, where feces were

The correlation between motor proficiency and physical activity in ...

African Journals Online (AJOL)

Lizl-Louise van Niekerk

eOseretsky Test of Motor Proficiency 2 (BOT-2) for motor proficiency, and the International. Physical Activity ... Pienaar, 2007; Malina, 2012). ..... sults of Hardy, Reinten-Reynolds, Espinel, Zask, and Okely .... Journal of Psychiatric and Mental.

Caracterização do secretoma de Aspergillus niger crescido em bagaço de cana e purificação de xilanases de interesse biotecnológico

OpenAIRE

Martins, Pedro Alves

2012-01-01

Os fungos filamentosos quando submetidos ao crescimento em meios contendo material lignocelulósico, são capazes de produzir diferentes tipos de enzimas extracelulares responsáveis por catalisar a hidrólise de celulose, hemicelulose e outros polissacarídeos encontrados nas paredes celulares vegetais. Algumas dessas enzimas podem ser aplicadas comercialmente em processos industriais, como nas indústrias alimentícia, têxtil, papeleira e também na produção de bioetanol. O fungo filamentoso mesofí...

Purificación y caracterización de poligalacturonasas de Aspergillus kawachii

OpenAIRE

Contreras Esquivel, Juan Carlos

2003-01-01

El desarrollo de nuevas aplicaciones de las pectinasas depende de la búsqueda de enzimas que catalizen reacciones novedosas o que presenten propiedades diferentes a las ya estudiadas. Considerando la capacidad de A. kawachii de producir enzimas activas en condiciones de alta acidez y al escaso conocimiento de su sistema pectolítico, se inició un proyecto de investigación dirigido a estudiar las pectinasas de este microorganismo. El presente trabajo de Tesis se enmarcó en los objetivos de dich...

Efeito da adição de enzimas em dietas de frangos de corte à base de milho e farelo de soja sobre a digestibilidade ileal de nutrientes Effect of enzymes supplementation in corn soybean meal broiler diets on ileal digestibility of nutrients

Directory of Open Access Journals (Sweden)

Anel Atencio Tejedor

2001-06-01

Full Text Available Dois experimentos, utilizando pintos de corte Avian Farm, no período de 8 a 18 dias de idade, foram realizados para se avaliar o efeito de enzimas microbianas sobre os coeficientes de digestibilidade (CD ileal aparente da matéria seca (MS, da proteína bruta (PB, da energia bruta (EB, do fósforo (P e do cálcio (Ca de dietas à base de milho e farelo de soja com diferentes níveis de Ca e P disponível (Pd. Óxido crômico (0,5% foi adicionado às dietas a fim de se estimar o fator de indigestibilidade. Todas as aves foram abatidas aos 19 dias, para a coleta da digesta. O primeiro experimento foi realizado em arranjo fatorial 2 x 2, com seis repetições contendo 10 aves. As dietas foram formuladas contendo dois níveis de Ca e Pd normal (1%Ca/0,45%Pd e baixo (0,70%Ca/0,32%Pd x dois níveis de enzima fitase (0 e 1 kg/t de ração. O segundo experimento foi realizado em arranjo fatorial 2 x 3, sendo dois níveis de Ca e Pd normal (1%Ca/0,45%Pd e baixo (0,70%Ca/0,32Pd% e três combinações de enzimas: complexo multienzimático (CM com fitase (Fit: 0 kg/t para o controle, 2 kg/t (CM e 2 kg/t mais 1 kg/t de fitase (CM+Fit, com seis repetições, contendo 10 aves cada. No experimento 1, a adição de fitase melhorou a digestibilidade ileal da MS (5,2%, PB (2,4% e EB (3,8%. Interação significativa foi observada na digestibilidade do Ca e P, sendo que adição da fitase melhorou a digestibilidade do Ca e P em ambos níveis de Ca e P. No segundo experimento, a adição do CM melhorou o CD da PB, P e Ca em 3; 4,7; e 7,8%, respectivamente. A associação do CM+fitase mostrou efeito aditivo no aproveitamento do Ca e P. Interação foi observada para o CD da EB, a adição do CM melhorou a digestibilidade em ambos níveis de Ca e Pd, entretanto o CM+fitase melhorou o CD da EB apenas no nível baixo de Ca e P.Two experiments, using 8 to 18 days old Avian -Farm male broiler chicks, were conducted to evaluate the effect of microbial enzymes on the ileal

Phi Class of Glutathione S-transferase Gene Superfamily Widely Exists in Nonplant Taxonomic Groups.

Science.gov (United States)

Munyampundu, Jean-Pierre; Xu, You-Ping; Cai, Xin-Zhong

2016-01-01

Glutathione S-transferases (GSTs) constitute a superfamily of enzymes involved in detoxification of noxious compounds and protection against oxidative damage. GST class Phi (GSTF), one of the important classes of plant GSTs, has long been considered as plant specific but was recently found in basidiomycete fungi. However, the range of nonplant taxonomic groups containing GSTFs remains unknown. In this study, the distribution and phylogenetic relationships of nonplant GSTFs were investigated. We identified GSTFs in ascomycete fungi, myxobacteria, and protists Naegleria gruberi and Aureococcus anophagefferens. GSTF occurrence in these bacteria and protists correlated with their genome sizes and habitats. While this link was missing across ascomycetes, the distribution and abundance of GSTFs among ascomycete genomes could be associated with their lifestyles to some extent. Sequence comparison, gene structure, and phylogenetic analyses indicated divergence among nonplant GSTFs, suggesting polyphyletic origins during evolution. Furthermore, in silico prediction of functional partners suggested functional diversification among nonplant GSTFs.

Effects of cerium dioxide nanoparticles in Oncorhynchus mykiss liver after an acute exposure: assessment of oxidative stress, genotoxicity and histological alterations

Directory of Open Access Journals (Sweden)

Ana Cristina Nunes

2015-12-01

Full Text Available At present cerium oxide nanoparticles (CeO2 NP have numerous applications ranging from industry to the household, leading to its wide distribution namely in the aquatic environment. The hereby study aimed to assess the toxic effects of CeO2 NPs in Oncorhynchus mykiss liver following an acute exposure (96h to three different concentrations (0.25, 2.5 and 25 mg/L in terms of the genotoxicity (comet assay, oxidative stress response (Catalase CAT; Glutathione S-Transferases GSTs; Thiobarbituric Acid Reactive Substances TBARS and histopathology. CeO2 NP exposure resulted in genotoxic damage in all exposure treatments, inhibition of CAT in the highest concentration and histopathological changes in all exposure concentrations with predominance of progressive and circulatory alterations. However TBARS and GSTs showed no significant differences comparatively to the control (unexposed group. The results suggest that CeO2 NP are able to cause genotoxicity, biochemical impairment and histological alterations in the liver of rainbow trout.

AJDAS vol 8 No 1.indd

African Journals Online (AJOL)

cognitive-behaviour therapy among male patients, self efficacy and religiosity were found to be effective components of cognitive behaviour therapy in substance use disorders as this led to ..... sult of parents and peers drug use, advertising.

Collocations and grammatical patterns in a Multilingual Online Term ...

African Journals Online (AJOL)

user

equivalents for key concepts in the African languages, but also additional con- ... for, inter alia, computational identification and extraction of collocations exist; .... sult' is to be followed by a prepositional phrase in which the preposition is.

Technological and chemical properties of heat-treated Anatolian ...

African Journals Online (AJOL)

STORAGESEVER

2009-06-03

Jun 3, 2009 ... heat treatment temperature and time justifies these re- sults. Cellulose ..... properties of light-irradiated wood with heat treatment: Part 1. Effect ... Norway spruce (Picea abies) and birch (Betula pubescens) subjected to heat ...

Determinación in vitro de los patrones de inducción de una endo-xilanasa (E.C.3.2.1.8 secretada por Fusarium oxysporum f.sp. dianthi. Purificación y caracterización parcial

Directory of Open Access Journals (Sweden)

Sixta Tulia Martinez

2014-04-01

Full Text Available Al hacer un ensayo in vitro, la enzimaxilanasa secretada por el patógeno Fusariumoxysporum f.sp. dianthi se induceen presencia de pared celular de tallo yraíz de clavel de variedades tolerantesy resistentes. Se observa un comportamientodiferencial según el órgano y lavariedad, que demuestra la influencia,en la inducción, de las características delhospedero.La enzima se purifica parcialmenteutilizando cromatografías de intercambiocatiónico, aniónico y tamiz molecular. Lacaracterización bioquímica presenta lossiguientes valores: masa molecular 38,8kDa, temperatura y pH óptimo 35 0Cy 10, respectivamente, Kmap = 0,2 mg/mL, Vmaxap = 500 U/mg.La presencia de oligogalacturónidoscomo productos de hidrólisis detectadoscon una cromatografía en capa delgadapermite clasificar la enzima como unaendoxilanasa.La enzima retiene el 50 % de su estabilidada pH muy alcalino, y presentaalta termoestabilidad en un intervalo detemperatura entre 30 y 80 0C.

Estudo da produção do suco clarificado de cajá (Spondias lutea L.

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SILVA Antonio de Pádua Valença da

1999-01-01

Full Text Available Cajás ( Spondias lutea L., oriundos do município de Maranguape, Ceará, Brasil, foram processados em nível de escala-piloto para obtenção de suco clarificado através do emprego de enzimas pectinolíticas e agentes clarificantes. Aplicaram-se 120ppm de enzima pectinolítica (Pectinex Ultra SP-L na polpa para obtenção do suco polposo e, em seguida, 500ppm de enzima pectinolítica (Pectinex-AR no suco extraído, utilizando-se a "Prova do álcool" como indicador da presença de pectina. Para o processo de clarificação do suco foram utilizados, como agentes clarificantes, 400ppm de gelatina e 500ppm de sílica sol. A "Prova do excesso e/ou da insuficiência de clarificação", bem como a "Prova de estabilidade" mostraram resultados negativos, indicando a eficiência do processo de clarificação.

Purificación y determinación de actividad enzimática de la catalasa en Staphylococcus aureus

Directory of Open Access Journals (Sweden)

Iván Marin Arguello

2000-02-01

Full Text Available Ampliamente distribuida en células animales, vegetales y micro- organismos, la catalasa es una enzima crucial para la vida. Su función catalítica es la descomposición del peróxido de hidrógeno (HOY, compuesto altamente tóxico y generado en los procesos metabólicos de la célula. Esta enzima en Staphylococcus aureus es una proteína tetramérica presentando grupos hemos y moléculas de NADPH en su estructura. La determinación espectrofotométrica de los parámetros cinéticos (Km, Vmax. y otros en dependencia del pH y temperatura nos podría permitir regular su actividad enzimática. La enzima manifiesta un Km de 50mM y la presencia de dos aminoácidos (Histidina y Cisteína en el sitio activo aparentemente son fundamentales en el proceso de catálisis.

ORIGINAL ARTICLE

African Journals Online (AJOL)

User

UDS Publishers Limited All Right Reserved 2026-6294. ORIGINAL ... Reproductive development and function in human and other ... sulting solution was filtered and left to stand for three days to ..... male rat brain and pituitary. Brain Res 164,.

Alpha amylase from a fungal culture grown on oil cakes and its properties

Directory of Open Access Journals (Sweden)

Sumitra Ramachandran

2004-06-01

Full Text Available Solid-state fermentation was carried out for the production of alpha-amylase using Aspergillus oryzae. Different oil cakes such as coconut oil cake (COC sesame oil cake (SOC, groundnut oil cake (GOC, palm kernel cake (PKC and olive oil cake (OOC were screened to be used as substrate for the enzyme production and also compared with wheat bran (WB. GOC was found to be the best producer of the enzyme among these. Combination of WB and GOC (1:1 resulted higher enzyme titres than the individual substrates. Maximum amount of enzyme (9196 U/gds was obtained when SSF was carried out using WB + GOC, having initial moisture of 64% and supplemented with lactose and ammonium nitrate (1% each at 30ºC for 72h using 2 mL spore suspension (6x10(7spores/ml. Partial purification of the enzyme using ammonium sulphate fractionation resulted in 2.4-fold increase in the activity. The enzyme showed molecular weight of 68 KDa by SDS-PAGE. Except Mn, all other metal ions such as Ca, K, Na, Mg were found to be inhibitory for the enzyme activity. The enzyme was optimally active at 50(0C and pH 5.0.Fermentação no Estado Sólido foi empregada na produção de alfa-amilase usando Aspergillus niger. Diferentes tipos de torta foram utilizadas, como torta de óleo de coco (COC, torta de de óleo de amendoim (GOC torta de óleo de sesamo (SOC, torta de palma (PKC e torta de óleo de oliva (OOC foram selecionadas para serem usadas como substratos para produção de enzima e comparadas com o farelo de trigo (WB, GOC foi escolhido por ser o que produziu maiores concentrações de enzima. A combinação WB e GOC (1:1 resultou em maiores títulos da enzima quando em comparação com os substratos individuais. A máxima concentração de enzima (9196 U/ gms foi obtida quando a FES foi conduzida utilizando WB + GOC, com umidade de 64% e suplementada com lactose e nitrato de amônia (1% cada a 300C por 72 horas utilizando 2 mL de uma suspensão de esporo (6x107sporos/ml. A purifica

Enzyme activity of β-galactosidase from Kluyveromyces lactis and Aspergillus oryzae on simulated conditions of human gastrointestinal system

Directory of Open Access Journals (Sweden)

Alessandra Bosso

2015-09-01

 capacidade de hidrólise de β-galactosidase produzida por Kluyveromyces lactis e Aspergillus oryzae simulando as condições do trato gastrintestinal humano. O teste foi realizado nas temperaturas ótimas de ação para cada enzima, 40 e 55°C, respectivamente, e na temperatura corpórea humana (37°C, nas concentrações de 1,5; 3,0 e 5,0 g/L para a enzima de Aspergillus oryzae ou mL/L para a de Kluyveromyces lactis. Na simulação da condição estomacal humana (pH 2, ambas enzimas foram totalmente inativadas. Quando as enzimas foram submetidas às condições simuladas do intestino delgado (pH 7,4, observou-se hidrólise da lactose, porém, a 37°C, a porcentagem foi menor do que a observada nas temperaturas ótimas de cada enzima. A enzima de K. lactis nas concentrações de 1,5; 3,0 e 5,0 mL/L apresentou hidrólise de 76,63%, 88,91% e 94,80% a 40°C e 55,99%, 80,91% e 81,53%, a 37°C, respectivamente. Nas concentrações 1,5; 3,0 e 5,0 g/L, a porcentagem de hidrólise pela enzima de A. oryzae a 55°C foi de 7,11%, 16,18% e 21,29%. Para esta enzima, nessas concentrações, a hidrólise obtida a 37°C foi 8,4%, 11,85% e de 16,43%. Sob condições intestinais simuladas, a enzima de K. lactis apresentou maior eficiência na hidrólise da lactose quando comparada à enzima de A. oryzae. Considerando-se as etapas avaliadas neste estudo, observa-se que é extremamente necessário o uso de um revestimento entérico em cápsulas de β-galactosidase, para que esta enzima seja liberada somente no intestino delgado, seu local de ação, não sofrendo, portanto, a ação do pH estomacal.Palavras-chave: Lactase. Hidrólise. Intolerância à lactose. Trato gastrintestinal.

Detecção e caracterização de complexos multi-enzimáticos em secretoma de fungos filamentosos

OpenAIRE

Silva, Adelson Joel da

2013-01-01

As enzimas que degradam parede celular vegetal produzidas por micro-organismos possuem aplicações biotecnológicas importantes, incluindo a produção de bioetanol. Algumas batérias anaeróbicas são capazes de produzir complexos multi-enzimáticos chamados de celulosomos, enquanto os fungos filamentosos normalmente secretam enzimas hidrolíticas individuais que atuam sinergisticamente no processo de degradação de polissacarídeo. Neste trabalho, nós mostramos que os fungos filamentosos Trichoderm...

Clonación, expresión y caracterización de una nueva esterasa derivada de metagenomas de suelos agrícolas colombianos

OpenAIRE

Carolina Villamil; Patricia Del Portillo; Alvaro Monguí

2016-01-01

El presente trabajo tuvo como objetivo la bioprospección de ADN metagenómico derivado de comunidades microbianas asociadas a un agroecosistema de importancia nacional. Este análisis permitió realizar la producción, expresión, purificación y caracterización de una enzima novedosa con actividad esterasa. Esta enzima, denominada LipM, había sido previamente identificada en clones metagenómicos derivados de suelos dedicados al cultivo de papa criolla (Solanum pureja), mediante secuencia de nueva ...

The effects of mutating Tyr9 and Arg15 on the structure, stability, conformational dynamics and mechanism of GSTA3-3

CSIR Research Space (South Africa)

Robertson, GJ

2017-03-01

Full Text Available activity of the steroid isomerase reaction; however, Arg15 is more important for lowering the pKa of GSH. Lowering the pKa of GSH being how GSTs catalyse their reactions. Additionally, there is evidence to suggest that Arg15 is integral to allowingGSTA3...

Conducting Polymers

Indian Academy of Sciences (India)

discovery of dopingl led to a further dramatic increase in ..... sulted in a further drop in forward voltage to about 5 V, and an increase ... microamperometric glucose and galactose blosensors based on a silicon chip employing polypyrrole have.

Does maternal exposure to artificial food coloring additives increase oxidative stress in the skin of rats?

Science.gov (United States)

Başak, K; Başak, P Y; Doğuç, D K; Aylak, F; Oğuztüzün, S; Bozer, B M; Gültekin, F

2017-10-01

Glutathione-S-transferase (GST) and cytochrome P450 family 1 subfamily A polypeptide 1 (CYP1A1) metabolize and detoxify carcinogens, drugs, environmental pollutants, and reactive oxygen species. Changes of GST expression in tissues and gene mutations have been reported in association with many neoplastic skin diseases and dermatoses. Widely used artificial food coloring additives (AFCAs) also reported to effect primarily behavioral and cognitive function and cause neoplastic diseases and several inflammatory skin diseases. We aimed to identify the changes in expression of GSTs, CYP1A1, and vascular endothelial growth factor (VEGF) in rat skin which were maternally exposed AFCAs. A rat model was designed to evaluate the effects of maternal exposure of AFCAs on skin in rats. "No observable adverse effect levels" of commonly used AFCAs as a mixture were given to female rats before and during gestation. Immunohistochemical expression of GSTs, CYP1A1, and VEGF was evaluated in their offspring. CYP1A1, glutathione S-transferase pi (GSTP), glutathione S-transferase alpha (GSTA), glutathione S-transferase mu (GSTM), glutathione S-transferase theta (GSTT), and VEGF were expressed by epidermal keratinocytes, dermal fibroblasts, sebaceous glands, hair follicle, and subcutaneous striated muscle in the normal skin. CYP1A1, GSTA, and GSTT were expressed at all microanatomical sites of skin in varying degrees. The expressions of CYP1A1, GSTA, GSTT, and VEGF were decreased significantly, while GSTM expression on sebaceous gland and hair follicle was increased. Maternal exposure of AFCAs apparently effects expression of the CYP1A1, GSTs, and VEGF in the skin. This prominent change of expressions might play role in neoplastic and nonneoplastic skin diseases.

Efeito da adição de aminoácidos essenciais à dieta sobre a secreção de enzimas digestivas de jundiá Rhamdia quelen (Siluriformes, Pimelodidae = Effect of dietary essential amino acids addition on digestive enzime secretion in silver catfish Rhamdia quelen (Siluriformes, Pimelodidae

Directory of Open Access Journals (Sweden)

Alexandre Bernardini Ungar

2009-01-01

Full Text Available Em estudos realizados com variação no teor de proteína na ração para peixes, foi demonstrado o aumento das atividades de protease alcalina, protease ácida, tripsina e quimiotripsina do trato digestório de jundiá (Rhamdia quelen. Esses fatos sugerem que produtos liberados pela digestão na luz desse trato podem influenciar a síntese e a secreção de enzimas digestivas. A secreção destas enzimas pelo pâncreas, em vertebrados, responde à influência neurale humoral. No presente trabalho, foram testadas quatro concentrações (0, 3, 6 e 12% de uma mistura de aminoácidos (treonina, fenilalanina, leucina, valina, arginina e triptofano em iguaisproporções em dietas isocalóricas contendo 3.500 kcal de energia digestível kg-1 de ração, e dietas isoproteicas contendo 20% de proteína, em juvenis de jundiá. Foram utilizados dez animais portratamento, sendo os peixes estocados num sistema fechado com recirculação de água e temperatura controlada. Os peixes submetidos a estes tratamentos apresentaram atividade da protease alcalina na região anterior do intestino, responsiva aos diferentes níveis aminoácidos da alimentação. Todavia, as atividades protease alcalina do fígado e amilase do fígado e intestino anterior não foram detectadas. Estes resultados sugerem que a atividade protease alcalina do intestino anterior seja induzida por aminoácidos liberados na luz do trato digestório. Studies carried out with variation of protein concentration in the fish feed have shown an increase in the activity of alkaline proteases, acid proteases, trypsin and chimotrypsin from the digestive tract of silver catfish (Rhamdia quelen. These facts suggest that products released by digestion in the lumen of this tract can influenced the synthesis and secretion of digestive enzymes. The secretion of these enzymes by the pancreas in vertebrate responses to neural and humoral influences. In the present study, four concentration (0, 3, 6 and 12% of

Cultivo de Penicillium spp. em resíduos da colheita de soja para produção de celulase, protease e amilase

Directory of Open Access Journals (Sweden)

Joice Raísa Barbosa Cunha

Full Text Available RESUMO O objetivo deste estudo foi avaliar a produção de enzimas amilolíticas, celulolíticas e proteolíticas pela linhagem Penicillium spp. LEMI A8221 cultivada em estado sólido em resíduos da colheita de soja, ao longo de quatro dias, em diferentes condições de pH (5,0 e 6,0, temperatura (30 e 35°C e concentrações de substrato (70 e 90% p/v. As atividades máximas obtidas para α-amilase, β-amilase, CMCase e protease foram de 0,20; 0,13; 0,65 e 147 U.mg-1, respectivamente. As condições de fermentação influenciaram a atividade das enzimas, sendo a concentração de substrato, a variável mais significativa para o processo. O tempo de fermentação exerceu efeito apenas para as atividades de amilase sacarificante e CMCase, sendo registrados os menores valores de atividade para essas enzimas, nas primeiras 24 e 48 horas de fermentação, respectivamente. Conclui-se que o isolado Penicillium spp. LEMI A8221 pode ser considerado promissor agente biológico, com aplicação industrial, e o resíduo de soja apresentou-se como fonte de carbono alternativa, no cultivo em estado sólido, para produção de enzimas por esta linhagem microbiana.

An action-learning model to assist Circuit Teams to support School ...

African Journals Online (AJOL)

EAOSA

2016-11-17

Nov 17, 2016 ... If the role of the Education District and Circuit Officers in South Africa is to work collaboratively with schools to improve educational ..... sulted in the Chief Curriculum Advisor not allowing ..... 2008/09 financial year. Zwelitsha ...

Matrix-derived combination effect and risk assessment for estragole from basil-containing plant food supplements (PFS)

NARCIS (Netherlands)

Berg, van den S.J.P.L.; Klaus, V.; Alhusainy, W.; Rietjens, I.

2013-01-01

Basil-containing plant food supplements (PFS) can contain estragole which can be metabolised into a genotoxic and carcinogenic 1'-sulfoxymetabolite. This study describes the inhibition of sulfotransferase (SULT)-mediated bioactivation of estragole by compounds present in basil-containing PFS.

Antioxidant role of glutathione S-transferases: 4-Hydroxynonenal, a key molecule in stress-mediated signaling

International Nuclear Information System (INIS)

Singhal, Sharad S.; Singh, Sharda P.; Singhal, Preeti; Horne, David; Singhal, Jyotsana; Awasthi, Sanjay

2015-01-01

4-Hydroxy-2-trans-nonenal (4HNE), one of the major end products of lipid peroxidation (LPO), has been shown to induce apoptosis in a variety of cell lines. It appears to modulate signaling processes in more than one way because it has been suggested to have a role in signaling for differentiation and proliferation. It has been known that glutathione S-transferases (GSTs) can reduce lipid hydroperoxides through their Se-independent glutathione-peroxidase activity and that these enzymes can also detoxify LPO end-products such as 4HNE. Available evidence from earlier studies together with results of recent studies in our laboratories strongly suggests that LPO products, particularly hydroperoxides and 4HNE, are involved in the mechanisms of stress-mediated signaling and that it can be modulated by the alpha-class GSTs through the regulation of the intracellular concentrations of 4HNE. We demonstrate that 4HNE induced apoptosis in various cell lines is accompanied with c-Jun-N-terminal kinase (JNK) and caspase-3 activation. Cells exposed to mild, transient heat or oxidative stress acquire the capacity to exclude intracellular 4HNE at a faster rate by inducing GSTA4-4 which conjugates 4HNE to glutathione (GSH), and RLIP76 which mediates the ATP-dependent transport of the GSH-conjugate of 4HNE (GS-HNE). The balance between formation and exclusion promotes different cellular processes — higher concentrations of 4HNE promote apoptosis; whereas, lower concentrations promote proliferation. In this article, we provide a brief summary of the cellular effects of 4HNE, followed by a review of its GST-catalyzed detoxification, with an emphasis on the structural attributes that play an important role in the interactions with alpha-class GSTA4-4. Taken together, 4HNE is a key signaling molecule and that GSTs being determinants of its intracellular concentrations, can regulate stress-mediated signaling, are reviewed in this article. - Highlights: • GSTs are the major

Efeito do cobre na atividade da enzima pirogalol peroxidase em plantas de Myriophyllum aquaticum cultivadas em solução nutritiva Effect of copper on the activity of pirogalol peroxidase in Myriophyllum aquaticum plants cultivated in nutritives solution

Directory of Open Access Journals (Sweden)

V.D. Domingos

2005-06-01

Full Text Available Myriophyllum aquaticum é uma planta perene, herbácea, que pode se desenvolver totalmente submersa ou com a porção terminal dos ramos acima da superfície da água. É também considerada uma planta daninha que possui elevado potencial de colonização, o qual, dependendo da densidade populacional, pode causar aumento no teor de matéria orgânica e redução de oxigênio na água, comprometendo a qualidade da água e seus usos múltiplos. O objetivo do presente trabalho foi verificar a influência do cobre na atividade da pirogalol peroxidase de plantas de M. aquaticum submetidas à solução nutritiva contendo concentrações de cobre de 1,2; 11,2; 21,2; 31,2; e 41,2 µg L-1. O experimento foi conduzido em um delineamento experimental inteiramente casualizado, com quatro repetições e cinco tratamentos, aos quais as plantas foram submetidas durante 21 dias. Aos 81 dias após a instalação das mudas em solução nutritiva contendo os diferentes níveis de cobre, as folhas foram colhidas a partir do ápice da planta até o final do ramo, que não estavam em contato com a solução. Esse material fresco foi envolvido por plástico transparente e papel-alumínio e, a seguir, congelado em nitrogênio líquido, sendo armazenado em freezer a -20 ºC até o momento da determinação da atividade da enzima pirogalol peroxidase. A atividade da enzima foi progressiva com o aumento das doses de cobre. As plantas cultivadas com 40 µg L-1 de Cu2+ após três semanas, com base em avaliação visual, apresentaram redução no desenvolvimento.Myriophyllum aquaticum is a perennial, herbaceous plant which grows totally submerged or with the terminal portion of the branches above the surface of the water. It is also considered to be a harmful plant, possessing a high colonization potential which, depending on population density, can increase the organic matter content and reduce oxygen in the water, compromising the quality of water and its multiple uses

Cloning and characterization of a glutathione S-transferase homologue from the plant pathogenic fungus Botrytis cinerea

NARCIS (Netherlands)

Prins, T.W.; Wagemakers, L.; Schouten, A.; Kan, van J.A.L.

2000-01-01

A gene was cloned from Botrytis cinerea that encodes a protein homologous to glutathione S-transferase (GST). The gene, denominated Bcgst1, is present in a single copy and represents the first example of such a gene from a filamentous fungus. The biochemical function of GSTs is to conjugate toxic

26 CFR 26.2642-5 - Finality of inclusion ratio.

Science.gov (United States)

2010-04-01

... 26 Internal Revenue 14 2010-04-01 2010-04-01 false Finality of inclusion ratio. 26.2642-5 Section...-5 Finality of inclusion ratio. (a) Direct skips. The inclusion ratio applicable to a direct skip...) Other GSTs. With respect to taxable distributions and taxable terminations, the inclusion ratio for a...

Implementing a High School Level Geospatial Technologies and Spatial Thinking Course

Science.gov (United States)

Nielsen, Curtis P.; Oberle, Alex; Sugumaran, Ramanathan

2011-01-01

Understanding geospatial technologies (GSTs) and spatial thinking is increasingly vital to contemporary life including common activities and hobbies; learning in science, mathematics, and social science; and employment within fields as diverse as engineering, health, business, and planning. As such, there is a need for a stand-alone K-12…

Avaliação da atividade da superóxido dismutase e catalase de Candida albicans e Candida dubliniensis expostas a antineoplásicos, íons metálicos e antifúngicos

OpenAIRE

Carlos Eduardo Blanco Linares

2009-01-01

A atividade de catalase em Candida albicans tem sido sugerida como um mecanismo de resistência ao antifúngico anfotericina B. Neste contexto, poucos são os estudos de enzimas como catalase e superóxido dismutase em leveduras do gênero Candida expostas a diferentes situações. Assim, este estudo teve por objetivo investigar o efeito da exposição de Candida a antineoplásicos, íons metálicos e antifúngicos como fluconazol e anfotericina B sobre a atividade dessas enzimas. Os resultados apontaram ...

Sensores de zinc para aplicaciones en neurociencias

OpenAIRE

Estela Falla, Aldo David; Estela Falla, Aldo David; Estela Falla, Aldo David

2011-01-01

En los últimos años, los estudios realizados con respecto a la química involucrada en los procesos neurológicos han logrado dar una idea de la importancia del zinc en las funciones nerviosas, así como también en el estudio de enfermedades neurodegenerativas tales como la Enfermedad de Alzheimer. A nivel biológico, el zinc tiene representantes en los seis grupos de enzimas que existen, lo cual demuestra su gran importancia en los sistemas biológicos. En estas enzimas, el zinc como ion metál...

Tipos de pasteurização e agentes coagulantes na fabricação do queijo tipo prato

OpenAIRE

Ramos, Thaís de Melo

2013-01-01

O presente trabalho foi realizado com os objetivos de avaliar a influência do tipo de pasteurização pelos sistemas de aquecimento indireto high temperature short time, ou HTST, e injeção direta de vapor (IDV), e avaliar a influência de duas diferentes enzimas coagulantes na fabricação do queijo tipo prato. O delineamento experimental utilizado neste experimento foi um fatorial 2 x 2 x 5 inteiramente casualizado, sendo dois sistemas de pasteurização (HTST e IDV), duas enzimas coagulantes e cin...

Aplicación de la Transglutaminasa como ingrediente funcional para mejorar las propiedades tecnológicas en yogur y queso fresco

OpenAIRE

Miró Fuertes, Elisabet

2017-01-01

La transglutaminasa es una enzima que se encuentra de forma natural en la mayoría de tejidos animales y está relacionada con varios procesos biológicos. Esta enzima actúa únicamente en proteínas catalizando la reacción de formación de enlaces covalentes entre residuos de glutamina y lisina entre proteínas de distinto tipo y origen. El añadir transglutaminasa en nuevas formulaciones confiere al producto nuevas propiedades en la estructura y acabado de los mismos aumentando su valor añadido. En...

Dgroup: DG02549 [KEGG MEDICUS

Lifescience Database Archive (English)

Full Text Available DG02549 Chemical ... DGroup Rotigotine ... D05768 ... Rotigotine (JAN/USAN/INN) ... ATC code: N04BC09 Antiparkinson...ian, Dopamine D2 receptor agonist DRD2 [HSA:1813] [KO:K04145] Enzyme: SULT1A1 [HSA:

Treinamento aeróbio de natação aumenta a atividade de enzimas antioxidantes e o conteúdo de glicogênio no musculoesquelético de ratos

Directory of Open Access Journals (Sweden)

Roberto Carlos Vieira Junior

2013-06-01

Full Text Available INTRODUÇÃO: São escassos os estudos os quais verificaram os efeitos do exercício físico sobre os biomarcadores de estresse oxidativo e sua relação com o glicogênio muscular. OBJETIVO: O presente estudo verificou o efeito do treinamento aeróbio de natação (TAN sobre os biomarcadores de estresse oxidativo, conteúdo de glicogênio e crescimento celular no músculo esquelético de ratos. MÉTODOS: Dezoito ratos Wistar macho (60 dias foram divididos em dois grupos: Grupo Sedentário (SG; n = 10: ratos sedentários; e Grupo Treinado (TG; n = 8: ratos submetidos ao TAN (5,0 % do peso corporal, 1h/dia, 5X/semana, durante oito semanas. A atividade das enzimas antioxidantes (AEA superóxido dismutase (SOD; U/ml, catalase (CAT; µmol/min/100 mg e glutationa peroxidase (GPx; nmol/min/100 mg, bem como a concentração das substâncias que reagem ao ácido tiobarbitúrico (TBARs; nmolMDA/mg proteína foram determinadas no músculo gastrocnêmio direito. Os conteúdos de glicogênio (mg/100 mg, proteína (g/100 g e DNA (g/100 g foram mensurados no músculo gastrocnêmio esquerdo. RESULTADOS: A AEA foi maior no TG (CAT: 0,87 ± 0,04; SOD: 6,49 ± 0,045; GPX: 6,49 ± 0,52 quando comparados com SG (CAT: 0,52 ± 0,03; SOD: 4,1 ± 0,37; GPx: 2,94 ± 0,56. Os níveis de TBARs foram menores em TG (TG: 2,35 ± 0,41; SG: 8,90 ± 0,47. O conteúdo de glicogênio muscular (SG: 0,108 ± 0,013; TG: 0,320 ± 0,012 e a razão proteína/DNA (SG: 24,94 ± 3,25; TG: 41,68 ± 4,02 foram maiores no TG. CONCLUSÃO: Em conjunto, estes dados confirmam que o TAN melhorou a defesa antioxidante, a qual pode estar associada ao aumento do conteúdo de glicogênio no músculo esquelético dos animais.

Optimizing conditions for enzymatic extraction of sunflower oil

Directory of Open Access Journals (Sweden)

Badr, F. H.

1992-10-01

Full Text Available Sunflower seed oil was extracted with an enzymatic processes using different hydrolytic enzymes: cellulase, hemicellulase, animal proteinase, acid proteinase, pectinase and pectinex, as compared to enzyme - free aqueous extraction. All the hydrolytic enzymes enhanced oil extraction from sunflower seeds. The most optimal conditions for oil extraction from sunflower seeds were: 2% enzyme concentration, 30% substrate concentration and 3 hrs period. Using Boganov and Buchkov equation showed that time must be prolonged to get higher yields. The maximum yield during 3 hrs extraction with enzymatic process ranged between 44,5%-57,1% of the soxhlet extractable oil. The potency of the investigated enzymes in extracting oil was in the following order: acid proteinase > cellulase > hemicellulase > animal proteinase > pectinex > pectinase when compared at the previous optimal conditions.

Aceite de semilla de girasol fue extraído mediante un proceso enzimático usando diferentes enzimas hidrolíticos: celulasa, hemicelulasa, proteinasa animal, proteinasa acida, pectinasa y pectinex, comparando con la extracción acuosa libre de enzima. Todos los enzimas hidrolíticos incrementan la extracción de aceites de semilla de girasol. Las condiciones óptimas para la extracción de aceite a partir de semillas de girasol fueron: 2% de concentración de enzima, 30% de concentración de sustrato y un período de 3 horas. La ecuación de Boganov y Buchkov mostró que el tiempo debe ser prolongado para alcanzar altos rendimientos. El máximo rendimiento durante tres horas de extracción con proceso enzimático osciló entre el 44,5%-57,1% del aceite extraído con soxhlet. La potencia de los enzimas investigados en la extracción de aceite siguió el orden: proteinasa acida > celulasa > hemicelulasa > proteinasa animal > pectinex > pectinasa cuando fue previamente comparado con las condiciones óptimas.

BpuAmI: a novel SacI neoschizomer from Bacillus pumilus discovered in an isolate from Amazon Basin, recognizing 5'-GAG¯CTC-3' BpuAmI: um novo neoesquisomero de SacI de Bacillus pumilus descoberta em um isolado oriundo da Bacia Amazônica, reconhecendo 5'-GAG¯CTC-3'

Directory of Open Access Journals (Sweden)

Jocelei M. Chies

2006-03-01

Full Text Available A strain of Bacillus pumilus was isolated and identified from water samples collected from a small affluent of the Amazon River. Type II restriction endonuclease activity was detected in these bacteria. The enzyme was purified and the molecular weight of the native protein estimated by gel filtration and SDS-PAGE. The optimum pH, temperature and salt requirements were determined. Quality control assays showed the complete absence of "nonspecific nucleases." Restriction cleavage analysis and DNA sequencing of restriction fragments allowed the unequivocal demonstration of 5´GAG¯CTC3´ as the recognition sequence. This enzyme was named BpuAmI and is apparently a neoschizomer of the prototype restriction endonuclease SacI. This is the first report of an isoschizomer and/or neoschizomer of the prototype SacI identified in the genus Bacillus.Uma linhagem de Bacillus pumilus foi isolada e identificada de amostras de águas coletadas em um pequeno Igarapé do Rio Amazonas. Foi detectada atividade de restrição do tipo II nesta bactéria. A enzima foi purificada e o peso molecular da proteína nativa foi estimado por gel filtração e por eletroforese em gel de poliacrilamida. Foram determinados, o pH e temperatura ótimos e as necessidades de sais. Os ensaios do controle de qualidade mostraram uma ausência completa de "nucleases não específicas". As analises das clivagens e o seqüenciamento do DNA dos fragmentos de restrição permitiram uma demonstração inequívoca de que 5´GAG¯CTC 3´ é a seqüência de reconhecimento da enzima. Esta enzima foi denominada de BpuAmI e aparentemente é um neoesquisômero da enzima protótipo SacI. Este é o primeiro relato de um isoesquisômero e/ou neoesquisômero da enzima protótipo SacI identificada no gênero Bacillus.

Chitosanase production by Paenibacillus ehimensis and its application for chitosan hydrolysis

Directory of Open Access Journals (Sweden)

Maria Giovana Binder Pagnoncelli

2010-12-01

Full Text Available The chitosanase production by Paenibacillus ehimensis was studied in submerged cultures and the chitosan hydrolysis was evaluated by using these enzymes without purification. The bacterium produced inducibles enzymes after 12 h of growth in a culture medium containing 0.2% (w/v of soluble chitosan as carbon source. The enzyme production was strongly repressed by the presence of glucose. The production started as soon as the available sugars finished in the culture medium. The maximum level of chitosanase activity was 500 U.L-1 at 36°C after 36 h incubation. The crude enzyme was optimally active at pH 6.0 and 55°C and in these conditions, the enzyme presented good stability (6 days. The enzyme without purification was used to hydrolyze the chitosan which resulted chitooligosaccharides between 20 and 30 min of reaction.A produção de quitosanases pelo Paenibacillus ehimensis foi estudada em culturas submersas e a hidrólise da quitosana foi realizada utilizando essas enzimas sem purificação. As enzimas foram obtidas após 12 horas de crescimento desta bactéria em meio de cultivo contendo 0,2% (p/v de quitosana solúvel como fonte de carbono. A produção das enzimas foi fortemente reprimida na presença de glicose, sendo obtida após o consumo total dos açúcares disponibilizados no referido meio de cultivo. A máxima atividade quitosanolítica foi obtida após 36 horas de cultivo a 36ºC, atingindo valores de 500 U.L-1. As enzimas utilizadas no extrato bruto apresentaram melhores atividades quando submetidas a condições de pH e temperatura de 6,0 e 55ºC, respectivamente, e nessas condições permaneceram estáveis durante 6 dias. Essas enzimas sem serem submetidas aos processos de purificação foram utilizadas para hidrolisar a quitosana, obtendo os quito-oligossacarídeos entre 20 e 30 minutos de reação.

Relationship of socio-economic status and childhood cancer: an in ...

African Journals Online (AJOL)

2017-11-30

Nov 30, 2017 ... Social and economic factors are known to affect health and well-being in several ... -seeking attitudes in childhood cancer; this usually re- sults in late ..... reported in guardians of 5.0% in Kenya and 19% in. Uganda.20 This ...

evaluation of a modified passive solar housing system for poultry

African Journals Online (AJOL)

User

The hourly efficiency of the solar brick passive system was estimated at about 78.42% in a day of May and ... to high cost and unavailability of kerosene in most developing .... sulted in intermittent rainfall, cloud cover and sunshine. From the ...

Estudo e determinação teórica de propriedades físico-químicas de fármacos anti-hipertensivos inibidores da Enzima Conversora de Angiotensina

Directory of Open Access Journals (Sweden)

LUíZA REIS CRUZ

2010-12-01

Full Text Available O pacote de softwares SpartanTM vs 04 foi utilizado para calcular algumas propriedades físico-químicas de uma série de inibidores da Enzima Conversora de Angiotensinogênio ECA. Simultaneamente, o pacote MarvinSketch 5.0.0 foi empregado para calcular o Coeficiente de Partição P dos inibidores. Através da análise dos resultados, conclui-se que: a o arranjo termodinâmico mais estável desses inibidores mimetiza aquele dos resíduos pré-terminais da angiotensina II, b há grande similaridade entre as cargas dos átomos que se ligam à macromolécula e c os cálculos mostram diferenças significativas entre os valores de P para os inibidores. Portanto, as diferenças farmacológicas existentes entre os inibidores estão mais intimamente relacionadas ao coeficiente de partição do que à capacidade destes de inibir o sítio ativo da ECA. Palavras-chave: ECA. Ab initio. Equilíbrio conformacional. Lipofilicidade. ABSTRACT Theoretical study and determination of physicochemical properties of antihypertensive angiotensin-converting enzyme inhibitors The software package SpartanTM 4.0 was employed to calculate some physicochemical properties of a series of available ACE inhibitors. Simultaneously, the program MarvinSketch 5.0.0 was employed to calculate the partition coefficients (P of the same compounds. After analyzing the results, we conclude that: a from a thermodynamic point of view, the most stable conformer of each inhibitor resembles, as expected, the most stable spatial arrangement of the preterminal residues of angiotensin II; b there is great similarity among the charge profiles of the potential binding sites of all the inhibitors; c there are large differences in P among these compounds. Summing up, the pharmacological differences reported between the inhibitors are more closely linked to their lipophilic properties than to their capacity to block the ACE active center. Keywords: ACE. Ab initio. Conformational

INFLUENCE OF MOTOR CAPaBILITIES AND COGNITiVE CHARACTERISTICS ON SPEED OF LEADING THE BALL AND ACCURACY IN BASKETBALL

Directory of Open Access Journals (Sweden)

Rašid Hadžić

2007-05-01

Full Text Available In goal of for ti fi ca tion the sca le of in fl u en ce of mo tor ca pa bi li ti es and cog ni ti ve cha rac te ri stics on re sult suc cess in ball le a ding speed and si tu a te ac cu racy in ba sket ball re sults de ri ved from sam ple of 105 boys bet we en 14 and 16 years ha ve been analyzed. Re gres si ve analyze of cri te ria va ri a ble SMKVLS and va ri a ble SMKBVL gi ves suf fi ci ent in for ma tion abo ut in fl u en ce of ap plied mo tor and cog ni ti ve on suc cess of per for ming of tre a ted cri te ria va ri a ble

Impacts on silkworm larvae midgut proteomics by transgenic Trichoderma strain and analysis of glutathione S-transferase sigma 2 gene essential for anti-stress response of silkworm larvae.

Science.gov (United States)

Li, Yingying; Dou, Kai; Gao, Shigang; Sun, Jianan; Wang, Meng; Fu, Kehe; Yu, Chuanjin; Wu, Qiong; Li, Yaqian; Chen, Jie

2015-08-03

Lepidoptera is a large order of insects that have major impacts on humans as agriculture pests. The midgut is considered an important target for insect control. In the present study, 10 up-regulated, 18 down-regulated, and one newly emerged protein were identified in the transgenic Trichoderma-treated midgut proteome. Proteins related to stress response, biosynthetic process, and metabolism process were further characterized through quantitative real-time PCR (qPCR). Of all the identified proteins, the glutathione S-transferase sigma 2 (GSTs2) gene displayed enhanced expression when larvae were fed with Trichoderma wild-type or transgenic strains. Down regulation of GSTs2 expression by RNA interference (RNAi) resulted in inhibition of silkworm growth when larvae were fed with mulberry leaves treated with the transgenic Trichoderma strain. Weight per larva decreased by 18.2%, 11.9%, and 10.7% in the untreated control, ddH2O, and GFP dsRNA groups, respectively, at 24h, while the weight decrease was higher at 42.4%, 28.8% and 32.4% at 72 h after treatment. Expression of glutathione S-transferase omega 2 (GSTo2) was also enhanced when larvae were fed with mulberry leaves treated with the transgenic Trichoderma strain. These results indicated that there was indeed correlation between enhanced expression of GSTs2 and the anti-stress response of silkworm larvae against Trichoderma. This study represents the first attempt at understanding the effects of transgenic organisms on the midgut proteomic changes in silkworm larvae. Our findings could not only broaden the biological control targets of insect at the molecular level, but also provide a theoretical foundation for biological safety evaluation of the transgenic Trichoderma strain. Copyright © 2015 Elsevier B.V. All rights reserved.

Genome-Wide Identification, Characterization, and Expression Profiling of Glutathione S-Transferase (GST) Family in Pumpkin Reveals Likely Role in Cold-Stress Tolerance

Science.gov (United States)

Abdul Kayum, Md.; Nath, Ujjal Kumar; Park, Jong-In; Choi, Eung Kyoo; Song, Jae-Young; Kim, Hoy-Taek; Nou, Ill-Sup

2018-01-01

Plant growth and development can be adversely affected by cold stress, limiting productivity. The glutathione S-transferase (GST) family comprises important detoxifying enzymes, which play major roles in biotic and abiotic stress responses by reducing the oxidative damage caused by reactive oxygen species. Pumpkins (Cucurbita maxima) are widely grown, economically important, and nutritious; however, their yield can be severely affected by cold stress. The identification of putative candidate genes responsible for cold-stress tolerance, including the GST family genes, is therefore vital. For the first time, we identified 32 C. maxima GST (CmaGST) genes using a combination of bioinformatics approaches and characterized them by expression profiling. These CmaGST genes represent seven of the 14 known classes of plant GSTs, with 18 CmaGSTs categorized into the tau class. The CmaGSTs were distributed across 13 of pumpkin’s 20 chromosomes, with the highest numbers found on chromosomes 4 and 6. The large number of CmaGST genes resulted from gene duplication; 11 and 5 pairs of CmaGST genes were segmental- and tandem-duplicated, respectively. In addition, all CmaGST genes showed organ-specific expression. The expression of the putative GST genes in pumpkin was examined under cold stress in two lines with contrasting cold tolerance: cold-tolerant CP-1 (C. maxima) and cold-susceptible EP-1 (Cucurbita moschata). Seven genes (CmaGSTU3, CmaGSTU7, CmaGSTU8, CmaGSTU9, CmaGSTU11, CmaGSTU12, and CmaGSTU14) were highly expressed in the cold-tolerant line and are putative candidates for use in breeding cold-tolerant crop varieties. These results increase our understanding of the cold-stress-related functions of the GST family, as well as potentially enhancing pumpkin breeding programs. PMID:29439434

Case-only study of interactions between metabolic enzymes and smoking in colorectal cancer

International Nuclear Information System (INIS)

Fan, Chunhong; Jin, Mingjuan; Chen, Kun; Zhang, Yongjing; Zhang, Shuangshuang; Liu, Bing

2007-01-01

Gene-gene and gene-environment interactions involved in the metabolism of carcinogens may increase the risk of cancer. Our objective was to measure the interactions between common polymorphisms of P450 (CYP1A2, CYP1B1, CYP2E1), GSTM1 and T1, SULT1A1 and cigarette smoking in colorectal cancer (CRC). A case-only design was conducted in a Chinese population including 207 patients with sporadic CRC. Unconditional logistic regression analysis was performed adjusting for age, gender, alcohol consumption, and cigarette smoking. The interaction odds ratio (COR) for the gene-gene interaction between CYP1B1 1294G and SULT1A1 638A allele was 2.68 (95% CI: 1.16–6.26). The results of the gene-environment analyses revealed that an interaction existed between cigarette smoking and the CYP1B1 1294G allele for CRC (COR = 2.62, 95%CI: 1.01–6.72), the COR for the interaction of CYP1B1 1294G and smoking history > 35 pack-years was 3.47 (95%CI: 1.12–10.80). No other significant gene-gene and gene-environment interactions were observed. Our results showed that the interaction between polymorphisms in CYP1B1 1294G and SULT1A1*2 may play a significant role on CRC in the Chinese population. Also, it is suggested that the association between cigarette smoking and CRC could be differentiated by the CYP1B1 1294G allele

Effects of curcumin on cytochrome P450 and glutathione S-transferase activities in rat liver.

NARCIS (Netherlands)

Oetari, S.; Sudibyo, M.; Commandeur, J.N.M.; Samhoedi, R.; Vermeulen, N.P.E.

1996-01-01

The stability of curcumin, as well as the interactions between curcumin and cytochrome P450s (P450s) and glutathione S-transferases (GSTs) in rat liver, were studied. Curcumin is relatively unstable in phosphate buffer at pH 7.4. The stability of curcumin was strongly improved by lowering the pH or

Antioxidant and chemoprotective properties of Momordica charantia ...

African Journals Online (AJOL)

We have studied the effect of M. charantia, collected from Kazdaglari (Mount Ida) in Balikesir, fruit extract on glutathione S-transferases (GSTs), cytochrome P450s (CYPs), and antioxidant enzymes in rats. Male Wistar rats, aged 12 weeks and weighing 200-250 g, were given 200 mg M. charantia fruit extract per kg body ...

Ativação da enzima conversora de angiotensina no coração após infarto do miocárdio e suas repercussões no remodelamento ventricular

Directory of Open Access Journals (Sweden)

Mill José Geraldo

1997-01-01

Full Text Available OBJETIVO: Determinar as alterações de atividade da enzima conversora de angiotensina (ECA no coração com infarto do miocárdio (IM e comparar os efeitos do captopril e losartan em parâmetros morfológicos e funcionais de ratos com IM. MÉTODOS: O IM foi produzido em ratos Wistar por ligadura de ramos da artéria coronária esquerda. Os controles (Con foram submetidos a uma cirurgia fictícia. Animais com IM e Con foram tratados com captopril (30mg/kg/dia ou losartan (15mg/kg/dia e estudados 30 dias após, determinando-se a atividade da ECA nos ventrículos direito (VD e esquerdo (VE, as alterações hemodinâmicas e as concentrações de hidroxiprolina (OH-Pro e proteína total no VD e VE. RESULTADOS: A atividade da ECA aumentou no VD (+25% e VE (+70% após IM. A maior atividade foi observada na cicatriz fibrótica, onde atingiu cerca de 4,5 vezes a do músculo do VE que sobreviveu ao IM (420±68 vs 94±8nmoles/g/min; P<0,01. O IM determinou aumento da pressão diastólica final e hipertrofia do VD e VE. Captopril e losartan foram igualmente eficazes em atenuar a hipertrofia e o aumento da pré-carga. O captopril também atenuou o aumento de OH-Pro no VD e VE após IM. O IM reduziu a concentração de proteína principalmente no músculo de VE, efeito esse acentuado pelo captopril. CONCLUSÃO: A grande atividade da ECA na cicatriz deve produzir altas concentrações de angiotensina II (AII no sangue que drena da cicatriz. Os efeitos dos inibidores da ECA seriam decorrentes, principalmente, da redução de geração local de AII, e não de aumento de cininas, uma vez que captopril e losartan exerceram efeitos similares no remodelamento pós-infarto.

Molecular evolution of Theta-class glutathione transferase for enhanced activity with the anticancer drug 1,3-bis-(2-chloroethyl)-1-nitrosourea and other alkylating agents.

Science.gov (United States)

Larsson, Anna-Karin; Shokeer, Abeer; Mannervik, Bengt

2010-05-01

Glutathione transferase (GST) displaying enhanced activity with the cytostatic drug 1,3-bis-(2-chloroethyl)-1-nitrosourea (BCNU) and structurally related alkylating agents was obtained by molecular evolution. Mutant libraries created by recursive recombination of cDNA coding for human and rodent Theta-class GSTs were heterologously expressed in Escherichia coli and screened with the surrogate substrate 4-nitrophenethyl bromide (NPB) for enhanced alkyltransferase activity. A mutant with a 70-fold increased catalytic efficiency with NPB, compared to human GST T1-1, was isolated. The efficiency in degrading BCNU had improved 170-fold, significantly more than with the model substrate NPB. The enhanced catalytic activity of the mutant GST was also 2-fold higher with BCNU than wild-type mouse GST T1-1, which is 80-fold more efficient than wild-type human GST T1-1. We propose that GSTs catalyzing inactivation of anticancer drugs may find clinical use in protecting sensitive normal tissues to toxic side-effects in treated patients, and as selectable markers in gene therapy. Copyright 2010 Elsevier Inc. All rights reserved.

Deterioro del pescado: Guía didáctica

OpenAIRE

Oliveira, C.; Dragonetti Saucero, J.P.; Friss, C.

2001-01-01

Después de la captura y muerte del pescado, éste sufre inmediatamente un deterioro, la velocidad de degradación es más elevada que la de otros tipos de carnes. Este proceso de degradación es llevado a cabo en una primera etapa, por enzimas propias del músculo del pescado y posteriormente por enzimas producidas por los microrganismos que ingresan al músculo. La velocidad de deterioro varía según las especies dependiendo de diversos factores, tales como tamaño, estado fisiológico, alimentación ...

Expresión de β-1,3-Glucanasas de Beauveria bassiana en cultivo con extracto de los fitopatógenos Peronospora variabilis y Fusarium oxysporum

OpenAIRE

Montoya Espinoza, W. Jhoel; Nolasco Cárdenas, Oscar P; Acuña Payano, Rosalyn K; Gutiérrez, Ana I. F

2016-01-01

El hongo Beauveria bassiana Vuill. es muy conocido por su capacidad entomopatógena, pero también existe referencias de ser un hongo antagonista, una de las posibles formas de su antagonismo es la antibiosis debido a la presencia de enzimas hidrolíticas en su genoma. Las principales enzimas expresadas contra fitopatógenos son las β-1,3-glucanasas, ya que la pared celular de los fitopatógenos como hongos y oomycetes está constituida en su mayoría por polímeros de β-1,3-glucanos. Se evaluó por q...

Efecto del grado de saturación de ácidos grasos en la dieta sobre diferentes actividades aminopeptidasas solubles y unidas a membrana en suero, tejido nervioso y otros tejidos periféricos

OpenAIRE

Segarra-Robles, Ana-Belén

2013-01-01

[ES]El nivel circulante o tisular de un determinado péptido, es el resultado de un equilibrio entre su secreción y degradación. Así, el estado funcional de estos péptidos bioactivos depende no sólo de su secreción sino también de su inactivación o procesamiento mediante la acción de enzima s proteolíticos tales como las aminopeptidasas (AP). Estos enzimas juegan un importante papel en la regulación del Sistema Renina Angiotensina (SRA), de las hormonas oxitocina y vasopresina y de la TRH. Es ...

Importância do zinco na nutrição humana

OpenAIRE

Mafra Denise; Cozzolino Sílvia Maria Franciscato

2004-01-01

Recentes pesquisas experimentais e clínicas têm reforçado a importância do zinco na saúde humana. O zinco possibilita várias funções bioquímicas, pois é componente de inúmeras enzimas, dentre estas, álcool desidrogenase, superóxido dismutase, anidrase carbônica, fosfatase alcalina e enzimas do sistema nervoso central. Participa na divisão celular, expressão genética, processos fisiológicos como crescimento e desenvolvimento, na transcrição genética, na morte celular, age como estabilizador de...

Descoloração de efluentes têxteis sintéticos por catálise enzimática

OpenAIRE

Silva, Maria Cristina; Corrêa, Angelita D.; Amorim, M. T. Pessoa de; Oliveira, Sara L. C.; Duarte, António A. L. Sampaio; Torres, Juliana A.

2010-01-01

Peroxidases podem ser utilizadas em processos de descoloração no tratamento de efluentes têxteis. Há um interesse crescente por novas fontes desta enzima, e por processos de obtenção viáveis economicamente.1Neste trabalho o potencial de descoloração de efluentes têxteis sintéticos por peroxidase extraída de nabo (Brassica campestre ssp. rapifera) é avaliado. Para a obtenção da enzima de baixa pureza utiliza-se um processo de baixo custo, Os efluentes utilizados nos ensaios são efluentes sinté...

Estudo de produção enzimatica da dextrana clinica

OpenAIRE

Juan Heraldo Viloche Bazan

1993-01-01

Resumo: Este trabalho consiste na obtenção "in vitro" de dextrana clínica a partir da sacarose utilizando a enzima dextrana-sacarase obtida pela fermentação do Leuconostoc mesenteroides linhagem NRRL B512-F. A dextrana clínica, de peso molecular médio 40.000 daltons, tem sua aplicação consagrada como expansor volumétrico de sangue humano. A enzima foi produzida por fermentação e purificada usando ultrafiltração, separada com o uso de polietilenoglicol de peso molecular 1500 daltons e estocada...

Estudo das condições de hidrolise acida para obtenção de dextrana clinica

OpenAIRE

Vanessa Mendes Santos

1996-01-01

Resumo: A dextrana é um biopolímero formado por resíduos de glicose unidos por ligação et-1,6 maioritariamente. É sintetizada peia enzima dextrana-sacarase, enzima esta produzida extracelularmente por algumas bactérias, principalmente do genêro Leuconosíoa A apiicacão deste polímero está diretamente relacionada com o seu peso molecular- As dextranas de peso molecular na faixa de 20,000 a 100,000 daltons (dextrana clínica) possui notáveis aplicações no ramo da medicina como expansor do plasma ...

Inmovilización de transaminasas y amonio liasas y su aplicación en síntesis de compuestos aminados

OpenAIRE

Cárdenas Fernández, Anthony Max

2012-01-01

El objetivo del presente trabajo de tesis es establecer métodos biocatalíticos para la síntesis de los aminoácidos L-fenilalanina, L-aspartato, β−aminobutirato y de las aminas aromáticas 1-feniletilamina y 3-amino-1-fenilbutano, usando biocatalizadores con actividad transaminasa y amonio liasa inmovilizados por técnicas de formación de enlaces covalentes (en el caso de enzimas) y por atrapamiento (tanto para enzimas como para células). Se establecieron dos métodos enzimáticos para la síntesis...

Uso da papaina como potencializadora da penetração cutanea de diclofenaco dietilamonio em pomada

OpenAIRE

Maria Lucia Reque

2006-01-01

Resumo: A barreira dérmica é representada por uma estrutura lipoprotéica e constitui um ambiente bioquímico altamente complexo. Trata-se de uma barreira muito eficiente ao ingresso de agentes químicos. Não há na literatura nenhum relato do uso de enzimas proteolíticas como agente potencializador da permeação de fármacos presentes em formulações para uso tópico. Neste trabalho desenvolvemos uma formulação de pomada contendo a enzima para potencializar a penetração de diclofenaco dietilamônio n...

Sacarificación enzimática para la producción de jarabe glucosado

OpenAIRE

Flórez Franco, Olga Elena; Uribe Restrepo, Nora Dabeiba

2001-01-01

En este trabajo se evaluó la producción de jarabe glucosado a partir de la sacarificación de maltodextrina con enzima glucoamilasa libre, para tres concentraciones de maltodextrina (20, 40 y 60 g/l) y tres concentraciones de enzima glucoamilasa (0,75000; 0,43125 y 0,11250 AGU/ml), con un diseño experimental tres factorial, a temperatura de 60 ºC y pH 4,3. El propósito era encontrar las concentraciones óptimas de producción de glucosa, teniendo como variables de respuesta el equivalente de dex...

Hemo-oxigenasa 1: un promisorio blanco terapéutico

OpenAIRE

Sánchez, Carlos; Rodeiro, Idania; Garrido-Garrido, Gabino; Delgado, René

2005-01-01

La enzima hemo-oxigenasa es la principal enzima implicada en el catabolismo del grupo hemo y da lugar a tres productos fundamentales: biliverdina, el hierro libre y el monóxido de carbono. Fue descubierta a principios de la década del 60, pero no fue hasta mediado de los años ´80 donde empezó a estudiarse con detenimiento y se determinó que existía una isoforma inducible, denominada hemo oxigenasa-1. Esta proteína juega un papel muy importante en la modulación de procesos inflamatorios y eso ...

Two new polyoxovanadate clusters templated through cysteamine

Indian Academy of Sciences (India)

Unknown

variably formed.6–10 These may be salts between organic cations and discrete ... Vanadium pentoxide (V2O5), sodium hydroxide. (NaOH) and cysteamine were .... sulting in 2D layers and other nitrogen (N1) forming. H-bonding to one water ...

Using FML and fuzzy technology in adaptive ambient intelligent environments

NARCIS (Netherlands)

Acampora, G.; Loia, V.

2005-01-01

Ambient Intelligence (AmI, shortly) gathers best re-sults from three key technologies, Ubiquitous Computing, Ubiq-uitous Communication, and Intelligent User Friendly Inter-faces. The functional and spatial distribution of tasks is a natu-ral thrust to employ multi-agent paradigm to design and

Production and properties of alpha-amylase from thermophilic Bacillus sp. Produção e propriedades de alfa-amilase de Bacillus sp. termofílico

Directory of Open Access Journals (Sweden)

Carlos Alberto Martins Cordeiro

2002-01-01

Full Text Available alpha-amylase (1,4-alpha-D-glucan glucanohydrolase, EC 3.2.1.1 production by thermophilic Bacillus sp strain SMIA-2 cultivated in liquid media containing soluble starch reached a maximum at 48h, with levels of 57U/mL. Studies on the a-amylase characterization revealed that the optimum temperature for activity was 70ºC. The enzyme was stable for 2h at 50ºC, while at 60ºC, 70ºC and 90ºC, 4%, 13% and 38% of the original activities were lost, respectively. The optimum pH of the enzyme was 7.5. After incubation of crude enzyme solution for 24h at pH 7.5, a decrease of about 5% of its original activity was observed. The enzyme was strongly inhibited by Co2+, Cu2+ and Ba2+, but less affected by Ca2+, Mg2+, Ni2+, Sr2+ and Mn2+. The enzyme in 1M and 5M NaCl solutions the enzyme retained 70% and 47% of the original activity after 24h of incubation at 4ºC, respectively.A produção de alfa-amilase (1,4-alfa-D-glicano glicanohidrolase, EC 3.2.1.1 por um Bacillus sp cepa SMIA-2 cultivado em meios líquidos contendo amido solúvel, alcançou o máximo em 48h com níveis de 57U/mL. Estudos sobre a caracterização de alfa-amilase revelaram que a temperatura ótima de atividade desta enzima foi 70ºC. A enzima foi estável por 2h a 50ºC, enquanto que a 60ºC, 70ºC e 90ºC, 4%, 13% e 38% da atividade original foram perdidas, respectivamente. O pH ótimo da enzima foi 7,5. Após a incubação da enzima bruta por 24h a pH 7,5 observou-se um decréscimo em torno de 5% de sua atividade original. A enzima foi fortemente inibida por Co2+, Cu2+ e Ba2+, mas foi menos afetada por Ca2+, Mg2+, Ni2+, Sr2+ e Mn2+. Em solução de NaCl 1M e 5M, a enzima reteve 70% e 47% da sua atividade original após 24h a 4ºC, respectivamente.

Navodnenija v umah zactroishtshikov / Maris Kuuda

Index Scriptorium Estoniae

Kuuda, Maris

2007-01-01

Kohalikel omavalitsustel pole piisavalt hoobasid sundida arendajaid üleujutusohuga arvestama. Pärnusse hotelli September rajajad (arhitektid Emil Urbel, Andrus Mark) arvestavad ujutusohuga. Arvamust avaldavad Rene Reisner, Mari Sepp, Ülo Sults, Karri Tiigisoon, Indrek Rannik, Mark Soosaar, Tiiu Pärn, Merle Looring, Rita Sepp, Kaspar Alles

Análise da atividade da enzima conversora da angiotensina na hipertrofia aguda do ventrículo direito em modelo experimental de estenose endovascular ajustável do tronco pulmonar

Directory of Open Access Journals (Sweden)

RABELLO Renato Rocha

2001-01-01

Full Text Available INTRODUÇÃO: A bandagem do tronco pulmonar (TP tem sido aplicada para treinamento do ventrículo esquerdo (VE em pacientes portadores de transposição das grandes artérias (TGA com septo íntegro. Este procedimento, além de apresentar alta morbi-mortalidade, pode ocasionar alterações da função ventricular a longo prazo. Com o objetivo de analisar a hipertrofia aguda do ventrículo direito (VD, foi implantado um cateter balão no TP de seis cabritos jovens. MATERIAL E MÉTODOS: A sobrecarga sistólica foi aplicada através de insuflações progressivas do balão, durante 96 horas. Esta hipertrofia foi acompanhada por medidas hemodinâmicas diárias, através de cateteres implantados na aorta, VD e TP, além de ecocardiogramas seriados a cada 24 horas, com medidas das espessuras do septo interventricular e dos ventrículos. Ao final das 96 horas, os animais foram mortos para remoção dos corações. Os ventrículos e o septo foram pesados separadamente. Foram colhidas biópsias musculares de cada câmara para análise da atividade da enzima conversora da angiotensina (ECA. Oito cabritos (idade e peso semelhantes foram utilizados como controle para os pesos dos ventrículos e para a atividade da ECA. RESULTADOS: Observou-se um aumento do gradiente VD/TP (p=0,001, com conseqüente aumento da razão VD/VE (p=0,005 durante o tempo de sobrecarga sistólica. Ao fim do protocolo, a parede livre do VD apresentou aumento de espessura (p=0,002 e, conseqüentemente, um aumento do peso indexado (p=0,002. A análise da atividade da ECA revelou aumento somente no músculo do VD hipertrofiado (p=0,002. CONCLUSÃO: O cateter balão foi eficiente em induzir a hipertrofia aguda do VD através do protocolo utilizado. Conseqüentemente, um aumento expressivo da atividade da ECA está associado ao processo de hipertrofia miocárdica induzida por sobrecarga pressórica.

Properties of an amylase from thermophilic Bacillus SP. Propriedades de uma amilase de um termofílico Bacillus sp

Directory of Open Access Journals (Sweden)

Raquel Vieira de Carvalho

2008-03-01

Full Text Available alpha-Amylase production by thermophilic Bacillus sp strain SMIA-2 cultivated in liquid cultures containing soluble starch as a carbon source and supplemented with 0.05% whey protein and 0.2% peptone reached a maximum activity at 32 h, with levels of 37 U/mL. Studies on the amylase characterization revealed that the optimum temperature of this enzyme was 90ºC. The enzyme was stable for 1 h at temperatures ranging from 40-50ºC while at 90ºC, 66% of its maximum activity was lost. However, in the presence of 5 mM CaCl2, the enzyme was stable at 90ºC for 30 min and retained about 58% residual activity after 1 h. The optimum pH of the enzyme was found to be 8.5. After incubation of enzyme for 2 h at pH 9.5 and 11.0 was observed a decrease of about 6.3% and 16.5% of its original activity. At pH 6.0 the enzyme lost about 36% of its original activity. The enzyme was strongly inhibited by Co2+, Cu2+ and Ba2+, but less affected by Mg2+, Na+ and K+. In the presence of 2.0 M NaCl, 63% of amylase activity was retained after 2 h incubation at 45ºC. The amylase exhibited more than 70% activity when incubated for 1 h at 50ºC with sodium dodecyl sulphate. However, very little residual activity was obtained with sodium hypochlorite and with hydrogen peroxide the enzyme was completely inhibited. The compatibility of Bacillus sp SMIA-2 amylase with certain commercial detergents was shown to be good as the enzyme retained 86%, 85% and 75% of its activity after 20 min incubation at 50ºC in the presence of the detergent brands Omo®, Campeiro® and Tide®, respectively.A produção de alfa-amilase por um termofilico, Bacillus sp SMIA-2, cultivado em meio líquido contendo amido solúvel como fonte de carbono, alcançou uma atividade máxima de 37 U/mL em 32 horas. Estudos sobre a caracterização da amilase revelaram que a temperatura ótima desta enzima foi 90ºC. A enzima foi estável por 1 hora a temperaturas de 40 e 50ºC enquanto a 90ºC, 66% da atividade

Quality assessment of Moringa concanensis seed oil extracted through solvent and aqueous-enzymatic techniques

Directory of Open Access Journals (Sweden)

Anwar, Farooq

2008-03-01

Full Text Available The composition and quality of the M. concanensis seed oil extracted through an aqueous-enzyme-assisted technique, using three commercial enzyme-mixtures (Natuzyme, Kemzyme, and Feedzyme was compared to those of the control-, (without enzymes and solvent-extracted oils. Aqueous enzyme-extracted M.concanensis seed oil content ranged from 23.54 to 27.46% and was significantly (P 0.05 variation in the contents of fiber and ash within the three extraction methods. However, the protein content of the meal obtained through the aqueous-enzyme and control methods was significantly (P M. concanensis seed oils extracted using the three methods. The specific extinctions at 232 and 270 nm, peroxide value, p-anisidine, free fatty acid contents and color values of the aqueous-enzyme-extracted oil were found to be lower than that of solvent-extracted oil and thus revealed good quality. The oils extracted through the three methods exhibited no significant (P En este estudio se compara la composición y la calidad del aceite de semilla de M. concanensis extraído mediante enzimas, utilizando tres enzimas comerciales (Natuzyme, Kemzyme, y Feedzyme con las de un control extraído sin enzimas y con las del aceite extraído con disolvente. El contenido en aceites de las semilla extraídas con enzimas osciló entre 23,54 a 27,46% y fue significativamente más elevado (P 0,05 en el contenido de fibra y ceniza para los tres métodos de la extracción. Sin embargo, el contenido proteínico de la harina obtenido por métodos enzimáticos y el control sin enzimas fue significativamente menor (P < 0,05 que el de la harina obtenida después de la extracción por disolvente. Las diferencias en el índice de yodo (67.1-68.0 g /100 g of oil, densidad en 24 °C (0,865-0,866 g/mL, índice de refracción a 40 °C (1,4622-1,4627 y fracción insaponificable (0,69-0,76 % no fueron significativamente diferentes para ninguna de las técnicas de extracción. Las extinciones espec

Influência do estrógeno e do interferon γ sobre a expressão da indoleamina-2,3-dioxigenase em cultura de células de placenta e embriões de ratas Wistar

Directory of Open Access Journals (Sweden)

Maria Letícia Baptista Salvadori

2015-09-01

Full Text Available Resumo: A indoleamina 2,3-dioxigenase (IDO é uma enzima responsável por catabolizar o aminoácido triptofano. Sua presença no ambiente uterino placentário está relacionada à tolerância imunológica ao semi-aloenxerto, pois impede a proliferação de células imunológicas maternas, seja pela falta do aminoácido, ou pela ação de alguns catabólitos oriundos da quebra do triptofano, como o ácido quinolínico, que é tóxico principalmente para os linfócitos T. Pouco se conhece sob a influência de substâncias (hormônios e citocinas presentes na interface materno fetal e a expressão dessa enzima. Por esta razão, formulou-se a hipótese de que hormônios e interleucinas presentes na região uteroplacentária poderiam exercer algum efeito na expressão da IDO. Células oriundas da interface materno fetal de ratas Wistar foram mantidas em cultivo, onde receberam suplementação com estradiol e interferon-γ. A expressão da enzima foi avaliada pela técnica de citometria de fluxo nos períodos de 4, 24 e 48 horas e confirmação da presença proteica por imuno-histoquímica. Os resultados mostraram um aumento na expressão de IDO após a adição de estrógeno (9,03±0,81/11,25±0,25 e interferon-γ (9,03±0,81/20,43±0,60. O efeito do interferon-γ já era esperado como relatado na literatura, contudo, a elevação da expressão da IDO pela adição do estrógeno constitui nova informação sobre possíveis mecanismos envolvidos na ativação da enzima. O melhor esclarecimento desses achados poderia contribuir para uma melhor compreensão da participação dessa enzima na tolerância materno-fetal e para uma futura modulação terapêutica da mesma.

Production of CGTase by a Bacillus alkalophilic CGII strain isolated from wastewater of a manioc flour industry Produção de CGTase por Bacillus alkalophilic CGII isolado de água residuária de uma fecularia de mandioca

Directory of Open Access Journals (Sweden)

Telma Luisa de Freitas

2004-09-01

Full Text Available GCTase production by a new strain of Bacillus alkalophilic CGII isolated from Brazilian wastewater of manioc flour industry was examined. The growth medium used was composed by 1.5% starch, 1.5% nitrogen and 1% Na2CO3. Higher activity was obtained with starch, maltodextrin and galactose. When glucose was added to the medium, no enzyme production was observed. High enzyme activity and growth were reached when aeration was increased (88.6 U/mL. The enzyme characterization showed an optimum pH and temperature 8.0 and 55ºC for starch hydrolyses, respectively. Mg+ and Ca++ showed small activation; however, Hg+ and Cu+ showed a strong enzyme inhibition.Estudou-se a produção de CGTase por uma nova cepa de Bacillus alkalophilic CGII, isolada de água residuária de uma fecularia de mandioca, durante cultivo em meio composto de 1,5% de amido, 1,5% de fonte de nitrogênio e 1% Na2CO3. A atividade enzimática foi alta quando se utilizou amido, maltodextrina e galactose como fontes de carbono. Quando se utilizou glicose no meio de cultivo não se observou produção da enzima. Atividade enzimática alta (88,6 U/mL e melhor crescimento foram obtidos quando se aumentou a aeração. A caracterização da enzima mostrou um pH ótimo de 8,0 e temperatura ótima de 55ºC sendo que a enzima sofreu uma pequena ativação por Mg+ e Ca++. A enzima foi fortemente inibida por Hg+ e Cu+.

OS AVANÇOS DO USO DA BROMELINA NA ÁREA DE ALIMENTAÇÃO E SAÚDE

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Lenice Freiman de OLIVEIRA

2009-07-01

Full Text Available

Ao longo de muitas décadas, o estudo da produção e aplicação de enzimas vem despertando interesse de muitos pesquisadores, sendo a bromelina uma enzima proteolítica, proveniente do abacaxi, de fácil extração e de muita utilidade na área de alimentação e na medicina. A extração dessa enzima pode ser feita em todas as partes do abacaxizeiro (polpa, caule, folha, casca e coroa, por meio de processo extrativo com precipitação em álcool ou acetona. Na alimentação, as utilidades mais comuns são no amaciamento de carnes, aproveitamento tecnológico de farinha de trigo de alto teor protéico para produção de pães e biscoitos, clarificação de cerveja, na indústria de óleos vegetais, de ovos desidratados e de leite de soja e na medicina como vermífugos, na cicatrização de ferimentos, como antiinflamatório vegetal, na inibição de células cancerígenas e outros .

RESPOSTAS DA ATIVIDADE ENZIMÁTICA HEPÁTICA E CEREBRAL EM FUNÇÃO DE DIFERENTES NÍVEIS DE RESTRIÇÃO ALIMENTAR: I – FOSFATASE

Directory of Open Access Journals (Sweden)

Olga Maria Mascarenhas Faria de Oliveira

2007-01-01

Full Text Available O estudo foi feito para determinar a influência de restrição alimentar estabelecida em vários níveis sobre a atividade enzimática de algumas enzimas. Foram alimentados ratos machos com dieta comercial ad libitum (grupo controle e com restrição de 30% e 40% (grupos experimentais. Após o período experimental que teve duração de 28 dias, os animais foram sacrificados e os fígados, cérebros e rins removidos, homogeneizados e submetidos a diferentes velocidades de centrifugação e, em cada etapa foram realizados estudos cinético-enzimáticos das referidas enzimas e dosagem protéica. Os resultados mostraram que a restrição alimentar afeta a quantidade das enzimas fosfatase alcalina e amilase, além do conteúdo protéico, levando a significativo aumento ou diminuição de acordo com o grau de restrição e com a fase do fracionamento centrífugo estudada. O mesmo observou-se para o exercício físico.

Image Analysis and Modeling

Science.gov (United States)

1975-05-01

place "subgraphs" with more complicated subgraphs. There have beer many re- sults reported which extend string-grammar theorems to web grammar...W. Bacus and E. E. Gose , "Leukocyte Pattern Recognition," IEEE SMC-2. No. 2, pp. 513-536, September 1972. [4] J. K. Hawkins

Notre Continent, Notre Avenir: Perspectives africaines sur l ...

International Development Research Centre (IDRC) Digital Library (Canada)

On ne sait pas très bien si la non-application effective résulte du refus ...... La main-d'œuvre est moins mobile au plan international que le capital, et donc ...... Auty, R., 1995, « Economic development and the resource curse thesis », dans ...

Some Elementary Examples from Newton's Principia -R-ES ...

Indian Academy of Sciences (India)

ing both differential and integral calculus. Newton used many geometrical methods extensively to derive the re- sults in spite of his having discovered calculus. Geome- try, judiciously used with limiting procedures, was one principal strategy used by Newton in the Principia. The Principia presents an enormous range of ...

Alterations of the Antioxidant Enzyme Activities are not General Characteristics of the Colonization Process by Arbuscular Mycorrhizal Fungi Alteraciones de las Actividades de Enzimas Antioxidantes no son Características Generales del Proceso de Colonización por Hongos Micorrízicos Arbusculares

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Yakelin Rodríguez

2012-09-01

antioxidante durante la colonización radical por los hongos micorrízicos arbusculares, se evaluaron las actividades de enzimas antioxidantes claves en raíces de tomate (Solanum lycopersicum L. inoculadas con seis cepas diferentes: dos Glomus mosseae, Glomus cubense, Glomus intraradices, Glomus sp. y A. scrobiculata. Las cepas G. cubense y A. scrobiculata alcanzaron los niveles de infectividad superiores con valores máximos de colonización e intensidad de 29-10,88% y 18-9,20%, respectivamente; las cepas G. mosseae mostraron una infectividad intermedia, ambas con 15% de colonización e intensidades máximas de 7,64-7,06%, respectivamente; mientras que los niveles de infectividad de las cepas Glomus sp. y G. intraradices fueron inferiores con colonización del 13% e intensidades de 5,07 y 5,41, respectivamente. Algunos patrones de actividad de las enzimas peroxidasa, superóxido dismutasa y polifenol oxidasa no resultaron específicos para los niveles y estadios de colonización, temprano o tardío, ni tipo de cepa. No obstante, la única banda de superóxido dismutasa presente en la colonización temprana y el bajo nivel de actividad guayacol-peroxidasa en estadio tardío presente en todos los tratamientos micorrizados indican que estas respuestas antioxidantes son independientes de la cepa y el grado de colonización. En conjunto, los datos sugieren que las alteraciones de las actividades enzimáticas antioxidantes no son características generales del proceso de colonización por los hongos micorrízicos arbusculares, teniendo probablemente el rol clave sobre estas respuestas las características específicas de cada cepa más que la colonización per se.

Produção, caracterização bioquímica e aplicação de L-asparaginase fúngica

OpenAIRE

Fernanda Furlan Gonçalves Dias

2016-01-01

Resumo: L-Asparaginases são enzimas que catalisam a hidrólise da L-asparagina a ácido L-aspártico e amônia. As L-asparaginases microbianas têm recebido atenção devido ao seu papel no tratamento da leucemia linfoblástica aguda e devido à sua crescente aplicação na indústria de alimentos visando à redução da formação de acrilamida durante o processamento de alimentos. Um número crescente de pesquisas tem estudado a aplicação dessa enzima em diferentes matrizes alimentares comprovando seus efeit...

Estudio de pacientes colombianos con homocistinuria

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M. Bermúdez

2001-07-01

Full Text Available La homocistinuria es una alteración bioquímica producida por causas genéticas como son la deficiencia de alguna de las enzimas involucradas en el metabolismo de metionina u homocisteína: Cistationina b sintasa, 5-10 Metilentetrahidrofolato reductasa, o Metionina sintasa al igual que a deficiencias nutricionales y asocio a otras patologías como son la enfermedad renal y problemas  vasculares. El objetivo de este trabajo es la identificación de la deficiencia bioquímica en pacientes con homocistinuria, mediante la cuantificación de las enzimas involucradas, utilizando como muestras fibroblastos y linfocitos.

Purificación y determinación de actividad enzimática de la catalasa en Staphylococcus aureus

OpenAIRE

Iván Marin Arguello; lan Roustan-Espinoza.

2000-01-01

Ampliamente distribuida en células animales, vegetales y micro- organismos, la catalasa es una enzima crucial para la vida. Su función catalítica es la descomposición del peróxido de hidrógeno (HOY), compuesto altamente tóxico y generado en los procesos metabólicos de la célula. Esta enzima en Staphylococcus aureus es una proteína tetramérica presentando grupos hemos y moléculas de NADPH en su estructura. La determinación espectrofotométrica de los parámetros cinéticos (Km, Vmax. y otros) en ...

Mecanismos que analizan a nivel hepático el destino de los ácidos grasos del plasma y su relación con la resistencia insulínica en dislipemia experimental. Estrategias de prevención y/o mejoramiento por dietas

OpenAIRE

Hein, Gustavo Juan

2011-01-01

Los resultados alcanzados demuestran que la administración crónica de una dieta rica en sacarosa (DRS) indujo en hígado: incremento en la masa proteica del receptor nuclear LXR alfa y disminución en la masa del receptor PPAR alfa. Esto se relacionó con la activación de las enzimas lipogénicas y menor actividad de las enzimas de la oxidación de ácidos grasos, indicando un destino preferencial de éstos hacia la vía de reesterificación y síntesis de triglicéridos hepáticos. Los animales dislipid...

Estudo da sintese enzimatica de dextrana na presença de maltose como aceptor

OpenAIRE

Sueli Rodrigues

2003-01-01

Resumo: O principal objetivo do trabalho foi o estudo de um processo para obtenção de dextrana de massa molar controlada, via síntese enzimática, para com isso reduzir os custos da obtenção de dextrana clínica. Para isso foi considerada a modelagem de um reator contínuo com re-aproveitamento da enzima através da adsorção em resinas trocadoras de íons. Entretanto, os ensaios de adsorção demonstraram que a presença de dextrana impede a adsorção da enzima e seu re-aproveitamento a...

Estudo de sistema bifasico duplo estagio para produção de dextrana

OpenAIRE

Isabel Cristina Baddini Curralero

2000-01-01

Resumo: A dextrana, um homopolissacarídeo de origem bacteriana, possui grande número de aplicações na indústria química, alimentícia e farmacêutica. Neste trabalho foi estudado um processo contínuo, de produção de dextrana, que permite o reciclo da enzima dextranasacarase. Neste sistema a enzima é adsorvida em resina de troca iônica (DEAE CELULOSE) e mantida em recirculação entre dois estágios. No primeiro, se dá a dessorção da dextranasacarase, pela alimentação de solução de NaCl, e a reação...

Extracción del ADN de Fusarium Oxysporum f.sp. Dianthi

OpenAIRE

Sixta T. Martínez; Carlos Y. Soto

2010-01-01

Se estudia la utílización de dos métodos para la extracción del ADN del Fusarium o.xy.sporum f.sp. Dianthi. En los dos métodos la pared del hongo se rompió con nitrógeno líquido, uno de ellos empleó como solución extractora bromuro de cetiltrimetil amonio (BCTA) y el otro una solución de sacarosa con altas concentraciones de protcasa y EDTA. Para la desproteinización ambos métodos utilizaron soluciones de fcnol-clorofonno y enzimas proteolítícas. El ADN obtenido se digirió con enzimas d...

Isolamento e caracterização de um mutante de saccharomyces cerevisiae com características fenotípicas opostas à cepa pkc

OpenAIRE

Gomes, Katia das Neves

2004-01-01

Em leveduras, a proteína quinase C participa da regulação da via bioquímica responsável pela transcrição de uma subunidade da enzima glucano sintase, a qual está envolvida na síntese da parede celular. A via PKC MAP quinase consiste das enzimas Bck1, Mkk1/2 e Mpk1 que são ativadas por fosforilação. Recentemente, nós descobrimos que o mutante pkc1 D, contrariamente aos demais mutantes da cascata Map quinase, exibe dois principais defeitos no controle do metabolismo de carbono. A cepa pkc1 D ap...

Enzimas carboidrases na dieta de porcas lactantes e suas leitegadas Carbohydrase enzymes in diets for lactating sows and their litters

Directory of Open Access Journals (Sweden)

Débora Cristina Nichelle Lopes

2009-12-01

Full Text Available O experimento foi realizado para se avaliar o uso de um complexo enzimático (CE contendo carboidrases na dieta de porcas lactantes e de suas leitegadas. Foram utilizadas 264 fêmeas cruzadas geneticamente homogêneas, que foram bloqueadas pela ordem de parto e distribuídas ao acaso em quatro tratamentos, resultantes de um arranjo fatorial 2x2 (dieta lactação (DL x dieta pré-inicial (DPI, com dois níveis cada fator (com e sem CE. Nas porcas, foram avaliados: peso ao desmame (PDESM; variação do peso corporal; espessura de toucinho (ET; variação de ET; escore corporal (EC; consumo de ração (CRF e composição do leite e nas leitegadas: consumo de ração (CRL; peso da leitegada ao nascer e ao desmame (PLDESM; e peso médio do leitão ao desmame (PMLDESM. Não houve interação significativa entre DL e DPI para nenhuma variável resposta. O PDESM, a ET, a variação de ET, o EC e a composição do leite (teores de gordura, proteína bruta, lactose e matéria seca das porcas não foram influenciados (P>0,10 pela adição de enzimas na DL e na DPI. O CRF e o CRL não foram influenciados (P>0,10 pelos tratamentos. O PLDESM e o PLMDESM foram significativamente maiores quando as porcas receberam DL suplementada com CE, em comparação àquelas que não receberam CE na dieta (74,6 vs 72,9kg e 6,32 vs 6,47kg, respectivamente. Houve um aumento de 1,7kg no PLDESM e de 0,15kg no PLMDESM (P=0,06 e P=0,07, respectivamente, resultando em um incremento estimado de 2,27kg de peso vivo quando os animais chegam ao peso de abate. Porcas alimentadas com rações suplementadas com complexo enzimático contendo carboidrases apresentam leitegadas mais pesadas ao desmame.This research was conducted to evaluate the effect of adding enzymatic complex (EC containing carbohydrases to diets for lactating sows and their litters. A total of 264 genetically homogeneous crossbred sows were used. They were blocked by farrowing order and randomly allotted to four

Utilização de enzimas proteolíticas para produção de hidrolisados proteicos a partir de carcaças de frango desossadas manualmente

Directory of Open Access Journals (Sweden)

Mari Silvia Rodrigues de OLIVEIRA

2015-09-01

Full Text Available Resumo O emprego de hidrolisados proteicos, oriundos de fontes animais e vegetais, em formulações específicas, é uma área de crescente interesse. O objetivo deste trabalho foi desenvolver diferentes hidrolisados liofilizados com alto valor proteico, obtidos a partir da hidrólise enzimática de carcaças de frango manualmente desossadas (CMD, um subproduto da indústria avícola, que normalmente é utilizado para a fabricação de carne mecanicamente separada (CMS. A matéria-prima utilizada foram carcaças de frango desossadas manualmente e congeladas (CMD, provenientes de animais abatidos com aproximadamente 42 dias de vida e com peso médio de 2,5 kg, adquiridas em um abatedouro da região sul do Brasil. Antes de serem processadas, foram descongeladas sob temperatura de refrigeração e cortadas em pedaços menores com faca de aço inox para facilitar sua homogeneização durante o tempo de hidrólise. Foram utilizadas três enzimas comerciais, Papaína®, Flavourzyme® e Protamex®. A hidrólise ocorreu em banho termostatizado com temperatura, tempo e pH controlados. Foi realizada a composição proximal da matéria-prima e dos hidrolisados liofilizados, atividade de água dos hidrolisados liofilizados e foram feitas as seguintes análises de controle da hidrólise: grau de hidrólise, teores de proteínas, sólidos totais, cinzas, caracterização de aminoácidos dos hidrolisados, rendimento, percentual de hidrólise e cor dos hidrolisados. Os resultados foram submetidos à análise de variância e teste de Tukey para comparação de médias. O grau de hidrólise maior foi com a Protamex, seguido da Papaína e da Flavourzyme. O teor de proteínas após os 120 minutos de hidrólise não variou estatisticamente (p>0,05 entre a Papaína e a Flavourzyme. A composição de aminoácido demonstra que o hidrolisado obtido da Papaína possui uma composição mais próxima da recomendada pelos órgãos de controle. Concluiu-se que os

Theoretical Modeling of Ultrashot Laser Pulse Interaction With Dielectric and Semiconductor Materials

Science.gov (United States)

2010-05-24

of linearly polarized electromagnetic radiation re- sults in an anisotropic electronic transition rate characterized by field dependence via a Bessel...where Ω ( ~k, ~r ) is the renormalized Rabi frequency defined by: ~Ω ( ~k, ~r ) = µ ( ~k ) ~E (~r, t) + ∑ ~k′ p ( ~k′, ~r ) V s~k−~k′ . (12) The second

Fulltext PDF

Indian Academy of Sciences (India)

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So you should expect to ¯nd,say,a certain kind of duck on one ... w as beaten by a great m any people on various notions like the tree of ... sults that I think he w as truly original in are: 1. ..... appealing to `forerunners', heroic and successful, one.

Genetic Determinants of Gastric Cancer

NARCIS (Netherlands)

S. Boccia (Stefania)

2009-01-01

textabstractResults show that gastric cancer risk is increased by the inheritance of the variant alleles of the metabolic genes SULT1A1 and CYP2E1 *6, especially among smokers and drinkers, respectively. An additional increased risk is conferred by the inheritance of GSTT1 null variant, especially

Efeito da peletização e adição de enzimas e vitaminas sobre o desempenho e aproveitamento da energia e nutrientes em frangos de corte de 1 a 21 dias de idade Effect of pelleting and addition of enzymes and vitamins on the performance and advantage of energy and nutrients in broiler chickens from 1 to 21 days old

Directory of Open Access Journals (Sweden)

José Laureano Barbosa Leite

2008-08-01

Full Text Available Conduziu-se este experimento para avaliar o efeito da peletização e adição de enzimas e vitaminas sobre o desempenho e aproveitamento da energia e nutrientes, em frangos de corte de 1 a 21 dias de idade. Foram utilizados 350 pintos de corte, COBB, machos, com peso inicial médio de 47g ± 2,5g, em um delineamento inteiramente casualizado, com os tratamentos em arranjos fatorial 3 x 2 (três dietas, com e sem enzimas sendo que as dietas consistiram de uma ração farelada e duas rações peletizadas,uma com adição de vitaminas e minerais após a peletização e a outra com adição de vitaminas e minerais antes da peletização. O tempo e a temperatura da peletizadora variaram de 05 a 09 minutos e 58ºC a 62ºC, respectivamente. Após esse processo, as rações peletizadas foram trituradas, buscando-se manter a mesma granulometria das rações fareladas. O ensaio de desempenho foi realizado com 10 repetições e o de metabolismo com 5 repetições cada tratamento, em uma densidade de 5 aves por parcela (gaiola. As rações foram formuladas com 95% das recomendações nutricionais e fornecidas às aves com uma restrição alimentar de, aproximadamente, 10% do consumo ad libitum, o qual foi calculado diariamente a partir do consumo médio de um grupo de aves controle, que foram submetidas ao consumo à vontade de uma ração formulada para atender 100% dos níveis nutricionais recomendados. O complexo enzimático utilizado foi o Allzyme Vegpro®, contendo em sua composição as enzimas protease, amilase e celulase, adicionado aos tratamentos seguindo as recomendações do fabricante, de 0,5 kg por tonelada de ração. Aos 21 dias de idade avaliou-se o ganho de peso e a conversão alimentar. Também foram coletadas as excretas, pela metodologia de coleta total de excretas, nos dias 20, 21 e 22, para determinação dos valores energéticos das rações e digestibilidade da gordura e da proteína. Pelos resultados obtidos, observou-se que o

As enzimas no quotidiano: contextos de aprendizagem

OpenAIRE

Nunes, Ana Margarida Martins

2010-01-01

O presente trabalho propõe-se divulgar um conjunto de actividades sobre o tema “Fermentação e Actividade Enzimática”, desenvolvidas na disciplina de Biologia do 12ºano de escolaridade do Curso Científico-humanístico de Ciências e Tecnologias. Procurou-se demonstrar as vantagens da utilização do Jogo como oportunidade educativa de motivação e aprendizagem, através de actividades de Role-play, especialmente aplicadas ao trabalho prático de cariz experimental, no ensino das Ciências, uma área...

Novel function of N,N-bis(2-chloroethyl)docos-13-enamide for reversal of multidrug resistance in tongue cancer.

Science.gov (United States)

Qin, Qing; Ma, Peng-Fei; Kuang, Xiao-Cong; Gao, Ming-Xing; Mo, De-Huan; Xia, Shuang; Jin, Ning; Xia, Jun-Jie; Qi, Zhong-Quan; Lin, Cui-Wu

2013-12-05

Multidrug resistance (MDR) is a key element in the failure of chemotherapies, and development of agents to overcome MDR is crucial to improving cancer treatments. The overexpression of glutathione-S-transferases (GSTs) is one of the major mechanisms of MDR. Because some agents used in traditional Chinese medicine have strong antitumor effects coupled with low toxicity; we investigated the ability of N,N-bis(2-chloroethyl)docos-13-enamide (compound J), the synthesized analog of a highly unsaturated fatty acid from Isatis tinctoria L., to reverse the MDR induced by adriamycin (ADM) in TCA8113/ADM cells. We found that compound J significantly increased the cytotoxicity of ADM in TCA8113/ADM cells, with a reversal fold of 2.461. Analysis of the mechanisms through which compound J reversed MDR indicated that compound J significantly decreased the activity of GSTs and enhanced the depletion of GSH in TCA8113/ADM cells, but did not affect the P-glycoprotein (P-gp) efflux. Taken together, our data suggested that compound J was an excellent candidate for reversing MDR in cancer therapy. © 2013 Published by Elsevier B.V.

The Association between Gene-Environment Interactions and Diseases Involving the Human GST Superfamily with SNP Variants

Directory of Open Access Journals (Sweden)

Antoinesha L. Hollman

2016-03-01

Full Text Available Exposure to environmental hazards has been associated with diseases in humans. The identification of single nucleotide polymorphisms (SNPs in human populations exposed to different environmental hazards, is vital for detecting the genetic risks of some important human diseases. Several studies in this field have been conducted on glutathione S-transferases (GSTs, a phase II detoxification superfamily, to investigate its role in the occurrence of diseases. Human GSTs consist of cytosolic and microsomal superfamilies that are further divided into subfamilies. Based on scientific search engines and a review of the literature, we have found a large amount of published articles on human GST super- and subfamilies that have greatly assisted in our efforts to examine their role in health and disease. Because of its polymorphic variations in relation to environmental hazards such as air pollutants, cigarette smoke, pesticides, heavy metals, carcinogens, pharmaceutical drugs, and xenobiotics, GST is considered as a significant biomarker. This review examines the studies on gene-environment interactions related to various diseases with respect to single nucleotide polymorphisms (SNPs found in the GST superfamily. Overall, it can be concluded that interactions between GST genes and environmental factors play an important role in human diseases.

Glutathione transferase (GST) as a candidate molecular-based biomarker for soil toxin exposure in the earthworm Lumbricus rubellus

International Nuclear Information System (INIS)

LaCourse, E. James; Hernandez-Viadel, Mariluz; Jefferies, James R.; Svendsen, Claus; Spurgeon, David J.; Barrett, John; John Morgan, A.; Kille, Peter; Brophy, Peter M.

2009-01-01

The earthworm Lumbricus rubellus (Hoffmeister, 1843) is a terrestrial pollution sentinel. Enzyme activity and transcription of phase II detoxification superfamily glutathione transferases (GST) is known to respond in earthworms after soil toxin exposure, suggesting GST as a candidate molecular-based pollution biomarker. This study combined sub-proteomics, bioinformatics and biochemical assay to characterise the L. rubellus GST complement as pre-requisite to initialise assessment of the applicability of GST as a biomarker. L. rubellus possesses a range of GSTs related to known classes, with evidence of tissue-specific synthesis. Two affinity-purified GSTs dominating GST protein synthesis (Sigma and Pi class) were cloned, expressed and characterised for enzyme activity with various substrates. Electrospray ionisation mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) following SDS-PAGE were superior in retaining subunit stability relative to two-dimensional gel electrophoresis (2-DE). This study provides greater understanding of Phase II detoxification GST superfamily status of an important environmental pollution sentinel organism. - This study currently provides the most comprehensive view of the Phase II detoxification enzyme superfamily of glutathione transferases within the important environmental pollution sentinel earthworm Lumbricus rubellus.

Glutathione transferase (GST) as a candidate molecular-based biomarker for soil toxin exposure in the earthworm Lumbricus rubellus

Energy Technology Data Exchange (ETDEWEB)

LaCourse, E. James, E-mail: james.la-course@liverpool.ac.u [Institute of Biological, Environmental, and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA (United Kingdom); Hernandez-Viadel, Mariluz; Jefferies, James R. [Institute of Biological, Environmental, and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA (United Kingdom); Svendsen, Claus; Spurgeon, David J. [Centre for Ecology and Hydrology, Huntingdon PE28 2LS (United Kingdom); Barrett, John [Institute of Biological, Environmental, and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA (United Kingdom); John Morgan, A.; Kille, Peter [Biosciences, University of Cardiff, Cardiff CF10 3TL (United Kingdom); Brophy, Peter M. [Institute of Biological, Environmental, and Rural Sciences, Aberystwyth University, Aberystwyth SY23 3DA (United Kingdom)

2009-08-15

The earthworm Lumbricus rubellus (Hoffmeister, 1843) is a terrestrial pollution sentinel. Enzyme activity and transcription of phase II detoxification superfamily glutathione transferases (GST) is known to respond in earthworms after soil toxin exposure, suggesting GST as a candidate molecular-based pollution biomarker. This study combined sub-proteomics, bioinformatics and biochemical assay to characterise the L. rubellus GST complement as pre-requisite to initialise assessment of the applicability of GST as a biomarker. L. rubellus possesses a range of GSTs related to known classes, with evidence of tissue-specific synthesis. Two affinity-purified GSTs dominating GST protein synthesis (Sigma and Pi class) were cloned, expressed and characterised for enzyme activity with various substrates. Electrospray ionisation mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) following SDS-PAGE were superior in retaining subunit stability relative to two-dimensional gel electrophoresis (2-DE). This study provides greater understanding of Phase II detoxification GST superfamily status of an important environmental pollution sentinel organism. - This study currently provides the most comprehensive view of the Phase II detoxification enzyme superfamily of glutathione transferases within the important environmental pollution sentinel earthworm Lumbricus rubellus.

Structures of a putative ζ-class glutathione S-transferase from the pathogenic fungus Coccidioides immitis

International Nuclear Information System (INIS)

Edwards, Thomas E.; Bryan, Cassie M.; Leibly, David J.; Dieterich, Shellie H.; Abendroth, Jan; Sankaran, Banumathi; Sivam, Dhileep; Staker, Bart L.; Van Voorhis, Wesley C.; Myler, Peter J.; Stewart, Lance J.

2011-01-01

The pathogenic fungus C. immitis causes coccidioidomycosis, a potentially fatal disease. Here, apo and glutathione-bound crystal structures of a previously uncharacterized protein from C. immitis that appears to be a ζ-class glutathione S-transferase are presented. Coccidioides immitis is a pathogenic fungus populating the southwestern United States and is a causative agent of coccidioidomycosis, sometimes referred to as Valley Fever. Although the genome of this fungus has been sequenced, many operons are not properly annotated. Crystal structures are presented for a putative uncharacterized protein that shares sequence similarity with ζ-class glutathione S-transferases (GSTs) in both apo and glutathione-bound forms. The apo structure reveals a nonsymmetric homodimer with each protomer comprising two subdomains: a C-terminal helical domain and an N-terminal thioredoxin-like domain that is common to all GSTs. Half-site binding is observed in the glutathione-bound form. Considerable movement of some components of the active site relative to the glutathione-free form was observed, indicating an induced-fit mechanism for cofactor binding. The sequence homology, structure and half-site occupancy imply that the protein is a ζ-class glutathione S-transferase, a maleylacetoacetate isomerase (MAAI)

The influence of metabolic gene polymorphisms on urinary 1-hydroxypyrene concentration in Thai bus drivers.

Science.gov (United States)

Petchpoung, Krittaya; Kaojarern, Sming; Yoovathaworn, Krongtong; Sura, Thanyachai; Sirivarasai, Jintana

2011-01-01

Polycyclic aromatic hydrocarbons (PAHs) are associated with an increased cancer risk. CYP1A1 and GSTs enzymes are important in metabolism of PAHs. Genetic polymorphisms of these enzymes are responsible for enzyme activity and concentration variation. The objectives of this study were to evaluate association of 1-OHP concentration with genetic polymorphisms of CYP1A1 and GSTs in Thai bus drivers. The results showed that 1-OHP levels in bus drivers were significantly higher than that in the control group. Significant difference in 1-OHP was found between smokers and non-smokers, in only bus drivers. Significantly increasing of 1-OHP levels were observed in bus drivers with CYP1A1 MspI and exon 7 variants. Whereas, bus drivers with GSTP1 Val and GSTM1 null genotypes showed decreasing in excretion of 1-OHP. No association between 1-OHP and polymorphisms of GSTT1 was found. This study indicated that 1-OHP concentrations were associated with exposure to air pollution, cigarette smoking and polymorphisms of CYP1A1, GSTM1 and GSTP1 genes. Copyright © 2010 Elsevier B.V. All rights reserved.

Actividad enzimática del suelo -Deshidrogenasa , -ß Glucosidasa , Fosfatasa y Ureasa - bajo diferentes cultivos.

Directory of Open Access Journals (Sweden)

Carlos Henríquez

2014-01-01

Full Text Available Se analizó la actividad de 4 enzimas del suelo en fincas bajo diferentes manejos agronómi - cos y en diferentes tipos de suelos. Las enzimas evaluadas fueron la Fosfatasa, β-Glucosidasa, Deshidrogenasa y Ureasa. Se tomaron muestras de suelo de los primeros 20 cm de profundidad con el fin de analizarlas químicamente y determinar la actividad de las 4 enzimas a evaluar. Se encontró que la actividad de la Deshidrogenasa varió de 0,13 a 4,46 con un promedio de 1,17 (μg INTF.g - 1 .h -1 ; la β-Glucosidasa varió de 31,9 a 208,1 con un promedio de 108,1 (μg PNP.g -1 .h -1 . Por otro lado la Fosfatasa tuvo valores entre 413,4 y 3043,6 con un promedio de 1521,5 μg PNP.g -1 .h -1 , mientras que la Ureasa varió de 12,5 a 52,8 con un promedio de 38,3 μg N-NH4.g -1 .h -1 . La Fos - fatasa correlacionó con el porcentaje de materia orgánica y de carbono en el suelo (0,75 y 0,76 respectivamente, p<0,01; se encontró una rela - ción inversa entre la actividad de esta enzima y el contenido de P disponible (r de -0,53, p<0,10. La Ureasa correlacionó con la Fosfatasa (r de 0,61 p<0,05 y con la Deshidrogenasa (r de -0,77 p<0,01. El análisis multivariado de conglomera - dos a partir de la actividad enzimática permitió hacer grupos similares según el tipo de cultivo, lo que evidencia el potencial de utilización que tiene esta propiedad para realizar estudios más detallados.

Enzymatic oxidation of volatile malodorous organosulfur compounds in a two-phase reactor

Directory of Open Access Journals (Sweden)

Julio C. Cruz

2015-01-01

Full Text Available En este trabajo reportamos la oxidación de una serie de compuestos organoazufrados volátiles (COV catalizada por la enzima cloroperoxidasa obtenida del hongo Caldariomyces fumago . Los COV se consideran contaminantes atmosféricos debido a su olor desagradable y a su bajo umbral de detección. El sulfuro de etilo, disulfuro de dimetilo, propanotiol, butanotiol y hexanotiol fueron transformados por la enzima en un medio de reacción acuoso a pH 6 y en presencia de peróxido de hidrógeno. El análisis de los productos demostró que los sulfuros fueron oxidados a sus respectivos sulfóxidos, mientras que los tioles fueron oxidados a sus correspondientes disulfuros. Los productos identificados tienen una presión de vapor significativamente menor que los compuestos originales, por lo que son mucho menos volátiles y por tanto no se consideran contaminantes atmosféricos. Se ensambló un reactor de dos fases de 70 mL de volumen con el fin de determinar la eficiencia del tratamiento enzimático. La fase líquida, compuesta por 85% de amortiguador y 15% de solvente orgánico, se puso en contacto con la fase gaseosa, compuesta por aire enriquecido con el sustrato. Usando disulfuro de metilo como sustrato modelo, encontramos únicamente reacción enzimática en este sistema; al controlar la concentración de enzima y de peróxido en la fase líquida se logró transferir el sustrato a la fase acuosa en donde 1 mol de enzima convirtió aproximadamente 12,400 moles de sustrato, resaltando el potencial de los tratamientos enzimáticos para las corrientes gaseosas con mal olor por COV.

Identificación molecular y actividad sobre sustratos cromogénicos de la venombina A del veneno de la serpiente peruana Bothrops atrox

Directory of Open Access Journals (Sweden)

Gustavo A. Sandoval

2011-05-01

Full Text Available En el presente trabajo se ha realizado la identificación molecular de la enzima similar a trombina (EST del veneno de Bothrops atrox y se ha evaluado su actividad enzimática sobre diversos sustratos sintéticos. La enzima fue purificada utilizando tres pasos cromatrográficos, sobre Sephadex G-75, CM-Sephadex C-50 y Agarosa-PAB, determinándose su peso molecular por PAGE-SDS. La identificación molecular de la enzima aislada se realizó por la técnica de peptide mass fingerprinting basada en espectrometría de masas MALDI-TOF y posterior análisis in silico. Las actividades fibrinocoagulante y amidolítica fueron ensayadas sobre fibrinó- geno bovino y BApNA, respectivamente, así como la hidrólisis sobre los sustratos cromogénicos específicos S-2238, S-2251 y S-2266. Como resultado de los ensayos bioquímicos y estructurales, la EST del veneno de B. atrox, presentó un peso molecular de 29,6 kDa. El análisis mediante espectrometría de masas de los péptidos obtenidos, permitió identificar a esta enzima como una venombina A, presentando una identidad del 75%. Del análisis de actividad enzimática, se obtuvo que la EST de B. atrox produjo coagulación del fibrinógeno bovino y presentó actividad sobre BApNA, S-2238 y S-2266, siendo incapaz de hidrolizar el sustrato S-2251. El empleo de estas aproximaciones estructurales y funcionales ha permitido lograr la identificación molecular del principal componente del veneno de B. atrox relacionado con su acción coagulante, así como evaluar en detalle la naturaleza de su actividad enzimática sobre diversos sustratos.

AISLAMIENTO DE CLONES CON ACTIVIDAD ENDO-b-1,4-GLUCANASA A PARTIR DE UN SEGMENTO DE ADN DE 13KB DE Clostridium sp IBUN22A

Directory of Open Access Journals (Sweden)

I. Roncancio-Sánchez

2006-06-01

Full Text Available La producción de combustibles, solventes y algunos productos químicos a partir de sustratos celulósicos usando microorganismos ofrece una ventaja frente a los de origen fósil. Un acercamiento prometedor ha sido la implementación de ingeniería genética utilizando genes de enzimas involucradas en la degradación de desechos celulósicos. En la última década se han generado bibliotecas genéticas para proveer enzimas celulolíticas, que hagan el proceso más rentable, lo cual permitiría aprovechar mejor residuos celulósicos disponibles. Este trabajo describe el aislamiento de dos fragmentos de ADN que expresan actividad endo-β-1,4-glucanasa, a partir de un segmento ADN de 13Kb (clon 02080-25 aislado de una biblioteca genómica de la cepa nativa Clostridium sp IBUN22A. El aislamiento de la región codificadora se realizó a través de pruebas de inducción de la actividad, análisis por restricción del segmento y de una sub-biblioteca con la enzima Sau3A I. 325 clones fueron obtenidos, de los cuales 271 tenían inserto. El tamizaje molecular de estos últimos mostró que siete clones presentaron tamaños entre 3500pb y 7000pb y el tamizaje enzimático con carboximetilcelulosa como sustrato permitió el aislamiento de los clones pBSh-37 y pBSh-26 con la actividad endo-β-1,4-glucanasa original, de tamaños de inserto de 627pb y 879pb respectivamente. Este trabajo es el punto de partida para el aislamiento de enzimas de alto potencial biotecnológico.

Mitigation of Shore Damage Attributed to the Federal Navigation Structures at Port Sanilac Harbor, Michigan.

Science.gov (United States)

1975-09-01

produces waves with nealioible wace heights (i.e., 0.5 feet); (b) all waves were deep water waves with no correction for shallow water; and, (c) the...at that harbor. This would re- sult in a loss to the local economy . Ini addition, littoral drift would resume southward thus affecting the littoral

Selective activation around the left occipito-temporal sulcus for words relative to pictures: Individual variability or false positives?

NARCIS (Netherlands)

Wright, Nicholas D.; Mechelli, Andrea; Noppeney, Uta; Veltman, Dick J.; Rombouts, Serge A. R. B.; Glensman, Janice; Haynes, John-Dylan; Price, Cathy J.

2008-01-01

We used high-resolution fMRI to investigate claims that learning to read r !sults in greater left occipito-temporal (OT) activation for written words relative to pictures of objects. In tl e first experiment, 9/16 subjects performing a one-back task showed activation in >= 1 left OT voxel for word:

The Effect of Lexicographical Information Costs on Dictionary Making and Use

DEFF Research Database (Denmark)

Nielsen, Sandro

2008-01-01

­tinction is proposed between two general types of lexicographical information costs. Firstly, search-related costs are the efforts required by the look-up activities users have to perform when con­sulting a dictionary to find access to the data they are searching for. It is argued that the access route, article...

Hvordan kan man sige nej til GMO?

DEFF Research Database (Denmark)

Heðinsdóttir, Katla; Gjerris, Mickey

2015-01-01

Modstandere af genteknologi udtrykker ofte deres bekymring med, at teknologien er “unaturlig”. Men hvad mener de egentlig med det? Og er det et synspunkt, der skal tages seriøst, hvis genmodificering af fødevarevarer kan være med til at eliminere sult og fejlernæring i verden?...

Permanent neonatal diabetes mellitus - a case report of a rare cause ...

African Journals Online (AJOL)

sulting from a mutation in the KCNJ11 gene encoding the Kir6.2 subunit. Despite the rarity of permanent neonatal diabetes, this diagnosis should be considered in infants with persis- tent hyperglycaemia requiring insulin therapy. Children with an ATP-sensitive potassium channel defect in the pancreatic beta cell have an ...

EVALUACIÓN DEL USO DE ENZIMAS Y FILTRACIÓN POR GRAVEDAD PARA LA CLARIFICACIÓN DE UNA MEZCLA DILUIDA DE PULPA DE FRUTOS DE CACTUS (OPUNTIA BOLDINGHII BRITTON & ROSE, JUGOS DE NARANJA Y TORONJA EVALUATION OF THE USE OF ENZYMES AND GRAVITY FILTRATION FOR CLARIFICATION OF DILUTED MIXTURE OF CACTUS (OPUNTIA BOLDINGHII BRITTON & ROSE FRUITS PULP, ORANGE AND GRAPEFRUIT JUICES

Directory of Open Access Journals (Sweden)

Carlos Alberto Padrón Pereira

2010-06-01

Full Text Available En Venezuela los frutos de cactus (Opuntia boldinguii son de escasa utilidad comercial. Para su aprovechamiento se evaluó el efecto del uso combinado de enzimas fibrolíticas y filtración por gravedad en la clarificación de una mezcla diluida de pulpa de frutos de cactus, jugos de naranja y toronja. Se compararon los valores de las curvas de flujo de filtrado de las mezclas hidrolizada y sin hidrolizar mediante la prueba t de Welch. La concentración óptima de enzimas fue 0,76% (v/v. La hidrólisis enzimática previa a la pasteurización no afectó el color de la mezcla. Durante los ensayos de pasteurización (62 ± 1 ºC por 30 min hubo grandes cambios de coloración, solo en la mezcla hidrolizada, por degradación de las betalaínas y la adición de ácido ascórbico (0,5% p/v permitió preservar el color. Hubo diferencias altamente significativas entre los valores de las curvas de flujo de filtrado (PIn Venezuela the fruits of cactus (Opuntia boldinguii are of scarce commercial usefulness. For its utilization the effect of combined use of fibrolytic enzymes and gravity filtration for clarification of diluted mixture of cactus fruits pulp, orange and grapefruit juices was evaluated. The values of filtered-flow curves of hydrolyzed and not hydrolyzed mixtures were compared by means of Welch’s t-test. The optimal enzymes concentration was 0.76% (v/v. The prior enzymatic hydrolysis to the pasteurization does not affect the color of the mixture. During the pasteurization assays (62 ± 1 ºC for 30 min there were large changes of coloring, only in the hydrolyzed mixture, by betalains degradation and the addition of ascorbic acid (0.5% w/v permitted to preserve the color. There were highly significant differences between the filtered-flow curves values (P<0.01. The filtered-flow of the hydrolyzed mixture elapsed 110 min (0.159 mL·min-1 was greater to the not hydrolyzed mixture (control flow to the 8 min (0.100 mL·min-1. The juice production

Alterações metabólicas de plantas de milho submetidas à aplicação de glyphosate e fosfito

OpenAIRE

Gomes, Giovanna Larissa Gimenes Cotrick [UNESP

2011-01-01

O glyphosate é um herbicida de largo espectro de controle, não seletivo, e seu sítio de ação é a inibição da enzima 5-enolpiruvilchiquimato 3-fosfato sintase (EPSPs). Com a inibição da enzima, e o bloqueio da rota do ácido chiquímico pelo glyphosate, ocorre o acúmulo de alguns compostos como os ácidos chiquímico e quínico, além de outras alterações metabólicas e fisiológicas nas plantas. O objetivo deste trabalho foi avaliar as alterações metabólicas e fisiológicas de plantas de milho submeti...

ESTANDARIZACIÓN DE UN ENSAYO DE ACTIVIDAD PARA LA ATPasa DE CALCIO DEL ERITROCITO

Directory of Open Access Journals (Sweden)

Claudia P. Tinjacá

2010-09-01

Full Text Available Se describe la estandarización de un ensayo de actividad para la ATPasa de calcio del eritrocito. De acuerdo con el ensayo, la actividad máxima de la enzima corresponde a 3.79 micromoles de fosfato por miligramo de proteína de membrana por hora y la actividad basal representa aproximadamente el 10% de la actividad máxima. El comportamiento de la curva de actividad contra el tiempo es lineal durante los primeros 30 minutos y se presenta una alta correlación entre aumento en la actividad de la enzima y concentración de Calmodulina. Gracias al ensayo es posible cuantificar Calmodulina en concentraciones menores a I^iM.

Are glutathione S transferases involved in DNA damage signalling? Interactions with DNA damage and repair revealed from molecular epidemiology studies

International Nuclear Information System (INIS)

Dusinska, Maria; Staruchova, Marta; Horska, Alexandra; Smolkova, Bozena; Collins, Andrew; Bonassi, Stefano; Volkovova, Katarina

2012-01-01

Glutathione S-transferases (GSTs) are members of a multigene family of isoenzymes that are important in the control of oxidative stress and in phase II metabolism. Acting non-enzymically, GSTs can modulate signalling pathways of cell proliferation, cell differentiation and apoptosis. Using a molecular epidemiology approach, we have investigated a potential involvement of GSTs in DNA damage processing, specifically the modulation of DNA repair in a group of 388 healthy adult volunteers; 239 with at least 5 years of occupational exposure to asbestos, stone wool or glass fibre, and 149 reference subjects. We measured DNA damage in lymphocytes using the comet assay (alkaline single cell gel electrophoresis): strand breaks (SBs) and alkali-labile sites, oxidised pyrimidines with endonuclease III, and oxidised purines with formamidopyrimidine DNA glycosylase. We also measured GST activity in erythrocytes, and the capacity for base excision repair (BER) in a lymphocyte extract. Polymorphisms in genes encoding three GST isoenzymes were determined, namely deletion of GSTM1 and GSTT1 and single nucleotide polymorphism Ile105Val in GSTP1. Consumption of vegetables and wine correlated negatively with DNA damage and modulated BER. GST activity correlated with oxidised bases and with BER capacity, and differed depending on polymorphisms in GSTP1, GSTT1 and GSTM1. A significantly lower BER rate was associated with the homozygous GSTT1 deletion in all asbestos site subjects and in the corresponding reference group. Multifactorial analysis revealed effects of sex and exposure in GSTP1 Ile/Val heterozygotes but not in Ile/Ile homozygotes. These variants affected also SBs levels, mainly by interactions of GSTP1 genotype with exposure, with sex, and with smoking habit; and by an interaction between sex and smoking. Our results show that GST polymorphisms and GST activity can apparently influence DNA stability and repair of oxidised bases, suggesting a potential new role for these

Are glutathione S transferases involved in DNA damage signalling? Interactions with DNA damage and repair revealed from molecular epidemiology studies

Energy Technology Data Exchange (ETDEWEB)

Dusinska, Maria, E-mail: Maria.DUSINSKA@nilu.no [CEE-Health Effects Group, NILU - Norwegian Institute for Air Research, Kjeller (Norway); Staruchova, Marta; Horska, Alexandra [Department of Experimental and Applied Genetics, Slovak Medical University, Bratislava (Slovakia); Smolkova, Bozena [Laboratory of Cancer Genetics, Cancer Research Institute of the Slovak Academy of Sciences, Bratislava (Slovakia); Collins, Andrew [Department of Nutrition, Faculty of Medicine, University of Oslo (Norway); Bonassi, Stefano [Unit of Clinical and Molecular Epidemiology, IRCCS San Raffaele Pisana, Rome (Italy); Volkovova, Katarina [Department of Experimental and Applied Genetics, Slovak Medical University, Bratislava (Slovakia)

2012-08-01

Glutathione S-transferases (GSTs) are members of a multigene family of isoenzymes that are important in the control of oxidative stress and in phase II metabolism. Acting non-enzymically, GSTs can modulate signalling pathways of cell proliferation, cell differentiation and apoptosis. Using a molecular epidemiology approach, we have investigated a potential involvement of GSTs in DNA damage processing, specifically the modulation of DNA repair in a group of 388 healthy adult volunteers; 239 with at least 5 years of occupational exposure to asbestos, stone wool or glass fibre, and 149 reference subjects. We measured DNA damage in lymphocytes using the comet assay (alkaline single cell gel electrophoresis): strand breaks (SBs) and alkali-labile sites, oxidised pyrimidines with endonuclease III, and oxidised purines with formamidopyrimidine DNA glycosylase. We also measured GST activity in erythrocytes, and the capacity for base excision repair (BER) in a lymphocyte extract. Polymorphisms in genes encoding three GST isoenzymes were determined, namely deletion of GSTM1 and GSTT1 and single nucleotide polymorphism Ile105Val in GSTP1. Consumption of vegetables and wine correlated negatively with DNA damage and modulated BER. GST activity correlated with oxidised bases and with BER capacity, and differed depending on polymorphisms in GSTP1, GSTT1 and GSTM1. A significantly lower BER rate was associated with the homozygous GSTT1 deletion in all asbestos site subjects and in the corresponding reference group. Multifactorial analysis revealed effects of sex and exposure in GSTP1 Ile/Val heterozygotes but not in Ile/Ile homozygotes. These variants affected also SBs levels, mainly by interactions of GSTP1 genotype with exposure, with sex, and with smoking habit; and by an interaction between sex and smoking. Our results show that GST polymorphisms and GST activity can apparently influence DNA stability and repair of oxidised bases, suggesting a potential new role for these

Antioxidant role of glutathione S-transferases: 4-Hydroxynonenal, a key molecule in stress-mediated signaling.

Science.gov (United States)

Singhal, Sharad S; Singh, Sharda P; Singhal, Preeti; Horne, David; Singhal, Jyotsana; Awasthi, Sanjay

2015-12-15

4-Hydroxy-2-trans-nonenal (4HNE), one of the major end products of lipid peroxidation (LPO), has been shown to induce apoptosis in a variety of cell lines. It appears to modulate signaling processes in more than one way because it has been suggested to have a role in signaling for differentiation and proliferation. It has been known that glutathione S-transferases (GSTs) can reduce lipid hydroperoxides through their Se-independent glutathione-peroxidase activity and that these enzymes can also detoxify LPO end-products such as 4HNE. Available evidence from earlier studies together with results of recent studies in our laboratories strongly suggests that LPO products, particularly hydroperoxides and 4HNE, are involved in the mechanisms of stress-mediated signaling and that it can be modulated by the alpha-class GSTs through the regulation of the intracellular concentrations of 4HNE. We demonstrate that 4HNE induced apoptosis in various cell lines is accompanied with c-Jun-N-terminal kinase (JNK) and caspase-3 activation. Cells exposed to mild, transient heat or oxidative stress acquire the capacity to exclude intracellular 4HNE at a faster rate by inducing GSTA4-4 which conjugates 4HNE to glutathione (GSH), and RLIP76 which mediates the ATP-dependent transport of the GSH-conjugate of 4HNE (GS-HNE). The balance between formation and exclusion promotes different cellular processes - higher concentrations of 4HNE promote apoptosis; whereas, lower concentrations promote proliferation. In this article, we provide a brief summary of the cellular effects of 4HNE, followed by a review of its GST-catalyzed detoxification, with an emphasis on the structural attributes that play an important role in the interactions with alpha-class GSTA4-4. Taken together, 4HNE is a key signaling molecule and that GSTs being determinants of its intracellular concentrations, can regulate stress-mediated signaling, are reviewed in this article. Copyright © 2015 Elsevier Inc. All rights

Large-scale determination of sequence, structure, and function relationships in cytosolic glutathione transferases across the biosphere.

Science.gov (United States)

Mashiyama, Susan T; Malabanan, M Merced; Akiva, Eyal; Bhosle, Rahul; Branch, Megan C; Hillerich, Brandan; Jagessar, Kevin; Kim, Jungwook; Patskovsky, Yury; Seidel, Ronald D; Stead, Mark; Toro, Rafael; Vetting, Matthew W; Almo, Steven C; Armstrong, Richard N; Babbitt, Patricia C

2014-04-01

The cytosolic glutathione transferase (cytGST) superfamily comprises more than 13,000 nonredundant sequences found throughout the biosphere. Their key roles in metabolism and defense against oxidative damage have led to thousands of studies over several decades. Despite this attention, little is known about the physiological reactions they catalyze and most of the substrates used to assay cytGSTs are synthetic compounds. A deeper understanding of relationships across the superfamily could provide new clues about their functions. To establish a foundation for expanded classification of cytGSTs, we generated similarity-based subgroupings for the entire superfamily. Using the resulting sequence similarity networks, we chose targets that broadly covered unknown functions and report here experimental results confirming GST-like activity for 82 of them, along with 37 new 3D structures determined for 27 targets. These new data, along with experimentally known GST reactions and structures reported in the literature, were painted onto the networks to generate a global view of their sequence-structure-function relationships. The results show how proteins of both known and unknown function relate to each other across the entire superfamily and reveal that the great majority of cytGSTs have not been experimentally characterized or annotated by canonical class. A mapping of taxonomic classes across the superfamily indicates that many taxa are represented in each subgroup and highlights challenges for classification of superfamily sequences into functionally relevant classes. Experimental determination of disulfide bond reductase activity in many diverse subgroups illustrate a theme common for many reaction types. Finally, sequence comparison between an enzyme that catalyzes a reductive dechlorination reaction relevant to bioremediation efforts with some of its closest homologs reveals differences among them likely to be associated with evolution of this unusual reaction

Large-scale determination of sequence, structure, and function relationships in cytosolic glutathione transferases across the biosphere.

Directory of Open Access Journals (Sweden)

Susan T Mashiyama

2014-04-01

Full Text Available The cytosolic glutathione transferase (cytGST superfamily comprises more than 13,000 nonredundant sequences found throughout the biosphere. Their key roles in metabolism and defense against oxidative damage have led to thousands of studies over several decades. Despite this attention, little is known about the physiological reactions they catalyze and most of the substrates used to assay cytGSTs are synthetic compounds. A deeper understanding of relationships across the superfamily could provide new clues about their functions. To establish a foundation for expanded classification of cytGSTs, we generated similarity-based subgroupings for the entire superfamily. Using the resulting sequence similarity networks, we chose targets that broadly covered unknown functions and report here experimental results confirming GST-like activity for 82 of them, along with 37 new 3D structures determined for 27 targets. These new data, along with experimentally known GST reactions and structures reported in the literature, were painted onto the networks to generate a global view of their sequence-structure-function relationships. The results show how proteins of both known and unknown function relate to each other across the entire superfamily and reveal that the great majority of cytGSTs have not been experimentally characterized or annotated by canonical class. A mapping of taxonomic classes across the superfamily indicates that many taxa are represented in each subgroup and highlights challenges for classification of superfamily sequences into functionally relevant classes. Experimental determination of disulfide bond reductase activity in many diverse subgroups illustrate a theme common for many reaction types. Finally, sequence comparison between an enzyme that catalyzes a reductive dechlorination reaction relevant to bioremediation efforts with some of its closest homologs reveals differences among them likely to be associated with evolution of this

Producción in vitro de pectinasas por Colletotrichum acutatum

Directory of Open Access Journals (Sweden)

Carlos Patiño Torres

2010-01-01

Full Text Available Los hongos del género Colletotrichum son reconocidos productores de sustancias fifitotóxicas de bajo peso molecular y de enzimas que juegan papeles clave en su interacción patogénica con varios cultivos. Este estudio tuvo como objetivo determinar la naturaleza de las sustancias fitotóxicas producidas por C. acutatum en los medios de cultivo líquido Czapeck-Dox, Fries y MS, que estuviesen implicados en su interacción patogénica con el tomate de árbol (Solanum betaceae. Los procedimientos y resultados de la evaluación permitieron descartar que la actividad fitotóxica observada sobre los frutos de tomate inoculados con los extractos de los medios de cultivo se debiera a metabolitos de bajo peso molecular. Por el contrario, utilizando la prueba de placa de pozos (Dingle et al., 1953 se estableció que C. acutatum produce en los medios de cultivo evaluados enzimas con actividad pectinasa, tanto liasas como hidrolasas, las cuales podrían estar implicadas en la enfermedad de la antracnosis del tomate de árbol. Los resultados demostraron además que la producción de las enzimas se ve influenciada por el pH del medio de cultivo.

IDENTIFICACIÓN MORFOLÓGICA, MORFOMÉTRICA Y MOLECULAR DE Meloidogyne incognita EN HIGUERA (Ficus carica L. EN COSTA RICA

Directory of Open Access Journals (Sweden)

Walter Peraza-Padilla

2013-01-01

Full Text Available El objetivo de este estudio fue identificar la especie de Meloidogyne asociada a dos plantaciones de higuera en Costa Rica. En marzo de 2012, en el distrito de Pacayas, provincia de Cartago, se detectaron agallas en los sistemas radicales de plantas de higuera (Ficus carica L. de dos fincas. De las raíces agalladas se extrajeron hembras, masas de huevos y juveniles (J2 de Meloidogyne sp. Se examinaron los patrones perineales de las hembras y los segundos estadios infectivos se analizaron morfológica y molecularmente mediante PCR-RFLP. Se amplificó la región intergénica (IGS del genoma mitocondrial, delimitada por el gen de la subunidad II de la citocromo oxidasa (COII y el gen ribosomal 16S. La población de nematodos se identificó como M. incognita. El tamaño de los productos de PCR generados con los imprimadores C2F3 y 1108 fue de 1,7 kb. Al tratar los productos de PCR con enzimas de restricción, se generaron cuatro fragmentos de 850, 450, 250 y 150 pb con la enzima AluI y dos fragmentos de 1300 y 400 pb con la enzima HinfI.

Producción in vitro de pectinasas por Colletotrichum acutatum

Directory of Open Access Journals (Sweden)

Patiño Torres Carlos

2010-03-01

Full Text Available Los hongos del género Colletotrichum son reconocidos productores de sustancias fifitotóxicas de bajo peso molecular y de enzimas que juegan papeles clave en su interacción patogénica con varios cultivos. Este estudio tuvo como objetivo determinar la naturaleza de las sustancias fitotóxicas producidas por C. acutatum en los medios de cultivo líquido Czapeck-Dox, Fries y MS, que estuviesen implicados en su interacción patogénica con el tomate de árbol (Solanum betaceae. Los procedimientos y resultados de la evaluación permitieron descartar que la actividad fitotóxica observada sobre los frutos de tomate inoculados con los extractos de los medios de cultivo se debiera a metabolitos de bajo peso molecular. Por el contrario, utilizando la prueba de placa de pozos (Dingle et al., 1953 se estableció que C. acutatum produce en los medios de cultivo evaluados enzimas con actividad pectinasa, tanto liasas como hidrolasas, las cuales podrían estar implicadas en la enfermedad de la antracnosis del tomate de árbol. Los resultados demostraron además que la producción de las enzimas se ve influenciada por el pH del medio de cultivo.

Response surface methodology to evaluation the recovery of amylases by hollow fiber membrane

Directory of Open Access Journals (Sweden)

João Baptista Severo Júnior

2007-07-01

Full Text Available This work aimed to study the pH and the transmembrane pressure effects during the recovery of alpha and beta amylases enzymes from corn malt (Zea mays by hollow fiber membrane. The optimal condition was obtained for a statistical model, established by response surface methodology (RSM. The response surface analysis showed that the best operation condition for amylolitics enzymes recovery by hollow fiber membrane was 0.05 bar and pH 5.00, while the enzymes were purified about of 26 times.Este trabalho objetivou estudar o efeito do pH e da pressão trans-membrana durante a recuperação das enzimas alfa e beta amilases do malte de milho (Zea mays por membranas de fibras ocas, a obtenção das condições ótimas foi feita por um modelo estatístico, estabelecido pela metodologia de superfície de resposta (RSM. A análise da superfície de resposta mostrou que as melhores condições operacionais para a recuperação das enzimas amiloliticas por membranas de fibras ocas foi 0,05 bar e pH 5,00; onde as enzimas foram purificadas cerca de 26 vezes.

CATALASA, PEROXIDASA Y POLIFENOLOXIDASA DE PITAHAYA AMARILLA (Acanthocereus pitajaya

Directory of Open Access Journals (Sweden)

Jhon Alexander Castro Rivera

2008-03-01

Full Text Available Las enzimas catalasa (CAT, peroxidasa (POD y polifenoloxidasa (PFO fueron extraídas de la corteza de frutos de pitaya amarilla (Acanthocereus pitajaya y caracterizadas parcialmente. Para CAT se halló que su actividad fue máxima a pH entre 6,8 y 7,5 y temperatura entre 30 a 50 °C y un KM de 442 mM con H2O2 como sustrato. Para POD se encontró un pH de máxima actividad entre 5,0 a 5,5, temperatura de máxima actividad entre 20 a 25 °C y valores de KM de 10,6 mM para guayacol y 5,1 mM para H2O2. Para PFO las actividades máximas se obtuvieron a pH 7,0 y a temperaturas entre 30 a 40 °C; para esta enzima se obtuvo un KM de 5,5 mM con L-DOPA como sustrato. Las características encontradas para POD y PFO indican que estas enzimas pueden jugar un papel importante en el pardeamiento de la corteza de pitaya amarilla. Además, se evidenció el papel complementario que tienen CAT y POD ante diversas concentraciones celulares de H2O2.

Forfatteren som eksperimentator

DEFF Research Database (Denmark)

Hedin, Gry

2011-01-01

Det kan overraske, at man i Hamsun-forskningen ikke har skænket den første version af Sult mere opmærksomhed. Hamsuns publicering i tidsskriftet Ny Jord af det, der skulle blive bogens andet stykke, er med enkelte undtagelser overset i forskningen. En analyse af teksten og dens kontekst kan imidl...

l04------------------------------~------------RESONANCEI--N-ov-ember--19 ...

Indian Academy of Sciences (India)

Needless to say, I strongly recom- mend that all students of mathematics read this book. For the lay person, this is an excellent exposure to mathematics. For the undergraduate student this book serves as an introduction to many beautiful re- sults in mathematics and also an opportu- nity to learn about the rich history and.

The Coast Artillery Journal. Volume 75, Number 2, March-April 1932

Science.gov (United States)

1932-04-01

after three years’ study of the Japanese language-study which has re- sulted not in my gaining easy fluency in the language of the Samurai but in...eight and a half feet thick, using 850 pounds of cement to ite cubic yard. In standard European fortifications the proportion of cement to the cubic yard

Atividade do sistema antioxidante e desenvolvimento de aerênquima em raízes de milho 'Saracura' Antioxidant system activity and aerenchyma formation in 'Saracura' maize roots

Directory of Open Access Journals (Sweden)

Fabricio José Pereira

2010-05-01

Full Text Available Este trabalho teve como objetivo avaliar a influência de sucessivos ciclos de seleção do milho 'Saracura' na atividade das enzimas do sistema antioxidante, e a relação dessas enzimas com a capacidade dessa variedade em desenvolver aerênquima. Sementes de 18 ciclos de seleção intercalados do milho 'Saracura' e da cultivar BR 107, sensível à hipoxia, foram semeadas em vasos e em casa de vegetação. As plantas foram submetidas ao alagamento intermitente de dois em dois dias. As amostras de raízes foram coletadas após 60 dias e analisaram-se as atividades das enzimas peroxidase do guaiacol, peroxidase do ascorbato e catalase, além da capacidade das plantas de cada ciclo desenvolverem aerênquima. Ao longo dos ciclos, as plantas apresentaram modificações na atividade das enzimas, com aumento na de peroxidase do ascorbato e diminuição na de catalase e de peroxidase do guaiacol. Observou-se, ainda, maior capacidade de desenvolver aerênquima nos últimos ciclos de seleção. A redução na atividade das enzimas do sistema antioxidante parece estar relacionada a um desbalanço na decomposição de H2O2.This work aimed to assess the influence of successive selection cycles in 'Saracura' maize on the enzyme activity of the antioxidant system and the relationship of these enzymes with the aerenchyma development capacity of this variety. Seeds of 18 intercalated selection cycles of the 'Saracura' maize and of the cultivar BR 107, sensitive to hipoxia, were sown in pots in the greenhouse. Plants were submitted to intermittent soil flooding each two days. After 60 days, the roots were sampled and analysis were done for the guaiacol peroxidase, ascorbate peroxidase, and catalase activities and for the capacity of the plants of each cycle to develop aerenchyma. The plants showed modifications in enzyme activity along the cycles, increasing the ascorbate peroxidase activity and decreasing the catalase and guaiacol peroxidase ones. A greater

Produção de galactooligossacarídeo por Scopulariopis sp. Production of galactooligosaccharide by Scopulariopis sp.

Directory of Open Access Journals (Sweden)

Rosângela dos Santos

2009-09-01

Full Text Available Os galactooligossacarídeos (GOS são um grupo de oligossacarídeos não digeríveis (NDOs, resistentes às enzimas digestivas do intestino com efeitos similares ao da fibra. Sua ingestão aumenta seletivamente o crescimento das Bifidobacterium e dos Lactobacilus no intestino. O benéfico da ingestão de galactooligossacarídeos ocorre pelo aumento dessas populações de bifidobactérias no cólon, suprimindo a atividade de bactérias putrefativas e reduzindo a formação de produtos tóxicos por fermentação. A cepa de Scopulariopsis sp. apresentou boa produtividade de β-galactosidase quando crescida em meio de fermentação semissólida. O objetivo deste trabalho foi extrair a enzima β-galactosidase produzida por uma linhagem de Scopulariopsis sp. e avaliar o efeito da temperatura, tempo de reação, concentração de enzima e lactose para produção de GOS. Foram analisadas as temperaturas 35, 45 e 60 °C, os tempos de reação 12, 24 e 48 horas, concentrações de enzima 0,5 a 10 U.mL-1 e 10, 25 e 40% (p/v de solução de lactose (em 0,1 M de tampão acetato de sódio, pH 5.0. As condições ótimas foram de 40% (p/v, a 45 °C, 10 U.mL-1 de enzima e o melhor tempo foi de 12 horas de reação. Nessas condições, a enzima converteu 20% de lactose em oligossacarídeos (80,8 mg.mL-1 de 4'galactosyl-lactose.The galactooligosaccharides (GOS, a group of oligosaccharides, are not digerible carbohydrates (NDOs and are resistant to hydrolysis by intestine digestive enzymes with similar dietary fiber physiological effect. The benefits of galactooligosaccharides ingestion arise from a population of bifidobacteria in the colon that suppress the activity of putrefactive bacteria and reduce the formation of toxic fermentation products. Scopulariopsis strains showed good productivity of β-galactosidase when grown in a semi-solid fermentation medium. The aim of this work was to extract the β-galactosidase from Scopulariopsis sp. and to evaluate the

Low Cost Technical Solutions to Jump Start an Insider Threat Program

Science.gov (United States)

2016-05-11

from the OpenDLP server. The OpenDLP server manages the re- sults of the scans. OpenDLP also has the ability to scan Microsoft SQL Server and MySQL data...Learning Group at the University of Waikato, "Weka 3: Data Mining Software in Java ," [Online]. Available: http://www.cs.waikato.ac.nz/ml/weka

Parallelization of TMVA Machine Learning Algorithms

CERN Document Server

Hajili, Mammad

2017-01-01

This report reflects my work on Parallelization of TMVA Machine Learning Algorithms integrated to ROOT Data Analysis Framework during summer internship at CERN. The report consists of 4 impor- tant part - data set used in training and validation, algorithms that multiprocessing applied on them, parallelization techniques and re- sults of execution time changes due to number of workers.

Identification of Mutations in SDR9C7 in 6 Families with Autosomal Recessive Congenital Ichthyosis

DEFF Research Database (Denmark)

Hotz, A; Fagerberg, C; Vahlquist, A

2018-01-01

Autosomal recessive congenital ichthyosis (ARCI) is a heterogeneous group of disorders of keratinization. To date, ARCI has been associated with following genes: ABCA12, ALOX12B, ALOXE3, CERS3, CYP4F22, NIPAL4, TGM1, PNPLA1 and recently SDR9C7 and SULT2B1.(1-6) Furthermore, seven patients from...

The influence of farmer perception on pesticide usage for ...

African Journals Online (AJOL)

A case study conducted in eastern Uganda revealed that: (a) pesticide usage depended on farmer production goal, (b) change from one pesticide to another depended on ... Ces résults ont fermément montré que les recommendations de la gestion des pestes aux fermiers doivent prendre en considération la perception des ...

Sult er en sjælden årsag til metabolisk acidose

DEFF Research Database (Denmark)

Vestergaard, Thea; Aaen, Jeppe Frøkjær; Bruun, Jens Meldgaard

2014-01-01

of infection, dehydration and normoglycaemic metabolic acidosis with elevated P-3-hydroxybutyrate. The case presents a starvation- (weight loss ~ 13 kg) and infection-induced non-diabetic metabolic ketoacidosis treated with intravenous supplementation of isotonic saline, potassium, bicarbonate and insulin....

Estonian household forms and motives for saving and borrowing / Marge Sults

Index Scriptorium Estoniae

Sults, Marge

2003-01-01

Kui 1990-ndate alguses mõjutas majapidamissektori säästmiskäitumist inflatsioon, siis nüüd on oluliseks saanud nii sissetulekute kasv, madalad laenuintressid kui ka uued laenutooted ja väiksemad laenupiirangud. Siia lisandub Eestis läbiviidav pensionireform, mis peaks säästmisele mõjuma positiivselt. Tabelid

Caractérisation biochimique et fonctionnelle de glutathion-S-transferases (GSTs) chez Phanerochaete chrysosporium

OpenAIRE

Anak-Ngadin, Andrew

2011-01-01

Phanerochaete chrysosporium est un champignon ligninolytique largement étudié pour ses capacités à dégrader la lignine et certains xénobiotiques grâce à un important système d'enzymes extracellulaires. Son génome est entièrement séquencé et constitue un inventaire de séquences protéiques prédites qui a permis la description de nombreuses superfamilles de protéines. Parmi elles, les Glutathion S-transférases sont essentiellement impliquées dans le métabolisme secondaire du champignon. Cependan...

Hipertensión arterial mineralocorticoidea

Directory of Open Access Journals (Sweden)

Carlos E. Fardella B., Dr.

2013-09-01

Full Text Available La hipertensión arterial (HTA dependiente de mineralocorticoides representa actualmente una de las formas secundarias de hipertensión más prevalentes. Entre las causas más conocidas está el hiperaldosteronismo primario (HAP. A nivel renal, la aldosterona reabsorbe sodio y agua aumentando el volumen intravascular y la presión arterial. Actualmente la prevalencia de HAP, detectada por la razón aldosterona/actividad renina plasmática (ARR que es considerado el mejor test de screening, es cercana al 10% en población hipertensa. Por otra parte, defectos congénitos o adquiridos en la enzima 11β-Hidroxiesteroide deshidrogenasa tipo 2 (11β-HSD2 resultan en una ineficiente inactivación de cortisol a cortisona favoreciendo la aparición de hipertensión por activación del receptor mineralocorticoideo. La actividad de esta enzima se evalúa midiendo la razón cortisol/cortisona en suero o en orina de 24 horas. Recientemente, hemos observado déficit parciales de la actividad de la enzima 11β-HSD-2 en alrededor del 15% de los pacientes hipertensos esenciales los que podrían ser tratados con bloqueadores específicos del receptor mineralocorticoide y/o con corticoides de acción prolongada sin actividad mineralocorticoide como dexametasona o betametasona.

Aspectos toxicológicos do bissulfito e outros agentes tóxicos sobre a síntese de glicerol-3-fosfato desidrogenase citoplasmática de levedura de panificação

Directory of Open Access Journals (Sweden)

E. A.L. GATTáS

2009-01-01

Full Text Available

A síntese de glicerol-3-fosfato desidrogenase intracelular (EC 1.1.1.8 foi investigada a partir de fermento de panificação em cultivos submersos, contendo glicose ou glicerol como únicas fontes de carbono. Agentes inibitórios da via glicolítica, do ciclo de Krebs e da Cadeia Respiratória inibiram a síntese da enzima quando adicionados em baixas concentrações até 7,5 x 10^-4 mol/L. A repressão exercida pela glicose sobre a síntese da glicerol-3-fosfato desidrogenase em meio YP-glicose foi reduzida, com a adição de produtos de fermentação e de bissulfito de sódio. Observou-se aumentos de 22-110% na síntese da enzima. Entretanto, em meio YP-glicerol, a adição de bissulfito de sódio 0,06 % (p/v reduziu a síntese da enzima em 29%, enquanto, o acetaldeído 0,012 % (v/v estimulou a síntese de glicerol-3-fosfato desidrogenase em 12%. Palavras-chave: fermento de panificação; glicerol-3-fosfato desidrogenase; inibidores.

Comité Scientifique de la Revue des Sciences et de la Technologie ...

African Journals Online (AJOL)

MICROSIM

du Turonien enregistrent les variations lithologiques et microfaunistique relatives à l'événement anoxique océanique 2 (EAO-2). Les calcaires ... depuis des bassins océaniques profonds à des mers peu profondes [2, 3]. Cette crise résulte ..... tectonique, de la subsidence: une mise au point- Bulletin de la société géologique ...

Le CRDI au Bhoutan

International Development Research Centre (IDRC) Digital Library (Canada)

Il en résulte des solutions locales, novatrices et durables, qui offrent des choix aux personnes qui en ont le plus besoin et font changer les choses. Centre de recherches pour le développement international. CP 8500, Ottawa (Ontario) Canada K1G 3H9. Pour en savoir plus, consulter la page. Web du Bureau régional de ...

Le CRDI au Kenya

International Development Research Centre (IDRC) Digital Library (Canada)

afin d'y favoriser la croissance et le développement. Il en résulte des solutions novatrices et durables qui ont pour but d'améliorer les conditions de vie et les moyens de subsistance. Centre de recherches pour le développement international. CP 8500, Ottawa (Ontario) Canada K1G 3H9. Pour en savoir plus, consulter.

Le CRDI en Égypte

International Development Research Centre (IDRC) Digital Library (Canada)

afin d'y favoriser la croissance et le développement. Il en résulte des solutions locales, novatrices et durables, qui offrent des choix aux personnes qui en ont le plus besoin et font changer les choses. Centre de recherches pour le développement international. CP 8500, Ottawa (Ontario) Canada K1G 3H9. 1012 alexandrie.

AcEST: DK944467 [AcEST

Lifescience Database Archive (English)

Full Text Available sults ■■ - Swiss-Prot (release 56.9) Link to BlastX Result : Swiss-Prot sp_hit_id O24339 Definition sp|O24339|CATA_SOLAP........................done Score E Sequences producing significant alignments: (bits) Value sp|O24339|CATA_SOLAP...TA_IPOBA Catalase OS=Ipomoea batatas PE=2 SV=1 248 2e-65 >sp|O24339|CATA_SOLAP Ca

Honduras | CRDI - Centre de recherches pour le développement ...

International Development Research Centre (IDRC) Digital Library (Canada)

Honduras. Le CRDI appuie des chercheurs honduriens depuis 1978. Les premiers travaux ont porté sur l'amélioration des cultures et des systèmes culturaux. Les techniques qui en ont résulté ont accru les rendements des cultures vivrières de base et des cultures de rapport, et les agriculteurs les ont rapidement adoptées.

Asie | CRDI - Centre de recherches pour le développement ...

International Development Research Centre (IDRC) Digital Library (Canada)

... défis agricoles, environnementaux, technologiques, sociaux et économiques de l'Asie. Grâce à nos investissements stratégiques, nous aidons les acteurs régionaux à participer plus efficacement à résolution des problèmes régionaux. Qu'en résulte-t-il ? Des dirigeants régionaux forts, pour aujourd'hui et pour demain.

FONTES E APLICAÇÕES DE TANINOS E TANASES EM ALIMENTOS SOURCE AND APPLICATION OF TANNIN AND TANNASE IN FOODS.

Directory of Open Access Journals (Sweden)

V. BATTESTIN

2008-08-01

Full Text Available

Os taninos ocorrem em uma ampla variedade de vegetais, são classificados em hidrolisáveis e condensados. O ácido tânico é um tanino hidrolisável, que é quebrado por enzimas. São utilizados para estabilização da cerveja, curtimento de pele e produção de resinas. Atualmente, são empregados em processos biotecnológicos para produção de enzimas como a tanase, que hidrolisa ésteres e ligações laterais de taninos hidrolisáveis produzindo ácido gálico e glicose. A maior aplicação desta enzima tem sido para produção de ácido gálico, que é utilizado na produção de trimetropim e sínteses de ésteres como propil galato, usado como antioxidante na indústria de alimentos. A enzima também é aplicada no processamento de cerveja e clarificação de sucos, processamento de chás instantâneos e tratamento de efluentes contaminados com compostos fenólicos. Tannins occur in an ample variety of vegetables and are classified in condensed and hydrolysable tannins. The tannic acid is a hydrolysable tannin that is broken by enzymes. Tannins are used for stabilization of the beer and resin production. Currently, they are used in biotechnological processes for enzyme production as tannase which hydrolyses ester and depside bonds of hydrolysable tannins releasing gallic acid and glucose. The major applications of this enzyme are in the production of gallic acid, which is used for the manufacture of an trimethoprim and synthesis of esters, such as propyl gallate, used as antioxidants in the food industry. The enzyme also has its applications in the production of beer and juices as a clarifier, in the manufacture of instant tea, and the treatment of wastewater contaminated with polyphenolic compounds.

Construcción de un vector de expresión derivado de virus adenoasociados para corregir in vitro el defecto genético de la enfermedad de Morquio A

Directory of Open Access Journals (Sweden)

Luis Alejandro Barrera

2008-09-01

Full Text Available Introducción. La mucopolisacaridosis IV A (Morquio A es una enfermedad de depósito lisosómico causada por la deficiencia en la actividad de la enzima N-acetil-galactosamina- 6-sulfato-sulfatasa que produce la acumulación intralisosómica de queratán y condroitín-6-sulfato. Hasta el momento, su manejo es paliativo, por lo que las investigaciones se han enfocado en establecer una terapia que pueda aplicarse tempranamente y garantice la expresión estable de la enzima. En este sentido, la terapia génica se presenta como una de las potenciales alternativas terapéuticas para corregir el defecto genético en la mucopolisacaridosis IV A. Objetivo. Construir vectores de expresión derivados de virus adenoasociados para corregir in vitro la deficiencia enzimática en la mucopolisacaridosis IV A. Materiales y métodos. Se produjeron vectores derivados de virus adenoasociados que portaban el gen humano de la enzima N-acetil-galactosamina-6-sulfato-sulfatasa dirigido por el promotor temprano del citomegalovirus humano, empleando un sistema libre de adenovirus. Se transfectaron células HEK293 y fibroblastos humanos Morquio A con los virus recombinantes, y se determinó la actividad enzimática en el lisado celular a las 24 y 48 horas después de la transfección. Resultados. Se obtuvieron virus adenoasociados recombinantes, libres de adenovirus, con títulos hasta de 2,08 x 1010 cápsides/ml. Tanto en células HEK293 como en fibroblastos Morquio A transfectados, se obtuvieron actividades enzimáticas hasta de 3,05 nmoles/mg por hora, 48 horas después de la transfección. Conclusión. Los virus recombinantes producidos expresaron in vitro la enzima GALNS en las células transfectadas. Estos resultados constituyen el paso inicial para el desarrollo de una terapia génica para la enfermedad de Morquio A empleando vectores derivados de virus adenoasociados.

Efecto de la aplicación de tratamientos combinados de aditivos sobre la inhibición del pardeamiento enzimático en manzanas cv. Granny Smith mínimamente procesadas

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DENOYA, G. I.

2012-10-01

Full Text Available El pardeamiento enzimático, catalizado principalmente por la Enzima Polifenol Oxidasa (PPO, es uno de los principales problemas que afectan la calidad y limitan la vida útil de frutas y hortalizas mínimamente procesadas.Los compuestos tradicionalmente utilizados para inhibir la PPO, son los sulfitos. Sin embargo, se ha desalentado su utilización en la industria alimentaria debido a que se han registrado casos de reacciones alérgicas, especialmente en individuos asmáticos. Como consecuencia, en la actualidad, se evalúa la utilización de otros compuestos como potenciales inhibidores de la enzima, para garantizar productos frescos y naturales. En el presente trabajo, se analizó la evolución de la actividad de la enzima PPO y las características cromáticas de la pulpa de rodajas de manzanas cv. Granny Smith, tratadas por inmersión en una solución de aditivos. Los tratamientos utilizados fueron: I. 2% ácido ascórbico + 1% ácido cítrico + 0,5% EDTA, II. 1% ácido ascórbico + 0,5 % ácido cítrico + 0,25% EDTA y III. agua, empleada como control. Durante el almacenamiento, se evaluaron las coordenadas de color del espacio CIE L*a*b* de las rodajas y se demostró que los tratamientos I y II fueron efectivos en evitar el pardeamiento de la fruta. La evaluación espectrofotométrica de la actividad enzimática de la PPO, en extractos de manzanas sometidas a los distintos tratamientos, mostró que el más severo (I fue el que produjo el mayor grado de inhibición de la enzima, en todos los tiempos analizados. Se propone evaluar a futuro, la efectividad de estos inhibidores “in vitro”, a efectos de compararlos con los resultados obtenidos en rodajas de manzana.

Evaluación de una esterasa de ácido ferúlico (Fae1A aislada del hongo del rumen Anaeromyces mucronatus sobre la degradación de la pared celular del heno de alfalfa y paja de trigo: Ensayo In vitro

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José C. López O

2015-01-01

Full Text Available Las esterasas de ácido ferúlico son capaces de liberar compuestos fenólicos de la pared celular vegetal, aumentando el acceso a los polisacáridos. Se realizaron determinaciones In vitro con la finalidad de determinar el potencial de una esterasa de ácido ferúlico (Fae1A aislada del hongo del rumen Anaeromyces mucronatus. Esta estearasa se obtuvo de un trabajo previo realizado en el Centro de Investigación de Lethbridge, Alberta Canadá, a través de procesos de clonación y hetero-expresión del gen Fae1aA en una cepa de E. coli BL21. Tres niveles (0, 9 y 18 mg de proteína contenida en la enzima se combinaron con liquido ruminal e incubaron en 2 diferentes fuentes de fibra (heno de alfalfa y paja de trigo utilizando 15 repeticiones por tratamiento. La inclusión de la enzima incrementó (P <0.05 la desaparición de la materia seca en el heno de alfalfa, liberándose más cantidad (P <0.01 de azúcares reducidos (AR a las 4 y 12 h. Fae1A liberó más compuestos fenólicos del heno de alfalfa vs. la paja de trigo. Los compuestos fenólicos totales e individuales liberados de la alfalfa se incrementaron con el nivel de enzima añadida. En la paja de trigo, el ácido ferúlico se duplicó en comparación con el ácido p-cumárico. En conclusión, la adición de la Fae1A mejoró la cantidad de azucares reducidos y se liberaron compuestos fenólicos en el heno de alfalfa y la paja de trigo, lo que indica su potencial para ser usada como enzima fibrolítica y mejorar la digestibilidad de la fibra en los rumiantes.

Actividad inhibitoria de dihidroxifenilpropenona sobre β-lactamasa de Enterobacter cloacae: estudio preliminar en el desarrollo de fármacos para enfrentar la resistencia bacteriana

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Cristina Lucía Mora

2014-04-01

Full Text Available Introducción. Microorganismos patógenos como Enterobacter cloacae producen enzimas β-lactamasas que les confieren resistencia a los antibioticos β-lactámicos y actualmente se ha identificado la actividad limitada de los inhibidores enzimáticos, de modo que las únicas posibilidades para enfrentar la problemática son el diseño de nuevos fármacos y el uso racional de los mismos. Objetivo. Evaluar el efecto de la chalcona dihidroxifenilpropenona sobre un aislamiento clínico de E. cloacae y sobre la enzima β-lactamasa aislada a partir de este microorganismo resistente como un aporte hacia la búsqueda de compuestos inhibidores de β-lactamasa. Materiales y métodos. Se sintetizó la chalcona dihidroxifenilpropenona y se evaluó sobre el aislamiento clínico de E. cloacae para determinar la concentración mínima inhibitoria mediante el método de microdilución en caldo y con la β-lactamasa purificada mediante cromatografía de afinidad se realizaron estudios espectrofotométricos de cinética enzimática. Resultados. La concentración mínima inhibitoria de la dihidroxifenilpropenona sobre E. cloacae fue de 35 μg/ml, el porcentaje de recuperación de la enzima β-lactamasa a partir del microorganismo fue de 31,75% y en el estudio cinético se evidenció actividad inhibitoria de acuerdo con los parámetros cinéticos de Vmax= 1,7 x 10-3 μM/min y KM´= 2330 μM. Conclusión. La chalcona dihidroxifenilpropenona ejerce su actividad inhibitoria a través de la interacción con la enzima β-lactamasa y de esta manera protege la integridad estructural de los antibióticos β-lactámicos, este efecto sinérgico lo convierte en un compuesto promisorio en la búsqueda de alternativas para enfrentar la problemática de resistencia bacteriana.

Enzymatic antioxidant responses to biostimulants in maize and soybean subjected to drought Respostas de enzimas antioxidantes a bioestimulantes em plantas de milho e de soja sob estresse hídrico

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Ana Carolina Feitosa de Vasconcelos

2009-06-01

Full Text Available Water stress is one of the most important environmental factors inducing physiological changes in plants, such as decrease in the water potential of the cells, the stomatal closure; and the development of oxidative processes mediated by reactive oxygen species (ROS. Antioxidant enzymes superoxide dismutase (SOD, catalase (CAT, and ascorbate peroxidase (APX are efficient scavengers of ROS. The aim of this research was to examine how the application of biostimulant based on humic substances and aminoacids may affect activity levels of SOD, CAT, and APX of maize and soybean plants under well-watered or drought stress conditions. Pots (4.5 L were filled with a Typic Hapludult soil where the biostimulants doses were applied. It was taken leaf samples in order to analyze SOD, CAT, and APX activities in plants. SOD and APX activity levels were increased by application of biostimulant 1 in maize subjected to stress. Catalase activity was not enhanced in plants by using the biostimulants. The composition of the biostimulants was not able to enhance stress tolerance in maize and soybean plants subjected to water stress.O estresse hídrico é um dos mais importantes fatores ambientais que induz mudanças fisiológicas, como diminuição do potencial de água na célula, o fechamento dos estômatos e o desenvolvimento de processos oxidativos mediante a formação das espécies reativas de oxigênio (ROS. As enzimas antioxidantes superóxido dismutase (SOD, catalase (CAT e ascorbato peroxidase (APX são eficientes eliminadores das ROS. O objetivo deste estudo foi examinar como a aplicação de bioestimulantes com substâncias húmicas e aminoácidos em sua composição afeta os níveis de SOD, CAT e APX nos tecidos das folhas de plantas de milho e de soja cultivadas com ou sem estresse hídrico. Amostras de um Argissolo foram colocadas em vasos (4,5 L onde foram adicionadas as doses dos bioestimulantes. Foram retiradas amostras de folhas para análise da

Avaliação de métodos para manutenção e preservação de bactéria esporulada produtora da enzima CGTase - DOI: 10.4025/actascihealthsci.v31i2.6910 Evaluation of methods for maintenance and preservation of sporulating bacteria producer of CGTase enzyme- DOI: 10.4025/actascihealthsci.v31i2.6910

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Cristiane Moriwaki

2009-09-01

Full Text Available A conservação de células sem mudanças morfológicas, fisiológicas ou genéticas é uma necessidade da biotecnologia. Bacillus firmus cepa 37 é uma bactéria esporulada produtora da enzima ciclodextrina glicosiltransferase (CGTase, que transforma o amido em ciclodextrinas (CDs. O objetivo deste estudo foi avaliar a manutenção e preservação de B. firmus cepa 37 estocada em meio de cultivo sólido, solo estéril e em glicerol a baixa temperatura (-70ºC. Para avaliação do melhor método de manutenção da bactéria foram utilizados procedimentos de imobilização das células em matrizes inorgânicas. As células imobilizadas foram submetidas ao teste do efeito da biomassa inicial e à microscopia eletrônica de varredura (MEV. O repique não foi um método adequado, pois a cepa diminuiu a produção de CGTase. A estocagem em solo estéril mostrou-se eficaz e a produção da enzima mantida constante. A conservação a baixas temperaturas também foi satisfatória, com contagem de células praticamente a mesma após 360 dias. A imobilização, avaliada por MEV, não mostrou diferença na adsorção das células conservadas pelos diferentes métodos. O mesmo ocorreu para o teste do efeito da biomassa inicial, que apresentou maior produção de beta-CD quando do uso de 1,5 g de células.The conservation of cells without morphologic, physiologic or genetic changes is a biotechnology necessity. Bacillus firmus strain 37 is a sporulating bacteria that produces the cyclodextrin glycosyltransferase (CGTase enzyme, which transforms starch into cyclodextrins (CDs. This study aimed to evaluate the maintenance and preservation of B. firmus strain 37 stored in a solid medium, sterile soil and in glycerol at low temperature (-70ºC. In order to evaluate the best bacteria maintenance method, cell immobilization procedures were used on inorganic matrices. The immobilized cells were submitted to the initial biomass effect test and scanning electron

Simultaneous enzymatic hydrolysis and lactic fermentation to obtain a yogurt with low lactose content Hidrólise enzimática e fermentação lática simultâneas na obtenção de um iogurte com baixo teor de lactose

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André Rosa Martins

2012-10-01

Full Text Available In a single-stage process with the simultaneous addition of β-galactosidase and lactic culture, the lactose conversion, the processing time, viscosity and syneresis were evaluated. Fermentation was promoted by lactic culture containing two probiotic microorganisms, Bifidobacterium animalis and Lactobacillus acidophilus, associated with the typical microorganisms of yogurt. An enzymatic preparation containing β-galactosidases from Kluyveromyces lactis and Aspergillus niger was used. A central composite design (2³ trials plus three central points was proposed in order to evaluate the effects of initial lactose concentration, enzyme concentration and the time of addition of the enzyme. The following conditions were established: initial lactose concentration of 91 g L-1, enzyme concentration of 0.5 g L-1 and enzyme addition at the beginning of fermentation. In these conditions, a decrease in processing time (from 4.55 to 3.68 h and an increase in lactose conversion (from 15.2 to 97.9% were observed in relation to the fermentation without enzyme addition, and no detrimental changes in some physical properties of yogurt (viscosity, and syneresis.No processo em um único estágio com a adição simultânea de β-galactosidase e de cultura lática, foram avaliadas a conversão da lactose, o tempo de processamento, a viscosidade e a sinérese. A fermentação foi promovida por cultura láctica, contendo dois micro-organismos probióticos, Bifidobacterium animalis e Lactobacillus acidophilus, associados aos micro-organismos característicos do iogurte. Foi utilizado um preparado enzimático contendoβ-galactosidases obtidas de Kluyveromyces lactis e de Aspergillus niger. Os efeitos da concentração inicial de lactose, concentração de enzima e tempo de adição da enzima foram avaliados por meio de um planejamento experimental (2³ ensaios mais três pontos centrais. As seguintes condições foram estabelecidas: concentração inicial de lactose de

Production of xylanase and CMCase on solid state fermentation in different residues by Thermoascus aurantiacus miehe Produção de xilanase e CMCase por fermentação em estado sólido em diferentes resíduos pelo fungo termofílico Thermoascus aurantiacus miehe

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Roberto da Silva

2005-09-01

Full Text Available The use of waste as raw material is important for government economy and natural balance. The purpose of this work was to study the production of CMCase and xylanase by a Brazilian strain of Thermoascus aurantiacus in solid state fermentation (SSF using different agricultural residues (wheat bran, sugarcane bagasse, orange bagasse, corncob, green grass, dried grass, sawdust and corn straw as substrates without enrichment of the medium and characterize the crude enzymes.The study of the extracellular cellulolytic and hemicellulolytic enzymes showed that T. arantiacus is more xylanolytic than cellulolytic. The highest levels of enzymes were produced in corncob, grasses and corn straw. All the enzymes were stable at room temperature by 24 h over a broad pH range (3.0-9.0 and also were stable at 60ºC for 1 h. The optimum pH and temperature for xylanase and CMCase were 5.0-5.5 and 5.0 and 75ºC, respectively. The microorganism grew quickly in stationary, simple and low cost medium. The secreted extracellular enzymes presented properties that match with those frequently required in industrial environment.O emprego de residuos como matéria prima é importante como estrategia governamental e para o balanço ambiental. O propósito deste trabalho foi estudar a produção de CMCase e xilanase de uma linhagem de Thermoascus aurantiacus isolado de solo brasileiro em fermentação em estado sólido (SSF usando diferentes resíduos agrícolas (farelo de trigo, bagaço de cana, bagaço de laranja, sabugo de milho, grama verde, grama seca, serragem de eucalipto e palha de milho como substratos sem enriquecimento dos meios e caracterizar as enzimas. O estudo das enzimas hemiceluloliticas extracelulares mostrou que o fungo T. arantiacus é mais xilanolítico do que celulolítico. Ele produziu maiores níveis das enzimas em meios contendo sabugo de milho, grama e palha de milho. Todas as enzimas foram estáveis por 24 h à temperatura ambiente numa ampla faixa

Studies on the stability of protease from Bacillus sp. and its compatibility with commercial detergent Estudos sobre a estabilidade de uma protease de Bacillus sp. e sua compatibilidade com detergentes comerciais

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Wellingta Cristina Almeida do Nascimento

2006-09-01

Full Text Available Enzymes, and particularly proteases, have become an important and indispensable part of industrial processes such as laundry detergents, pharmaceuticals and food products. Detergents such as Tide®, Ariel® and Biz® contain proteolytic enzymes, most of them produced by members of the genus Bacillus. This paper describes the compatibility of protease produced by the thermophilic Bacillus sp, with commercial laundry detergent. Stability studies indicated that this enzyme retained about 95% and 74% of its maximum activity after 1h at 60ºC in the presence of glycine in combination with MnSO4 and CaCl2, respectively. No inhibitory effect was observed at 1.0-5.0 mM of EDTA. Triton X-100 inhibited the enzyme in all the concentrations tested. The enzyme was unstable in a 5% (v/v concentration of peroxide solution. The protease retained more than 80% and 65% of its activity after 30 min incubation at 60ºC in the presence of Tide® and Cheer® detergents, respectively. After supplementation of CaCl2 (10 mM and glycine (1 mM, the enzyme in Tide® detergent retained more than 85% of its activity after 1h. Based on these findings, Bacillus sp. protease shows a good potential for application in laundry detergents.As enzimas, principalmente as proteases, têm uma participação importante e indispensável em muitos processos industriais tais como na indústria farmacêutica, de alimentos e de detergentes. Alguns detergentes como Tide®, Ariel® e Biz® contem enzimas proteolíticas em sua formulação, sendo a maioria produzida por bactérias do gênero Bacillus sp. Neste artigo, foi avaliada a compatibilidade de uma protease produzida por um microrganismo termofílico, Bacillus sp., com alguns detergentes comerciais. Estudos sobre a estabilidade mostraram que a enzima reteve cerca de 95% e 74% de sua máxima atividade após 1h a 60ºC na presença de glicina em combinação com MnSO4 e CaCl2 respectivamente. A enzima não foi inibida em presença de 1

Predicción computacional de la estructura terciaria de la iduronato 2-sulfato sulfatasa humana

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Homero Sáenz

2007-03-01

Conclusión. Los valores de RMS y la correlación genotipo-fenotipo en los pacientes analizados sugieren el modelo puede usarse para predecir ciertos aspectos del comportamiento biológico de la enzima.

CORRELATION OF MORPHOLOGICAL CHA RACTERISTICS STUDENTS WITH SUCCESS IN PERFORMAnCE OF FOLK DANCES

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Amra Nožinović Mujanović

2007-05-01

Full Text Available On the sam ple of 121stu dent from se cond and third gra de of Fa culty of physi cal edu ca tion and sport, is be ing me a su red by using 20 va ri a bles of morp ho lo gi cal cha racte ri stics and 4 va ri a bles for esti ma tion of the suc cess in per for ming the folk dan ces, with the aim to de ter mi ne the ir con nec tion to the re sults of the esti ma ted va lue from the practi cal part of Folk dan ce lec tu ring. Ba sed on pre sen ted re sults, by ca no nic cor re la tion analysis in ma ni fe sted spa ce, ma jor part of pre dic tion gro up is in vol ved in va ri a bi lity of suc cess of cri te ria va ri a ble what con fi rms one im por tant ca no nic par of fac tors (Ca noni cal R .40.

Hipotensão arterial em cirurgia de revascularização do miocárdio: influência dos inibidores da enzima conversora de angiotensina Hipotensión arterial en cirugía de revascularización del miocardio: influencia de los inhibidores de la enzima conversora de angiotensina Arterial hypotension in myocardial revascularization surgery: influence of angiotensin-converting enzyme inhibitors

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Míriam Gomes Jordão

2002-04-01

Full Text Available JUSTIFICATIVA E OBJETIVOS: Os inibidores da enzima conversora da angiotensina (IECA são drogas muito utilizadas em estados hipertensivos e insuficiência cardíaca. Seu uso prolongado pode acarretar instabilidade hemodinâmica com episódios hipotensivos durante a indução anestésica. O objetivo deste estudo é comparar a incidência de hipotensão arterial em pacientes cronicamente tratados com IECA com pacientes não tratados com IECA, quando submetidos à anestesia para cirurgia de revascularização do miocárdio. MÉTODO: Participaram do estudo 50 pacientes, estado físico ASA II, III e IV, divididos em dois grupos: Grupo 1 - pacientes tratados com IECA por mais de dois meses e Grupo 2 - pacientes que não fazem uso de IECA. Os parâmetros avaliados foram pressão arterial média (PAM, freqüência cardíaca (FC, sendo anotados os menores valores da PAM e FC verificados em diferentes períodos da anestesia, e análise do segmento ST em D II e V5. Durante a CEC, foi determinada a resistência vascular sistêmica. RESULTADOS: A incidência de hipotensão arterial em pacientes anestesiados em uso de IECA foi maior do que no grupo controle em vários períodos da anestesia, mas principalmente na indução anestésica. Neste grupo foi necessário o uso de dopamina por tempo mais prolongado. Dos 26 pacientes tratados previamente com IECA, 23% necessitaram de drogas para correção da hipotensão desde a indução até a CEC e 19,1% em outros períodos da anestesia, perfazendo um total de 42,3%. No grupo controle nenhum paciente necessitou infusão contínua de drogas para aumentar a pressão arterial sistêmica, da indução até a CEC. Porém, 21% dos pacientes deste grupo necessitaram dopamina ou araminol em um ou mais períodos da anestesia. CONCLUSÕES: Neste estudo, os pacientes tratados com IECA, por tempo prolongado, apresentam maior incidência de hipotensão arterial na indução anestésica, necessitando, com maior freqüência, de

Influência da adubação potássica na produção e na atividade de enzimas pós-colheita em escarola (Cichorium endivia L. / Influence of potassium fertilization in endive (Cichorium endiviaL. production and enzyme post-harvest activity

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Paulo Sérgio Pavinato

2009-12-01

Full Text Available ResumoO potássio é um dos nutrientes mais abundantes no citoplasma das células vegetais, e sua alta contribuição no metabolismo das plantas está relacionada ao controle osmótico das células e à ativação de inúmeras enzimas. Sendo de conhecimento que a produção e manutenção da qualidade pós-colheita de sementes, frutos e hortaliças depende da boa nutrição potássica, o presente trabalho teve por objetivo avaliar o efeito de doses de potássio na produção e qualidade pós-colheita de escarola. A escarola, variedade Malan, foi cultivada em vasos com capacidade de 10 kg de solo, em casa de vegetação, na Fazenda Experimental Lajeado, Departamento de Produção Vegetal/Horticultura da FCA-UNESP, Botucatu-SP. Os tratamentos consistiram em cinco doses de potássio, correspondentes a 0, 75, 150, 300 e 600 kg ha-1 de K2O, aplicadas 1/3 no plantio e o restante em três coberturas (8, 16 e 28 dias após o transplante. Por ocasião do transplante, aplicou-se a adubação nitrogenada e fosfatada recomendadas para a cultura. Aos 35 dias após o transplante, as plantas foram colhidas e se efetuaram as avaliações de diâmetro de cabeça, massa fresca e área foliar. Ainda utilizando plantas frescas, determinou-se a atividade das enzimas polifenol oxidase e peroxidase. Em plantas secas, em estufa a 60°C por 72 h, foram determinadas a massa seca, o teor de K no tecido e o acúmulo total de proteínas. De maneira geral, a aplicação de 150 kg ha-1 de K2O, correspondente à dose recomendada pelos boletins de adubação para a escarola, resultou em máxima produção e qualidade pós-colheita, não respondendo a doses mais elevadas.AbstractPotassium is one of the nutrients that is present in higher levels in the cytoplasm of plant cells, and its contribution in plant metabolism is related to cell osmotic regulation and the activation of many enzymes. Given the knowledge that production and post-harvest quality maintenance of seeds, fruit

Mapping insecticide resistance and characterization of resistance mechanisms in Anopheles arabiensis (Diptera: Culicidae) in Ethiopia.

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Alemayehu, Eba; Asale, Abebe; Eba, Kasahun; Getahun, Kefelegn; Tushune, Kora; Bryon, Astrid; Morou, Evangelia; Vontas, John; Van Leeuwen, Thomas; Duchateau, Luc; Yewhalaw, Delenasaw

2017-09-02

The emergence and spread of insecticide resistance in the major African malaria vectors Anopheles gambiae (s.s.) and An. arabiensis may compromise the current vector control interventions and threatens the global malaria control and elimination efforts. Insecticide resistance was monitored in several study sites in Ethiopia from 2013 to 2015 using papers impregnated with discriminating concentrations of DDT, deltamethrin, bendiocarb, propoxur, malathion, fenitrothion and pirimiphos-methyl, following the WHO insecticide susceptibility test procedure. Mosquitoes sampled from different localities for WHO bioassay were morphologically identified as An. gambiae (s.l.) using standard taxonomic keys. Samples were identified to species using species-specific polymerase chain reaction (PCR) and screened for the presence of target site mutations L1014F, L1014S and N1575Y in the voltage gated sodium channel (VGSC) gene and G119S in the acethylcholinesterase (AChE) gene using allele-specific PCR. Biochemical assays were performed to assess elevated levels of acetylcholinesterases, carboxylcholinesterases, glutathione-S-transferases (GSTs) and cytochrome P450s monooxygenases in wild populations of An. arabiensis, compared to the fully susceptible Sekoru An. arabiensis laboratory strain. Populations of An. arabiensis were resistant to DDT and deltamethrin but were susceptible to fenitrothion in all the study sites. Reduced susceptibility to malathion, pirimiphos-methyl, propoxur and bendiocarb was observed in some of the study sites. Knockdown resistance (kdr L1014F) was detected in all mosquito populations with allele frequency ranging from 42 to 91%. Elevated levels of glutathione-S-transferases (GSTs) were detected in some of the mosquito populations. However, no elevated levels of monooxygenases and esterases were detected in any of the populations assessed. Anopheles arabiensis populations from all surveyed sites in Ethiopia exhibited resistance against DDT and pyrethroids

Characterizing the insecticide resistance of Anopheles gambiae in Mali.

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Cisse, Moussa B M; Keita, Chitan; Dicko, Abdourhamane; Dengela, Dereje; Coleman, Jane; Lucas, Bradford; Mihigo, Jules; Sadou, Aboubacar; Belemvire, Allison; George, Kristen; Fornadel, Christen; Beach, Raymond

2015-08-22

The impact of indoor residual spraying (IRS) and long-lasting insecticide nets (LLINs), key components of the national malaria control strategy of Mali, is threatened by vector insecticide resistance. The objective of this study was to assess the level of insecticide resistance in Anopheles gambiae sensu lato populations from Mali against four classes of insecticide recommended for IRS: organochlorines (OCs), pyrethroids (PYs), carbamates (CAs) and organophosphates (OPs). Characterization of resistance was done in 13 sites across southern Mali and assessed presence and distribution of physiological mechanisms that included target-site modifications: knockdown resistance (kdr) and altered acetycholinesterase (AChE), and/or metabolic mechanisms: elevated esterases, glutathione S-transferases (GSTs), and monooxygenases. The World Health Organization (WHO) tube test was used to determine phenotypic resistance of An. gambiae s.l. to: dichlorodiphenyltrichloroethane (DDT) (OC), deltamethrin (PY), lambda-cyhalothrin (PY), bendiocarb (CA), and fenitrothion (OP). Identification of sibling species and presence of the ace-1 (R) and Leu-Phe kdr, resistance-associated mutations, were determined using polymerase chain reaction (PCR) technology. Biochemical assays were conducted to detect increased activity of GSTs, oxidases and esterases. Populations tested showed high levels of resistance to DDT in all 13 sites, as well as increased resistance to deltamethrin and lambda-cyhalothrin in 12 out of 13 sites. Resistance to fenitrothion and bendiocarb was detected in 1 and 4 out of 13 sites, respectively. Anopheles coluzzii, An. gambiae sensu stricto and Anopheles arabiensis were identified with high allelic frequencies of kdr in all sites where each of the species were found (13, 12 and 10 sites, respectively). Relatively low allelic frequencies of ace-1 (R) were detected in four sites where this assessment was conducted. Evidence of elevated insecticide metabolism, based on oxidase

Semiannual Report to the Congress: April 1, 2013 to September 30, 2013

Science.gov (United States)

2013-09-30

providers’ selection and utilization of Aranesp within nursing homes, skilled nursing facilities and long-term care settings. reSult: On April 4... nurse case managers, are appropriate to meet the mission for effective management and support of soldiers during their healing and transition...needs to perform additional audit procedures to support the audit conclusions and overall audit opinion. DoD IG will reschedule its review once the

2741-IJBCS-Proof-Njoyim Estella Buleng T

African Journals Online (AJOL)

hp

Néanmoins, le chrome et fer étaient fortement concentrés dans presque tous les échantillons au mois de janvier (0,14 à ...... d'eau, augmentant ainsi les infections observées en ces mois. Ce cycle résulte en l'occurrence des infections d'origine hydrique tout au long de l'année. Conclusion. A l'issue des analyses physiques,.

Serological study of Helicobacter pylori infection in patients with Polycystic Ovary Syndrome

Directory of Open Access Journals (Sweden)

Farnaz Sohrabvand

2014-01-01

Conclusion: This study showed no significant difference in serologic examination re-sults in PCOS versus non PCOS patients. The finding of high prevalence of H.Pylori IgG and IgA positive levels in both PCOS and non PCOS patients can be probably re-lated to the high prevalence of H.Pylori infection or exposure in Iranian population and therefore suggest an issue for further investigation.

Case report

African Journals Online (AJOL)

raoul

21 déc. 2011 ... Abstract. La thrombasthenie de Glanzmann est une anomalie plaquettaire qualitative, à transmission autosomale récessive, due à un déficit en glycoprotéine. IIb/IIIa (GPIIb/IIIa), avec un défaut d'agrégation plaquettaire. Il en résulte une perturbation de l'hémostase primaire, à l'origine d'hémorragie plus.

Use of GERAM as Basis for a Virtual Enterprise Framework Model

DEFF Research Database (Denmark)

Vesterager, Johan; Bernus, Peter; Larsen, Lars B

2001-01-01

In the IMS-project Globeman21, the enterprise reference architecture GERAM was used as basis for creation of a virtual enterprise framework model. The model was used to map different industrial pilot projects, to classify virtual en-terprise concepts, and as underlying structure for a virtual...... enterprise manage-ment methodology. The paper gives a survey of the use of GERAM and the re-sults obtained....

Enfants travailleurs domestiques a cotonou : Entre contraintes ...

African Journals Online (AJOL)

Le décalage de perception sur le travail des enfants résulte d'abord de la définition du concept d'enfant perçu différemment selon qu'il s'agit des normes juridiques ou des codes sociaux ou communautaires. ... Par conséquent, la domesticité enfantine ressemble davantage à un projet élaboré par un ensemble d'acteurs.

Proceedings of the Conference on the Stability and Dynamic Response of Rotors with Squeeze Film Bearings, 8-10 May 79.

Science.gov (United States)

1979-01-01

in flexible engine rotors and structures. 2. Small physical size, especially in frontal area and wheel diameter. 3. Increasingly higher shaft speeds...requirements for high power and small wheel diameter re- sults in extremely small blade tip clearances. This limits the acceptable rotor disk excursions...installation, alignment of the damper center with the bearing centers should be accurately maintained, lest runout error would cause excitations at rota

De nouvelles variétés de pomme de terre pour lutter contre la ...

International Development Research Centre (IDRC) Digital Library (Canada)

C'est la principale culture à laquelle s'adonnent les membres de 67 collectivités autochtones dans le département de Nariño, la région qui affiche le deuxième plus fort taux de malnutrition en Colombie. Pourtant, cette culture est fort susceptible au mildiou et donne, par conséquent, de faibles rendements. Il en résulte une ...

Le CRDI en Jordanie

International Development Research Centre (IDRC) Digital Library (Canada)

... dans les pays en développement afin d'y favoriser la croissance et le développement. Il en résulte des solutions novatrices et durables qui ont pour but d'améliorer les conditions de vie et les moyens de subsistance. Centre de recherches pour le développement international. CP 8500, Ottawa (Ontario) Canada K1G 3H9.

Mathematics in Ancient India

Indian Academy of Sciences (India)

In this series of articles, we intend to have a glimpse of some of the landmarks ... then the complete solution due to J ayadeva and .... on the numerical and not so much on the proper geo- .... sult of which children all over the world learn it even at a tender age .... own right as well as an indispensable tool in other fields. In fact ...

Study of association between genetic polymorphisms of phospholipase A2 enzymes and Alzheimer's disease Associação entre polimorfismos das enzimas fosfolipases A2 e doença de Alzheimer

Directory of Open Access Journals (Sweden)

Quirino Cordeiro

2010-04-01

Full Text Available Several genes have been related to late-onset Alzheimer's disease (LOAD. Phospholipases A2 (PLA2 influence the processing and secretion of the amyloid precursor protein, which gives rise to the beta-amyloid peptide, the major component of the amyloid plaque in AD. Hence, in the present study, polymorphisms of three genes encoding PLA2 enzymes group (cytosolic PLA2: BanI cPLA2 polymorphism; calcium-independent PLA2: AvrII iPLA2 polymorphism; PAFAH: Val279Phe PAFAH polymorphism were analysed in a case-control sample using 58 patients with LOAD and 107 matched healthy controls. There was a genotypic association between the BanI cPLA2 polymorphism and LOAD (χ2=6.25, 2df, p=0.04, however there was no allelic association. There were no associations between AvrII iPLA2 and Val279Phe PAFAH polymorphisms and LOAD. These data suggest that the BanI cPLA2 polymorphism may play a role in the susceptibility for LOAD in our Brazilian sample.Vários genes têm sido investigados como fatores de risco para o desenvolvimento da doença de Alzheimer (DA de início tardio. As fosfolipases A2 (PLA2 influenciam o processamento e secreção da proteína precursora do amilóide, que dá origem ao peptídeo meta-amilóide, o principal componente da placa amilóide na DA. Assim, no presente estudo, foram analisados três polimorfismos genéticos que codificam enzimas do grupo das PLA2 (PLA2 citosólica: polimorfismo BanI cPLA2; PLA2 cálcio-independente: polimorfismo AvrII iPLA2; PAFAH: polimorfismo Val279Phe PAFAH em 58 pacientes com DA de início tardio e 107 controles saudáveis pareados. Houve associação genotípica entre o polimorfismo BanI cPLA2 e DA de início tardio (χ2=6,25, 2df, p=0,04; no entanto não foi observada associação alélica. Não houve associação entre os polimorfismos AvrII iPLA2 e Val279Phe PAFAH com a doença. Tais dados sugerem que o polimorfismo BanI cPLA2 pode estar envolvido como fator de susceptibilidade para DA de início tardio em

Potential hazards to embryo implantation: A human endometrial in vitro model to identify unwanted antigestagenic actions of chemicals

International Nuclear Information System (INIS)

Fischer, L.; Deppert, W.R.; Pfeifer, D.; Stanzel, S.; Weimer, M.; Hanjalic-Beck, A.; Stein, A.; Straßer, M.; Zahradnik, H.P.; Schaefer, W.R.

2012-01-01

Embryo implantation is a crucial step in human reproduction and depends on the timely development of a receptive endometrium. The human endometrium is unique among adult tissues due to its dynamic alterations during each menstrual cycle. It hosts the implantation process which is governed by progesterone, whereas 17β-estradiol regulates the preceding proliferation of the endometrium. The receptors for both steroids are targets for drugs and endocrine disrupting chemicals. Chemicals with unwanted antigestagenic actions are potentially hazardous to embryo implantation since many pharmaceutical antiprogestins adversely affect endometrial receptivity. This risk can be addressed by human tissue-specific in vitro assays. As working basis we compiled data on chemicals interacting with the PR. In our experimental work, we developed a flexible in vitro model based on human endometrial Ishikawa cells. Effects of antiprogestin compounds on pre-selected target genes were characterized by sigmoidal concentration–response curves obtained by RT-qPCR. The estrogen sulfotransferase (SULT1E1) was identified as the most responsive target gene by microarray analysis. The agonistic effect of progesterone on SULT1E1 mRNA was concentration-dependently antagonized by RU486 (mifepristone) and ZK137316 and, with lower potency, by 4-nonylphenol, bisphenol A and apigenin. The negative control methyl acetoacetate showed no effect. The effects of progesterone and RU486 were confirmed on the protein level by Western blotting. We demonstrated proof of principle that our Ishikawa model is suitable to study quantitatively effects of antiprogestin-like chemicals on endometrial target genes in comparison to pharmaceutical reference compounds. This test is useful for hazard identification and may contribute to reduce animal studies. -- Highlights: ► We compare progesterone receptor-mediated endometrial effects of chemicals and drugs. ► 4-Nonylphenol, bisphenol A and apigenin exert weak

Potential hazards to embryo implantation: A human endometrial in vitro model to identify unwanted antigestagenic actions of chemicals

Energy Technology Data Exchange (ETDEWEB)

Fischer, L.; Deppert, W.R. [Department of Obstetrics and Gynecology, University Hospital Freiburg (Germany); Pfeifer, D. [Department of Hematology and Oncology, University Hospital Freiburg (Germany); Stanzel, S.; Weimer, M. [Department of Biostatistics, German Cancer Research Center, Heidelberg (Germany); Hanjalic-Beck, A.; Stein, A.; Straßer, M.; Zahradnik, H.P. [Department of Obstetrics and Gynecology, University Hospital Freiburg (Germany); Schaefer, W.R., E-mail: wolfgang.schaefer@uniklinik-freiburg.de [Department of Obstetrics and Gynecology, University Hospital Freiburg (Germany)

2012-05-01

Embryo implantation is a crucial step in human reproduction and depends on the timely development of a receptive endometrium. The human endometrium is unique among adult tissues due to its dynamic alterations during each menstrual cycle. It hosts the implantation process which is governed by progesterone, whereas 17β-estradiol regulates the preceding proliferation of the endometrium. The receptors for both steroids are targets for drugs and endocrine disrupting chemicals. Chemicals with unwanted antigestagenic actions are potentially hazardous to embryo implantation since many pharmaceutical antiprogestins adversely affect endometrial receptivity. This risk can be addressed by human tissue-specific in vitro assays. As working basis we compiled data on chemicals interacting with the PR. In our experimental work, we developed a flexible in vitro model based on human endometrial Ishikawa cells. Effects of antiprogestin compounds on pre-selected target genes were characterized by sigmoidal concentration–response curves obtained by RT-qPCR. The estrogen sulfotransferase (SULT1E1) was identified as the most responsive target gene by microarray analysis. The agonistic effect of progesterone on SULT1E1 mRNA was concentration-dependently antagonized by RU486 (mifepristone) and ZK137316 and, with lower potency, by 4-nonylphenol, bisphenol A and apigenin. The negative control methyl acetoacetate showed no effect. The effects of progesterone and RU486 were confirmed on the protein level by Western blotting. We demonstrated proof of principle that our Ishikawa model is suitable to study quantitatively effects of antiprogestin-like chemicals on endometrial target genes in comparison to pharmaceutical reference compounds. This test is useful for hazard identification and may contribute to reduce animal studies. -- Highlights: ► We compare progesterone receptor-mediated endometrial effects of chemicals and drugs. ► 4-Nonylphenol, bisphenol A and apigenin exert weak

Differential Gene Expression across Breed and Sex in Commercial Pigs Administered Fenbendazole and Flunixin Meglumine.

Directory of Open Access Journals (Sweden)

Jeremy T Howard

Full Text Available Characterizing the variability in transcript levels across breeds and sex in swine for genes that play a role in drug metabolism may shed light on breed and sex differences in drug metabolism. The objective of the study is to determine if there is heterogeneity between swine breeds and sex in transcript levels for genes previously shown to play a role in drug metabolism for animals administered flunixin meglumine or fenbendazole. Crossbred nursery female and castrated male pigs (n = 169 spread across 5 groups were utilized. Sires (n = 15 of the pigs were purebred Duroc, Landrace, Yorkshire or Hampshire boars mated to a common sow population. Animals were randomly placed into the following treatments: no drug (control, flunixin meglumine, or fenbendazole. One hour after the second dosing, animals were sacrificed and liver samples collected. Quantitative Real-Time PCR was used to measure liver gene expression of the following genes: SULT1A1, ABCB1, CYP1A2, CYP2E1, CYP3A22 and CYP3A29. The control animals were used to investigate baseline transcript level differences across breed and sex. Post drug administration transcript differences across breed and sex were investigated by comparing animals administered the drug to the controls. Contrasts to determine fold change were constructed from a model that included fixed and random effects within each drug. Significant (P-value <0.007 basal transcript differences were found across breeds for SULT1A1, CYP3A29 and CYP3A22. Across drugs, significant (P-value <0.0038 transcript differences existed between animals given a drug and controls across breeds and sex for ABCB1, PS and CYP1A2. Significant (P <0.0038 transcript differences across breeds were found for CYP2E1 and SULT1A1 for flunixin meglumine and fenbendazole, respectively. The current analysis found transcript level differences across swine breeds and sex for multiple genes, which provides greater insight into the relationship between flunixin

Gene expression profiling in chemoresistant variants of three cell lines of different origin

DEFF Research Database (Denmark)

Johnsson, Anders; Vallon-Christensson, Johan; Strand, Carina

2005-01-01

lines (K562 leukemia, MCF-7 breast cancer and S1 colon cancer) with acquired resistance against five cytostatic drugs; daunorubicin (DNR), doxorubicin (DOX), vincristine (VCR), etoposide (VP) and mitoxantrone (MX). RESULTS: The resistant cell lines clustered together based on their type of origin...... was also seen in, e.g., GSTs, topoisomerases, caveolins, annexins and CD44. CONCLUSION: These results will constitute a platform for further studies on specific pathways and biological processes involved in chemotherapy resistance....

Thermostabilisation of human serum butyrylcholinesterase for detection of its inhibitors in water and biological fluids

Directory of Open Access Journals (Sweden)

Lakshmanan Jaganathan

1999-01-01

Full Text Available The ability of gelatine-trehalose to convert the normally fragile, dry human serum BChE into a thermostable enzyme and its use in the detection of cholinesterase inhibitors in water and biological fluids is described. Gelatine or trehalose alone is unable to protect the dry enzyme against exposure to high temperature, while a combination of gelatine and trehalose were able to protect the enzyme activity against prolonged exposure to temperature as high as +50°C. A method for rapid, simple and inexpensive means of screening for cholinesterase inhibitors such as carbamates and organophosphates in water, vegetables and human blood has been developed.A capacidade da gelatina-trehalose em converter a frágil BChE do soro humano em uma enzima termoestável e seu uso na descoberta de inibidores de colinesterase em água e fluidos biológicos é apresentado. A Gelatina ou trehalose são incapazes de proteger a enzima seca BchE do soro humano contra exposição a elevadas temperaturas, enquanto que uma combinação de gelatina e trehalose são capazes de proteger a atividade de enzima contra exposição prolongada a temperaturas elevadas e da ordem de 50° C. Um método barato, simples e rápido de screening para inibidores de colinesterase tal como carbamatos e organofosfatos em água, verduras e sangue humano foi desenvolvido.

Activity-Based Probes for Isoenzyme- and Site-Specific Functional Characterization of Glutathione S -Transferases

Energy Technology Data Exchange (ETDEWEB)

Stoddard, Ethan G. [Chemical Biology and Exposure; Killinger, Bryan J. [Chemical Biology and Exposure; Nair, Reji N. [Chemical Biology and Exposure; Sadler, Natalie C. [Chemical Biology and Exposure; Volk, Regan F. [Chemical Biology and Exposure; Purvine, Samuel O. [Chemical Biology and Exposure; Shukla, Anil K. [Chemical Biology and Exposure; Smith, Jordan N. [Chemical Biology and Exposure; Wright, Aaron T. [Chemical Biology and Exposure

2017-11-01

Glutathione S-transferases (GSTs) comprise a highly diverse family of phase II drug metabolizing enzymes whose shared function is the conjugation of reduced glutathione to various endo- and xenobiotics. Although the conglomerate activity of these enzymes can be measured by colorimetric assays, measurement of the individual contribution from specific isoforms and their contribution to the detoxification of xenobiotics in complex biological samples has not been possible. For this reason, we have developed two activity-based probes that characterize active glutathione transferases in mammalian tissues. The GST active site is comprised of a glutathione binding “G site” and a distinct substrate binding “H site”. Therefore, we developed (1) a glutathione-based photoaffinity probe (GSH-ABP) to target the “G site”, and (2) a probe designed to mimic a substrate molecule and show “H site” activity (GST-ABP). The GSH-ABP features a photoreactive moiety for UV-induced covalent binding to GSTs and glutathione-binding enzymes. The GST-ABP is a derivative of a known mechanism-based GST inhibitor that binds within the active site and inhibits GST activity. Validation of probe targets and “G” and “H” site specificity was carried out using a series of competitors in liver homogenates. Herein, we present robust tools for the novel characterization of enzyme- and active site-specific GST activity in mammalian model systems.

Analyses of Genetic Variations of Glutathione S-Transferase Mu1 and Theta1 Genes in Bangladeshi Tannery Workers and Healthy Controls

Directory of Open Access Journals (Sweden)

Jobaida Akther

2016-01-01

Full Text Available Glutathione S-transferases (GSTs belong to a group of multigene detoxification enzymes, which defend cells against oxidative stress. Tannery workers are at risk of oxidative damage that is usually detoxified by GSTs. This study investigated the genotypic frequencies of GST Mu1 (GSTM1 and GST Theta1 (GSTT1 in Bangladeshi tannery workers and healthy controls followed by their status of oxidative stress and total GST activity. Of the 188 individuals, 50.0% had both GSTM1 and GSTT1 (+/+, 12.2% had GSTM1 (+/−, 31.4% had GSTT1 (−/+ alleles, and 6.4% had null genotypes (−/− with respect to both GSTM1 and GSTT1 alleles. Among 109 healthy controls, 54.1% were double positive, 9.2% had GSTM1 allele, 32.1% had GSTT1 allele, and 4.6% had null genotypes. Out of 79 tannery workers, 44.3% were +/+, 16.8% were +/−, 30.5% were −/+, and 8.4% were −/−. Though the polymorphic genotypes or allelic variants of GSTM1 and GSTT1 were distributed among the study subjects with different frequencies, the differences between the study groups were not statistically significant. GST activity did not vary significantly between the two groups and also among different genotypes while level of lipid peroxidation was significantly higher in tannery workers compared to controls irrespective of their GST genotypes.

Analyses of Genetic Variations of Glutathione S-Transferase Mu1 and Theta1 Genes in Bangladeshi Tannery Workers and Healthy Controls

Science.gov (United States)

Akther, Jobaida; Ebihara, Akio; Nakagawa, Tsutomu; Islam, Laila N.; Suzuki, Fumiaki; Hosen, Md. Ismail; Hossain, Mahmud; Nabi, A. H. M. Nurun

2016-01-01

Glutathione S-transferases (GSTs) belong to a group of multigene detoxification enzymes, which defend cells against oxidative stress. Tannery workers are at risk of oxidative damage that is usually detoxified by GSTs. This study investigated the genotypic frequencies of GST Mu1 (GSTM1) and GST Theta1 (GSTT1) in Bangladeshi tannery workers and healthy controls followed by their status of oxidative stress and total GST activity. Of the 188 individuals, 50.0% had both GSTM1 and GSTT1 (+/+), 12.2% had GSTM1 (+/−), 31.4% had GSTT1 (−/+) alleles, and 6.4% had null genotypes (−/−) with respect to both GSTM1 and GSTT1 alleles. Among 109 healthy controls, 54.1% were double positive, 9.2% had GSTM1 allele, 32.1% had GSTT1 allele, and 4.6% had null genotypes. Out of 79 tannery workers, 44.3% were +/+, 16.8% were +/−, 30.5% were −/+, and 8.4% were −/−. Though the polymorphic genotypes or allelic variants of GSTM1 and GSTT1 were distributed among the study subjects with different frequencies, the differences between the study groups were not statistically significant. GST activity did not vary significantly between the two groups and also among different genotypes while level of lipid peroxidation was significantly higher in tannery workers compared to controls irrespective of their GST genotypes. PMID:27294127

Targeting Glutathione-S Transferase Enzymes in Musculoskeletal Sarcomas: A Promising Therapeutic Strategy

Directory of Open Access Journals (Sweden)

Michela Pasello

2011-01-01

Full Text Available Recent studies have indicated that targeting glutathione-S-transferase (GST isoenzymes may be a promising novel strategy to improve the efficacy of conventional chemotherapy in the three most common musculoskeletal tumours: osteosarcoma, Ewing's sarcoma, and rhabdomyosarcoma. By using a panel of 15 drug-sensitive and drug-resistant human osteosarcoma, Ewing's sarcoma, and rhabdomyosarcoma cell lines, the efficay of the GST-targeting agent 6-(7-nitro-2,1,3-benzoxadiazol-4-ylthiohexanol (NBDHEX has been assessed and related to GST isoenzymes expression (namely GSTP1, GSTA1, GSTM1, and MGST. NBDHEX showed a relevant in vitro activity on all cell lines, including the drug-resistant ones and those with higher GSTs levels. The in vitro activity of NBDHEX was mostly related to cytostatic effects, with a less evident apoptotic induction. NBDHEX positively interacted with doxorubicin, vincristine, cisplatin but showed antagonistic effects with methotrexate. In vivo studies confirmed the cytostatic efficay of NBDHEX and its positive interaction with vincristine in Ewing's sarcoma cells, and also indicated a positive effect against the metastatisation of osteosarcoma cells. The whole body of evidence found in this study indicated that targeting GSTs in osteosarcoma, Ewing's sarcoma and rhabdomyosarcoma may be an interesting new therapeutic option, which can be considered for patients who are scarcely responsive to conventional regimens.

Fort Hood Army Internal Review Team: Final Report

Science.gov (United States)

2010-08-04

lead to or re- sult in violent acts. 2. Finding 2.3 - DoD standards for denying requests for recognition as an ecclesiastical endorser of chaplains may...review and pos- sible revision to ensure the supporting architecture and capabilities exist to support the mission. No cost estimate is necessary to...enterprise architecture . The goal is to field interoperable installation physi- cal access control systems that can access the Defense Manpower Data

2011 Agile (Scrum) Workshop Held in Baltimore, Maryland on November 14-15, 2011

Science.gov (United States)

2011-11-15

ment melded with CMMI Level 3 discipline. Her programs put the end-user in control of priorities and delivered capabilities re- sulting in a...OPS PEX Team: Internal •Devs/ SMEs /Testers •Architecture Committee •Etc. none Future implementation, not Current release Big R/Little r I n...Refine Pre-release Planning Release Planning Release Execution Post- release POT: ( SMEs and Developers) •Based on Approved Roadmap •Decomposes

Rendre les villes plus sûres grâce à l'emploi et à la rémunération

International Development Research Centre (IDRC) Digital Library (Canada)

crimes contre les biens (et non des crimes « payants » pour les criminels). Certains analystes croient que le taux de criminalité élevé de l'Afrique du. Sud résulte de la fracture psychologique et sociale qui survient lorsqu'un grand nombre de personnes est laissé pour compte au sein d'une société d'abondance. « L'inégalité ...

Model-Driven Engineering: Automatic Code Generation and Beyond

Science.gov (United States)

2015-03-01

herein to any specific commercial product, process, or service by trade name, trade mark, manufacturer , or otherwise, does not necessarily constitute or...export of an Extensible Markup Language (XML) representation of the model. The XML Metadata Interchange (XMI) is an OMG standard for representing...overall company financial results for the past 3 years. What financial re- sults are you projecting for the next year? 1.2.5.2 Percentage of Gross

Structural Metadata Research in the Ears Program

Science.gov (United States)

2005-01-01

detecting structural information in the word stream (the so-called “structural MDE” portion of the EARS program); other MDE efforts on speaker ... diarization are overviewed in [13]. The rest of this paper is organized as follows. We describe the structural MDE tasks, performance measurement, and corpora...tems have only recently been introduced, with NIST reporting re- sults with the Wilcoxon signed rank test for speaker -level average score differences

Wind pressure testing of tornado safe room components made from wood

Science.gov (United States)

Robert Falk; Deepak Shrestha

2016-01-01

To evaluate the ability of a wood tornado safe room to resist wind pressures produced by a tornado, two safe room com-ponents were tested for wind pressure strength. A tornado safe room ceiling panel and door were static-pressure-tested according to ASTM E 330 using a vacuum test system. Re-sults indicate that the panels had load capacities from 2.4 to 3.5 times that...

Assessing two different peroxidases´ potential for application in recalcitrant organic compound bioremediation Evaluación del potencial de dos peróxidas para su aplicación en biorremedación de compuestos orgánicos recalcitrantes

Directory of Open Access Journals (Sweden)

Torres Rodrigo

2001-12-01

Full Text Available This work shows the promising future presented by the following enzymes: Chloroperoxidase (CPO from Caldariomyces fumago and royal palm peroxidase (Roystonea regia, PPR. These peroxidases were obtained from different sources (microbial and vegetable and used as biocatalysts for applicating them in bioremediation of recalcitrant organic compounds. Each one of the enzymes' peroxidase catalytic activity was evaluated in organic phase systems, using different model compounds such as: PAHs (pyrene and anthracene, organic-nitrogenated compounds (diphenylamine, monoaromatic phenolic molecules (guayacol and dyes (methyl orange and ABTS. The reaction systems were composed of mono-phase water mixtures and organic miscible solvent (methanol, ethanol, isopropanol, acetonitrile, tetrahydrofuran, dimethyl sulfoxide and dimethyl formamide, on which both peroxidases' catalytic activity was evaluated. The two enzymes' catalytic activity was observed on the evaluated substrates in most of these assays. However, PPR did not show biocatalytic oxidation for methyl orange dye and some PAHs. This enzyme did show the best tolerance to the evaluated solvents. Its catalytic activity was appreciably enhanced when low hydrophobic solvents were used. The kcat was calculated from this experimental data (as kinetic parameter leading to each enzyme's biocatalytic performance on substrates being compared.El presente trabajo demuestra el promisorio futuro que presentan las enzimas cloroperoxidasa del hongo Caldariomyces fumago (CPO y la peroxidasa de palma real (Roystonea regia, PPR, dos peroxidasas de diferente fuente (microbiana y vegetal, usadas como biocatalizadores para aplicación en biorremediación de compuestos orgánicos recalcitrantes. Para este estudio, se evaluó la actividad catalítica tipo peroxidasa, en sistemas de fase orgánica, de cada una de las enzimas empleando diferentes compuestos modelos tales como: PAH's (pireno y antraceno, órgano nitrogenados

Inmovilización covalente de glucosa oxidasa y peroxidasa

Directory of Open Access Journals (Sweden)

Yuria Bilbao Abraham

2000-08-01

Full Text Available Las enzimas por su función catalítica tienen amplia aplicación en infinidad de procesos tecnológicos y en los últimos 15 a han marcado avances significativos en la industria. Dentro de la Industria Farmacéutica y Biológica, la dedicada a los medios diagnósticos ha recibido también el impacto de la introducción de este tipo de productos, soportando en la actualidad tecnologías tan importantes como el inmunoensayo enzimático, el diagnóstico en química clínica y la química seca, donde las técnicas de inmovilización alcanzan un desarrollo cada vez mayor por el incremento de la estabilidad que se logra con estos sistemas. Se presentan ensayos de inmovilización covalente de las enzimas glucosa oxidasa y peroxidasa sobre papel de filtro Whatman No. 2. Fueron ensayadas 2 variantes: la inmovilización covalente de un polímero soluble de las enzimas y la inmovilización covalente de una solución de las enzimas libres. Los resultados del proceso se evaluaron frente a soluciones de referencia de glucosa en concentraciones entre 2,0 y 55,0 mmol/L. Las mejores respuestas se encontraron con el más bajo porcentaje de inmovilización en el caso del enlazamiento del polímero, y para la solución de las enzimas libres correspondió al más alto grado de inmovilización logrado.Due to their catalytic function, enzimes have a wide application in a considerable number of technological processes and during the last l5 years there have been significant advances in industry. Within the Pharmaceutical and Biological Industry, that one devoted to diagnostic tools have also received the impact of the introduction of this type of products, supporting at present technologies as important as the enzime immunoassay, the diagnosis in clinical chemistry and dry chemistry, where the immobilization techniques attain an increasingly higher development as a result of the increase of stability achieved with these systems. Covalent immbolization assays of

Production, characterization and application of inulinase from fungal endophyte CCMB 328

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Diego S. Nascimento

2012-06-01

Full Text Available Inulinase (β-2,1-D- fructan fructanohydrolase, EC 3.2.1.7, targets the β-2,1 linkage of inulin, a polyfructan consisting of linear β-2,1 linked fructose, and hydrolyzes it into fructose. This use provides an alternative to produce fructose syrup through the hydrolysis of inulin. The objective of this work was to study the production, characterization and applications of inulinases from the fungal endophyte CCMB 328 isolated from the Brazilian semi-arid region. Response Surface Methodology (RSM was employed to evaluate the effect of variables (concentration of glucose and yeast extract, on secreted inulinase activities detected in the culture medium and also in the inulin hydrolysis. The results showed that the best conditions for inulinase production by CCMB 328 are 9.89 g / L for glucose and 1.09 g / L for yeast extract. The concentration of 0.20 mol/L of NaCl and KCl increased the activity of inulinase from CCMB 328 by approximately 63% and 37%, respectively. The results also showed that the inulinase has potential for inulin hydrolysis, whose conversion yields roughly 72.48 % for an initial concentration of inulin at 1% (w/v.A enzima inulinase (EC 3.2.1.7, β-D-frutano frutanohidrolase atua sobre as ligações β-2,1 da inulina, um polifrutano consistindo de frutose unida por ligações β-2,1. A hidrólise de inulina através do uso de inulinase é uma alternativa viável para a obtenção de xarope de frutose. O objetivo deste trabalho foi estudar a produção, caracterização e aplicação de inulinase obtidas a partir do fungo endofítico CCMB 328, isolado do semi-árido brasileiro. A metodologia de Superfície de Resposta (MSR foi empregado para avaliar os efeitos das variáveis (concentração de glicose e extrato de levedura na atividade da enzima inulinase produzida em meio de cultura líquido e também para avaliar a atividade da enzima na hidrólise de inulina. Os resultados mostraram que as melhores condições para a produ

Observational tests of the Electro-Magnetic Black Hole Theory in Gamma-Ray Bursts

OpenAIRE

Ruffini, Remo

2002-01-01

The Relative Space-Time Transformation (RSTT) Paradigm and the Interpretation of the Burst Structure (IBS) Paradigm are applied to the analysis of the structure of the burst and afterglow of Gamma-Ray Bursts within the theory based on the vacuum polarization process occurring in an Electro-Magnetic Black Hole, the EMBH theory. This framework is applied to the study of the GRB991216 which is used as a prototype. The GRB-Supernova Time Sequence (GSTS) Paradigm, which introduces the concept of i...

Immobilization of alpha-amylase produced by Bacillus circulans GRS 313

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Gargi Dey

2003-03-01

Full Text Available A maltooligosaccharide-forming amylase from B circulans GRS 313 was immobilized by entrapment in calcium alginate beads. The immobilized activity was affected by the size of the bead and bead size of 2mm was found to be most effective for hydrolysis. Kinetics constants, Km and Vmax were estimated and were found to be affected by the bead size. The catalytic activity of the enzyme was studied in presence of various starchy residues and metal ions. HgCl2, CuSO4 and FeCl3 caused inhibition of the enzyme. The reaction conditions, pH and temperature, was optimized using response surface methodology. At the optimum pH and temperature of 4.9 and 57ºC, the apparent activity was 25.6U/g of beads, resulting in almost 2-fold increase in activity. The immobilized enzyme showed a high operational stability by retaining almost 85% of the initial activity after seventh use.Um maltooligossacarideo obtido a partir de amilase produzida por B. circulans GRS 313 foi imobilizada em alginato de sódio. A atividade enzimática foi afetada pelo tamanho da partícula. Partículas com 2mm foram as mais efetivas na hidrólise. Constantes cinética Km e Vmax foram estimadas e afetadas pelo tamanho das partículas. A atividade catalítica da enzima foi estuda na presença de diferentes tipos de amido e íons metálicos. HgCl2, CuSO4 e FeCl3 provocaram inibição na enzima. As condições de reação (temperatura e pH foram otimizadas utilizando a metodologia da superfície de resposta. Em pH ótimo de 4,9 e temperatura de 57 ºC, a atividade aparente foi de 25.6 U/g de partículas, resultando num acréscimo de mais de 2 vezes na atividade da enzima. A imobilização da enzima mostrou uma alta estabilidade operacional pela retenção de 85% de sua atividade inicial após sete ciclos de utilização.

Aislamiento y caracterización bioquímica de la α-glucosidasa II del hongo patógeno Candida albicans Aislamiento y caracterización bioquímica de la α-glucosidasa II del hongo patógeno Candida albicans

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Arturo Flores Carreón

2012-02-01

Full Text Available Alpha-glucosidase II participates in N-linked glycosylation of proteins. A soluble 47 kDa α-glucosidase II has been previously isolated from C. albicans; however, bioinformatics analysis indicate that native enzyme has a molecular mass of 100 kDa. In this study we assessed the effect of protease inhibitors on intracellular distribution of α-glucosidase II. Despite there was not a significant change in the enzyme distribution, α-glucosidase II activity was associated to a 83 or 47 kDa polypeptide in absence or presence of inhibitors, respectively. Soluble 83-kDa protein was purified by conventional methodology and its biochemical characteristics were similar to those reported for the 47 kDa enzyme. Thus, these results indicated the 83 kDa protein is an α-glucosidase II and also suggested it is a precursor of the 47 kDa enzyme previously reported. La α-glucosidasa II participa en la ruta de la N-glicosilación de proteínas. En Candida albi­cans se ha aislado un polipéptido soluble de 47 kDa con actividad de α-glucosidasa II; sin embargo, análisis bioinformáticos indican que la enzima nativa pudiera tener un peso mo­lecular de 100 kDa. En este trabajo se estudió el efecto de inhibidores de proteasas sobre la distribución intracelular de la α-glucosidasa II. Se demostró que la distribución intracelular no fue afectada significativamente, pero la actividad de la α-glucosidasa II estuvo asociada a una proteína de 83 ó 47 kDa en ausencia o presencia de inhibidores de proteasas, res­pectivamente. La enzima soluble de 83 kDa se purificó por métodos convencionales y se demostró que presenta características bioquímicas similares a la enzima de 47 kDa. Estos datos confirmaron que la proteína de 83 kDa es una α-glucosidasa II y sugieren que es precursora de la enzima de 47 kDa previamente descrita.

Efectos de extractos de Neopetrosia rosariensis sobre mieloperoxidasa y elastasa con la utilización del modelo neutrófilos

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Marlene Durán Lengua

Full Text Available Introducción: los neutrófilos son la primera línea de defensa, los cuales liberan su contenido granular, constituido entre otros por enzimas generadoras de especies reactivas del oxígeno como mieloperoxidasa y enzimas proteolíticas como la elastasa, que degradan elastina y colágeno. Objetivo: evaluar la toxicidad de los extractos de la esponja marina Neopetrosia rosariensis y su acción como inhibidores de la desgranulación del neutrófilo, e inhibición de la actividad catalítica de la elastasa de neutrófilos humanos. Métodos: se determinó la toxicidad por el método de exclusión del colorante azul de tripano. Se evaluó la desgranulación por la disminución del porcentaje de liberación de la mieloperoxidasa, y se determinó la inhibición de la actividad catalítica de la elastasa de neutrófilos humanos por la reducción de la actividad de la enzima de un sustrato específico. Resultados: los extractos presentaron baja toxicidad a las concentraciones evaluadas, con porcentaje de viabilidad celular superior al 80 %, incluso a la concentración más alta utilizada (25 µg/mL. También presentaron actividad inhibitoria de la liberación de mieloperoxidasa, concentrándose el efecto en los extractos más polares (metanólico total y parcial y la inhibición de la actividad de la elastasa en los de más baja polaridad (hexano y diclorometano parcial, a concentraciones de 10 y 25 µg/mL, acorde con la naturaleza lipofílica del sitio activo de la enzima. Conclusiones: los resultados en los extractos evaluados son promisorios, teniendo en cuenta que cada muestra representa una mezcla compleja de compuestos activos entre 10 y 25 µg/mL, lo que resulta de mucho interés para continuar con su fraccionamiento como agentes potenciales para la posible terapia de enfermedades que presentan procesos inflamatorios.

Producción de lípidos estructurados por transesterificación enzimática del aceite de soja y aceite de palmiste en reactor de lecho empacado

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Perea Villamil, Aide

2008-12-01

Full Text Available Enzymatic synthesis of structured lipids by transesterification of soybean oil with palm kernel oil was evaluated in a packed-bed reactor with a capacity for 500g of enzyme loading. Lipozyme RM-IM was used as catalyst. Substrate blends were passed through the enzyme bed at different flow rates. Transesterification reached a level of 19.6 %, with a maximum calculated productivity of 2344 kg of transesterified oil/kg of immobilized enzyme, a flow rate of 9,36 kg oil/kg enzyme/h. The triacylglycerols formed in major proportion were C40:2, C42:2, C42:3, C44:2, C44:3, C50:3 and C50:4. Stereoespecific analysis of the fat before and after transesterification shows a slight migration of acyl groups. The products obtained by this technology can be applied in the formulation of lipid emulsions for enteral and parenteral nutrition and the food industry.Se evaluó la síntesis enzimática de lípidos estructura-dos por transesterificación de aceite de soja con aceite de palmiste en un reactor de lecho empacado (PBR con capacidad para 500 gramos de enzima, utilizando como catalizador Lipozyme RM-IM. La mezcla de sustratos se hizo pasar a través del lecho de enzima a 70 °C y diferentes flujos de aceite. A un flujo de 9.36 kg aceite/kg enzima/h se alcanzó un grado transesterificación de 19.6 % con una productividad máxima calculada de 2344 kg aceite/kg enzima. Los triacilgliceroles que se formaron en mayor proporción fueron el C40:2, C42:2, C42:3, C44:2, C44:3, C50:3 y C50:4. El análisis estereoespecífico de la mezcla grasa antes y después de la transesterificación indicó baja migración de grupos acilo. Los productos obtenidos pueden tener aplicación en la formulación de emulsiones lípidicas para nutrición enteral y parenteral y en la industria de alimentos.

Regulación de la acción de la Aldosterona al nivel del receptor mineralocorticoide

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Roberto Franco Saenz

2001-08-01

Full Text Available

Se revisan nuevos conceptos acerca de la secreción de aldosterona y de la interacción de la aldosterona con el receptor mineralocorticoide así como el papel de la enzima 11b-hidroxisteroid dehidrogenasa tipo 2 (11b-HSD-2 en la protección del receptor mineralocorticoides contra la acción de los glucocorticoides endógenos. Alteraciónes en la actividad de esta enzima causan hipertensión arterial en humanos y animales de experimentación. En vista del papel crítico que esta enzima juega en la reabsorción de sodio y el volumen sanguíneo en este estudio se investiga la regulación del gen de la 11b-HSD-2 en el riñón de la rata Dahl, un modelo experimental de hipertensión genética sensible al sodio dietético y se muestra que el sodio dietético aumenta la expresión del gen en el riñón de estas ratas.

Introducción
La aldosterona es una hormona mineralocorticoide producida por las células glomerulosas de la corteza adrenal. La aldosterona actúa en el riñón, en el túbulo convoluto distal causando retención de sodio y eliminación de potasio y iones de hidrógeno. La aldosterona juega un papel principal en el mantenimiento del volumen sanguíneo y de la presión arterial. En este manuscrito se revisan nuevos conceptos en la regulación de la secreción de aldosterona y el papel de la enzima 11b-hidroxisteroid dehydrogenasa (11b-HSD en la acción de la aldosterona y en la protección del receptor mineralocorticoide contra los glucocorticoides.

También se reportan estudios de la regulación del gen de la 11b-HSD-2 en el riñón de la rata Dahl, un modelo experimental de hipertensión genética con sensibilidad al sodio dietético.

Mecanismo de acción de las fosfatasas ácidas de bajo peso molecular

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Emilio Guija

2007-12-01

Full Text Available Las fosfatasas ácidas son enzimas ampliamente distribuidas en la naturaleza y tienen la propiedad de hidrolizar fosfomonoésteres a pH 5,0, liberando como productos de la reacción un alcohol y fosfato inorgánico. Cuando se compara los valores de kcat/Km de las fosfatasas de hígado, de bovino, alpaca y porcino, nos permite sugerir que estas enzimas tienen elevada afinidad por el sustrato p-nitrofenil fosfato. Los productos que se liberan durante la catálisis por fosfatasa ácida, muestran que el fenol o el p-nitrofenol se comportan como inhibidores de tipo no competitivo, mientras que el fosfato inorgánico muestra una inhibición de tipo competitiva. Para evidenciar la probable formación de un complejo covalente en la secuencia catalítica, se utilizó diversos nucleófilos más eficientes que el agua, tales como metanol, etanol y glicerol. Las modificaciones que produjeron en los valores Km, Vmax y los productos liberados en la reacción sugieren la formación de un complejo enzima-fosfato. El pH afecta los valores de Km y Vmax de las fosfatasas ácidas, un análisis del comportamiento de estas enzimas en un rango de pH comprendido entre 3,8 y 6,8 sugieren que en el sitio activo existe un residuo de aminoácido con un valor pKa de 6,0. El uso del dietilpirocarbonato, un compuesto que selectivamente reacciona con el residuo histidina en las proteínas, inhibe completamente a estas fosfatasas. Así mismo, un experimento de cinética de inhibición múltiple realizado en presencia de fosfato inorgánico y dietilpirocarbonato permite calcular un valor Ki que es igual al obtenido en otras condiciones experimentales. En tal sentido, las fosfatasas ácidas de bajo peso molecular catalizarían las reacciones a través de un modelo de tipo uni biordenado, con la formación de un complejo covalente intermedio, y que el residuo histidina participaría directamente en la catálisis.

Utilização do trigo e do centeio na alimentação do frango de carne

OpenAIRE

Mourão, J.L

2001-01-01

Estudo realizado a fim de conhecer os efeitos do centeio e da sua suplementação com xilanases nas performances de crescimento do frango de carne. A centeio afetou negativamente os resultados mas os enzimas não tiveram efeitos evidentes.

Differential activation of diverse glutathione transferases of Clonorchis sinensis in response to the host bile and oxidative stressors.

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Young-An Bae

Full Text Available BACKGROUND: Clonorchis sinensis causes chronic cumulative infections in the human hepatobiliary tract and is intimately associated with cholangiocarcinoma. Approximately 35 million people are infected and 600 million people are at risk of infections worldwide. C. sinensis excretory-secretory products (ESP constitute the first-line effector system affecting the host-parasite interrelationship by interacting with bile fluids and ductal epithelium. However, the secretory behavior of C. sinensis in an environment close to natural host conditions is unclear. C. sinensis differs from Fasciola hepatica in migration to, and maturation in, the hepatic bile duct, implying that protein profile of the ESP of these two trematodes might be different from each other. METHODOLOGY/PRINCIPAL FINDINGS: We conducted systemic approaches to analyze the C. sinensis ESP proteome and the biological reactivity of C. sinensis glutathione transferases (GSTs, such as global expression patterns and induction profiles under oxidative stress and host bile. When we observed ex host excretion behavior of C. sinensis in the presence of 10% host bile, the global proteome pattern was not significantly altered, but the amount of secretory proteins was increased by approximately 3.5-fold. Bioactive molecules secreted by C. sinensis revealed universal/unique features in relation to its intraluminal hydrophobic residing niche. A total of 38 protein spots identified abundantly included enzymes involved in glucose metabolism (11 spots, 28.9% and diverse-classes of glutathione transferases (GSTs; 10 spots, 26.3%. Cathepsin L/F (four spots, 10.5% and transporter molecules (three spots, 7.9% were also recognized. The universal secretory proteins found in other parasites, such as several enzymes involved in glucose metabolism and oxygen transporters, were commonly detected. C. sinensis secreted less cysteine proteases and fatty acid binding proteins compared to other tissue-invading or

Differential Activation of Diverse Glutathione Transferases of Clonorchis sinensis in Response to the Host Bile and Oxidative Stressors

Science.gov (United States)

Bae, Young-An; Ahn, Do-Whan; Lee, Eung-Goo; Kim, Seon-Hee; Cai, Guo-Bin; Kang, Insug; Sohn, Woon-Mok; Kong, Yoon

2013-01-01

Background Clonorchis sinensis causes chronic cumulative infections in the human hepatobiliary tract and is intimately associated with cholangiocarcinoma. Approximately 35 million people are infected and 600 million people are at risk of infections worldwide. C. sinensis excretory-secretory products (ESP) constitute the first-line effector system affecting the host-parasite interrelationship by interacting with bile fluids and ductal epithelium. However, the secretory behavior of C. sinensis in an environment close to natural host conditions is unclear. C. sinensis differs from Fasciola hepatica in migration to, and maturation in, the hepatic bile duct, implying that protein profile of the ESP of these two trematodes might be different from each other. Methodology/Principal Findings We conducted systemic approaches to analyze the C. sinensis ESP proteome and the biological reactivity of C. sinensis glutathione transferases (GSTs), such as global expression patterns and induction profiles under oxidative stress and host bile. When we observed ex host excretion behavior of C. sinensis in the presence of 10% host bile, the global proteome pattern was not significantly altered, but the amount of secretory proteins was increased by approximately 3.5-fold. Bioactive molecules secreted by C. sinensis revealed universal/unique features in relation to its intraluminal hydrophobic residing niche. A total of 38 protein spots identified abundantly included enzymes involved in glucose metabolism (11 spots, 28.9%) and diverse-classes of glutathione transferases (GSTs; 10 spots, 26.3%). Cathepsin L/F (four spots, 10.5%) and transporter molecules (three spots, 7.9%) were also recognized. The universal secretory proteins found in other parasites, such as several enzymes involved in glucose metabolism and oxygen transporters, were commonly detected. C. sinensis secreted less cysteine proteases and fatty acid binding proteins compared to other tissue-invading or intravascular

A conservative region of the mercuric reductase gene (merA as a molecular marker of bacterial mercury resistance Região conservada do gene da mercúrio redutase (merA como marcador molecular da resistência bacteriana ao mercúrio

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Adriana Sotero-Martins

2008-06-01

Full Text Available The most common bacterial mercury resistance mechanism is based on the reduction of Hg(II to Hg0, which is dependent of the mercuric reductase enzyme (MerA activity. The use of a 431 bp fragment of a conservative region of the mercuric reductase (merA gene was applied as a molecular marker of this mechanism, allowing the identification of mercury resistant bacterial strains.O mecanismo de resistência bacteriana ao mercúrio mais comum é baseada na redução do Hg(II a Hg0, através da atividade da enzima mercúrio redutase (MerA. O uso do fragmento de 431 pb amplificado de uma região conservada do gene merA, que codifica a enzima MerA,foi utilizado como marcador molecular deste mecanismo, permitindo a identificação de bactérias resistentes ao mercúrio.

Electromagnetic Simulations of an Embedded BPM in Collimator Jaws

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Boccard, C; Nosych, A

2011-01-01

Next gen­er­a­tion of the LHC col­li­ma­tors will be equipped with but­ton beam po­si­tion mon­i­tors (BPMs) em­bed­ded into the col­li­ma­tor jaws. Such a so­lu­tion will im­prove the ac­cu­ra­cy of the jaw align­ment with respect to the beam and re­duce the beam time nec­es­sary for the collima­tor setup. This paper de­scribes re­sults of elec­tro­mag­net­ic simu­la­tions of the jaw BPMs per­formed with the CST Par­ti­cle Stu­dio suite, aimed at char­ac­ter­i­sa­tion of the BPMs as well as the simulation soft­ware it­self. The re­sults are com­pared to the mea­sure­ments ob­tained with beam on a pro­to­type sys­tem in­stalled in the CERN SPS.

Are the angiotensin-converting enzime gene and acticity risk factors for stroke? São fatores de risco para acidente vascular cerebral o gene e a atividade da enzima conversora de angiotensina ?

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Miris Dikmen

2006-06-01

Full Text Available Stroke is a multifactorial disease in which genetic factors play an important role. This study was carried out to determine angiotensin-converting enzyme (ACE gene polymorphism in Turkish acute stroke patients and to establish whether there is an association of angiotensin-converting enzyme gene I/D polymorphism with clinical parameters. In this study 185 patients and 50 controls were recruited. We have investigated the association among the allelic distribution of the insertion/deletion (I/D polymorphism of the ACE gene identified by polymerase chain reaction. Distribution of ACE gene I/D genotypes and allele frequencies in patients were not significantly different from controls. D allele frequencies were 57.8% in patients versus 53.0% in controls and I allele 42.2% versus 47% respectively. History of hypertension, stroke, renal, heart and vessel diseases incidence and age, gender, systolic-diastolic blood pressures and creatinine levels were significantly high in patients. But these results and ACE activities had no significant differences among the ACE genotypes in patients and controls. Our results suggest that the ACE gene polymorphism is not associated with the pathogenesis of stroke in Turkish stroke patients.O acidente vascular cerebral (AVC é doença multifatorial em que fatores genéticos desempenham papel importante. Este estudo foi desenvolvido para verificar o polimorfismo do gene da enzima conversora da angiotensina (ECA em pacientes turcos com AVC agudo e estabelecer se existe associação do gene I/D da ECA com parâmetros clínicos. O estudo foi realizado com 185 pacientes e 50 controles. A associação entre a distribuição alélica da inserção / deleção (I/D do polimorfismo do gene da ECA foi estudada pela reação em cadeia da polimerase. A distribuição dos genótipos I/D do gene da ECA e suas freqüências não apresentaram significância estatística quando comparados os pacientes e controles. As freqüências dos

MECHANISMS OF RESISTANCE TO ANTIPARASITIC DRUGS IN TRYPANOSOMA CRUZI. CORRELATIONS BETWEEN GENOTYPE AND RESISTANCE

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John M Kelly

2013-01-01

Full Text Available El benznidazol y el nifurtimux conpuestos nitroheterocilicos son los medicamentos aprobados para el tratamiento de las infecciones por Trypanosoma cruzi. Ambos son profármacos y no tienen importantes propiedades tripanocidas hasta su activación intraparasitaria. La enzima responsable es una nitroreductasa (TcNTR , que inicia una cascada reductora que conduce a la generación de los metabolitos tóxicos que matan al parásito. Los procesos que actúan para regular a esta enzima conducen a la resis- tencia cruzada contra ambos fármacos. Estos incluyen la pérdida de uno de los cromosomas que contienen el gen TcNTR o mutaciones puntuales que inactivan la enzima. Los parásitos TcNTR heterocigotos son infecciosos, no muestran un fenotipo nocivo obvio y son hasta 5 veces más resistente a benznidazol y el nifurtimox. Sin embargo, la pérdida completa de la actividad TcNTR hace que T. cruzi no sea infeccioso, lo que sugiere que puede haber un límite para el nivel de resistencia por este mecanismo. En las poblaciones naturales de T. cruzi no se encontraron pruebas de que las amplias variaciones en la sensibilidad al benznidazol estén vinculadas a las mutaciones en TcNTR lo que, junto con la evidencia de que la resistencia a benznidazol y nifurtimox no siempre es conjunta, indica que existen otros mecanismos independientes de TcNTR. Los nuevos avances en tecnología ofrecen la oportunidad de explorar más a fondo esta cuestión.

Caracterização da agressividade e atividade enzimática de isolados de Colletotrichum spp. associados à antracnose do abacate

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Hugo José Tozze Júnior

Full Text Available RESUMO A antracnose é a principal doença pós-colheita do abacate. O presente trabalho teve como objetivos caracterizar a agressividade e a atividade enzimática de 30 isolados de Colletotrichum spp. de abacate. Abacates ‘Fuerte’ foram inoculados com disco de BDA contendo estruturas de cada isolado e após sete dias mensuraram-se os diâmetros das lesões. Dois grupos de isolados foram distinguidos quanto à agressividade pelo teste de teste de Scott-Knott (p=0,05, um com lesões entre 34,0 e 38,2 mm e outro com lesões entre 38,7 e 44,0 mm de diâmetro. Para caracterização enzimática, avaliaram-se as áreas dos halos formados em substratos específicos para detecção de amilase, celulase, lacase, lipase, pectato liase e protease e a atividade da enzima catalase. Os isolados não produziram amilase suficiente para formação de halo em meio de cultura. Todos os isolados produziram catalase, pectato liase e protease, enquanto 46% produziram lacase e 97% tiveram as atividades da celulase e lipase detectadas. Distinguiram-se dois grupos de isolados para as enzimas catalase e pectato liase, cinco grupos para a celulase, seis grupos para a lacase e sete grupos para as enzimas lipase e protease, pelo teste de teste de Scott-Knott (p=0,05. Não houve relação entre as atividades enzimáticas e a agressividade dos isolados.

Insectistatic and insecticide activity of Beauveria bassiana in Bradysia impatiens (Diptera: Sciaridae

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Víctor H. Marín-Cruz

2017-01-01

Full Text Available Introducción: Bradysia impatiens causa pérdidas significativas en viveros e invernaderos de México. Objetivo: El efecto insecticida e insectistático de Beauveria bassiana se evaluó sobre B. impatiens. Materiales y métodos: El efecto insecticida e insectistático de conidios (10 7 conidios·mL -1 , enzimas (10 000 ppm, metabolitos (10 000 ppm y extracto crudo de B. bassiana se evaluó a los 8 y 20 días. Los datos de mortalidad corregida de larvas y pupas de B. impatiens y emergencia relativa de adultos, transformados con la función arcoseno, se sometieron a un análisis de varianza y comparación de medias de Tukey ( P < 0.05. Resultados y discusión: En el día 8, los conidios tuvieron la mayor actividad insecticida con 31.1 % de mortalidad corregida, mientras que la actividad de las enzimas fue nula. A los 20 días, los tratamientos de metabolitos y conidios tuvieron el mayor efecto en la mortalidad, 47.5 y 42.1 %, respectivamente. Dichos tratamientos tuvieron la mayor actividad insectistática. La emergencia de adultos a los 20 días fue menor con los conidios (6 %, mientras que con las enzimas fue de 100 %. Los metabolitos provocaron que 65 % de los adultos mostraran malformaciones. Conclusión: Los metabolitos y conidios de B. bassiana podrían emplearse para el control de larvas y pupas de B. impatiens.

EVALUACIÓN DE ACTIVIDADES ENDOGLUCANASA, EXOGLUCANASA, LACASA Y LIGNINA PEROXIDASA EN DIEZ HONGOS DE PUDRICIÓN BLANCA

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SANDRA MONTOYA B.

Full Text Available Este trabajo presenta una vía de rastreo de producción de enzimas lignocelulolíticas en diez especies de hongos de pudrición blanca: Lentinula edodes, Schizophyllum commune, Trametes trogii, Coriolus versicolor, Pycnoporus sanguineus, Ganoderma applanatum, Ganoderma lucidum, Grifola frondosa, Pleurotus ostreatus y Auricularia delicata. Estas especies primero fueron rastreadas sobre medios de cultivo sólido que contienen carboximetil celulosa, celulosa cristalina, ABTS (2,2´-azino-bis(3-etilbenzotiazolina-6-sulfonato y azure B, los cuales evidenciaron la producción de las enzimas endoglucanasa, exoglucanasa, lacasa y lignina peroxidasa (LiP. Las actividades celulolíticas fueron detectadas a los cinco días de incubación con el indicador rojo congo, formándose un halo claro-blanco en las zonas donde se degrada la celulosa. Para las ligninasas, este rastreo consistió en el seguimiento a la formación de halos verdes por oxidación del ABTS para lacasa y halos de decoloración sobre el azure B para la LiP durante 14 días de incubación. De este rastreo cualitativo, se seleccionaron cuatro cepas (G. lucidum, L. edodes, C. versicolor y T. Trogii, como las mejores productoras de enzimas celulolíticas y ligninolíticas. Estas cuatro especies fueron inoculadas sobre un sustrato de aserrín de roble, obteniéndose 51,8% de lignina degradada por L. edodes y 22% de celulosa degrada por C. versicolor.

EFFECT OF A VITAMIN E AND SELENIUM ORAL SUPPLEMENTATION ON HEMATOLOGICAL PARAMETERS, MUSCULAR INJURIES MARKERS ENZYMES AND PEROXIDATION INDICES OF BIOMOLECULES IN SHOW JUMPING HORSES EFEITO DA SUPLEMENTAÇÃO COM VITAMINA E E SELÊNIO SOBRE O QUADRO HEMATOLÓGICO, ENZIMAS MARCADORAS DE LESÃO MUSCULAR E ÍNDICE DE PEROXIDAÇÃO DE BIOMOLÉCULAS EM EQUINOS SUBMETIDOS À ATIVIDADE DE SALTO

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Domingos Cachineiro Rodrigues Dias

2009-09-01

, vitamin E.
As alterações dos constituintes sanguíneos e o estresse oxidativo são mecanismos fisiopatológicos incriminados no aparecimento de lesões relacionadas ao exercício. Avaliou-se a influência da suplementação oral à base de ?-tocoferol (vitamina E e selênio sobre o quadro hematológico, bem como a atividade sérica de enzimas marcadoras de lesão muscular e o nível de substâncias derivadas da ação dos radicais livres sobre biomoléculas presentes na membrana de eritrócitos (TBARS de equinos normalmente treinados e submetidos à prova de hipismo clássico. Para isso, distribuíram-se vinte cavalos em dois grupos, sendo um suplementado com produto à base de vitamina E (2500 mg/dia e selênio (3 mg/dia por 45 dias (GT e outro mantido como controle (GC. Colheram-se amostras de sangue antes (TR e após (T0 a realização do exercício e seis (T6, doze (T12 e 24 horas (T24 após o término da atividade de esforço. Foram avaliados os constituintes do hemograma, a atividade sérica das enzimas CK, AST e LDH e a concentração TBARS na membrana de eritrócitos. A suplementação utilizada apresentou efeito sobre alguns parâmetros do hemograma, uma vez que no GT houve retorno aos níveis basais de número de hemácias e leucócitos, caracterizado por neutrofilia, em até seis horas. No GC, tais valores retornaram aos níveis de repouso entre 12 e 24 horas. Para a AST, o GT retornou ao valor do repouso seis horas após a realização do exercício. No entanto, não houve efeito sobre as enzimas CK e LDH, nem para o TBARS. Apesar de terem sido observadas diferenças na dinâmica de alguns parâmetros hematológicos entre os grupos, não foi possível determinar atividade antioxidante no GT no delineamento experimental utilizado, o que demonstra a necessidade de mais estudos para o entendimento do estresse oxidativo em cavalos atletas e a suplementação com antioxidantes na dieta desses animais.

PALAVRAS-CHAVES: Cavalo atleta, estresse

Strategy and Grand Strategy: What Students and Practitioners Need to Know

Science.gov (United States)

2015-12-01

Press publications may be downloaded free of charge from the SSI website. Hard copies of certain reports may also be obtained free of charge while...and means. Strategy rests on assump- tions; if assumptions go unexamined, then one risks building a strategic edifice on a foundation of sand .4 Often...Similar outcomes re- sult if civilians believe that military leaders are trying to game, stack, or stall a consequential decision. Wary of all this

2306-IJBCS-Article-Some Der

African Journals Online (AJOL)

hp

La gestion appropriée des sols cultivés peut permettre un stockage de carbone, en plus de l'avantage supplémentaire du maintien du niveau de fertilité qui en résulte. L'objectif de cette étude est d'appréhender l'influence réelle de pratiques culturales sous niébé ou sorgho dans le système zaï, sur l'évolution et.

MUSIC-CONTENT-ADAPTIVE ROBUST PRINCIPAL COMPONENT ANALYSIS FOR A SEMANTICALLY CONSISTENT SEPARATION OF FOREGROUND AND BACKGROUND IN MUSIC AUDIO SIGNALS

OpenAIRE

Papadopoulos , Hélène; Ellis , Daniel P.W.

2014-01-01

International audience; Robust Principal Component Analysis (RPCA) is a technique to decompose signals into sparse and low rank components, and has recently drawn the attention of the MIR field for the problem of separating leading vocals from accompaniment, with appealing re-sults obtained on small excerpts of music. However, the perfor-mance of the method drops when processing entire music tracks. We present an adaptive formulation of RPCA that incorporates music content information to guid...

Linear- and Repetitive-Feature Detection Within Remotely Sensed Imagery

Science.gov (United States)

2017-04-01

preprocessing image data, which has proven difficult and intensive for certain images. In many cases, the amount of user interaction needed to produce...a way that humans can easily understand. This data is represented as if there was a light source located to the northwest with an altitude angle of...them. Although there are some shortcomings to this methodology, the final re- sult is useful and would help a user or analyst distinguish the layout

Linear- and Repetitive Feature Detection Within Remotely Sensed Imagery

Science.gov (United States)

2017-04-01

preprocessing image data, which has proven difficult and intensive for certain images. In many cases, the amount of user interaction needed to produce...a way that humans can easily understand. This data is represented as if there was a light source located to the northwest with an altitude angle of...them. Although there are some shortcomings to this methodology, the final re- sult is useful and would help a user or analyst distinguish the layout

ÉTUDE DE CAS — mondiale : Repenser les stratégies de recherche ...

International Development Research Centre (IDRC) Digital Library (Canada)

24 déc. 2010 ... Le composite qui en a résulté -- appelé MKC, pour Maria Kaherero Composite -- a servi de base au programme national de sélection végétale du pays. Sperling fait une mise en garde : la non-participation des femmes à la sélection végétale pourrait avoir des conséquences néfastes, pas seulement ...

The Shock and Vibration Bulletin. Part 2. Isolation and Damping, Impact, Blast

Science.gov (United States)

1978-09-01

statistically distributed. "What is a reasonable value of the •- •- circumferential strain at failure, When a relationship is known be- efail ...of theinput design parameters. (These re- that is, the probability that the sults are presented in Table I). failure strain ( efail ) is exceeded...strain efail was set at 0.05 [cf..qua- 6, one has a frequency distribution of tion (5)].the number of occurrences vs. liner "strain, e. For the

Social Impacts Module (SIM) Transition

Science.gov (United States)

2012-09-28

or OABs. An agent can communicate about events, seek resources, or do nothing. This basic procedure repeats itself continuously. See Appendix C for a...still contains the ability to represent key leaders and social networks and there are no changes to the procedures described for SIM 2.0 regarding...of the study question, MAJ Ja- son Whipple of WSMR trav- eled to Monterey to sup- port designing the Nexus scenario. The meeting re- sulted in

Adénome de prostate révélé par des hypoglycémies profondes: A propos d'un cas

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M. Asseban

2013-03-01

Conclusion: L'insuffisance rénale obstructive peut constituer une complication de l'adénome de la prostate à un stade ultime. Elle est responsable d'accumulation dans le sang de médicaments y compris les antidiabétiques oraux. L'hypoglycémie qui en résulte est délétère pour la qualité de vie du patient et peut être responsable d'accidents neurologiques.

Effects of Anti-G Measures on Gas Exchange.

Science.gov (United States)

1981-05-01

position (+lGz), and the endotracheal tube was connected to a Rudolf valve arranged so that expired gas passed through a heated pneumotachograph and a... Steiner , 1960; Peterson, Bishop and Erickson, 1977). Data presented in Table 111-I suggest that application of the G-sult abdominal bladder tended to...accelerations. Aerospace Med. 31: 213-219, 1960. 18. Hershgold, E.J. and S.H. Steiner . Cardiovascular changes during acceleration stress in dogs. J

Photoaffinity labelling of the active site of the rat glutathione transferases 3-3 and 1-1 and human glutathione transferase A1-1.

OpenAIRE

Cooke, R J; Björnestedt, R; Douglas, K T; McKie, J H; King, M D; Coles, B; Ketterer, B; Mannervik, B

1994-01-01

The glutathione transferases (GSTs) form a group of enzymes responsible for a wide range of molecular detoxications. The photoaffinity label S-(2-nitro-4-azidophenyl)glutathione was used to study the hydrophobic region of the active site of the rat liver GST 1-1 and 2-2 isoenzymes (class Alpha) as well as the rat class-Mu GST 3-3. Photoaffinity labelling was carried out using a version of S-(2-nitro-4-azidophenyl)glutathione tritiated in the arylazido ring. The labelling occurred with higher ...

Presença de pesticidas anticolinérgicos (organofosforados e carbamatos em frutas e hortaliças no município de Cabo Frio, RJ

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Claudia Cristina Machado de Figueiredo de Oliveira

2010-12-01

Full Text Available No estudo em questão, utilizou-se a inibição da enzima acetilcolinesterase para detectar pesticidas organofosforados e carbamatos em frutas e hortaliças consumidas na cidade de Cabo Frio. Hortaliças e frutas ali coletadas, em anos distintos (1996 e 2007 apresentaram pesticidas em todas as amostras. As amostras de alface (1996 e agrião (2007 foram as que apresentaram maiores teores de equivalentes de metil-paration, 0,7 e 1,4 ppm, respectivamente. Os níveis mais baixos foram detectados nas uvas: 0,2 ppm. A solubilização dos compostos envolvidos no estudo em Triton X-100 foi eficiente sem inibir a atividade da enzima acetilcolinesterase. Entre os alimentos estudados, a alface, o agrião e a maçã são merecedores de atenção para monitoramento de resíduos de pesticidas organofosforados e carbamatos.

EFEITO DE MERCURIAIS NA ATIVIDADE DA PECTINAMETILESTERASE DE BERINJELA (Solanum melongena L.

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LEONARDO CARDELLO

2009-07-01

Full Text Available

RESUMO: Pectinametilesterase (E. C. 3.1.1.11, parcialmente purificada, teve sua atividade reduzida na presença de cloreto de mercúrio e p-cloromercuriobenzoato. A incubação prévia de enzima com cisteína protegeu-a da inibição causada pelo cloreto de mercúrio. Este reagente, no entanto, reverteu apenas parcialmente a atividade da PME previamente inibida pelo cloreto de mercúrio. Apesar disso, uma reversão total da inibição pelo cloreto de mercúrio foi obtida com NaCl 0,6 M. Os resultados sugerem que grupo(s SH essencial(is para a atividade da PME parece(m estar localizado(s fora do sítio ativo da enzima. PALAVRAS-CHAVE: Pectinametilesterase; berinjela; grupo sulfidrila; grupo tiol.

LIPASES PRODUZIDAS POR LEVEDURAS: CATALISADORES PODEROSOS PARA APLICAÇÕES INDUSTRIAIS

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Luiza Lux Lock

2007-12-01

Full Text Available O termo “enzimas lipolíticas” refere-se a lípases e hidrolases éster carboxílico. A produção de lipase é ampla entre as leveduras, mas poucas são capazes de produzir lipases com características interessantes e em quantidades suficientes para serem industrialmente úteis. A literatura relativa a lipases produzidas por Candida rugosa, Yarrowia (Candida lipolytica, Candida antarctica e outras leveduras produtoras de lípases é revisada. O uso de lípases recombinantes é discutido, com ênfase na utilização de sistemas de expressão heteróloga e desenho de quimeras. Finalmente, as três abordagens que visam à melhora da produção de lipase ou a modificação da seletividade do substrato da enzima (engenharia do meio, do biocatalisador e da proteína são discutidos.

de la fotosíntesis C4

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J. C. Raya-Pérez

2008-01-01

Full Text Available La fotosíntesis C4 surgió hace unos 7-5 millones de años y tiene un origen polifilético. La disminución en la concentración atmosférica de CO2 a menos de 500 partes por millón (ppm propició la aparición de un mecanismo para concentrar este gas en la zona donde actúa la Rubisco (ribulosa bifosfato carboxilasa/oxigenasa, evitando así su actividad de oxigenasa. El análisis de los genes que codifican para las enzimas usadas en la vía C4, así como la caracterización bioquímica de algunas de estas enzimas, permiten entrever algunos de los cambios que han sufrido a fin de adaptarse a una nueva función, la de concentrar el CO2 a fin de que sea utilizado por la Rubisco.

Influência do tratamento enzimatico sobre as características reológicas e microscópicas da polpa de acerola

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Luciana Balischi

2002-04-01

Full Text Available O consumo de sucos e bebidas à base de frutas tem aumentado consideravelmente nos últimos anos. No Brasil, destaca-se a produção da acerola (alto teor de vitamina C e de seus derivados. Objetivando melhoramentos na fabricação de sucos e derivados de frutas tropicais, surgiram pesquisas visando a encontrar o melhor tratamento enzimático. Neste trabalho, estudou-se a variação que ocorre na reologia da polpa integral de acerola submetida ao tratamento enzimático, bem como a variação no diâmetro médio das partículas, para as melhores condições de tratamento obtidas. Duas enzimas comerciais foram testadas: Citrozym Ultra L e Pectinex Ultra SP-L, variou-se as temperaturas, as concentrações das enzimas e os tempos de tratamento As características reológicas e microscópicas da polpa sofreram alterações com o tratamento enzimático.

Atividade enzimática, produção de slime e sensibilidade a antifúngicos de Candida sp Enzymatic activity, slime production and antifungal agent sensitivity of Candida sp

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Jaqueline Otero Silva

2007-06-01

Full Text Available A habilidade de Candida spp secretar enzimas extracelulares e slime tem sido associada como fatores de patogenicidade. Do total de 37 cepas de Candida sp, 100% foram produtoras de proteinase, 83,8% fosfolipase, 64,9% slime e 100% sensíveis ao fluconazol e itraconazol. Foram encontradas 17 tipagens (enzima/slime. Esta metodologia apresentou um bom índice discriminatório (D=0,93 podendo ser utilizado na caracterização fenotípica das leveduras.Abilith of Candida spp to secrete extracellular enzymes and slime has been associated as pathogenicity factors. Out of a total of 37 strains of Candida sp, 100% were proteinase producers, 83.8% were phospholipase producers, 64.9% were slime producers and 100% were sensitive to fluconazole and itraconazole. Seventeen typings (enzymes/slime were found. This methodology presented a good discrimination rate (D = 0.93 and could be used for phenotypic characterization of yeasts.

Extracción del ADN de Fusarium Oxysporum f.sp. Dianthi

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Sixta T. Martínez

2010-07-01

Full Text Available Se estudia la utílización de dos métodos para la extracción del ADN del Fusarium o.xy.sporum f.sp. Dianthi. En los dos métodos la pared del hongo se rompió con nitrógeno líquido, uno de ellos empleó como solución extractora bromuro de cetiltrimetil amonio (BCTA y el otro una solución de sacarosa con altas concentraciones de protcasa y EDTA. Para la desproteinización ambos métodos utilizaron soluciones de fcnol-clorofonno y enzimas proteolítícas. El ADN obtenido se digirió con enzimas de restricción EcoRI y Hindlll. Se corroboró que cl ADN estaba libre de los contaminantes más frecuentes en hongos, como proteínas y carbohidratos por medio de ultracentrifugación en cloruro de cesio (CsCI. La extracción con BCTA presentó los mejores rendimientos.

Characterization and mutational analysis of omega-class GST (GSTO1 from Apis cerana cerana, a gene involved in response to oxidative stress.

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Fei Meng

Full Text Available The Omega-class of GSTs (GSTOs is a class of cytosolic GSTs that have specific structural and functional characteristics that differ from those of other GST groups. In this study, we demonstrated the involvement of the GSTO1 gene from A. cerana cerana in the oxidative stress response and further investigated the effects of three cysteine residues of GSTO1 protein on this response. Real-time quantitative PCR (qPCR showed that AccGSTO1 was highly expressed in larvae and foragers, primarily in the midgut, epidermis, and flight muscles. The AccGSTO1 mRNA was significantly induced by cold and heat at 1 h and 3 h. The TBA (2-Thiobarbituric acid method indicated that cold or heat resulted in MDA accumulation, but silencing of AccGSTO1 by RNAi in honeybees increased the concentration of MDA. RNAi also increased the temperature sensitivity of honeybees and markedly reduced their survival. Disc diffusion assay indicated that overexpression of AccGSTO1 in E. coli caused the resistance to long-term oxidative stress. Furthermore, AccGSTO1 was active in an in vitro DNA protection assay. Mutations in Cys-28, Cys-70, and Cys-124 affected the catalytic activity and antioxidant activity of AccGSTO1. The predicted three-dimensional structure of AccGSTO1 was also influenced by the replacement of these cysteine residues. These findings suggest that AccGSTO1 plays a protective role in the response to oxidative stress.

Interactive effects of nutrition, reproductive state and pollution on molecular stress responses of mussels, Mytilus galloprovincialis Lamarck, 1819.

Science.gov (United States)

González-Fernández, Carmen; Albentosa, Marina; Sokolova, Inna

2017-10-01

Marine bivalves including mussels Mytilus galloprovincialis are commonly used as sentinels for pollution monitoring and ecosystem health assessment in the coastal zones. Use of biomarkers to assess the pollution effects assumes that the effects of pollutants on the biomarkers exceed the natural background variability; yet this assumption has rarely been tested. We exposed mussels at different reproductive stages and nutritive states to two concentrations of a polycyclic aromatic hydrocarbon (fluoranthene, 3 and 60 μg L -1 ) for three weeks. Expression levels of the molecular biomarkers related to the detoxification and general stress response [cytochrome P450 oxidase (CYP450), glutathione S-transferases (GST-α; GST-S1; GST-S2), the multixenobiotic resistance protein P-glycoprotein (PgP), metallothioneins (MT10 and MT20), heat shock proteins (HSP22, HSP70-2; HSP70-3; HSP70-4), as well as mRNA expression of two reproduction-related genes, vitellogenin (Vitel) and vitelline coat lysin M7 (VCLM7)] were measured. The mussels' nutrition and reproductive state affected the baseline mRNA levels of molecular biomarkers and modulated the transcriptional responses of biomarker genes to the pollutant exposure. Thus, mussel physiological state could act as a confounding factor in the evaluation of the response of pollution through molecular biomarkers. The biomarker baseline levels must be determined across a range of physiological states to enable the use of biomarkers in monitoring programs. Copyright © 2017 Elsevier Ltd. All rights reserved.

Comprehensive genome-wide analysis of Glutathione S-transferase gene family in potato (Solanum tuberosum L.) and their expression profiling in various anatomical tissues and perturbation conditions.

Science.gov (United States)

Islam, Md Shiful; Choudhury, Mouraj; Majlish, Al-Nahian Khan; Islam, Tahmina; Ghosh, Ajit

2018-01-10

Glutathione S-transferases (GSTs) are ubiquitous enzymes which play versatile functions including cellular detoxification and stress tolerance. In this study, a comprehensive genome-wide identification of GST gene family was carried out in potato (Solanum tuberosum L.). The result demonstrated the presence of at least 90 GST genes in potato which is greater than any other reported species. According to the phylogenetic analyses of Arabidopsis, rice and potato GST members, GSTs could be subdivided into ten different classes and each class is found to be highly conserved. The largest class of potato GST family is tau with 66 members, followed by phi and lambda. The chromosomal localization analysis revealed the highly uneven distribution of StGST genes across the potato genome. Transcript profiling of 55 StGST genes showed the tissue-specific expression for most of the members. Moreover, expression of StGST genes were mainly repressed in response to abiotic stresses, while largely induced in response to biotic and hormonal elicitations. Further analysis of StGST gene's promoter identified the presence of various stress responsive cis-regulatory elements. Moreover, one of the highly stress responsive StGST members, StGSTU46, showed strong affinity towards flurazole with lowest binding energy of -7.6kcal/mol that could be used as antidote to protect crop against herbicides. These findings will facilitate the further functional and evolutionary characterization of GST genes in potato. Copyright © 2017 Elsevier B.V. All rights reserved.

Comparative gene expression of intestinal metabolizing enzymes.

Science.gov (United States)

Shin, Ho-Chul; Kim, Hye-Ryoung; Cho, Hee-Jung; Yi, Hee; Cho, Soo-Min; Lee, Dong-Goo; Abd El-Aty, A M; Kim, Jin-Suk; Sun, Duxin; Amidon, Gordon L

2009-11-01

The purpose of this study was to compare the expression profiles of drug-metabolizing enzymes in the intestine of mouse, rat and human. Total RNA was isolated from the duodenum and the mRNA expression was measured using Affymetrix GeneChip oligonucleotide arrays. Detected genes from the intestine of mouse, rat and human were ca. 60% of 22690 sequences, 40% of 8739 and 47% of 12559, respectively. Total genes of metabolizing enzymes subjected in this study were 95, 33 and 68 genes in mouse, rat and human, respectively. Of phase I enzymes, the mouse exhibited abundant gene expressions for Cyp3a25, Cyp4v3, Cyp2d26, followed by Cyp2b20, Cyp2c65 and Cyp4f14, whereas, the rat showed higher expression profiles of Cyp3a9, Cyp2b19, Cyp4f1, Cyp17a1, Cyp2d18, Cyp27a1 and Cyp4f6. However, the highly expressed P450 enzymes were CYP3A4, CYP3A5, CYP4F3, CYP2C18, CYP2C9, CYP2D6, CYP3A7, CYP11B1 and CYP2B6 in the human. For phase II enzymes, glucuronosyltransferase Ugt1a6, glutathione S-transferases Gstp1, Gstm3 and Gsta2, sulfotransferase Sult1b1 and acyltransferase Dgat1 were highly expressed in the mouse. The rat revealed predominant expression of glucuronosyltransferases Ugt1a1 and Ugt1a7, sulfotransferase Sult1b1, acetyltransferase Dlat and acyltransferase Dgat1. On the other hand, in human, glucuronosyltransferases UGT2B15 and UGT2B17, glutathione S-transferases MGST3, GSTP1, GSTA2 and GSTM4, sulfotransferases ST1A3 and SULT1A2, acetyltransferases SAT1 and CRAT, and acyltransferase AGPAT2 were dominantly detected. Therefore, current data indicated substantial interspecies differences in the pattern of intestinal gene expression both for P450 enzymes and phase II drug-metabolizing enzymes. This genomic database is expected to improve our understanding of interspecies variations in estimating intestinal prehepatic clearance of oral drugs.

Some enzymatic properties of cholesterol oxidase produced by Brevibacterium sp Algumas propriedades enzimáticas da colesterol oxidase produzida por Brevibacterium sp.

Directory of Open Access Journals (Sweden)

Terezinha J.G. Salva

1999-12-01

Full Text Available In this study we determined some properties of the cholesterol oxidase from a Brevibacterium strain isolated from buffalo's milk and identified the cholesterol degradation products by the bacterial cell. A small fraction of the enzyme synthesized by cells cultured in liquid medium for 7days was released into the medium whereas a larger fraction remained bound to the cell membrane. The extraction of this fraction was efficiently accomplished in 1 mM phosphate buffer, pH 7.0, containing 0.7% Triton X-100. The enzyme stability under freezing and at 45oC was improved by addition of 20% glycerol. The optimum temperature and pH for the enzyme activity were 53°C and 7.5, respectively. The only steroidal product from cholesterol oxidation by the microbial cell and by the crude extract of the membrane-bound enzyme was 4-colesten-3-one. Chromatographic analysis showed that minor no steroidal compounds as well as 4-colesten-3-one found in the reaction media arose during fermentation process and were extracted together with the enzyme in the buffer solution. Cholesterol oxidation by the membrane-bound enzyme was a first order reaction type.Neste trabalho foram definidas algumas propriedades da enzima colesterol oxidase produzida por uma linhagem de Brevibacterium sp. isolada de leite de búfala e foram identificados os compostos resultantes da degradação do colesterol pela bactéria. Uma pequena fração da enzima sintetizada pelas células cultivadas em meio líquido por 7 dias foi liberada no meio de cultura e uma fração maior permaneceu ligada à membrana celular. A extração desta fração foi eficientemente efetuada em tampão fosfato 1mM, pH 7,0, contendo 0,7% de triton X-100. A estabilidade da enzima congelada e a 45oC foi aumentada pela adição de 20% de glicerol. A temperatura ótima para a atividade enzimática esteve ao redor de 53(0C e o pH ótimo esteve ao redor de 7,5. O único produto da degradação do colesterol, causada pela a

Qualidade fisiológica de sementes de milho na presença de bioestimulantes Physiological quality of corn seeds in the presence of biostimulants

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Tanismare Tatiana de Almeida Silva

2008-06-01

Full Text Available Os reguladores de crescimento podem influenciar na qualidade fisiológica das sementes assim como na atividade de enzimas envolvidas na germinação. Podem ainda apresentar em suas formulações micronutrientes que atuam principalmente como catalisadores de enzimas. Nesta pesquisa sementes de milho híbrido da cultivar GNZ 2004, e da linhagem Le 57 foram tratadas com os seguintes bioestimulantes e dosagens: 1.Stimulate® 12,5mL/kg de sementes, 2.Cellerate® 10,0mL/kg, 3.mistura de Cellerate®+ Stimulate® (10,0mL/kg + 12,5 mL/kg, 4.Booster® 100 mL/20kg e 5.testemunha. Logo após o tratamento, as sementes foram submetidas aos testes de germinação, envelhecimento acelerado, teste de frio e emergência de plântulas. Foram avaliadas ainda a massa de matéria seca de plântulas, parte aérea e raiz e as atividades das enzimas á-amilase, catalase, esterase e peroxidase. Nas sementes tratadas com os bioestimulantes não houve melhoria na qualidade de sementes e sob condições de estresses a utilização dos bioestimulantes Stimulate®+Cellerate® e Cellerate® reduziu a qualidade fisiológica de sementes de milho. Menor atividade da enzima peroxidase foi observada em sementes tratadas com Stimulate®, Cellerate® e Stimulate®+Cellerate® . Maior atividade da enzima esterase foi observada em sementes tratadas com Stimulate®+ Cellerate® indicando fitotoxidez dos mesmos.Growth regulators can influence the physiological quality of seeds as well as the activity of enzymes involved in germination. They may, further, present in their formulation micronutrients which act mainly as enzyme catalysts. In this research work, hybrid corn seeds of the cultivar GNZ 2004 and line Le 57 were treated with the following biostimulants and dosages: 1.Stimulate® 12.5mL/kg of seed, 2.Cellerate® 10,0mL/kg, 3.mixtue of Cellerate®+ Stimulate® (10.0mL/kg + 12.5 mL/kg, 4.Booster® 100 mL/20kg and 5.control. Soon after the treatment, the seeds were submitted to the

Polymorphism’s assessment of children’s candidate genes associated with low-level long-term exposure to strontium in drinking water

OpenAIRE

N.V. Zaitseva; O.V. Dolgilh; A.V. Krivtsov; K.G. Starkova; V.A. Luchnikova; O.A. Bubnov; E.A. Otavina; N.V. Bezruchenko; N.A. Vdovina

2015-01-01

A sequencing of the candidate genes of the pupils, exposed to strontium by the method of targeted resequencing has been performed. It is shown, that under conditions of increased revenues of strontium in drinking water the number of polymorphonuclear altered portions of candidate genes increases. As a result of the targeted resequencing in conditions of strontium exposure, the maximum polymorph modifications of the following genes are defined: sulfotransferase 1A1 (SULT1A1) and methylenetetra...

Combat-Related Heterotopic Ossification: Development of Animal Models for Identifying Mechanisms and Testing Therapeutics

Science.gov (United States)

2016-03-01

ulcerate, or envelop neurovascular structures causing pro- gressive deficits .10-14 As a result, up to 41% of patients with HO will ultimately require...in a logarithmic fashion under optimal culture and nutrient conditions re- sulting in a controlled cell growth rate. Next, 2 mL of the concentrated...Cooper RR, Saltzman CL. Bone and brain: a review of neural, hormonal , and muscu- loskeletal connections. Iowa Orthop J. 2004;24:123. 19. Mo IF, Yip KH

AcEST: DK958179 [AcEST

Lifescience Database Archive (English)

Full Text Available sult : TrEMBL tr_hit_id Q54CF8 Definition tr|Q54CF8|Q54CF8_DICDI Chromo (CHRomatin Organization...ments: (bits) Value tr|Q54CF8|Q54CF8_DICDI Chromo (CHRomatin Organization MOdifier) ... 41 0.059 tr|B1SAI8|B... protein OJ1249_F... 33 9.5 >tr|Q54CF8|Q54CF8_DICDI Chromo (CHRomatin Organization MOdifier) domain-containi

New Methods for the Computational Fabrication of Appearance

Science.gov (United States)

2015-06-01

constraint. The only variables are the values of u and the coecients ↵ i . In our current implementation, we chose a radial thin - plate spline basis h i (r...61] use thin - plate implicit functions to create a smooth morphing between two surfaces. Techniques more similar to our work include [4], who used...the thin - plate energy re- sulting in higher curvature in concentrated regions. On the left, minimizing the third derivative energy which results in

Modelling, Information, Processing, and Control

Science.gov (United States)

1989-01-15

have studied the use of transfer function methods to analyze closed - loop systems arising out of cer- tain linear feedback laws , use of transfer...classical Muntz- Szasz theory of real exponentials. As a re- sult it is seen that D has domain including that of A I / 2 in all cases. Further work is...approximation questions, we will discuss the use of transfer function methods to analyze closed - loop systems arising out of cer- tain linear feedback laws

Algunos aspectos de la hidroquímica del sistema acuífero Botucatú (Cuenca del Paraná, Estado de Sáo Paulo, Brasil)

OpenAIRE

Campos, H. C.; Cerón García, Juan Carlos

1997-01-01

The principal hydrochemical features of the Botucatú aquifer system in Sao Paulo State - SE Brasil - was studied from 67 groundwater chemical analysis. Westward, in the Botucatú aquifer system the progressive confining condition, with depth of the top of aquifer layer, increases the groundwater temperature slowly. Moreover, the residence time increases the water salt content towards the natural flow direction (E-W). As résults ,th'e recent calcium and magnesium bicarbonate groundwater cha...

Detonation Processes USSR

Science.gov (United States)

1960-06-06

second and quite definite for a given combustible mixture -- was an important. cientific re- sult of these researches discovered in 18881 by four...Consequently, the pulsating structure of the front of the "rnorr.ia]" detonation should be quite comrmon. (1) See K. i. Shchelkin, " Journal of...A1 "-൚ " 4 FOR REASONS OF SPEED AND ECONOMY THIS REPORT HAS BEEN REPRODUCED ELECTRONICALLY DIPWCTLY PROM OUR CONTRACTOR’S TYPESCRIPT THIS PUBLICATION

Chemical and structural order in silicon oxynitrides by methods of surface physics

Science.gov (United States)

Finster, J.; Heeg, J.; Klinkenberg, E.-D.

A large number of thin amorphous layers of SiO xN y and several (crystalline) reference compounds (SiO 2, Si 3N 4, Si 2N 2O) are studied. Although XANES and SEXAFS are well sulted to derive structural and chemical order, for these compounds many problems remain to be solved. We show how core level spectra (XPS, AES) can be used to gain such information (e.g. random bonding structure, N coordination, oxidation behaviour).