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.

Utilização da β-galactosidase para prevenção da cristalização em doce de leite Use of β-galactosidase in milk sweet: avoiding lactose crystallization

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Manuela Poletto Klein

2010-12-01

Full Text Available Este estudo avaliou o efeito da utilização da enzima β-galactosidase sobre a cristalização da lactose no doce de leite. As concentrações de lactase testadas variaram de 0 a 0,4 g/L. O grau de cristalização do produto foi avaliado sensorialmente (teste de ordenação em função de uma escala específica, após 30, 60, 90 e 180 dias de armazenamento à temperatura ambiente, por provadores previamente treinados. O teste de ordenação indicou não haver diferença estatística significativa (P>0,05 entre as amostras em relação à cristalização, tanto aos 30 quanto aos 60 dias de armazenamento, porém aos 90 e 180 dias essa diferença foi significativa entre o controle (T0 e os tratamentos. A utilização de 0,2 g/L de β-galactosidase (23,16% de hidrólise da lactose foi suficiente para que a arenosidade no doce de leite não fosse percebida sensorialmente pelos provadores, durante todo o período considerado.The effect of using β-galactosidase on the crystallization of lactose in milk sweet has been studied. The concentrations of lactase ranged from 0 to 0.4 g/L of processed milk. The product was subjected to sensorial analysis (crystallization degree ranking test using a specific scale by trained tasters after 30, 60, 90 e 180 days of storage. The ranking test showed no statistically significant difference between the samples in relation to crystallization, both at 30 and 60 days of storage, but at 90 and 180 days the difference was significant among control (T0 and the treatments. Usage of 0.2 g/L of β-galactosidase (23.16% lactose hydrolysis in milk was enough to prevent sandiness in milk sweet from being noticed throughout the studied period.

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.

Alternative method for quantification of alfa-amylase activity Alternativa para quantificação de alfa-amilase

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DF. Farias

2010-05-01

Full Text Available A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 °C. A linear relationship between the logarithm of enzyme activities and the area of clear zones was obtained. The method was validated by testing α-amylase from barley at the concentrations of 2.4; 60; 300 and 1,500 U.mL-1. The proposed method turned out to be simpler, faster, less expensive and able to determine on a macro-scale α-amylase over a wide range (2.4 to 7,500 U.mL-1 in scientific investigation as well as in teaching laboratory activities.Modificações foram propostas ao método sensível de difusão em ágar para a macrodeterminação de alfa-amilase. As modificações propostas diminuem os custos, com a utilização de amido como substrato e ágar como meio solidificante. Assim, foi construída uma curva padrão utilizando uma solução de alfa-amilase de Aspergillus oryzae com concentrações variando de 2,4 a 7.500 U.mL-1. Em seguida, as zonas claras de difusão radial foram mensuradas depois de 4 horas de incubação a 20 °C. Foi obtida uma relação linear entre o logaritmo da atividade enzimática e os diâmetros das zonas claras. O método foi validado utilizando-se soluções de alfa-amilase de cevada nas concentrações de 2,4; 60; 300 e 1.500 U.mL-1. O método tornou-se mais simples, rápido, com baixo custo e passível de ser utilizado para macrodeterminação de alfa-amilase em ampla faixa (2,4 a 7.500 U.mL-1 na investigação científica e para fins didáticos em aulas práticas.

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.

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

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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.

Purificación y caracterización de alfa-amilasa de penicillium commune producida mediante fermentación en fase sólida

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Espinel, Esperanza; López, Elizabeth

2010-01-01

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 acti...

Properties of a new fungal b-galactosidase with potential application in the dairy industry Propriedades de uma nova b-galactosidade fúngica com potencialidade para emprego na indústria de latícinios

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Rubens Cruz

1999-07-01

Full Text Available b-Galactosidase or b-D-galactoside-galactohydrolase (EC. 3.2.1.23 is an important enzyme industrially used for the hydrolysis of lactose from milk and milk whey for several applications. Lately, the importance of this enzyme was enhanced by its galactosyltransferase activity, which is responsible for the synthesis of transgalactosylated oligosaccharides (TOS that act as functional foods, with several beneficial effects on consumers. Penicillium simplicissimum, a strain isolated from soil, when grown in semi-solid medium showed good productivity of b-galactosidase with galactosyltransferase activity. The optimum pH for hydrolysis was in the 4.04.6 range and the optimum pH for galactosyltransferase activity was in the 6.07.0 range. The optimum temperature for hydrolysis and transferase activity was 55-60°C and 50°C, respectively, and the enzyme showed high thermostability for the hydrolytic activity. The enzyme showed a potential for several industrial applications such as removal of 67% of the lactose from milk and 84% of the lactose from milk whey when incubated at their original pH (4.5 and 6.34, respectively under optimum temperature conditions. When incubated with a 40% lactose solution in 150 mM McIlvaine buffer, pH 4.5, at 55°C the enzyme converted 86.5% of the lactose to its component monosaccharides. When incubated with a 60% lactose solution in the same buffer but at pH 6.5 and 50°C, the enzyme can synthetize up to 30.5% TOS, with 39.5% lactose and 30% monosaccharides remaining in the preparation.A b-galactosidase ou b-D-galactosídeo-galactohidrolase (EC. 3.2.1.23 é uma importante enzima utilizada industrialmente na hidrólise da lactose do leite e soro de leite para diversas aplicações. Ultimamente, sua importância foi realçada por sua atividade de galactosiltransferase responsável pela síntese de oligossacárides transgalactosilados (TOS que atuam como alimentos funcionais, trazendo diversos efeitos benéficos para seus

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

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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...

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.

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

Córnea verticilata - marcador clínico da doença de Fabry: relato de caso Cornea verticillata - a clinical marker of Fabry disease: case report

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Cynthia Azeredo Cordeiro

2007-08-01

Full Text Available A doença de Fabry é erro inato do metabolismo dos glicoesfingolipídeos (GL, resultante da atividade deficiente da enzima alfa-galactosidase A (a-Gal, com herança ligada ao cromossomo X. O acúmulo progressivo de GL nos tecidos resulta nas manifestações clínicas da doença, mais evidentes em homens hemizigotos, e incluem angioqueratomas, acroparestesias, córnea verticilata, hipo-hidrose, envolvimento cardíaco, renal e manifestações cerebrovasculares. Foi realizada avaliação em família acometida pela doença, sendo dois pacientes do sexo feminino e três do sexo masculino. Todos os pacientes foram submetidos a anamnese, exame oftalmológico completo e dosagem da atividade da enzima a-Gal. O único achado clínico presente em todos foi a córnea verticilata. Isto demonstra o importante papel que o exame oftalmológico apresenta no diagnóstico da doença, já que as alterações oculares são tão características.Fabry's disease is a rare X-linked lisosomal storage disorder of glycosphingolipid (GL metabolism, caused by a deficiency of alpha-galactosidase A activity. The progressive accumulation of GL in tissues results in the clinical manifestations of the disease, that are more evident in hemizygous males, and include angiokeratomas, acroparesthesia, cornea verticillata, cardiac and kidney involvement, cerebrovascular manifestations. A family with Fabry's disease including 2 female patients and 3 male patients is reported. The patients were submitted to complete medical history, ophthalmological examination and alpha-galactosidase activity test. Cornea verticillata was a constant finding in all patients. This demonstrates the important role of the ophtalmological examination for the diagnosis of Fabry's disease since the eye findings are so characteristic of the disease.

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.

Administration of follitropin alfa and lutropin alfa combined in a single injection: a feasibility assessment

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Agostinetto Rita

2009-05-01

Full Text Available Abstract Background Gonadotrophins are routinely administered in assisted reproductive technology (ART treatment protocols. Recombinant human follicle-stimulating hormone (r-hFSH; follitropin alfa and recombinant human luteinizing hormone (r-hLH; lutropin alfa can be administered individually or in a fixed combination. The ability to vary the FSH to LH dose ratio in a single injection without compromising the bioactivity of either gonadotrophin or generating losses of active principle is important for physicians and patients alike. Methods This study investigated whether follitropin alfa (GONAL-f (R, as lyophilized powder for reconstitution or solution from the GONAL-f (R (filled-by-mass [FbM] Prefilled Pen, could be used to reconstitute Pergoveris TM (follitropin alfa/lutropin alfa 150 IU/75 IU lyophilized powder. In Ratio Groups 1 and 2, the r-hFSH:r-hLH ratio was 3:1; in Ratio Groups 3 and 4, the ratios of r-hFSH:r-hLH were 5:1 and 8:1, respectively. The protein content and bioactivity of each mixed solution were evaluated. The r-hFSH and r-hLH content was determined using reverse-phase high performance liquid chromatography. The biological activity of r-hFSH and r-hLH was assessed using the Steelman-Pohley and Van Hell in vivo bioassays in rats, respectively. Results Follitropin alfa/lutropin alfa 150 IU/75 IU lyophilized powder could be successfully mixed with follitropin alfa 75 IU FbM solution that was either reconstituted from lyophilized powder or injected directly from the prefilled pen to create solutions with ratios of r-hFSH and r-hLH from 3:1 to 8:1. The measured content of r-hFSH and r-hLH corresponded favourably with the target protein content in Ratio Groups 1–4. The in vivo target and measured bioactivity of r-hFSH and r-hLH were also closely matched in all Ratio Groups. Conclusion Follitropin alfa lyophilized powder or solution can be accurately mixed with follitropin alfa/lutropin alfa 150 IU/75 IU lyophilized powder to

Eletroforese em acetato de celulose das proteínas do líquido cefalorraqueano: valores normais

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Izelino Caldas Filho

1977-09-01

Full Text Available Foi feita eletroforese das proteinas de LCR colhido por via lombar de 19 pacientes neurologicamente normais entre 10 e 54 anos. A proteinorraquia total variou entre 10 e 28 mg% (método de Meulemans. Sete pacientes eram normais e 12 apresentavam neuroses. As médias das frações separadas pela eletroforese foram: pré-albumina 4.3%; albumina 54.8%; globulina alfa-1 5.9%; globulina alfa-2 8.3%; globulinas beta 15.5%; globulinas gama 11.2%.

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.

PRODUCTION OF DISSOLVING GRADE PULP FROM ALFA

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Baya Bouiri

2010-02-01

Full Text Available Alfa, also known as Stipa tenacissimaI or “halfa”, is grown in North Africa and south Spain. Due to its short fiber length, paper made from alfa pulp retains bulk and takes block letters well. In this study alfa was evaluated for bleached pulp production. Two cellulose pulps with different chemical compositions were pulped by a conventional kraft process. One sample was taken from the original alfa material and another from alfa that had been pretreated by diluted acid. The pulp produced from the pretreated alfa was bleached by the elemental-chlorine-free sequences DEPD and DEDP. The yield, Kappa number, brightness, and α- cellulose content of bleached and unbleached pulps were evaluated. The results showed that during the chemical pulping process, treated alfa cooked more easily than the original alfa. The treated alfa pulp also showed very good bleaching, reaching a brightness level of 94.8% ISO with a yield of 93.6% at an α-cellulose content 96.8(% with a DEDP bleaching sequence, compared to 83.2% ISO brightness level, 92.8% yield, and 95.1% α-cellulose content for bleached pulp with a DEPD bleaching sequence. Therefore, this alfa material could be considered as a worthwhile choice for cellulosic fiber supply.

Farelo de trigo e complexo enzimático na alimentação de poedeiras semipesadas na fase de produção Wheat bran and enzymatic complex in the feeding of laying hens in the production phase

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Daniel de Magalhães Araujo

2008-05-01

Full Text Available O estudo foi conduzido com o objetivo de avaliar os efeitos da inclusão de farelo de trigo (FT na ração com ou sem a suplementação de um complexo enzimático (CE sobre o desempenho e a produção de ovos de frangas semipesadas na fase de produção. Foram utilizadas 288 frangas Lohmann Brown, distribuídas em delineamento inteiramente casualizado, em esquema fatorial 4 × 2, composto de quatro níveis de farelo de trigo (0, 3, 6 e 9% na ração e da suplementação de complexo enzimático (0 ou 100 g/100 kg de ração, com oito tratamentos e seis repetições de seis aves. O complexo enzimático continha as enzimas beta-galactosidase, galactomananase, xilanase e alfa-glucanase. O consumo de ração, o peso vivo final, a produção de ovos, o peso e a massa de ovos e a conversão por massa e por dúzia de ovos não foram afetados pela inclusão do farelo de trigo nas rações. A gravidade específica da casca dos ovos piorou com o aumento dos níveis de farelo de trigo nas dietas. Nenhuma característica foi afetada pela adição do complexo enzimático na ração, com exceção do peso do ovo, que melhorou de 62,74 para 64,28 g. Recomenda-se o uso de até 9% de farelo de trigo em rações para poedeiras em produção. A adição de alfa-galactosidase, galactomananase, xilanase e alfa-glucanase aumenta o peso dos ovos.The study was carried out with the objective to evaluate the effects of the inclusion of the wheat bran (WB with or without supplementation of an enzymatic complex (EC on the performance of semi-heavy hens in the egg-production phase. A total of 288 Lohmann Brown pullets were used, distributed to a completely randomized design in 4 × 2 factorial arrangement, composed by four WB levels (0, 3, 6 and 9% in the ration and enzymatic complex supplementation (0 or 100g/100 kg diet, with eight treatments and six replicates of six birds. The enzymatic complex contained the enzymes beta-galactosidase, galactomananase, xilanase and alpha

Drotrecogin alfa Eli Lilly

NARCIS (Netherlands)

de Jonge, E.

2002-01-01

Drotrecogin alfa (Xigris, recombinant activated protein C) is an anticoagulant developed and launched by Eli Lilly & Co for the treatment of sepsis [333781], [339372], [430133], [436271]. The FDA and the EMEA accepted the brand name Xigris for drotrecogin alfa in June 2001. This trade name had been

ALFA Detector Control System

CERN Document Server

Oleiro Seabra, Luis Filipe; The ATLAS collaboration

2015-01-01

ALFA (Absolute Luminosity For ATLAS) is one of the sub-detectors of ATLAS (A Toroidal LHC Apparatus). The ALFA system is composed by four stations installed in the LHC tunnel 240 m away from the ATLAS interaction point. Each station has a vacuum and ventilation system, movement control and all the required electronics for signal processing. The Detector Control System (DCS) provides control and monitoring of several components and ensures the safe operation of the detector contributing to good Data Quality. This paper describes the ALFA DCS system including a detector overview, operation aspects and hardware control through a SCADA system, WinCC OA.

ALFA Detector Control System

CERN Document Server

Oleiro Seabra, Luis Filipe; The ATLAS collaboration

2015-01-01

ALFA (Absolute Luminosity For ATLAS) is one of the sub-detectors of ATLAS/LHC. The ALFA system is composed by two stations installed in the LHC tunnel 240 m away from each side of the ATLAS interaction point. Each station has a vacuum and ventilation system, movement control and all the required electronic for signal processing. The Detector Control System (DCS) provides control and monitoring of several components and ensures the safe operation of the detector contributing to good Data Quality. This paper describes the ALFA DCS system including a detector overview, operation aspects and hardware control through a SCADA system, WinCC OA.

Dificuldades na identificação laboratorial da talassemia alfa Difficulty on laboratory identification of alpha thalassemia

Directory of Open Access Journals (Sweden)

Karlla Greick Batista Dias-Penna

2010-04-01

Full Text Available Introdução: Talassemia alfa é uma síndrome associada à redução da síntese de cadeias de globina do tipo alfa. A gravidade das manifestações clínicas está relacionada com a quantidade de globinas produzida e a estabilidade das cadeias beta presentes em excesso. A talassemia alfa mínima resulta da deleção de apenas um dos quatro genes a (-α/αα. Clinicamente apresenta anemia leve com microcitose ou ausência de anemia, sendo o diagnóstico realizado por meio de visualização da hemoglobina (Hb H por eletroforese alcalina em acetato de celulose ou por identificação de inclusões celulares de Hb H coradas pelo azul de crezil brilhante. Objetivo: Avaliar portadores de talassemia alfa e seus respectivos progenitores, correlacionando perfil hematológico e presença de Hb H, utilizando procedimentos laboratoriais clássicos em três diferentes amostragens. Discussão e conclusão: Os dados obtidos mostram que a presença de Hb H, indicativo de talassemia alfa, pode não ser confirmada em uma análise posterior. Entre os fatores que podem influenciar no não aparecimento de Hb H em pessoa comprovadamente com talassemia alfa está a deficiência de ferro. A talassemia alfa está associada a defeitos envolvendo os genes codificadores da cadeia alfa, mas também pode estar relacionada com desbalanciamento temporário na expressão dos genes globina, diminuição de alfa ou aumento de beta, o que poderia explicar o aparecimento de tetrâmeros de cadeia beta (Hb H, sugerindo diagnóstico de talassemia alfa mínima.Introduction: Alpha thalassemia is a syndrome with associated with the reduction of alpha globin chain synthesis. The severity of clinical manifestations is related to the amount of globins produced and the stability of beta chains that are present in excess. Alpha thalassemia minor is caused by the deletion of one of the four genes a (-α/αα. Clinically, it presents mild anemia with microcytosis or absence of anemia. The

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

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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

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.

Inhibidor de la enzima uracilo ADN glicosilasa y usos del mismo

OpenAIRE

Serrano-Heras, Gemma; Bravo, Alicia; Salas, Margarita

2005-01-01

Inhibidor de la enzima uracilo adn glicosilasa y usos del mismo Esta invención se refiere a una proteína que inhibe la enzima uracilo ADN glicosilasa (UDG) vírica, para su uso como un agente terapéutico, en particular como un agente antiví

Purification and characterization of α-galactosidase from ...

African Journals Online (AJOL)

α−Galactosidase (α-D-galactoside galactohydrolase [EC 3.2.1.22]) was obtained from Lactobacillus acidofillus which was grown in modified de Man, Rogosa and Sharpe (MRS) medium, supplemented with raffinose. α-Galactosidase was released from the cells by ultrasonic treatment, then precipitated by ...

Peginterferon alfa-2b and ribavirin: effective in patients with hepatitis C who failed interferon alfa/ribavirin therapy.

Science.gov (United States)

Poynard, Thierry; Colombo, Massimo; Bruix, Jordi; Schiff, Eugene; Terg, Ruben; Flamm, Steven; Moreno-Otero, Ricardo; Carrilho, Flair; Schmidt, Warren; Berg, Thomas; McGarrity, Thomas; Heathcote, E Jenny; Gonçales, Fernando; Diago, Moises; Craxi, Antonio; Silva, Marcelo; Bedossa, Pierre; Mukhopadhyay, Pabak; Griffel, Louis; Burroughs, Margaret; Brass, Clifford; Albrecht, Janice

2009-05-01

Treatment with peginterferon alfa and ribavirin produces a sustained virologic response (SVR) in approximately 60% of hepatitis C virus (HCV)-infected patients. Alternate options are needed for patients who relapse or do not respond to therapy. This prospective, international, multicenter, open-label study evaluated efficacy and safety of peginterferon alfa-2b (1.5 microg/kg/wk) plus weight-based ribavirin (800-1400 mg/day) in 2333 chronic HCV-infected patients with significant fibrosis/cirrhosis whose previous interferon alfa/ribavirin therapy failed. Patients with undetectable HCV-RNA at treatment week (TW) 12 received 48 weeks of therapy; patients with detectable HCV-RNA at TW12 could enter maintenance studies at TW18; 188 patients with low/detectable HCV-RNA at TW12 continued therapy at the investigator's request. Overall, 22% of the patients attained SVR (56% with undetectable HCV-RNA and 12% with low/detectable HCV-RNA at TW12). SVR was better in relapsers (38%) than nonresponders (14%), regardless of previous treatment, and in patients previously treated with interferon-alfa/ribavirin (25%) than peginterferon alfa-ribavirin (17%). Predictors of response in patients with undetectable HCV-RNA at TW12 were genotype (2/3 vs 1, respectively; odds ratio [OR] 2.4; P 600,000 IU/mL; OR, 1.4; P = .0223). These factors plus previous treatment and response were overall predictors of SVR. Safety was similar among fibrosis groups. Peginterferon alfa-2b plus weight-based ribavirin is effective and safe in patients who failed interferon alfa/ribavirin therapy. Genotype, baseline viral load, and fibrosis stage were predictors of response.

Drotrecogin alfa Eli Lilly.

Science.gov (United States)

de Jonge, E

2002-04-01

Drotrecogin alfa (Xigris, recombinant activated protein C) is an anticoagulant developed and launched by Eli Lilly & Co for the treatment of sepsis [333781], [339372], [430133], [436271]. The FDA and the EMEA accepted the brand name Xigris for drotrecogin alfa in June 2001. This trade name had been proposed by Lilly in place of the previous brand name, Zovant, which was deemed unacceptable by the EMEA due to concerns that the name could be confused with hospital-based drugs [412512]. Filings for sepsis were made in the US, EU and Australia in February 2001 [398514], [447870] and in March 2001, the US FDA assigned drotrecogin alfa Priority Review status [403435]. The FDA extended the action date from July 27 to October 27, 2001 for completion of its review of the biologics license application (BLA) for drotrecogin alfa to assess further supplementary data submitted by Lilly [412512]. At the October 16, 2001 meeting (postponed from September 12), the FDA Advisory Committee on Anti-Infective Drugs split 10 to 10 over whether to recommend approval [425873], [425940]. In late October 2001, Lilly received an approvable letter from the FDA for the treatment of severe sepsis. Approval was contingent upon successful negotiation of labeling, agreement on post-approval clinical trials, and successful completion of manufacturing inspections [427301]. In November 2001, the FDA approved drotrecogin alfa for the reduction of mortality in adult patients with severe sepsis who have a high risk of death [430133]; the product was launched onto the US market days later [436271]. Following the FDA committee's split decision in October 2001, Credit Suisse First Boston, which expected mid-2002 approval but with restrictive labeling, revised its predictions from $1.265 billion in 2004 sharply downwards to $543 million [425929].

β-Galactosidase Deficiency in Colombia

Directory of Open Access Journals (Sweden)

Alfredo Uribe PhD

2015-05-01

Full Text Available β-Galactosidase (BGal is the first enzyme involved in the catabolism of sphingolipids. Two pathologies have been directly associated with its deficiency: GM1 gangliosidosis and Morquio B. Morquio B is among the rarest types of mucopolysaccharidosis (MPS. We aim to document the β-galactosidase deficiency in Colombia. We evaluated leukocytes from 1492 healthy Colombian individuals and 923 patients, referred between 2005 and August 2014. Dried blood spot (DBS samples from the same number of patients were evaluated. β-Galactosidase was measured with 4-methylumbelliferyl-β- d -galactoside. As a control enzyme, the total hexosaminidase activity was also evaluated. We identified 14 patients with GM1 gangliosidosis, 5 patients with Morquio B, and 1 patient with I-cell disease. We could establish a reference value for Bgal in Colombian leukocyte samples. GM1 gangliosidosis is the main pathology associated with a direct deficiency of BGal. The high number of patients found with MPS IVB indicates that there are patients who could be misdiagnosed due to an unawareness of the disease.

Purification of a-galactosidase from seeds of Sesbania marginata

Directory of Open Access Journals (Sweden)

Falco A.L.P.

2000-01-01

Full Text Available Alpha-galactosidase taken from a raw extract of Sesbania marginata legume seeds was purified by partitioning in aqueous two-phase systems (ATPS. Initially, galactomannan/dextran 2,000,000 systems were used for the purification, and the partition coefficients of alpha -galactosidase varied from 1.5 to 4.0. However, mass transport in these systems was poor due to the high viscosity of the employed polymers. Therefore, partitioning in polyethyleneglycol (PEG/ sodium phosphate systems and the effect of sodium chloride upon the enzyme purification and the yield of alpha -galactosidase were also investigated. The purification achieved in a single-step was 5.7 with a recovery of 144% of alpha -galactosidase, possibly due to the removal of materials which inhibited alpha -galactosidase activity before the purification. The removal of the main protein contaminants and the highest yields were achieved in PEG 4,000/ sodium phosphate + 6% NaCl system at pH 5.0. Further purification by preparative on-exchange chromatography was also developed.

Implementation of Trigger Tiles for ALFA Simulation

CERN Document Server

Rehaag, Thomas Joseph

2017-01-01

The Absolute Luminosity For ATLAS (ALFA) experiment was designed to accurately measure the luminosity of the intersecting proton beams at the ATLAS interaction point [1]. However, the ALFA experiment has shifted its primary purpose from luminosity measurement to elastic and inelastic proton collisions. This change was the result of difficulty in fitting parameters in the region governed by Coulomb scattering. The operational principle for luminosity measurement with ALFA relied on detecting elastic proton collisions, so the detector is suited to its role in proton collision measurements. The ALFA detector consists of several sensitive components, including the main detector (MD), overlap detectors (ODs) and trigger tiles. A diagram of the ALFA detector is shown in Figure 1. The main detector is composed of layers of 0.5 × 0.5 mm2 cross section scintillating fibres with an active area of 0.48 × 0.48 mm2, which are directed diagonally across the detector with 64 fibres in each layer. The 20 total layers ar...

Functionality and performance of the ALFA_CTPIN module

CERN Document Server

Iwanski, Wieslaw; Korcyl, Krzysztof; Oechsle, Jan

2017-01-01

The ALFA_CTPIN module has been designed in response to increased internal processing time of the Central Trigger Processor (CTP) of the ATLAS experiment which resulted in reducing time left to the ALFA detector to deliver its own triggers to the CTP within specified latency. Accelerated extraction of ALFA triggers from encoded signals and the possibility to perform local triggers processing by this module allowed ALFA to contribute to global triggering of the ATLAS detector. A huge number of implemented scalers and flexibility in defining triggers processing criteria make also from this module a very attractive tool for in-depth analysis of properties of the LHC beam.

Optimization of β-galactosidase production from lactic acid bacteria

Directory of Open Access Journals (Sweden)

Carević Milica

2015-01-01

Full Text Available β-galactosidase, commonly known as lactase, represents commercially important enzyme that is prevalently used for lactose hydrolysis in milk and whey. To the date, it has been isolated from various sources. In this study different strains of lactic acid bacteria were assessed for their β-galactosidase productivity, and Lactobacillus acidophilus ATCC 4356 resulted with the highest production potential. Thereafter, optimal conditions for accomplishing high yields of β-galactosidase activity were determined. Maximal specific activity (1.01 IU mL-1 was accomplished after 2 days shake flask culture fermentation (150 rpm at 37ºC, with modified Man Rogosa Sharpe culture broth using lactose (2.5% as sole carbon source. Finally, in order to intensify release of intracellular β-galactosidase different mechanical and chemical methods were conducted. Nevertheless, vortexing with quartz sand (150 μm as abrasive was proven to be the most efficient method of cell disruption. The optimum temperature of obtained β-galactosidase was 45°C and the optimum range pH 6.5-7.5.

Análise comparativa do controle interno no contas a receber e a pagar nas empresas Beta Ltda e Alfa Ltda = A comparative internal control analysis of receivable and payable accounts in Beta and Alfa Ltd Companies

Directory of Open Access Journals (Sweden)

Cleyton de Oliveira Ritta

2010-07-01

Full Text Available A necessidade constante de um excelente desempenho empresarial leva à busca da qualidade e confiabilidade da informação para o auxílio na tomada de decisão. Neste contexto, a auditoria contábil tem um papel importante na análise das práticas administrativas adotadas pelas entidades. Desse modo, o objetivo deste artigo é comparar a estruturação dos controles internos no contas a receber e a pagar nas empresas Beta Ltda, a qual não contrata serviços de auditoria externa, e Alfa Ltda, que os contrata. Este estudo caracteriza-se como de caso comparativo de caráter descritivo por meio de questionário e entrevista. Os resultados apontam que a empresa Alfa Ltda tem melhores níveis de controle interno e, consequentemente, está menos exposta a erros e fraudes. Desse modo, entende-se que os seviços de auditoria atuam como um instrumento de auxílio para minimização dos riscos operacionais e servem de alerta aos gestores quanto à eficiência das transações. Contudo, percebe-se que a qualidade dos controles internos nas organizações depende diretamente da utilização de procedimentos de monitoramento.The constant need for excellent business performance triggers the quest for information quality and reliability to help in decision making. Considering this context, accounting auditing plays an important role in the analysis of administrative practices adopted by entities. Thus, the objective of this article is to compare the structure of the internal controls for receivable and payable accounts in Beta Ltd companies, which do not contract external auditing services, and Alfa Ltd, which does. This study is a descriptive comparative case study, carried out through questionnaire and interview. The results have showed that Alfa Ltd has better internal control levels and consequently is less exposed to errors and fraud. This way, auditing services are seen as auxiliary instruments to minimize operational risks and serve to alert

Inmovilización de enzimas en materiales mesoporosos ordenados

OpenAIRE

Serra, Elías M.

2011-01-01

En este trabajo se ha pretendido arrojar algo de luz sobre algunos aspectos que afectan a la inmovilización de enzimas en Materiales Mesoporosos Ordenados. La mayor parte del esfuerzo se ha centrado en la inmovilización de la lipasa, sin perder de vista la extensión de las técnicas más exitosas a otras enzimas de interés. En primer lugar se ha establecido la influencia de parámetros texturales y/o químicos de los MMO sobre la inmovilización, mostrando como tanto el tamaño como la forma de ...

Purification and properties of beta-galactosidase from Aspergillus nidulans.

Science.gov (United States)

Díaz, M; Pedregosa, A M; de Lucas, J R; Torralba, S; Monistrol, I F; Laborda, F

1996-12-01

Beta-Galactosidase from mycelial extract of Aspergillus nidulans has been purified by substrate affinity chromatography and used to obtain anti-beta-galactosidase polyclonal antibodies. A. nidulans growing in lactose as carbon source synthesizes one active form of beta-galactosidase which seems to be a multimeric enzyme of 450 kDa composed of monomers with 120 and 97 kDa. Although the enzyme was not released to the culture medium, some enzymatic activity was detected in a cell-wall extract, thus suggesting that it can be an extracellular enzyme. Beta-Galactosidase of A. nidulans is a very unstable enzyme with an optimum pH value of 7.5 and an optimum temperature of 30 degrees C. It was only active against beta-galactoside substrates like lactose and p-nitrophenyl-beta-D-galactoside (PNPG).

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 Reação da Desitratação da Gipsita Em Meio Aquoso Num Reator de Batelada (parr) Para Produção de Gesso Alfa

OpenAIRE

Pereira, Pedro Leiz de Mendonça

2012-01-01

A partir da reação de desidratação da gipsita (CaSO4.2H2O) pode-se obter o Gesso (CaSO4.1/2H2O) do tipo alfa e do tipo beta, dependendo das condições nas quais ocorre a reação. A produção do gesso alfa é mais complexa, por tratar-se de um produto com características de maior valor agregado e com aplicação diferenciada. O presente trabalho visa o estudo dos aspectos termocinéticos da reação de desidratação da gipsita para obtenção do gesso alfa em meio aquoso, e procura trazer maiores informaç...

Sequential combination of glucocorticosteroids and alfa interferon versus alfa interferon alone chronic hepatitis B. Protocol for a Cochrane Review

DEFF Research Database (Denmark)

Mellerup, M T; Krogsgaard, K; Mathurin, P

2000-01-01

Chronic hepatitis B has serious effects on morbidity and mortality. Alfa interferon has been shown to increase the rates of HBeAg-clearance as well as seroconversion to anti-HBe, but response rates are unsatisfactory. Glucocorticosteroid pretreatment may increase the response to alfa interferon....

Thermal inactivation kinetics of β-galactosidase during bread baking

NARCIS (Netherlands)

Zhang, L.; Chen, Xiao Dong; Boom, R.M.; Schutyser, M.A.I.

2017-01-01

In this study, β-galactosidase was utilized as a model enzyme to investigate the mechanism of enzyme inactivation during bread baking. Thermal inactivation of β-galactosidase was investigated in a wheat flour/water system at varying temperature-moisture content combinations, and in bread during

Nicotiana benthamiana α-galactosidase A1.1 can functionally complement human α-galactosidase A deficiency associated with Fabry disease.

Science.gov (United States)

Kytidou, Kassiani; Beekwilder, Jules; Artola, Marta; van Meel, Eline; Wilbers, Ruud H P; Moolenaar, Geri F; Goosen, Nora; Ferraz, Maria J; Katzy, Rebecca; Voskamp, Patrick; Florea, Bogdan I; Hokke, Cornelis H; Overkleeft, Herman S; Schots, Arjen; Bosch, Dirk; Pannu, Navraj; Aerts, Johannes M F G

2018-04-19

α-Galactosidases (EC 3.2.1.22) are retaining glycosidases that cleave terminal α-linked galactose residues from glycoconjugate substrates. α-Galactosidases take part in the turnover of cell wall-associated galactomannans in plants and in the lysosomal degradation of glycosphingolipids in animals. Deficiency of human α-galactosidase A (α-Gal A) causes Fabry disease (FD), a heritable, X-linked lysosomal storage disorder, characterized by accumulation of globotriaosylceramide (Gb3) and globotriaosylsphingosine (lysoGb3). Current management of FD involves enzyme-replacement therapy (ERT). An activity-based probe (ABP) covalently labeling the catalytic nucleophile of α-Gal A has been previously designed to study α-galactosidases for use in FD therapy. Here, we report that this ABP labels proteins in Nicotiana benthamiana leaf extracts, enabling the identification and biochemical characterization of an N. benthamiana α-galactosidase we name here A1.1 (gene accession GJZM-1660). The transiently overexpressed and purified enzyme was a monomer lacking N-glycans and was active toward 4-methylumbelliferyl-α-D-galactopyranoside substrate (Km = 0.17 mM) over a broad pH range. A1.1 structural analysis by X-ray crystallography revealed marked similarities with human α-Gal A, even including A1.1's ability to hydrolyze Gb3 and lysoGb3, which are not endogenous in plants. Of note, A1.1 uptake into FD fibroblasts reduced the elevated lysoGb3 levels in these cells, consistent with A1.1 delivery to lysosomes as revealed by confocal microscopy. The ease of production and the features of A1.1, such as stability over a broad pH range, combined with its capacity to degrade glycosphingolipid substrates, warrant further examination of its value as a potential therapeutic agent for ERT-based FD management. Published under license by The American Society for Biochemistry and Molecular Biology, Inc.

ALFA: an automated line fitting algorithm

Science.gov (United States)

Wesson, R.

2016-03-01

I present the automated line fitting algorithm, ALFA, a new code which can fit emission line spectra of arbitrary wavelength coverage and resolution, fully automatically. In contrast to traditional emission line fitting methods which require the identification of spectral features suspected to be emission lines, ALFA instead uses a list of lines which are expected to be present to construct a synthetic spectrum. The parameters used to construct the synthetic spectrum are optimized by means of a genetic algorithm. Uncertainties are estimated using the noise structure of the residuals. An emission line spectrum containing several hundred lines can be fitted in a few seconds using a single processor of a typical contemporary desktop or laptop PC. I show that the results are in excellent agreement with those measured manually for a number of spectra. Where discrepancies exist, the manually measured fluxes are found to be less accurate than those returned by ALFA. Together with the code NEAT, ALFA provides a powerful way to rapidly extract physical information from observations, an increasingly vital function in the era of highly multiplexed spectroscopy. The two codes can deliver a reliable and comprehensive analysis of very large data sets in a few hours with little or no user interaction.

OPPORTUNITIES OF DORNASE ALFA IN PEDIATRICS

Directory of Open Access Journals (Sweden)

O.I. Simonova

2011-01-01

Full Text Available Early treatment with Pulmozyme (dornase alfa and its prolonged use in combination with basis therapy results in decrease of exacerbation rate, prolongation of remission of bronchopulmonary process with normal ventilation functions of lungs and gives patients with cystic fibrosis an opportunity to live an active life. The drug is effective as a conservative treatment of chronic sinusitis polyposis; it’s frequent complication of cystic fibrosis. Dornase alfa has 3 clinical effects: strong mucolytic, anti-inflammatory and antibacterial effect. That is the reason for the treatment of other chronic bronchopulmonary diseases with this drug. Efficacy of dornase alpha depends on rough maintenance of the terms of drug use.Key words: children, dornase alfa, cystic fibrosis, chronic bronchopulmonary diseases, chronic rhinosinusitis, nasal polyposis, nebulizer therapy.(Voprosy sovremennoi pediatrii — Current Pediatrics. 2011; 10 (4: 145–152

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.

Mini review on role of β-galactosidase in lactose intolerance

Science.gov (United States)

A, Nivetha; V, Mohanasrinivasan

2017-11-01

This review mainly focuses on the role and properties of β-galactosidase in lactose intolerance and its industrial application. β-Galactosidase, hydrolyses the lactose into glucose and galactose and it is most commonly used in food based technology, particularly in the dairy manufacturing industry. This catalyst mainly focus for the improvement of new and novel products with hydrolyzed lactose, which can be appropriate for the lactose-intolerant persons, to improve the technological, texture and scientific properties of non-fermented dairy products. β-Galactosidase derived from the group of saccharides which is a converting enzymes in the family of hydrolases. They are broadly distributed in the several biological living systems. The enzymatic hydrolysis of lactose is also preferred in food based technology due to the low soluble range of lactose. The concentration lactose was found to be high in fermented dairy products such as ice cream, butter, cheese curd, yogurt, etc., can prompt extreme lactose crystallization bringing about items through a coarse, abrasive surface. Lactose hydrolysis in dairy products enhances adaptability also, richness altogether. These products are extra edible. Also for this purpose, the utilization of β-galactosidase enzyme prior to the condensing operation can reduce the lactose content to a point where lactose was no longer a problem industrial application of β-galactosidase. In Industries, due to the positive and constructive effect on intestinal bacterial microflora, different types of applications are possible in β-galactosidase enzyme.

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

Diagnóstico diferencial das distrofias musculares com referência especial às alterações enzimáticas

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H. Heick

1967-06-01

Full Text Available Depois de apresentar os dados clínicos e genéticos relativos a 221 casos de distrofia muscular progressiva (125 casos do tipo Duchenne, 63 do tipo cintura pélvica e 25 do tipo fácio-escápulo-umeral, os autores analisam os resultados das dosagens sôro-enzimáticas que fizeram; em alguns casos foram pesquisas também as atividades de 25 enzimas em homogeneizados de tecido muscular. Os mesmos exames foram feitos em casos de miopatias de outra etiologia e de distrofias musculares neuropáticas e mielopáticas. Foi verificado que o teor das sôro-enzimas aumenta em todas as formas de distrofia muscular progressiva hereditária. No tipo Duchenne as-atividades enzimáticas são elevadas desde o nascimento das crianças acometidas, precedendo o aparecimento da sintomatologia clínica. Com o progredir da moléstia e com o decorrer dos anos os teores sôro-enzimáticos tendem a diminuir, podendo reduzir-se aos valores normais nas fases finais da moléstia. No tecido muscular não existe diferença significante, no que respeita à composição enzimática, entre os diversos tipos de distrofia muscular progressiva e hereditária. Com exceção de poucas enzimas cuja atividade é maior nos tecidos conjuntivo e adiposo que progressivamente substituem o tecido muscular que entra em atrofia, o teor das mio-enzimas diminui com o progredir da moléstia, como expressão do crescente esgotamento do citoplasma da célula muscular. Nas distrofias musculares neuropáticas e mielopáticas as alterações são idênticas, embora de menor intensidade. Algumas diferenças quantitativas podem ser explicadas pela idade dos pacientes e pela intensidade do processo. A mesma diferença pode ser demonstrada mediante exames histopatológicos: nas formas que acometem crianças (Werdnig-Hoffmann as fibras musculares ainda não denervadas se apresentam com aspecto normal, ao passo que, nas formas do adulto (Kugelberg-Welander, possivelmente pelo desgaste provocado pelos esfor

ALFA: The new ALICE-FAIR software framework

Science.gov (United States)

Al-Turany, M.; Buncic, P.; Hristov, P.; Kollegger, T.; Kouzinopoulos, C.; Lebedev, A.; Lindenstruth, V.; Manafov, A.; Richter, M.; Rybalchenko, A.; Vande Vyvre, P.; Winckler, N.

2015-12-01

The commonalities between the ALICE and FAIR experiments and their computing requirements led to the development of large parts of a common software framework in an experiment independent way. The FairRoot project has already shown the feasibility of such an approach for the FAIR experiments and extending it beyond FAIR to experiments at other facilities[1, 2]. The ALFA framework is a joint development between ALICE Online- Offline (O2) and FairRoot teams. ALFA is designed as a flexible, elastic system, which balances reliability and ease of development with performance using multi-processing and multithreading. A message- based approach has been adopted; such an approach will support the use of the software on different hardware platforms, including heterogeneous systems. Each process in ALFA assumes limited communication and reliance on other processes. Such a design will add horizontal scaling (multiple processes) to vertical scaling provided by multiple threads to meet computing and throughput demands. ALFA does not dictate any application protocols. Potentially, any content-based processor or any source can change the application protocol. The framework supports different serialization standards for data exchange between different hardware and software languages.

Novel alpha-galactosidase A mutation in a female with recurrent strokes.

Science.gov (United States)

Tuttolomondo, Antonino; Duro, Giovanni; Miceli, Salvatore; Di Raimondo, Domenico; Pecoraro, Rosaria; Serio, Antonia; Albeggiani, Giuseppe; Nuzzo, Domenico; Iemolo, Francesco; Pizzo, Federica; Sciarrino, Serafina; Licata, Giuseppe; Pinto, Antonio

2012-11-01

Anderson-Fabry disease (AFD) is an X-linked inborn error of glycosphingolipid catabolism resulting from the deficient activity of the lysosomal exoglycohydrolase, a-galactosidase A. The complete genomic and cDNA sequences of the human alpha-galactosidase A gene have been determined and to date, several disease-causing alpha-galactosidase A mutations have been identified, including missense mutations, small deletions/insertions, splice mutations, and large gene rearrangements We report a case of a 56-year-old woman with recurrent cryptogenic strokes. Ophthalmological examination revealed whorled opacities of the cornea (cornea verticillata) and dilated tortuous conjunctival vessels. She did not show other typical signs of Fabry disease such as acroparesthesias and angiokeratoma. The patient's alpha-galactosidase A activity was 4.13 nmol/mL/h in whole blood. Alpha-galactosidase A gene sequence analysis revealed a heterozygous single nucleotide point mutation at nucleotide c.550T>A in exon 4 in this woman, leading to the p.Tyr184Asn amino acid substitution. Copyright © 2012 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.

Purification of beta-acetylglucosaminase and beta-galactosidase from ram testis.

Science.gov (United States)

Caygill, J C; Roston, C P; Jevons, F R

1966-02-01

1. The presence of beta-galactosidase (EC 3.2.1.23) in an acetic acid extract of ram testis is reported. Some properties of the crude enzyme preparation were studied. 2. The purification of beta-acetylglucosaminase (EC 3.2.1.30) and of beta-galactosidase from the ram-testis extract by ammonium sulphate precipitation and chromatography on a CM-cellulose column is described. 3. The final purifications of the separated enzymes achieved were for the beta-acetylglucosaminase 35 times and for the beta-galactosidase 99 times. 4. The possibility of using DEAE-cellulose and Sephadex G-200 to purify the enzymes was investigated.

Alternative method for quantification of alfa-amylase activity Alternativa para quantificação de alfa-amilase

OpenAIRE

DF. Farias; AFU. Carvalho; CC. Oliveira; NM. Sousa; LCB. Rocha-Bezerrra; PMP. Ferreira; GPG. Lima; DC. Hissa

2010-01-01

A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 °C. A linear relationship between the logarithm of...

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.

Curious Cases of the Enzymes / Neobiča Istorija Enzima

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Ulusu Nuriye Nuray

2015-07-01

Full Text Available : Život oko nas uglavnom se zasniva na biološkoj katalizi, pa bi se čak, u nekakvoj neromantičnoj priči, život mogao opisati kao niz hemijskih reakcija koje regulišu glavni principi termodinamike. U davnoj prošlosti, ku­canje srca bilo je znak dobrog zdravlja, međutim, za mene, najvažniju ulogu ima upravo prisustvo enzima... Mada ne obraćamo često pažnju na to, istorija enzimologije stara je koliko i sam ljudski rod i seže daleko u prošlost. CVaj rad posvećen je baš tim počecima u ovoj veoma zanimljivoj nauci, koja je u prošlosti ostavila traga na našim životima i koja će u budućnosti učiniti da život bude mnogo pro­duktivniji za čovečanstvo. Između čoveka i enzima postojao je oduvek jedan delikatan, suštinski važan odnos. Suočen s nepoznatom i neprijateljski nastrojenom prirodom punom grabljivaca, preistorijski čovek rano je otkrio lekovita svojstva biljaka, i to metodom probe i greške. Štaviše, slučajno su otkriveni inhibitori enzima, čime je, grubo rečeno, još tada otvorena nova oblast istraživanja. Oi biljni derivati koji su imali ulogu inhibitora enzima pomagali su preistorijskom čoveku u njegovoj borbi da opstane i zaštiti se od grabljiva­ca; prilikom lova i ribolova... Kasnije kroz istoriju, sa nepro- menjenom svrhom opstanka i poboljšanja kvaliteta života, enzimologija je iz korena transformisana, a metodologiju »probe i greške« iz praistorije zamenile su racionalne naučne teorije.

Study of β-Galactosidase Enzyme Activity Produced by Lactobacilli in Milk and Cheese

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J. Nowroozi

2008-04-01

Full Text Available Background and objectiveLactose intolerance is a discomfort state that occurs in some people after ingestion of milk and it is due to insufficient amount of beta galactosidase in the human gut to digest lactose. The aim of this study was to observe the presence of beta galactosidase enzyme produced by isolated lactobacilli from milk and cheese. Methods In this descriptive study, milk and cheese samples with different brand were bought from different shops. Lactobacilli were identified by plating samples on MRS medium, Gram staining and standard biochemical methods. β-galactosidase production by bacteria was assessed by X-Gal and ONPG methods. β-galactosidase was also detected by SDS-PAGE. ResultsFourteen genus of lactobacillus were isolated From 50 samples. All of the bacteria produced green color colonies on X-Gal plates (but in different times that indicated the presence of enzyme in the bacteria. All isolated lactobacilli were shown β-galactosidase activity in ONPG test. The highest enzymatic activity was seen in one strain of Lactobacillus Delbrueckii (1966 Miller unit /ml. In some bacteria (37% a strong β-galactosidase band(116-kDa was seen by SDS-PAGE.ConclusionAddition of beta galactosidase containing lactobacilli as a probiotic agent to milk, cheese, and other dairy products could ameliorate lactose intolerance. Meanwhile X-gal and ONPG methods which are simple, rapid and cheap can be used instead of SDS-PAGE.Keywords: Lactobacillus, Beta-Galactosidase, Nitrophenylgalactosids

Purification of β-acetylglucosaminase and β-galactosidase from ram testis

Science.gov (United States)

Caygill, J. C.; Roston, Christine P. J.; Jevons, F. R.

1966-01-01

1. The presence of β-galactosidase (EC 3.2.1.23) in an acetic acid extract of ram testis is reported. Some properties of the crude enzyme preparation were studied. 2. The purification of β-acetylglucosaminase (EC 3.2.1.30) and of β-galactosidase from the ram-testis extract by ammonium sulphate precipitation and chromatography on a CM-cellulose column is described. 3. The final purifications of the separated enzymes achieved were for the β-acetylglucosaminase 35 times and for the β-galactosidase 99 times. 4. The possibility of using DEAE-cellulose and Sephadex G-200 to purify the enzymes was investigated. PMID:5949569

Non-fusion and fusion expression of beta-galactosidase from Lactobacillus bulgaricus in Lactococcus lactis.

Science.gov (United States)

Wang, Chuan; Zhang, Chao-Wu; Liu, Heng-Chuan; Yu, Qian; Pei, Xiao-Fang

2008-10-01

To construct four recombinant Lactococcus lactis strains exhibiting high beta-galactosidase activity in fusion or non-fusion ways, and to study the influence factors for their protein expression and secretion. The gene fragments encoding beta-galactosidase from two strains of Lactobacillus bulgaricus, wch9901 isolated from yogurt and 1.1480 purchased from the Chinese Academy of Sciences, were amplified and inserted into lactococcal expression vector pMG36e. For fusion expression, the open reading frame of the beta-galactosidase gene was amplified, while for non-fusion expression, the open reading frame of the beta-galactosidase gene was amplified with its native Shine-Dalgarno sequence upstream. The start codon of the beta-galactosidase gene partially overlapped with the stop codon of vector origin open reading frame. Then, the recombinant plasmids were transformed into Escherichia coli DH5 alpha and Lactococcus lactis subsp. lactis MG1363 and confirmed by determining beta-galactosidase activities. The non-fusion expression plasmids showed a significantly higher beta-galactosidase activity in transformed strains than the fusion expression plasmids. The highest enzyme activity was observed in Lactococcus lactis transformed with the non-fusion expression plasmids which were inserted into the beta-galactosidase gene from Lactobacillus bulgaricus wch9901. The beta-galactosidase activity was 2.75 times as high as that of the native counterpart. In addition, beta-galactosidase expressed by recombinant plasmids in Lactococcus lactis could be secreted into the culture medium. The highest secretion rate (27.1%) was observed when the culture medium contained 20 g/L of lactose. Different properties of the native bacteria may have some effects on the protein expression of recombinant plasmids. Non-fusion expression shows a higher enzyme activity in host bacteria. There may be a host-related weak secretion signal peptide gene within the structure gene of Lb. bulgaricus beta-galactosidase

Thermomyces lanuginosus CBS 395.62/b Strain as Rich Source of α-Galactosidase Enzyme

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Quang D. Nguyen

2003-01-01

Full Text Available Seventeen Thermomyces lanuginosus strains, cultivated on raffinose and sucrose, were ranked on the basis of α-galactosidase activities. T. lanuginosus CBS 395.62/b strain showed the highest α-galactosidase activity on both investigated carbohydrates. Several carbon sources were tested as potential inducers for the α-galactosidase synthesis. On melibiose substrate α-galactosidase activity was higher in the intracellular fraction than in the filtrate of the fermentation broth, although both values were very low and did not reach the value of 1 U/mL. Raffinose, sucrose and Lactosucrose® proved to be inducers for α-galactosidase production. The highest titer (about 30 U/mL was achieved on 1 % sucrose and 0.45 % ammonium acetate. The optimum sucrose and ammonium acetate concentrations, at which about 90 U/mL α-galactosidase activity was reached during an 8-day fermentation, were 3 and 0.9 %, respectively.

O ENSINO DE ENZIMAS: UMA ABORDAGEM EXPERIMENTAL DE BAIXO CUSTO

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William de Azevedo Pinheiro

2011-07-01

Full Text Available Este trabalho descreve uma atividade prática de Bioquímica, a qual expõe alternativasdidáticas que visam facilitar processo ensino/aprendizado dos conceitos associados ao temaEnzimas. O estudo foi realizado em escolas de Ensino Médio da rede pública Estadual naregião Noroeste Fluminense do Estado do Rio de Janeiro. O trabalho consiste nodesenvolvimento de uma atividade prática, sobre Enzimas, utilizando materiais alternativose/ou de baixo custo. O trabalho objetiva, ainda, ser um agente de incentivo ao trabalho emequipes interdisciplinares dentro de uma visão holística multidisciplinar da construção doconhecimento, possível de ser explorada nas Ciências Biológicas e Exatas.

Potential applicaton of β-galactosidase in food science and nutrition

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Nika ŽIBRAT

2017-10-01

Full Text Available β-galactosidase is an enzyme with hydrolytic and transgalactosylation activity. The origin of the enzyme dictates the balance between both activities. Industrially used β-galactosidases are obtained with recombinant production from filamentus funghi Aspergillus sp. and yeasts Kluyveromyces sp. Recently thermostabile β-galactosidases have been subject of many research. The enzyme can be industrially used in free or immobilized form. Immobilization often provides better stability, reusability and lower expenses. Application of β-galactosidase is most common in food processing and nutrition, it is also used in medicine and ecology. Hydrolytic activity of the enzyme has long been used for reducing lactose content in milk, while transgalactosylitic activity is used for synthesis of products such as galactooligosaccharides, lactosucrose and others. The latter have a great potential in food industry for obtaining products with reduced lactose content and increasing of nutritional value by adding dietetic fibers such as galactooligosaccharides. Despite the potential it is vital that reaction mechanisms become better understood and optimization is in place in order to reach the usability of this enzyme at industrial level.

Lactose Hydrolysis in Milk and Dairy Whey Using Microbial β-Galactosidases

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Michele Dutra Rosolen

2015-01-01

Full Text Available This work aimed at evaluating the influence of enzyme concentration, temperature, and reaction time in the lactose hydrolysis process in milk, cheese whey, and whey permeate, using two commercial β-galactosidases of microbial origins. We used Aspergillus oryzae (at temperatures of 10 and 55°C and Kluyveromyces lactis (at temperatures of 10 and 37°C β-galactosidases, both in 3, 6, and 9 U/mL concentrations. In the temperature of 10°C, the K. lactis β-galactosidase enzyme is more efficient in the milk, cheese whey, and whey permeate lactose hydrolysis when compared to A. oryzae. However, in the enzyme reaction time and concentration conditions evaluated, 100% lactose hydrolysis was not reached using the K. lactis β-galactosidase. The total lactose hydrolysis in whey and permeate was obtained with the A. oryzae enzyme, when using its optimum temperature (55°C, at the end of a 12 h reaction, regardless of the enzyme concentration used. For the lactose present in milk, this result occurred in the concentrations of 6 and 9 U/mL, with the same time and temperature conditions. The studied parameters in the lactose enzymatic hydrolysis are critical for enabling the application of β-galactosidases in the food industry.

Strategicheskiy analiz predpriyatiya ZAO Alfa Motor Grupp

OpenAIRE

Antonova, Maria

2016-01-01

The subject of the bachelor work is the company ZAO Alfa Motor Group. The aim is to conduct strategic analysis of economic activity, evaluation of the effectiveness of current policies and develop recommendations and proposals to change the company's development strategy. ZAO Alfa Motor Group - a network of specialized service centers and car sales of spare parts, specialized in cars of Toyota and Lexus. Bachelor work has two main parts. In the first part of the work to uncover the meaning of...

Analysis of β-galactosidase production and their genes of two strains of Lactobacillus bulgaricus.

Science.gov (United States)

Zhang, Wen; Wang, Chuan; Huang, Cheng-Yu; Yu, Qian; Liu, Heng-Chuan; Zhang, Chao-Wu; Pei, Xiao-Fang; Xu, Xin; Wang, Guo-Qing

2012-06-01

A bacterial β-galactosidase delivery system is a potential therapy for lactose intolerance. Currently, two Lactobacillus bulgaricus strains with different biological characteristics are under consideration as potential sources. However, differences in these β-galactosidase genes and their resulting production levels are poorly characterized. The β-galactosidase ORF of L. bulgaricus yogurt isolate had high variability and was terminated at site 1924 due to a stop codon. However, the full 114 kDa β-galactosidase band was still resolved by SDS-PAGE, which may indicate that the interrupted ORF was translated into more than one peptide, and they together were folded into the complete enzyme protein that showed much higher β-galactosidase activity (6.2 U/mg protein) than the enzyme generated from L. bulgaricus reference strain (2.5 U/mg protein).

Hooked on α-d-galactosidases: from biomedicine to enzymatic synthesis.

Science.gov (United States)

Bakunina, Irina Yu; Balabanova, Larissa A; Pennacchio, Angela; Trincone, Antonio

2016-01-01

α-d-Galactosidases (EC 3.2.1.22) are enzymes employed in a number of useful bio-based applications. We have depicted a comprehensive general survey of α-d-galactosidases from different origin with special emphasis on marine example(s). The structures of natural α-galactosyl containing compounds are described. In addition to 3D structures and mechanisms of action of α-d-galactosidases, different sources, natural function and genetic regulation are also covered. Finally, hydrolytic and synthetic exploitations as free or immobilized biocatalysts are reviewed. Interest in the synthetic aspects during the next years is anticipated for access to important small molecules by green technology with an emphasis on alternative selectivity of this class of enzymes from different sources.

Interferon alfa with or without ribavirin for chronic hepatitis C

DEFF Research Database (Denmark)

Kjaergard, L L; Krogsgaard, K; Gluud, C

2001-01-01

To assess the efficacy and safety of interferon alfa with or without ribavirin for treatment of chronic hepatitis C.......To assess the efficacy and safety of interferon alfa with or without ribavirin for treatment of chronic hepatitis C....

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.

Adoption and de-adoption of drotrecogin alfa for severe sepsis in the United States.

Science.gov (United States)

Kahn, Jeremy M; Le, Tri Q

2016-04-01

Drotrecogin alfa was a landmark drug for treatment of severe sepsis, yet little is known about how it was adopted and de-adopted during its 10-year period of availability. We used hospitalization data on fee-for-service Medicare beneficiaries from 2002 to 2011 to characterize trends in the use of drotrecogin alfa in the United States. Drotrecogin alfa use peaked at 5.87 per 1000 severe sepsis hospitalizations in 2003 and then steadily declined to 0.94 administrations per 1000 severe sepsis hospitalizations in 2010. Large teaching hospitals were more likely to use drotrecogin alfa than small, nonteaching hospitals. The addition of "add-on payments" to hospitals for using drotrecogin alfa in 2002 was associated with significantly increased use (P publication of international sepsis guidelines with favorable drotrecogin alfa recommendations (in 2004 and 2008) nor the publication of a clinical trial focused on drotrecogin alfa (in 2005) were associated with consistent changes use (P > .05). Drotrecogin alfa use declined over time, with marked changes in use associated with drug-specific financial incentives but not the publication of clinical practice guidelines or clinical trials. Copyright © 2015 Elsevier Inc. All rights reserved.

Interferon alfa and ribavirin induced hair changes

International Nuclear Information System (INIS)

Amir, S.; Taj, A.; Muhamud, T.H.; Iqbal, Z.; Yaqub, F.

2007-01-01

Combination therapy of Interferon alfa and ribavirin in chronic hepatitis C has well documented cutaneous adverse effects. Most interesting of these has been reported on hair physiology. This study was conducted to determine the frequency and pattern of adverse effects involving hair in patients receiving combination of interferon alfa 2a and ribavirin for chronic hepatitis C. The study was conducted in Department of Dermatology, Division of Medicine Shaikh Zayed Hospital. Thirty Eight patients who completed treatment with interferon alfa (3 MIU subcutaneously thrice weekly) and 1200 mg ribavirin daily for 24 weeks were enrolled in this single-center study. The patient's response and examination finding particularly regarding involvement of hair was noted on a Proforma. Thirty Two out of thirty eight (84%) patients noted adverse effects involving hair. The most frequent was diffuse hair loss and occurred in 27 patients (71%). Hypertrichosis of eyelashes (trichomegaly) and eyebrows (synophyrs) was observed in 18 (47%) and 16 (42%) patients respectively. Graying of hair was noted in 4 patients (11%), while discoloration of moustache hair was seen in 2 patients (5%). Epilation at the site of subcutaneous injection was noted in 10 patients (26%). Alopecia areata was reported in 2 patients (5%). It is concluded that adverse effects involving hair are frequent and varied (hair loss to excess hair growth) during combination therapy with Interferon alfa-2a and Ribavirin for chronic hepatitis C. (author)

21 CFR 173.145 - Alpha-Galactosidase derived from Mortierella vinaceae var. raffinoseutilizer.

Science.gov (United States)

2010-04-01

... vinaceae var. raffinoseutilizer. 173.145 Section 173.145 Food and Drugs FOOD AND DRUG ADMINISTRATION... Alpha-Galactosidase derived from Mortierella vinaceae var. raffinoseutilizer. The food additive alpha-galactosidase and parent mycelial microorganism Mortierella vinaceae var. raffinoseutilizer may be safely used...

Determinação de proteína total e escore de aminoácidos de bromelinas extraídas dos resíduos do abacaxizeiro (Ananas comosus, (L. Merril

Directory of Open Access Journals (Sweden)

FREIMAN Lenice O.

1999-01-01

Full Text Available O abacaxi por longo tempo tem sido a fruta não cítrica mais popular nos países tropicais e sub-tropicais, principalmente pelo seu atrativo sabor e aroma, além do refrescante balanço açúcar-ácido que possui, o que certamente, determinou sua importância sócio-econômica. Além dessas características, é fonte da enzima proteolítica bromelina, um produto de alto valor comercial, que não é fabricado no Brasil. O presente trabalho objetivou a determinação do escore de aminoácidos de bromelinas extraídas das várias partes do abacaxizeiro (casca, folha, caule, coroa e polpa, com duas precipitações subseqüentes, utilizando-se como agente precipitante, o etanol p.a. Análises mostraram maior teor de proteínas na bromelina extraída da coroa. Nas duas precipitações a composição de aminoácidos das enzimas obtidas, apresentaram resultados inferiores aos da comercial.

Potential Applications of Immobilized β-Galactosidase in Food Processing Industries

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Parmjit S. Panesar

2010-01-01

Full Text Available The enzyme β-galactosidase can be obtained from a wide variety of sources such as microorganisms, plants, and animals. The use of β-galactosidase for the hydrolysis of lactose in milk and whey is one of the promising enzymatic applications in food and dairy processing industries. The enzyme can be used in either soluble or immobilized forms but the soluble enzyme can be used only for batch processes and the immobilized form has the advantage of being used in batch wise as well as in continuous operation. Immobilization has been found to be convenient method to make enzyme thermostable and to prevent the loss of enzyme activity. This review has been focused on the different types of techniques used for the immobilization of β-galactosidase and its potential applications in food industry.

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...

Structure of .beta.-galactosidase complexes

Czech Academy of Sciences Publication Activity Database

Dohnálek, Jan; Skálová, Tereza; Dušková, Jarmila; Petroková, Hana; Hašek, Jindřich; Lipovová, P.; Spiwok, V.; Strnad, Hynek; Králová, B.

2006-01-01

Roč. 13, č. 3 (2006), s. 137-138 ISSN 1211-5894. [Czech and Slovak Crystallographic Colloquium. 22.06.2006-24.06.2006, Grenoble] R&D Projects: GA AV ČR KJB500500512 Keywords : .beta.-galactosidase * X-ray diffraction * cold-active enzyme Subject RIV: EB - Genetics ; Molecular Biology http://www. xray .cz/ms/default.htm

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

Managing anemia with epoetin alfa in patients with rectal cancer

International Nuclear Information System (INIS)

Velenik, V.; Oblak, I.; Kodre, V.

2005-01-01

Background. Anemia is one of the most challenging problems in clinical oncology due to its high prevalence among the patients with malignant diseases. The purposes of our study were: (1) to assess the potential of epoetin alfa therapy to prevent the decline in Hb concentrations that typically accompanies chemotherapy/ radiotherapy (ChT/RT) of the patients with rectal cancer; (2) to test the hypothesis that the use of epoetin alfa significantly reduces the transfusion requirements in the patients with rectal cancer treated with ChT/RT after surgery, and (3) to evaluate the safety profile of the administration of epoetin alfa in the clinical setting. Methods. Sixty patients who underwent surgery for rectal cancer were prospectively enrolled. Group A consisted of 39 patients with Hb concentrations ≤13 g/dl at the start of ChT/RT following surgery, and group B of 17 patients with Hb concentrations ≥13 g/dl at the start of ChT/RT following surgery, but whose Hb concentrations fell below 13 g/dl during the ChT/RT protocol. The starting dose of epoetin alfa in both groups was 10,000 IU subcutaneously (sc) three times a week (tiw). The following major parameters were evaluated: (1) change in Hb concentrations relative to the baseline as measured at 4-week intervals, (2) allogenic blood transfusion requirements in relation to Hb concentrations, and (3) incidence and severity of adverse events and their potential relationship to epoetin alfa administration. Results. The study protocol was completed in 56/60 patients. In group A, a statistically significant increase in Hb concentration (p<0.001) was observed after the first 4 weeks of epoetin alfa treatment compared to the baseline values, with the mean increase of Hb concentration of 1.97 g/dl ± 0.91 g/dl and Hb concentrations remained significantly increased through the whole study (p=0.0017). In group B, a continuous decrease in Hb concentrations was observed during the first weeks of therapy, reaching the level of

Darbepoetin alfa once every 2 weeks for treatment of anemia in dialysis patients

DEFF Research Database (Denmark)

Mann, J; Kessler, M; Villa, Giulio Palludan

2007-01-01

AIM: Darbepoetin alfa has a longer half-life than epoetin-(EPO) alfa or beta, allowing administration at less frequent intervals for the treatment of renal anemia. The aim of the present analysis was to evaluate the efficacy and tolerability of an every-2-week (Q2W) schedule of darbepoetin alfa...

Purification of beta-galactosidase from Aspergillus Niger and studies of its kinetics

International Nuclear Information System (INIS)

Ali, M.A.A.

1989-01-01

The present study was carried out to extract purity and characterize B-galactosidase which was produced by a local strain of the fungus A. Niger. The obtained results were as follows: 1. The extraction of B-galactosidase by the preparation of acetone powder followed by extraction with 0.1M sodium acetate-2m M PMSC buffer, PH 5.0. 2. B-galactosidase was purified by a combination of techniques. The enzyme was purified 113.6 folds with 27% recovery. 3. The isoelectric point of the purified enzyme was 4.6. 7 tabs.; 37 figs.; 67 refs

Novel high-performance metagenome β-galactosidases for lactose hydrolysis in the dairy industry.

Science.gov (United States)

Erich, Sarah; Kuschel, Beatrice; Schwarz, Thilo; Ewert, Jacob; Böhmer, Nico; Niehaus, Frank; Eck, Jürgen; Lutz-Wahl, Sabine; Stressler, Timo; Fischer, Lutz

2015-09-20

The industrially utilised β-galactosidases from Kluyveromyces spp. and Aspergillus spp. feature undesirable kinetic properties in praxis, such as an unsatisfactory lactose affinity (KM) and product inhibition (KI) by galactose. In this study, a metagenome library of about 1.3 million clones was investigated with a three-step activity-based screening strategy in order to find new β-galactosidases with more favourable kinetic properties. Six novel metagenome β-galactosidases (M1-M6) were found with an improved lactose hydrolysis performance in original milk when directly compared to the commercial β-galactosidase from Kluyveromyces lactis (GODO-YNL2). The best metagenome candidate, called "M1", was recombinantly produced in Escherichia coli BL21(DE3) in a bioreactor (volume 35 L), resulting in a total β-galactosidase M1 activity of about 1100 μkatoNPGal,37 °C L(-1). Since milk is a sensitive and complex medium, it has to be processed at 5-10 °C in the dairy industry. Therefore, the β-galactosidase M1 was tested at 8 °C in milk and possessed a good stability (t1/2=21.8 d), a desirably low apparent KM,lactose,8 °C value of 3.8±0.7 mM and a high apparent KI,galactose,8 °C value of 196.6±55.5 mM. A lactose hydrolysis process (milk, 40 nkatlactose mLmilk,8 °C(-1)) was conducted at a scale of 0.5L to compare the performance of M1 with the commercial β-galactosidase from K. lactis (GODO-YNL2). Lactose was completely (>99.99%) hydrolysed by M1 and to 99.6% (w/v) by K. lactis β-galactosidase after 25 h process time. Thus, M1 was able to achieve the limit of lactose per litre milk, which is recommended for dairy products labelled as "lactose-free". Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.

Upgrade of ATLAS-ALFA for Run2

CERN Document Server

Jakobsen, Sune; The ATLAS collaboration

2016-01-01

Early experience with the operation of the ALFA detectors in the LHC environment has shown significant beam-induced heating. Comprehensive studies on the effects of heating with a spare detector module have furthermore revealed that heating effects could be disastrous in the case of the larger beam intensities foreseen for higher luminosity. A temporary solution was implemented and kept the detectors below the critical temperature in 2012. In LS1 all ALFA stations have been removed from LHC and are undergoing an upgrade of the geometry of Roman Pots to minimize the impedance losses. When the stations were re-installed ~4 m distance was added between them to gain level arm in the angular measurement between the stations. The updated geometry of the Roman Pot, together with a system to improve the internal heat transfer and an air cooling system, is expected to keep the temperatures of ALFA detectors below critical limits throughout the LHC Run2 and first results from the performance in 2016 will be presented. ...

Uso tópico do inteferon alfa-2b para neoplasia escamosa da superficie ocular em um caso de paciente com xeroderma pigmentoso

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Nayara Queiroz Cardoso Pinto

Full Text Available Resumo Relato de um caso clínico de Xeroderma Pigmentoso com carcinoma espinocelular de conjuntiva bilateral que apresentou regressão importante das dimensões tumorais com o uso de Interferon alfa-2b tópico. Relato de caso: Paciente feminina com Xeroderma Pigmentoso em estágio avançado, com ausência de pele sadia, tendo sido submetida a cerca de 60 exéreses de lesões de pele malignas. A paciente compareceu com tumoração conjuntival em ambos os olhos, correspondendo a carcinoma espinocelular de conjuntiva e neoplasia intraepitelial de conjuntiva em olho esquerdo. Devido as dificuldades cirúrgicas, alta taxa de recidiva e elevada probabilidade de formação de simbléfaro foi-se iniciado terapêutica com Interferon alfa-2beta 1.000.000 unidades tópico, obtendo-se bons resultados com importante regressão do tamanho da lesão e resolução dos sintomas. Conclusão: O uso tópico de interferon alfa-2beta em neoplasia escamosa de conjuntiva, mostrou-se uma boa opção terapêutica em situações de elevado risco cirúrgico e de complicações pós operatórias.

Avaliação do perfil hematológico de portadores de talassemia alfa provenientes das regiões Sudeste e Nordeste do Brasil Evaluation of alpha thalassemia carriers hematologic profile from Southeast and Northeast Brazilian regions

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Gislane L. V. Oliveira

2006-06-01

Full Text Available A talassemia alfa é uma anemia hereditária resultante da síntese deficiente de cadeias alfa, provocando um excesso relativo de cadeias beta, que vão formar tetrâmeros identificados como hemoglobina H (Hb H no indivíduo adulto. Para direcionar o diagnóstico laboratorial desta anemia, a análise dos índices eritrocitários, a eletroforese em acetato de celulose em pH neutro e a pesquisa de corpos de inclusão de Hb H são essenciais. O objetivo deste estudo foi traçar o perfil hematológico, por meio dos índices eritrocitários, dos portadores de talassemia alfa das regiões Sudeste e Nordeste do Brasil. Foram analisadas 1.010 amostras de sangue periférico após consentimento informado. Os índices eritrocitários como contagem de glóbulos vermelhos (RBC, dosagem de hemoglobina (HGB, hematócrito (HCT, volume corpuscular médio (VCM, hemoglobina corpuscular média (HCM e concentração de hemoglobina corpuscular média (CHCM foram fornecidos por aparelhos automatizados com controle de qualidade interno e externo. Para o diagnóstico de talassemia alfa foram utilizados testes de triagem e complementares para talassemias, como eletroforese em pH neutro e pesquisa de corpos de inclusão de Hb H com coloração de azul cresil brilhante. Comparando-se os valores hematológicos observados nos dois grupos, notou-se que, em ambas as regiões, os índices com valores discrepantes foram os níveis de HGB e HCT, sendo a maior freqüência de variação observada entre as mulheres. Nos portadores do fenótipo alfa talassêmico da região Nordeste, todos os índices eritrocitários estavam abaixo dos valores de normalidade. Estes resultados evidenciam a necessidade de melhor avaliação do perfil hematológico de talassemia alfa em diferentes regiões, considerando-se os interferentes ambientais para um diagnóstico mais preciso.Alpha thalassemia is a hereditary anaemia resulting from deficient synthesis of the alpha chains, causing a relative

Host-Pathogen Interactions: I. A Correlation Between α-Galactosidase Production and Virulence 1

Science.gov (United States)

English, Patricia D.; Albersheim, Peter

1969-01-01

Resistance or susceptibility of Red Kidney, Pinto and Small White beans (Phaseolus vulgaris) to the alpha, beta, and gamma strains of Colletotrichum lindemuthianum was either confirmed or established. These fungal strains secrete α-galactosidase, β-galactosidase and β-xylosidase when grown on cell walls isolated from the hypocotyls of any of the above bean varieties. These enzymes effectively degrade cell walls isolated from susceptible 5-day old hypocotyls but degrade only slightly the walls isolated from resistant 18-day old hypocotyls. The amounts of the β-galactosidase and β-xylosidase secreted by the 3 fungal strains are relatively low and are approximately equivalent. The secretion of these 2 enzymes is not dependent upon the bean variety from which the hypocotyl cell walls used as a carbon source were isolated. However, the fungal strains secrete greater amounts of α-galactosidase when grown on hypocotyl cell walls isolated from susceptible plants than when grown on walls from resistant plants. Virulent isolates of the fungus, when grown on hypocotyl cell walls isolated from a susceptible plant, secrete more α-galactosidase than do attenuated (avirulent) isolates of the same fungal strain grown under the same conditions. The α-galactosidase secreted by each of the fungal strains is capable of removing galactose from the hypocotyl cell walls of each bean variety tested. Galactose is removed from the cell walls of each variety at the same rate regardless of whether the cell walls were isolated from a susceptible or resistant plant. PMID:16657049

A trial of darbepoetin alfa in type 2 diabetes and chronic kidney disease

DEFF Research Database (Denmark)

Pfeffer, Marc A; Burdmann, Emmanuel A; Chen, Chao-Yin

2009-01-01

BACKGROUND: Anemia is associated with an increased risk of cardiovascular and renal events among patients with type 2 diabetes and chronic kidney disease. Although darbepoetin alfa can effectively increase hemoglobin levels, its effect on clinical outcomes in these patients has not been adequately...... tested. METHODS: In this study involving 4038 patients with diabetes, chronic kidney disease, and anemia, we randomly assigned 2012 patients to darbepoetin alfa to achieve a hemoglobin level of approximately 13 g per deciliter and 2026 patients to placebo, with rescue darbepoetin alfa when the hemoglobin...... assigned to darbepoetin alfa and 496 patients assigned to placebo (Pchronic kidney disease...

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.

Secretion and properties of a hybrid Kluyveromyces lactis-Aspergillus niger β-galactosidase

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Becerra Manuel

2006-12-01

Full Text Available Abstract Background The β-galactosidase from Kluyveromyces lactis is a protein of outstanding biotechnological interest in the food industry and milk whey reutilization. However, due to its intracellular nature, its industrial production is limited by the high cost associated to extraction and downstream processing. The yeast-system is an attractive method for producing many heterologous proteins. The addition of a secretory signal in the recombinant protein is the method of choice to sort it out of the cell, although biotechnological success is not guaranteed. The cell wall acting as a molecular sieve to large molecules, culture conditions and structural determinants present in the protein, all have a decisive role in the overall process. Protein engineering, combining domains of related proteins, is an alternative to take into account when the task is difficult. In this work, we have constructed and analyzed two hybrid proteins from the β-galactosidase of K. lactis, intracellular, and its Aspergillus niger homologue that is extracellular. In both, a heterologous signal peptide for secretion was also included at the N-terminus of the recombinant proteins. One of the hybrid proteins obtained has interesting properties for its biotechnological utilization. Results The highest levels of intracellular and extracellular β-galactosidase were obtained when the segment corresponding to the five domain of K. lactis β-galactosidase was replaced by the corresponding five domain of the A. niger β-galactosidase. Taking into account that this replacement may affect other parameters related to the activity or the stability of the hybrid protein, a thoroughly study was performed. Both pH (6.5 and temperature (40°C for optimum activity differ from values obtained with the native proteins. The stability was higher than the corresponding to the β-galactosidase of K. lactis and, unlike this, the activity of the hybrid protein was increased by the presence

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.

Optimization of drying process of Zea Mays malt to use as alternative source of amylolytics enzymes

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Joana Paula Menezes Biazus

2005-06-01

Full Text Available This work aimed to study the drying process optimization of maize (Zea Mays malt for obtaining maize malt, without affecting enzymatic activity of alpha e beta-amylases from maize malt. Results showed that dryer operation must occur in zone at 54°C and 5.18-6 h process time. The maize malt obtained had good enzymatic properties.Este trabalho objetivou a otimização da secagem do malte de milho (Zea Mays para obter um malte sem afetar a atividade das enzimas presentes neste, alfa e beta -amilases. Os resultados mostraram que a operação do secador deve ser feita a 54°C e entre 5,18-6 h de processo. O malte obtido possuiu boas propriedades enzimáticas.

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.

Kinetic Characterization of Galacto-Oligosaccharide (GOS) Synthesis by Three Commercially Important b-Galactosidases

NARCIS (Netherlands)

Warmerdam, A.; Zisopoulos, F.K.; Boom, R.M.; Janssen, A.E.M.

2014-01-01

Many b-galactosidases show large differences in galacto-oligosaccharide (GOS) production and lactose hydrolysis. In this study, a kinetic model is developed in which the effect of lactose, glucose, galactose, and oligosaccharides on the oNPG converting activity of various b-galactosidases is

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.

Role of Glutaraldehyde in Imparting Stability to Immobilized β-Galactosidase Systems

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Rukhsana Satar

2018-01-01

Full Text Available ABSTRACT This review article highlights the role of glutaraldehyde as a matrix activator/stabilizer in imparting higher operational and thermal stability to β-galactosidase (βG for biotechnological applications. Glutaraldehyde has been used extensively as a crosslinking agent as well as for functionalization of matrices to immobilize β-galactosidase. Immobilized β-galactosidase systems (IβGS obtained as a result of glutaraldehyde treatment has been employed to hydrolyze whey and milk lactose in batch reactors, continuous packed-bed and fluidized bed reactors under various operational conditions. Moreover, these IβGS have also been utilized for the production of galactooligosaccharides in food, dairy and fermentation industries. It was observed that glutaraldehyde provided remarkable stability to immobilize βG against various physical/chemical denaturants, thus enhancing thermal/operational stability and rendering it more suitable for repeated utilization in industrial scale operations.

The GlaA signal peptide substantially increases the expression and secretion of α-galactosidase in Aspergillus niger.

Science.gov (United States)

Xu, Yue; Wang, Yan-Hui; Liu, Tian-Qi; Zhang, Hui; Zhang, He; Li, Jie

2018-03-31

α-Galactosidases are widely used in many fields. It is necessary to improve the production of enzymes through microbiological processes. The aim of this study was to construct recombinant Aspergillus niger strains with high α-galactosidase production. Two recombinant A. niger strains were constructed: AB and AGB. The recombinant AB strain contained the α-galactosidase aglB gene from A. niger with its native AglB signal peptide regulated by the glucoamylase promoter. In the AGB recombinant strain, the AglB signal peptide was replaced with the glucoamylase (GlaA) signal peptide. The extracellular maximum α-galactosidase activity of the AGB strain was 215.7 U/ml and that of the AB strain was 9.8 U/mL. The optimal conditions for α-galactosidase were pH 3.5 and 35 °C. The GlaA signal peptide substantially increased the yield of secreted α-galactosidase in A. niger. This recombinant strain holds great potential for industrial applications.

O papel do Fator de Necrose Tumoral Alfa (TNF-alfa no processo de erosão óssea presente no colesteatoma adquirido da orelha média The role of Tumor Necrosis Factor -Alpha (TNF- alpha in bone resorption present in middle ear cholesteatoma

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Rodrigo Faller Vitale

2007-02-01

Full Text Available O colesteatoma adquirido da orelha média causa erosão óssea, com altas taxas de morbidade e mortalidade. O TNF-alfa (TNF-alfa lambda uma das principais citocinas envolvidas neste processo. OBJETIVO: Avaliar o papel do TNF-alfa na reabsorsão óssea e a ação dele no colesteatoma. MATERIAL E MÉTODOS: Foi realizado um levantamento e uma revisão crítica da literatura. RESULTADOS: Todos os autores estudados concordam com a importância do TNF-alfa no processo de reabsorção óssea presente no colesteatoma e com o grau de destruição observado. Diferentes trabalhos demonstraram que o TNF-alfa é capaz de provocar erosão óssea, através de diferentes vias de ação. Ele pode estimular a diferenciação e a maturação dos osteoclastos ou, ainda, agir na matriz óssea expondo-a à ação dos osteoclastos. Existe a possibilidade de inibir a ação do TNF-alfa, diminuindo seus efeitos e prevenindo a perda óssea em doenças como a artrite reumatóide. Não existe, entretanto, trabalhos específicos em colesteatoma. Não existe consenso sobre a sua localização. Estas diferenças, provavelmente, ocorrem devido à distribuição dos receptores. CONCLUSÃO: O TNF-alfa, presente no colesteatoma promove a reabsorsão óssea, juntamente com outras citocinas (RANKL e IL-1, estando relacionado com a presença de complicações.Cholesteatoma may cause bone erosion, with high morbidity and mortality rates. Tumor Necrosis Factor -Alpha (TNF-a is one of the main cytokines involved in this process. Our goal was to evaluate the role of TNF-a in Bone Resorption and its effect on cholesteatoma. MATERIAL AND METHODS: analysis and critical literature review. RESULTS: Different studies have demonstrated that TNF-a is capable of causing bone erosion. It may stimulate the differentiation and maturation of osteoclasts or it may act on the bone matrix, exposing it to the action of the osteoclasts. It is possible to inhibit TNF-a, reducing its effects and prevent

Cloning, purification and characterization of a thermostable β-galactosidase from Bacillus licheniformis strain KG9.

Science.gov (United States)

Matpan Bekler, F; Stougaard, P; Güven, K; Gül Güven, R; Acer, Ö

2015-06-28

A thermo— and alkalitolerant Bacillus licheniformis KG9 isolated from Taşlıdere hot water spring in Batman/Turkey was found to produce a thermostable β—galactosidase. Phylogenetic analysis showed that the 16S rRNA gene from B. licheniformis strain KG9 was 99.9% identical to that of the genome sequenced B. licheniformis strain DSM 13. Analysis of the B. licheniformis DSM 13 genomic sequence revealed four putative β—galactosidase genes. PCR primers based on the genome sequence of strain DSM 13 were used to isolate the corresponding β—galactosidase genes from B. licheniformis strain KG9. The calculated molecular weights of the β—galactosidases I, II, III, and IV using sequencing data were 30, 79, 74, and 79 kDa, respectively. The genes were inserted into an expression vector and recombinant β—galactosidase was produced in Escherichia coli. Of the four β—galactosidase genes identified in strain KG9, three of them were expressed as active, intracellular enzymes in E. coli. One of the recombinant enzymes, β—galactosidase III, was purified and characterized. Optimal temperature and pH was determined to be at 60 ºC and pH 6.0, respectively. Km was determined to be 1.3 mM and 13.3 mM with oNPG (ortho—nitrophenyl—β—D—galactopyranoside) and lactose as substrates, respectively, and Vmax was measured to 1.96 μmol/min and 1.55 μmol/min with oNPG and lactose, respectively.

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.

Synthesis of galacto-oligosaccharides with ß-galactosidases

NARCIS (Netherlands)

Warmerdam, A.

2013-01-01

Galacto-oligosaccharides (GOS) are generally enzymatically synthesized with β-galactosidases. GOS are of interest because of their prebiotic effects on human health. They are mainly applied in infant nutrition, because of their resemblance to human milk oligosaccharides, but they are also

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

a-galactosidase: general aspects and its application on soybean oligosaccharides/ a-galactosidase: aspectos gerais e sua aplicação em produtos a base de soja

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Mara Lucia Luiz Ribeiro

2002-05-01

Full Text Available This literature review has the aim to present topics related to a-galactosidase, considering its general characteristics, sources, functions and methods used in its determination. This enzyme, lacking in the human digestive tract, hydrolises µ-(1,6 bonds from oligosaccharides of the raffinose family. These carbohydrates, called flatulents, are found in leguminoses like soybean, a rich source of protein. However, due to the presence of these flatulents sugars, antinutritional factors are attributed to soyben, decreasing its consumption. a-galactosidase produced by some microorganisms and vegetables can be used in soybean based foods to reduce the interaction of the intestinal flora with the oligosaccharides, decreasing in this way the flatulence.This literature review has the aim to present topics related to a-galactosidase, considering its general characteristics, sources, functions and methods used in its determination. This enzyme, lacking in the human digestive tract, hydrolises µ-(1,6 bonds from oligosaccharides of the raffinose family. These carbohydrates, called flatulents, are found in leguminoses like soybean, a rich source of protein. However, due to the presence of these flatulents sugars, antinutritional factors are attributed to soyben, decreasing its consumption. a-galactosidase produced by some microorganisms and vegetables can be used in soybean based foods to reduce the interaction of the intestinal flora with the oligosaccharides, decreasing in this way the flatulence.

Adsorption of β-galactosidase on silica and aluminosilicate adsorbents

Science.gov (United States)

Atyaksheva, L. F.; Dobryakova, I. V.; Pilipenko, O. S.

2015-03-01

It is shown that adsorption of β-galactosidase of Aspergillus oryzae fungi on mesoporous and biporous silica and aluminosilicate adsorbents and the rate of the process grow along with the diameter of the pores of the adsorbent. It is found that the shape of the adsorption isotherms changes as well, depending on the texture of the adsorbent: the Michaelis constant rises from 0.3 mM for the enzyme in solution to 0.4-0.5 mM for the enzyme on a surface in the hydrolysis of o-nitrophenyl-β-D-galactopyranoside. It is concluded that β-galactosidase displays its maximum activity on the surface of biporous adsorbents.

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...

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

Active-site-directed inactivation of Aspergillus oryzae beta-galactosidase with beta-D-galactopyranosylmethyl-p-nitrophenyltriazene.

Science.gov (United States)

Mega, T; Nishijima, T; Ikenaka, T

1990-04-01

beta-D-Galactopyranosylmethyl-p-nitrophenyltriazene (beta-GalMNT), a specific inhibitor of beta-galactosidase, was isolated as crystals by HPLC and its chemical and physicochemical characteristics were examined. Aspergillus oryzae beta-galactosidase was inactivated by the compound. We studied the inhibition mechanism in detail. The inhibitor was hydrolyzed by the enzyme to p-nitroaniline and an active intermediate (beta-galactopyranosylmethyl carbonium or beta-galactopyranosylmethyldiazonium), which inactivated the enzyme. The efficiency of inactivation of the enzyme (the ratio of moles of inactivated enzyme to moles of beta-GalMNT hydrolyzed by the enzyme) was 3%; the efficiency of Escherichia coli beta-galactosidase was 49%. In spite of the low efficiency, the rate of inactivation of A. oryzae enzyme was not very different from that of the E. coli enzyme, because the former hydrolyzed beta-GalMNT faster than the latter did. A. oryzae beta-galactosidase was also inactivated by p-chlorophenyl, p-tolyl, and m-nitrophenyl derivatives of beta-galactopyranosylmethyltriazene. However, E. coli beta-galactosidase was not inactivated by these triazene derivatives. The results showed that the inactivation of A. oryzae and E. coli beta-galactosidases by beta-GalMNT was an enzyme-activated and active-site-directed irreversible inactivation. The possibility of inactivation by intermediates produced nonenzymatically was ruled out for E. coli, but not for the A. oryzae enzyme.

Biosensors based on β-galactosidase enzyme: Recent advances and perspectives.

Science.gov (United States)

Sharma, Shiv K; Leblanc, Roger M

2017-10-15

Many industries are striving for the development of more reliable and robust β-galactosidase biosensors that exhibit high response rate, increased detection limit and enriched useful lifetime. In a newfangled technological atmosphere, a trivial advantage or disadvantage of the developed biosensor may escort to the survival and extinction of the industry. Several alternative strategies to immobilize β-galactosidase enzyme for their utilization in biosensors have been developed in recent years in the quest of maximum utility by controlling the defects seen in the previous biosensors. The overwhelming call for on-line measurement of different sample constituents has directed science and industry to search for best practical solutions and biosensors are witnessed as the best prospect. The main objective of this paper is to serve as a narrow footbridge by comparing the literary works on the β-galactosidase biosensors, critically analyze their use in the construction of best biosensor by showing the pros and cons of the predicted methods for the practical use of biosensors. Copyright © 2017 Elsevier Inc. All rights reserved.

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

"Alfa" i "Võmpel" bez maski / Andrei Sharov

Index Scriptorium Estoniae

Sharov, Andrei

2004-01-01

Venemaa FSB keskuse, mille koosseisu kuuluvad ka eriüksused Alfa ja Võmpel, töötajad osalevad terroristide neutraliseerimise ja pantvangide vabastamise erioperatsioonides. Intervjuu keskuse staabi uue juhiga

The in vitro synthesis of {beta}-galactosidase induced in a subcellular structure of Escherichia coli (1961); Synthese in vitro de {beta}-galactosidase induite dans une structure subcellulaire d'Escherichia coli (1961)

Energy Technology Data Exchange (ETDEWEB)

Nisman, B; Kayser, A; Demailly, J; Genin, C [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires

1961-07-01

Isopropyl-thio-galactoside (IPTG), an inducer of 3-galactosidase, makes it possible to synthesise this enzyme in vitro with the subcellular structure (P{sub 1}). The enzyme is isolated from the bacteria Escherichia coli K 12 which are inductive but not induced. The incorporation of radioactive amino-acids, which is stimulated by the presence of an inducer, was studied during the course of the enzyme synthesis. Saccharose suppresses the induction of {beta}-galactosidase. The presence of a specific inhibitor in the structure studied is considered. (authors) [French] L'isopropylthiogalactoside (IPTG), inducteur de la 3-galactosidase, permet la synthese in vitro de cette enzyme dans la structure subcellulaire (P{sub 1}) isolee a partir des bacteries d'Escherichia coli K 12, inductibles mais non induites. L'incorporation d'acides amines radioactifs, stimulee par la presence d'inducteur, a ete etudiee au cours de la synthese de l'enzyme. Le saccharose supprime l'induction de la 3-galactosidase. La presence du represseur specifique dans la structure etudiee est consideree. (auteurs)

Purification and properties of a beta-galactosidase from carambola fruit with significant activity towards cell wall polysaccharides.

Science.gov (United States)

Balasubramaniam, Sumathi; Lee, Heng Chin; Lazan, Hamid; Othman, Roohaida; Ali, Zainon Mohd

2005-01-01

beta-Galactosidase (EC. 3.2.1.23) from ripe carambola (Averrhoa carambola L. cv. B10) fruit was fractionated through a combination of ion exchange and gel filtration chromatography into four isoforms, viz. beta-galactosidase I, II, III and IV. This beta-galactosidases had apparent native molecular masses of 84, 77, 58 and 130 kDa, respectively. beta-Galactosidase I, the predominant isoform, was purified to electrophoretic homogeneity; analysis of the protein by SDS-PAGE revealed two subunits with molecular masses of 48 and 36 kDa. N-terminal amino acid sequence of the respective polypeptides shared high similarities albeit at different domains, with the deduced amino acid sequence of certain plant beta-galactosidases, thus, explaining the observed low similarity between the two subunits. beta-Galactosidase I was probably a heterodimer that have glycoprotein properties and a pI value of 7.2, with one of the potential glycosylation sites appeared to reside within the 48-kDa-polypeptide. The purified beta-galactosidase I was substantially active in hydrolyzing (1-->4)beta-linked spruce and a mixture of (1-->3)beta- and (1-->6)beta-linked gum arabic galactans. This isoform also had the capability to solubilize and depolymerize structurally intact pectins as well as to modify alkaline-soluble hemicelluloses, reflecting in part changes that occur during ripening.

Treatment of anemia with darbepoetin alfa in systolic heart failure

DEFF Research Database (Denmark)

Swedberg, Karl; Young, James B; Anand, Inder S

2013-01-01

Patients with systolic heart failure and anemia have worse symptoms, functional capacity, and outcomes than those without anemia. We evaluated the effects of darbepoetin alfa on clinical outcomes in patients with systolic heart failure and anemia.......Patients with systolic heart failure and anemia have worse symptoms, functional capacity, and outcomes than those without anemia. We evaluated the effects of darbepoetin alfa on clinical outcomes in patients with systolic heart failure and anemia....

Resolution and t-spectrum for the ALFA detector

CERN Document Server

Guergouri, Hichem

2016-01-01

My report as Summer Student is in two parts. In the first part I report about my attendance to the run of the ALFA detector to test different collimation schemas and find a new way to get rid of the background. The second part is about the measurement of the resolution of the ALFA detector and comparing the real data with the Monte-Carlo (MC) simulations. First of all I tried to be more familiar with ROOT, worked on understanding the code used to access to the data in the trees inside the Ntuples, plotting histograms,… This was done using old Ntuples which contain MC data.

Entrappment of alkaline protease and β-galactosidase in radiation stitched together poly-N-vinylcaprolactam

International Nuclear Information System (INIS)

Davidenko, T.I.; Kravchenko, I.A.

1996-01-01

The gel formations by poly-N-vinylcaprolactam upon its γ-irradiation by the 20-25 kGy dose as a results of partial polymer stitching together is shown, which is confirmed by the CD-and thermogravimetric data. By the alkaline protease and β-galactosidase entrapment in poly-N- vinylcaprolactam stitched together by γ-irradiation, the active preparations are obtained with 90-98 % and 30-35 % activity retained for alkaline protease and β-galactosidase, respectively. The increased stability of alkaline protease at acidic pH values and higher temperature was noted, and for β-galactosidase - the possibility of repeated use of the obtained preparation for lactose hydrolysis

Nano-coating of beta-galactosidase onto the surface of lactose by using an ultrasound-assisted technique

DEFF Research Database (Denmark)

Genina, Natalja; Räikkönen, Heikki; Heinämäki, Jyrki

2010-01-01

We nano-coated powdered lactose particles with the enzyme beta-galactosidase using an ultrasound-assisted technique. Atomization of the enzyme solution did not change its activity. The amount of surface-attached beta-galactosidase was measured through its enzymatic reaction product D-galactose us......We nano-coated powdered lactose particles with the enzyme beta-galactosidase using an ultrasound-assisted technique. Atomization of the enzyme solution did not change its activity. The amount of surface-attached beta-galactosidase was measured through its enzymatic reaction product D......-galactose using a standardized method. A near-linear increase was obtained in the thickness of the enzyme coat as the treatment proceeded. Interestingly, lactose, which is a substrate for beta-galactosidase, did not undergo enzymatic degradation during processing and remained unchanged for at least 1 month....... Stability of protein-coated lactose was due to the absence of water within the powder, as it was dry after the treatment procedure. In conclusion, we were able to attach the polypeptide to the core particles and determine precisely the coating efficiency of the surface-treated powder using a simple approach....

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...

Dynamic behavior of reactive aluminum nanoparticle-fluorinated acrylic (AlFA) polymer composites

Science.gov (United States)

Crouse, Christopher A.; White, Brad; Spowart, Jonathan E.

2011-06-01

The dynamic behavior of aluminum nanoparticle-fluorinated acrylic (AlFA) composite materials has been explored under high strain rates. Cylindrical pellets of the AlFA composite materials were mounted onto copper sabots and impacted against a rigid anvil at velocities between 100 and 400 m/s utilizing a Taylor gas gun apparatus to achieve strain rates on the order of 104 /s. A framing camera was used to record the compaction and reaction events that occurred upon contact of the pellet with the anvil. Under both open air and vacuum environments the AlFA composites demonstrated high reactivity suggesting that the particles are primarily reacting with the fluorinated matrix. We hypothesize, based upon the compaction history of these materials, that reaction is initiated when the oxide shells on the aluminum nanoparticles are broken due an interparticle contact deformation process. We have investigated this hypothesis through altering the particle loading in the AlFA composites as well as impact velocities. This data and the corresponding trends will be presented in detail.

Thermal inactivation kinetics of β-galactosidase during bread baking.

Science.gov (United States)

Zhang, Lu; Chen, Xiao Dong; Boom, Remko M; Schutyser, Maarten A I

2017-06-15

In this study, β-galactosidase was utilized as a model enzyme to investigate the mechanism of enzyme inactivation during bread baking. Thermal inactivation of β-galactosidase was investigated in a wheat flour/water system at varying temperature-moisture content combinations, and in bread during baking at 175 or 205°C. In the wheat flour/water system, the thermostability of β-galactosidase increased with decreased moisture content, and a kinetic model was accurately fitted to the corresponding inactivation data (R 2 =0.99). Interestingly, the residual enzyme activity in the bread crust (about 30%) was hundredfold higher than that in the crumb (about 0.3%) after baking, despite the higher temperature in the crust throughout baking. This result suggested that the reduced moisture content in the crust increased the thermostability of the enzyme. Subsequently, the kinetic model reasonably predicted the enzyme inactivation in the crumb using the same parameters derived from the wheat flour/water system. However, the model predicted a lower residual enzyme activity in the crust compared with the experimental result, which indicated that the structure of the crust may influence the enzyme inactivation mechanism during baking. The results reported can provide a quantitative understanding of the thermal inactivation kinetics of enzyme during baking, which is essential to better retain enzymatic activity in bakery products supplemented with heat-sensitive enzymes. Copyright © 2017 Elsevier Ltd. All rights reserved.

Purification and chemical characterisation of a cell wall-associated β-galactosidase from mature sweet cherry (Prunus avium L.) fruit.

Science.gov (United States)

Gerardi, Carmela; Blando, Federica; Santino, Angelo

2012-12-01

Using four different chromatographic steps, β-galactosidase was purified from the ripe fruit of sweet cherry to apparent electrophoretic homogeneity with approximately 131-fold purification. The Prunus avium β-galactosidase showed an apparent molecular mass of about 100 kDa and consisted of four different active polypeptides with pIs of about 7.9, 7.4, 6.9 and 6.4 as estimated by native IEF and β-galactosidase-activity staining. The active polypeptides were individually excised from the gel and subjected to SDS-PAGE. Each of the four native enzymes showing β-galactosidase activity was composed of two polypeptides with an estimated mass of 54 and 33 kDa. Both of these polypeptides were subjected to N-terminal amino acid sequence analysis. The 54 kDa polypeptide of sweet cherry β-galactosidase showed a 43% identity with the 44 kDa subunit of persimmon and apple β-galactosidases and the 48 kDa subunit of carambola galactosidase I. The sweet cherry β-galactosidase exhibited a strict specificity towards p-nitrophenyl β-D-galactopyranoside, a pH optimum of 4.0 and K(m) and V(max) values of 0.42 mM and 4.12 mmol min(-1) mg(-1) of protein respectively with this substrate. The enzyme was also active towards complex glycans. Taken together the results of this study prompted a role for this class of enzymes on sweet cherry fruit ripening and softening. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

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

Directory of Open Access Journals (Sweden)

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

Future Applications of Apricot (Prunus Armeniaca Kaisa ß Galactosidase in Dairy Industry

Directory of Open Access Journals (Sweden)

Ansari Shakeel Ahmed

2014-09-01

Full Text Available The present study demonstrates the immobilization of β galactosidase from apricots (Prunus armeniaca kaisa on an inexpensive concanavalin A layered cellulose-alginate hybrid gel. Immobilized β galactosidase retained 78% of the initial activity after crosslinking by glutaraldehyde. It exhibited greater fraction of activity at both acidic and basic pH, and showed broad spectrum temperature optimum as compared to free enzyme. Moreover, immobilized enzyme exhibited higher thermal stability at 60°C and retained 80% of the original enzyme activity in presence of 3% galactose. The crosslinked immobilized enzyme showed improved hydrolysis of lactose from milk and whey in batch processes at 50°C as well as in continuous reactors operated at fl ow rate of 20 mL/h and 30 mL/h even after one month. Moreover, crosslinked adsorbed β galactosidase retained 76% activity even after its sixth repeated use, thereby promoting its use for lactose hydrolysis in various dairy products even for longer durations.

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...

Characterization of the activity of β-galactosidase from Escherichia coli and Drosophila melanogaster in fixed and non-fixed Drosophila tissues

Directory of Open Access Journals (Sweden)

Mizuki Tomizawa

2016-12-01

Full Text Available β-Galactosidase encoded by the Escherichia coli lacZ gene, is widely used as a reporter molecule in molecular biology in a wide variety of animals. β-Galactosidase retains its enzymatic activity in cells or tissues even after fixation and can degrade X-Gal, a frequently used colormetric substrate, producing a blue color. Therefore, it can be used for the activity staining of fixed tissues. However, the enzymatic activity of the β-galactosidase that is ectopically expressed in the non-fixed tissues of animals has not been extensively studied. Here, we report the characterization of β-galactosidase activity in Drosophila tissues with and without fixation in various experimental conditions comparing the activity of two evolutionarily orthologous β-galactosidases derived from the E. coli lacZ and Drosophila melanogaster DmelGal genes. We performed quantitative analysis of the activity staining of larval imaginal discs and an in vitro assay using larval lysates. Our data showed that both E. coli and Drosophila β-galactosidase can be used for cell-type-specific activity staining, but they have their own preferences in regard to conditions. E. coli β-galactosidase showed a preference for neutral pH but not for acidic pH compared with Drosophila β-galactosidase. Our data suggested that both E. coli and Drosophila β-galactosidase show enzymatic activity in the physiological conditions of living animals when they are ectopically expressed in a desired specific spatial and temporal pattern. This may enable their future application to studies of chemical biology using model animals.

Collaborative study for the validation of an improved HPLC assay for recombinant IFN-alfa-2.

Science.gov (United States)

Jönsson, K H; Daas, A; Buchheit, K H; Terao, E

2016-01-01

The current European Pharmacopoeia (Ph. Eur.) texts for Interferon (IFN)-alfa-2 include a nonspecific photometric protein assay using albumin as calibrator and a highly variable cell-based assay for the potency determination of the protective effects. A request was expressed by the Official Medicines Control Laboratories (OMCLs) for improved methods for the batch control of recombinant interferon alfa-2 bulk and market surveillance testing of finished products, including those formulated with Human Serum Albumin (HSA). A HPLC method was developed at the Medical Products Agency (MPA, Sweden) for the testing of IFN-alfa-2 products. An initial collaborative study run under the Biological Standardisation Programme (BSP; study code BSP039) revealed the need for minor changes to improve linearity of the calibration curves, assay reproducibility and robustness. The goal of the collaborative study, coded BSP071, was to transfer and further validate this improved HPLC method. Ten laboratories participated in the study. Four marketed IFN-alfa-2 preparations (one containing HSA) together with the Ph. Eur. Chemical Reference Substance (CRS) for IFN-alfa-2a and IFN-alfa-2b, and in-house reference standards from two manufacturers were used for the quantitative assay. The modified method was successfully transferred to all laboratories despite local variation in equipment. The resolution between the main and the oxidised forms of IFN-alfa-2 was improved compared to the results from the BSP039 study. The improved method even allowed partial resolution of an extra peak after the principal peak. Symmetry of the main IFN peak was acceptable for all samples in all laboratories. Calibration curves established with the Ph. Eur. IFN-alfa-2a and IFN-alfa-2b CRSs showed excellent linearity with intercepts close to the origin and coefficients of determination greater than 0.9995. Assay repeatability, intermediate precision and reproducibility varied with the tested sample within acceptable

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

Directory of Open Access Journals (Sweden)

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.

Kinetic Properties of α-Galactosidase and the Localization of Total Proteins in Erwinia chrysanthemi

Directory of Open Access Journals (Sweden)

John Morgan Brand

2004-01-01

Full Text Available Erwinia chrysanthemi is an enterobacterium that causes soft-rot in plants in general, resulting in enormous economic losses annually. For the pathogen to survive in the host plant, it has to use the readily assimilable compounds from the host fluids and degrade the host tissue. To accomplish this, E. chrysanthemi produces several extracellular and intracellular enzymes. Among the intracellular enzymes there is a special digestive class, the galactosidases, which can be either periplasmic or cytoplasmic. α-Galactosidase is known to degrade melibiose and raffinose into glucose and galactose, and into galactose and sucrose respectively. The aim of the present study was to investigate the kinetic properties of α-galactosidase in E. chrysanthemi, and the localization of total proteins, after culturing it in the presence of raffinose and melibiose. The α-galactosidase that degrades melibiose seems to be the same enzyme that is also responsible for the breakdown of raffinose in E. chrysanthemi. It is localized mainly in the cytoplasm with a fraction of between 2.4 and 5.4 % localized in the periplasm. The majority of E. chrysanthemi proteins have cytoplasmic localization.

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

Directory of Open Access Journals (Sweden)

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

A large, multicentre, observational, post-marketing surveillance study of the 2:1 formulation of follitropin alfa and lutropin alfa in routine clinical practice for assisted reproductive technology.

Science.gov (United States)

Bühler, Klaus; Naether, Olaf G J; Bilger, Wilma

2014-01-14

Follicle-stimulating hormone (FSH) and luteinizing hormone (LH) both have a role to play in follicular development during the natural menstrual cycle. LH supplementation during controlled ovarian stimulation (COS) for assisted reproductive technology (ART) is used for patients with hypogonadotropic hypogonadism. However, the use of exogenous LH in COS in normogonadotropic women undergoing ART is the subject of debate. The aim of this study was to investigate characteristics of infertile women who received the 2:1 formulation of follitropin alfa and lutropin alfa (indicated for stimulation of follicular development in women with severe LH and FSH deficiency) in German clinical practice. A 3-year, multicentre, open-label, observational/non-interventional, post-marketing surveillance study of women (21-45 years) undergoing ART. Primary endpoint: reason for prescribing the 2:1 formulation of follitropin alfa and lutropin alfa. Secondary variables included: COS duration/dose; oocytes retrieved; fertilization; clinical pregnancy; ovarian hyperstimulation syndrome (OHSS). In total, 2220 cycles were assessed; at least one reason for prescribing the 2:1 formulation was given in 1834/2220 (82.6%) cycles. Most common reasons were: poor ovarian response (POR) (39.4%), low baseline LH (17.8%), and age (13.8%). COS: mean dose of the 2:1 formulation on first day, 183.1/91.5 IU; mean duration, 10.8 days. In 2173/2220 (97.9%) cycles, human chorionic gonadotrophin was administered. Oocyte pick-up (OPU) was attempted in 2108/2220 (95.0%) cycles; mean (standard deviation) 8.0 (5.4) oocytes retrieved/OPU cycle. Fertilization (≥1 oocyte fertilized) rates: in vitro fertilization (IVF), 391/439 (89.1%) cycles; intracytoplasmic sperm injection (ICSI)/IVF + ICSI, 1524/1613 (94.5%) cycles. Clinical pregnancy rate: all cycles, 25.9%; embryo transfer cycles, 31.3%. OHSS: hospitalization for OHSS, 8 (0.36%) cycles, Grade 2, 60 (2.7%), and Grade 3, 1 (0.05%). In German routine clinical

Calibration, testing, commissioning and first data of ALFA at LHC

CERN Document Server

Jakobsen, S; The ATLAS collaboration

2011-01-01

The ALFA detector system (Absolute Luminosity For ATLAS) aims for measuring the absolute LHC luminosity with high precision by using pp-scattering under small angles. Scintillating fibers tracking detectors are positioned 240 m from LHC interaction point 1 inside Roman Pots at millimeter distance from the LHC beam axis. The detectors consist of layers of 64 scintillating square fibers of 500 µm size in a U-V configuration. The layers are staggered 10 times to improve the spatial resolution to about 30 micrometers. A total of 11680 scintillating fibers are read out on the 8 ALFA detectors through 184 Multi Anode Photomultiplier Tubes, MAPMTs, of type Hamamatsu R7600-00-M64, 64 channels. Each detector has dedicated scintillating trigger tiles read out by 8 mm Hamamatsu Photomultiplier Tubes (R7400P and R9880U-110) via clear fibers. The gain of the MAPMTs differs up to a factor 3 inside a MAPMT and an additional factor 2 between different MAPMTs. For compensation, ALFA has developed a front-end electronic based...

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...

Evaluating the transport layer of the ALFA framework for the Intel® Xeon Phi™ Coprocessor

Science.gov (United States)

Santogidis, Aram; Hirstius, Andreas; Lalis, Spyros

2015-12-01

The ALFA framework supports the software development of major High Energy Physics experiments. As part of our research effort to optimize the transport layer of ALFA, we focus on profiling its data transfer performance for inter-node communication on the Intel Xeon Phi Coprocessor. In this article we present the collected performance measurements with the related analysis of the results. The optimization opportunities that are discovered, help us to formulate the future plans of enabling high performance data transfer for ALFA on the Intel Xeon Phi architecture.

Release of β-galactosidase from poloxamine/α-cyclodextrin hydrogels

Directory of Open Access Journals (Sweden)

César A. Estévez

2014-12-01

Full Text Available All mammals lose their ability to produce lactase (β-galactosidase, the enzyme that cleaves lactose into galactose and glucose, after weaning. The prevalence of lactase deficiency (LD spans from 2 to 15% among northern Europeans, to nearly 100% among Asians. Following lactose consumption, people with LD often experience gastrointestinal symptoms such as abdominal pain, bowel distension, cramps and flatulence, or even systemic problems such as headache, loss of concentration and muscle pain. These symptoms vary depending on the amount of lactose ingested, type of food and degree of intolerance. Although those affected can avoid the uptake of dairy products, in doing so, they lose a readily available source of calcium and protein. In this work, gels obtained by complexation of Tetronic 90R4 with α-cyclodextrin loaded with β-galactosidase are proposed as a way to administer the enzyme immediately before or with the lactose-containing meal. Both molecules are biocompatible, can form gels in situ, and show sustained erosion kinetics in aqueous media. The complex was characterized by FTIR that evidenced an inclusion complex between the polyethylene oxide block and α-cyclodextrin. The release profiles of β-galactosidase from two different matrices (gels and tablets of the in situ hydrogels have been obtained. The influence of the percentage of Tetronic in media of different pH was evaluated. No differences were observed regarding the release rate from the gel matrices at pH 6 (t50 = 105 min. However, in the case of the tablets, the kinetics were faster and they released a greater amount of 90R4 (25%, t50 = 40–50 min. Also, the amount of enzyme released was higher for mixtures with 25% Tetronic. Using suitable mathematical models, the corresponding kinetic parameters have been calculated. In all cases, the release data fit quite well to the Peppas–Sahlin model equation, indicating that the release of β-galactosidase is governed by a

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...

Primary metabolite mobilization during germination in rosewood (Aniba rosaeodora Ducke seeds Mobilização de metabólitos primários durante a germinação de sementes de pau-rosa (Aniba rosaeodora Ducke

Directory of Open Access Journals (Sweden)

Renata Braga Souza Lima

2008-02-01

Full Text Available This study aimed to characterize protein, oil, starch and soluble sugar mobilization as well as the activity of alpha-amylase during rosewood seed germination. Germination test was carried out at 25°C and the following parameters were analyzed: percentage of germination, initial, average, and final germination time. Seed reserve quantification was monitored in quiescent seeds and during different stages of radicle growth. Starch mobilization was studied in function of a-amylase activity. Germination reached 87.5% at the initial, average, and final time of 16, 21 and 30 days, respectively. Oil mobilization showed a negative linear behavior, decreasing 40% between the first and the last stage analyzed, whereas protein levels increased 34.7% during the initial period of germination. Starch content (46.4% was the highest among those of the metabolites analyzed and starch mobilization occurred inversely to the observed for soluble sugars; alpha-amylase activity increased until the 15th day, a period before radicle emission and corresponding to the highest starch mobilization. The high percentage of rosewood seed germination may be related to the controlled condition used in the germination chamber as well as to high seed reserve mobilization, in special oil and starch.Este estudo teve por objetivo caracterizar a mobilização de metabólitos primários (proteínas, óleos, amido e açúcares solúveis e determinar a atividade da enzima alfa-amilase em sementes de pau-rosa durante diferentes períodos da germinação. Sementes quiescentes, após a assepsia, foram postas para germinar a 25 °C, analisando-se as seguintes variáveis: porcentagem de germinação e tempos inicial, médio e final de germinação. A quantificação das reservas orgânicas foi monitorada nas sementes quiescentes e nos diferentes estágios de crescimento da radícula. A mobilização do amido foi monitorada mediante a atividade da enzima alfa-amilase. A germinação atingiu

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...

Pilot study of dornase alfa (Pulmozyme) therapy for acquired ventilator-associated infection in preterm infants.

Science.gov (United States)

Scala, Melissa; Hoy, Deborah; Bautista, Maria; Palafoutas, Judith Jones; Abubakar, Kabir

2017-06-01

Evaluate the feasibility, safety, and efficacy of adjunctive treatment with dornase alfa in preterm patients with ventilator-associated pulmonary infection (VAPI) compared to standard care. We hypothesize that therapy with dornase alfa will be safe and well tolerated in the preterm population with no worsening of symptoms, oxygen requirement, or need for respiratory support. Prospective, randomized, blinded, pilot study comparing adjunctive treatment with dornase alfa to sham therapy. In addition to standard care, infants were randomized to receive dornase alfa 2.5 mg nebulized via endotracheal tube (ETT) every 12 hr for 7 days or sham therapy. ETT secretion gram stain and culture and chest X-ray (CXR) findings were evaluated. Respiratory support data were downloaded from the ventilator. Fourteen infants developed VAPI between 2012 and 2014; 11 enrolled in the study. Six received dornase alfa and five received sham therapy. Average gestational age at birth was 25 weeks and age at study entry was 31 days. There were no differences in demographics, ETT white blood cell count (WBC), CXR, or mean airway pressure (MAP) between the two groups. There was a trend towards decreased oxygen requirement (FiO2) in the treatment group that did not reach statistical significance. No side effects were observed in the treatment group. Treatment with dornase alfa is safe and treated infants had some improvement in FiO 2 requirement but no improvement in MAP. A larger randomized trial is needed to evaluate the efficacy of this therapy. Pediatr Pulmonol. 2017; 52:787-791. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Assessment of Relative Bioavailability of Moroctocog Alfa and Moroctocog Alfa (AF-CC) in Subjects With Severe Hemophilia A.

Science.gov (United States)

Korth-Bradley, Joan; Rupon, Jeremy; Plotka, Anna; Charnigo, Robert; Rendo, Pablo

2018-05-01

An open-label, single-dose, randomized, two-period, crossover study comparing the pharmacokinetics of factor VIII activity in plasma (FVIII:C) after administration of an albumin-free presentation of moroctocog alfa (test) and moroctocog alfa manufactured using the previous technique (reference) was conducted in 30 (25 evaluable) male subjects who had severe hemophilia A (FVIII:C < 1 IU/dL). Blood samples were collected for 48 h after administration of each dose. C was assayed using a chromogenic substrate assay. The FVIII:C pharmacokinetic parameters were calculated using noncompartmental analysis. The presentations would be bioequivalent if the 90% confidence limits of the ratio of the geometric mean values of AUC inf and recovery fell within the interval of 80-125%. The bioequivalence criteria were met. A total of 10 treatment-related adverse events were observed in a total of nine subjects. All were mild and none was determined to be related to administration of study medication. © 2018 The Authors. Clinical and Translational Science published by Wiley Periodicals, Inc. on behalf of American Society for Clinical Pharmacology and Therapeutics.

Hydrolysis of lactose with -D-galactosidase

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Vesna Stehlik-Tomas

2001-06-01

Full Text Available The conditions of lactose hydrolysis with enzyme preparation of D-galactosidase were investigated. The aim of this work was to considered the use of whey in fermentative processes with yeast Saccharomyces cerevisiae. Enzymatic hydrolysis was conducted at different temperatures, with different lactose concentrations in medium and different concentrations of added enzyme. The results show that optimal temperature for hydrolysis was 40°C. The optimal amount of enzyme preparation was 2 gL-1 in lactose medium with 5-10 % lactose.

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.

Generation and Characterization of Environmentally Sensitive Variants of the β-Galactosidase from Lactobacillus delbrueckii subsp. bulgaricus

Science.gov (United States)

Yoast, Sienna; Adams, Robin M.; Mainzer, Stanley E.; Moon, Keith; Palombella, Anthony L.; Schmidt, Brian F.

1994-01-01

A method is described for generating and screening variants of the β-galactosidase from Lactobacillus delbrueckii subsp. bulgaricus sensitive to several environmental stresses, with potential application in the food industry. Chemical mutagenesis with hydroxylamine or methoxylamine was performed on the β-galactosidase gene carried on an Escherichia coli expression vector. Mutants sensitive to cold, heat, low pH, low magnesium concentration, and the presence of urea were isolated by screening for reduced color development on β-galactosidase indicator plates. The mutations responsible for three variant β-galactosidases were localized, and the base substitutions were determined by DNA sequencing. The amino acid alterations associated with one low-pH-sensitive (pHs) and two urea-sensitive (Us) variants correspond to P584L (pHs1), G400S/R479Q (Us26), and G167E/E168K/E363K/V492M (Us17), respectively. Mutant pHs1 is also heat, cold, low magnesium, and urea sensitive; Us26 is also cold sensitive; and Us17 is also low-pH sensitive. PMID:16349230

Comportamiento de glicinina, beta-conglicinina y alfa-amilasa en semillas de soja deterioradas y no deterioradas Glycinin, beta-conglycinin and alpha-amylase behaviour in artificially deteriorated and not deteriorated soybean

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Adriana Rita Salinas

2002-08-01

Full Text Available El objetivo del trabajo fue estudiar el comportamiento de glicinina y beta-conglicinina y la actividad de alfa-amilasa en semillas deterioradas y no deterioradas de 10 cultivares de soja [Glycine max (L. Merr.]. Las semillas se sometieron a dos tratamientos: deterioradas por envejecimiento acelerado y no deterioradas. Se determinó la presencia de las proteínas de reserva a partir de semillas con 0, 3 y 8 días de germinadas por electroforesis en geles de poliacrilamida (SDS-PAGE. La actividad de la alfa-amilasa se determinó bioquímicamente en semillas con 0, 3, 8 y 12 días de germinadas. No hubo diferencias en la presencia de bandas de glicinina y beta-conglicinina en semillas no deterioradas hasta los 8 días de germinadas. Las semillas deterioradas se comportaron en forma similar a las no deterioradas. La actividad de la alfa-amilasa aumentó en semillas germinadas hasta 8 días y disminuyó a los 12 días. En semillas deterioradas la actividad enzimática disminuyó con respecto a las no deterioradas. El deterioro artificial no afectó la presencia de glicinina y beta-conglicinina pero alteró la actividad de la alfa-amilasa hasta los 12 días de germinación. Los cultivares estudiados mostraron comportamiento diferencial frente a la actividad de esta enzima.The objective of this research was to study the behaviour of the storage proteins glycinin and beta-conglycinin and the alpha-amylase activity in artificially deteriorated and not deteriorated soybean [Glycine max (L. Merr.] seeds of 10 cultivars. The seeds were submitted to two treatments: deteriorated by accelerated aging and not deteriorated seeds. The presence of glycinin and beta-conglycinin were determined in seeds with 0, 3 and 8 days of germination using polyacrylamid gel electrophoresis (SDS-PAGE. The alpha-amylase activity was biochemically determined in seeds with 0, 3, 8 and 12 days of germination. There were no differences in the presence of glycinin and beta

Methods in ALFA Alignment

CERN Document Server

Melendez, Jordan

2014-01-01

This note presents two model-independent methods for use in the alignment of the ALFA forward detectors. Using a Monte Carlo simulated LHC run at \\beta = 90m and \\sqrt{s} = 7 TeV, the Kinematic Peak alignment method is utilized to reconstruct the Mandelstam momentum transfer variable t for single-diractive protons. The Hot Spot method uses fluctuations in the hitmap density to pinpoint particular regions in the detector that could signal a misalignment. Another method uses an error function fit to find the detector edge. With this information, the vertical alignment can be determined.

Homodimeric β-galactosidase from Lactobacillus delbrueckii subsp. bulgaricus DSM 20081: expression in Lactobacillus plantarum and biochemical characterization.

Science.gov (United States)

Nguyen, Tien-Thanh; Nguyen, Hoang Anh; Arreola, Sheryl Lozel; Mlynek, Georg; Djinović-Carugo, Kristina; Mathiesen, Geir; Nguyen, Thu-Ha; Haltrich, Dietmar

2012-02-22

The lacZ gene from Lactobacillus delbrueckii subsp. bulgaricus DSM 20081, encoding a β-galactosidase of the glycoside hydrolase family GH2, was cloned into different inducible lactobacillal expression vectors for overexpression in the host strain Lactobacillus plantarum WCFS1. High expression levels were obtained in laboratory cultivations with yields of approximately 53000 U of β-galactosidase activity per liter of medium, which corresponds to ~170 mg of recombinant protein per liter and β-galactosidase levels amounting to 63% of the total intracellular protein of the host organism. The wild-type (nontagged) and histidine-tagged recombinant enzymes were purified to electrophoretic homogeneity and further characterized. β-Galactosidase from L. bulgaricus was used for lactose conversion and showed very high transgalactosylation activity. The maximum yield of galacto-oligosaccharides (GalOS) was approximately 50% when using an initial concentration of 600 mM lactose, indicating that the enzyme can be of interest for the production of GalOS.

Extraction, Purification and Thermodynamic Characterization of Almond (Amygdalus communis β-Galactosidase for the Preparation of Delactosed Milk

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Melita Lobo

2013-01-01

Full Text Available Buffer type, pH and ionic strength, as well as the fraction of polyvinylpyrrolidone were optimized for efficient extraction of β-galactosidase from almond seeds. The enzyme was purified up to electrophoretic homogeneity employing (NH42SO4 (15–60 % fractionation, size exclusion and ion-exchange chromatography. Molecular mass of β-galactosidase as estimated by gel filtration and SDS-PAGE was approx. 62 kDa, confirming its monomeric nature. The optimum activity of the enzyme was at pH=5.5, and it was stable within the range of pH=5.0–6.0. Various kinetic parameters of β-galactosidase thermal inactivation were calculated: ΔH°, ΔS° and ΔG° suggested that the enzyme undergoes significant processes of unfolding during denaturation. Using β-galactosidase from almond seed powder, lactose hydrolysis in milk up to approx. 50 % was observed. The findings indicate the potential use of almond seeds for the production of low/delactosed milk for lactose-intolerant population.

Hyporesponsiveness to Darbepoetin Alfa in Patients With Heart Failure and Anemia in the RED-HF Study (Reduction of Events by Darbepoetin Alfa in Heart Failure): Clinical and Prognostic Associations.

Science.gov (United States)

van der Meer, Peter; Grote Beverborg, Niels; Pfeffer, Marc A; Olson, Kurt; Anand, Inder S; Westenbrink, B Daan; McMurray, John J V; Swedberg, Karl; Young, James B; Solomon, Scott D; van Veldhuisen, Dirk J

2018-02-01

A poor response to erythropoiesis-stimulating agents such as darbepoetin alfa has been associated with adverse outcomes in patients with diabetes mellitus, chronic kidney disease, and anemia; whether this is also true in heart failure is unclear. We performed a post hoc analysis of the RED-HF trial (Reduction of Events by Darbepoetin Alfa in Heart Failure), in which 1008 patients with systolic heart failure and anemia (hemoglobin level, 9.0-12.0 g/dL) were randomized to darbepoetin alfa. We examined the relationship between the hematopoietic response to darbepoetin alfa and the incidence of all-cause death or first heart failure hospitalization during a follow-up of 28 months. For the purposes of the present study, patients in the lowest quartile of hemoglobin change after 4 weeks were considered nonresponders. The median initial hemoglobin change in nonresponders (n=252) was -0.25 g/dL and +1.00 g/dL in the remainder of patients (n=756). Worse renal function, lower sodium levels, and less use of angiotensin-converting enzyme inhibitors or angiotensin receptor blockers were independently associated with nonresponse. Although a low endogenous erythropoietin level helped to differentiate responders from nonresponders, its predictive value in a multivariable model was poor (C statistic=0.69). Nonresponders had a higher rate of all-cause death or first heart failure hospitalization (hazard ratio, 1.25; 95% confidence interval, 1.02-1.54) and a higher risk of all-cause mortality (hazard ratio, 1.30; 95% confidence interval, 1.04-1.63) than responders. A poor response to darbepoetin alfa was associated with worse outcomes in heart failure patients with anemia. Patients with a poor response were difficult to identify using clinical and biochemical biomarkers. URL: https://www.clinicaltrials.gov. Unique identifier: NCT00358215. © 2018 American Heart Association, Inc.

JSC “ALFA-BANK” marketing policy. problems and perspectives

OpenAIRE

Kirillov, A.; Kuznetcova, E.; Martirosian, M.

2013-01-01

The article is devoted to the results of JSC “Alfa-Bank” consumers’ segmentation and the following complex marketing research. The article suggests the ways of the bank’s marketing policy improvement.

Spotlight on taliglucerase alfa in the treatment of pediatric patients with type 1 Gaucher disease

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Gupta P

2017-06-01

Full Text Available Punita Gupta,1 Gregory M Pastores2 1Division of Genetics, Department of Pediatrics, St. Joseph’s Children’s Hospital, Paterson, New Jersey, USA; 2National Center for Inherited Metabolic Disorders, Mater Misericordiae University Hospital, University College Dublin, Dublin, Ireland Abstract: Gaucher disease (GD is a heritable storage disorder caused by functional defects of the lysosomal acid β-glucosidase and the accumulation of glucosylceramide within macrophages, resulting in multiple organ dysfunction. There are three commercially available enzyme replacement therapy (ERT products for the treatment of GD type 1 (GD1: imiglucerase, velaglucerase alfa, and taliglucerase alfa. Imiglucerase and velaglucerase alfa are produced in different mammalian cell systems; imiglucerase requires postproduction deglycosylation to expose terminal α-mannose residues, which are required for mannose receptor-mediated uptake by target macrophages. These steps are critical to the success of ERT for the treatment of visceral and hematologic manifestations of GD. Taliglucerase alfa is the first US Food and Drug Administration-approved plant-cell-expressed recombinant human protein, using carrot root cell cultures. Furthermore, it does not require postproduction glycosidic modifications. It is indicated for treatment of adults with GD1 in the US, Israel, Australia, Canada, Chile, Brazil, and other countries, and it is additionally approved for the treatment of pediatric patients in the US, Australia, and Canada and for the treatment of hematologic manifestations in pediatric patients with Type 3 GD in Canada and other countries. Our review focuses on the role of taliglucerase alfa in the pediatric population. A literature search through PubMed (from 1995 up till November 2016 of English language articles was performed with the following terms: Gaucher disease, lysosomal storage disease, taliglucerase. Secondary and tertiary references were obtained by reviewing

Drotrecogin alfa (activated) in adults with septic shock.

NARCIS (Netherlands)

Ranieri, V.M.; Thompson, B.T.; Barie, P.S.; Dhainaut, J.F.; Douglas, I.S.; Finfer, S.; Gardlund, B.; Marshall, J.C.; Rhodes, A.; Artigas, A.; Payen, D.; Tenhunen, J.; Al-Khalidi, H.R.; Thompson, V.; Janes, J.; Macias, W.L.; Vangerow, B.; Williams, M.D.; Pickkers, P.; Raemaekers, J.M.; et al.,

2012-01-01

BACKGROUND: There have been conflicting reports on the efficacy of recombinant human activated protein C, or drotrecogin alfa (activated) (DrotAA), for the treatment of patients with septic shock. METHODS: In this randomized, double-blind, placebo-controlled, multicenter trial, we assigned 1697

ATLAS-ALFA measurements on the total cross section and diffraction

CERN Document Server

Mortensen, Simon Stark; The ATLAS collaboration

2015-01-01

The measurement of the total pp cross section at the LHC at $\\sqrt{s}=7$ TeV with the ALFA subdetector of ATLAS is presented in this talk. In a special run with $\\beta^*=90$ m beam optics corresponding to an integrated luminosity of 80 $\\text{mb}^{-1}$ the differential elastic cross section is measured in the range from $-t=0.0025\\text{ GeV}^2$ to $-t=0.38\\text{ GeV}^2$. The total cross section $\\sigma(pp\\rightarrow X)$ is extracted using the Optical Theorem by extrapolation of the differential elastic cross section to $t=0\\text{ GeV}^2$. Prospects for diffractive measurements using ALFA to detect the intact proton(s) is also discussed.

The in vitro mucolytic effect of xylitol and dornase alfa on chronic rhinosinusitis mucus.

Science.gov (United States)

Hardcastle, Tim; Jain, Ravi; Radcliff, Fiona; Waldvogel-Thurlow, Sharon; Zoing, Melissa; Biswas, Kristi; Douglas, Richard

2017-09-01

The overproduction and stagnation of purulent mucus impair mucociliary clearance and exacerbate the symptoms of chronic rhinosinusitis (CRS). There is a clinical need for effective topical mucolytic agents to facilitate removal of mucus and improve postoperative outcomes. The effects of xylitol (5%) and dornase alfa (1 mg/mL) on mucus and mucus crusts were investigated. Viscoelasticity and viscosity of wet mucus derived from 30 CRS patients was measured with a plate rheometer. Postoperative dried mucus crust dissolution was measured by examining peripheral transparency, central transparency, and border definition of treated crust samples from 17 CRS patients. Xylitol and dornase alfa reduced wet mucus viscoelasticity at a frequency of 0.1 Hz significantly more than the saline control. Treatments also produced significantly lower viscosities than saline at a shear rate of 10 and 100 seconds -1 . Xylitol and dornase alfa significantly decreased mucus crust border definition relative to saline. Xylitol and dornase alfa may be efficacious mucolytics, encouraging the breakdown of postoperative mucus crusts and the reduction of viscoelasticity and viscosity of wet mucus. In vivo study is required to evaluate the potential of these agents in treating recalcitrant CRS. © 2017 ARS-AAOA, LLC.

Espectroscopia Mössbauer e refinamento estrutural no sistema cerâmico isomórfico (alfa-Fe2O3-(alfa-Al2O 3 Mössbauer spectroscopy and structural refinement in the isomorphic (alpha-Fe2O3-( alpha-Al2O3 ceramic system

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J. A. Moreto

2007-06-01

Full Text Available Os multiferróicos são materiais em que duas ou três propriedades tais como ferroeletricidade, ferromagnetismo e ferroelasticidade são observadas em uma mesma fase. Em particular, os multiferróicos magnetoelétricos representam os materiais que são simultaneamente (antiferromagnéticos e ferroelétricos, com ou sem ferroelasticidade. Especificamente, o sistema hematita (alfa-Fe2O3-alumina (alfa-Al2O3 tem sido estudado, principalmente devido às suas potencialidades para aplicações em metalurgia e como catalisador na síntese de amônia. Contudo, compostos deste sistema também podem apresentar efeitos magnetoelétricos. Neste trabalho um estudo estrutural do composto (alfa-Fe2O30,25(alfa-Al2O 30,75 submetido à moagem em altas energias e tratamento térmico pós-moagem é apresentado. As amostras foram caracterizadas por difração de raios X, refinamento estrutural Rietveld e espectroscopia Mössbauer. A determinação e identificação das fases cristalográficas, parâmetros de rede e volume de celas unitárias nas amostras moída e tratada termicamente permitiu verificar as mudanças dos parâmetros, a formação de espinélios e possíveis deformações relativas advindas do processo de moagem em altas energias.Multiferroics are materials in which two or all three of the properties, ferroelectricity, ferromagnetism, and ferroelasticity occur in the same phase. In particular, multiferroic magnetoelectrics represent the materials that are simultaneously ferromagnetic and ferroelectric, with or without ferroelasticity. Specifically, the hematite (alpha-Fe2O3-alumina (alpha-Al2O3 system has been studied, mainly due to its potential applications in metallurgy and as catalyst of ammonia synthesis. However, compounds of this system also may present magnetoelectric effects. In this work, a structural study of the high-energy ball milled and annealed (alpha-Fe2O3 0.25(alpha-Al2O30.75 compound was carefully conducted. The samples were

Alternative method for quantification of alfa-amylase activity.

Science.gov (United States)

Farias, D F; Carvalho, A F U; Oliveira, C C; Sousa, N M; Rocha-Bezerrra, L C B; Ferreira, P M P; Lima, G P G; Hissa, D C

2010-05-01

A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 A degrees C. A linear relationship between the logarithm of enzyme activities and the area of clear zones was obtained. The method was validated by testing alpha-amylase from barley at the concentrations of 2.4; 60; 300 and 1,500 U.mL-1. The proposed method turned out to be simpler, faster, less expensive and able to determine on a macro-scale alpha-amylase over a wide range (2.4 to 7,500 U.mL-1) in scientific investigation as well as in teaching laboratory activities.

Alternative method for quantification of alfa-amylase activity

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DF. Farias

Full Text Available A modification of the sensitive agar diffusion method was developed for macro-scale determination of alfa-amylase. The proposed modifications lower costs with the utilisation of starch as substrate and agar as supporting medium. Thus, a standard curve was built using alfa-amylase solution from Aspergillus oryzae, with concentrations ranging from 2.4 to 7,500 U.mL-1. Clear radial diffusion zones were measured after 4 hours of incubation at 20 °C. A linear relationship between the logarithm of enzyme activities and the area of clear zones was obtained. The method was validated by testing α-amylase from barley at the concentrations of 2.4; 60; 300 and 1,500 U.mL-1. The proposed method turned out to be simpler, faster, less expensive and able to determine on a macro-scale α-amylase over a wide range (2.4 to 7,500 U.mL-1 in scientific investigation as well as in teaching laboratory activities.

Influence of cultivating conditions on the alpha-galactosidase biosynthesis from a novel strain of Penicillium sp. in solid-state fermentation.

Science.gov (United States)

Wang, C L; Li, D F; Lu, W Q; Wang, Y H; Lai, C H

2004-01-01

The work is intended to achieve optimum culture conditions of alpha-galactosidase production by a mutant strain Penicillium sp. in solid-state fermentation (SSF). Certain fermentation parameters involving incubation temperature, moisture content, initial pH value, inoculum and load size of medium, and incubation time were investigated separately. The optimal temperature and moisture level for alpha-galactosidase biosynthesis was found to be 30 degrees C and 50%, respectively. The range of pH 5.5-6.5 was favourable. About 40-50 g of medium in 250-ml flask and inoculum over 1.0 x 10(6) spores were suitable for enzyme production. Seventy-five hours of incubation was enough for maximum alpha-galactosidase production. Substrate as wheat bran supplemented with soyabean meal and beet pulp markedly improved the enzyme yield in trays. Under optimum culture conditions, the alpha-galactosidase activity from Penicillium sp. MAFIC-6 indicated 185.2 U g(-1) in tray of SSF. The process on alpha-galactosidase production in laboratory scale may have a potentiality of scaling-up.

Drotrecogin alfa (activated in severe sepsis: a systematic review and new cost-effectiveness analysis

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Brophy James M

2007-06-01

Full Text Available Abstract Background Activated drotrecogin alfa (human activated protein C, rhAPC, is produced by recombinant DNA technology, and purports to improve clinical outcomes by counteracting the inflammatory and thrombotic consequences of severe sepsis. Controversy exists around the clinical benefits of this drug and an updated economic study that considers this variability is needed. Methods A systematic literature review was performed using Medline, Embase and the International Network of Agencies for Health Technology Assessment (INAHTA databases to determine efficacy, safety and previous economic studies. Our economic model was populated with systematic estimates of these parameters and with population life tables for longer term survival information. Monte Carlo simulations were used to estimate the incremental cost-effectiveness ratios (ICERs and variance for the decision analytic models. Results Two randomized clinical trials (RCTS of drotrecogin alfa in adults with severe sepsis and 8 previous economic studies were identified. Although associated with statistical heterogeneity, a pooled analysis of the RCTs did not show a statistically significant 28-day mortality benefit for drotrecogin alfa compared to placebo either for all patients (RR: 0.93, 95% CI: 0.69, 1.26 or those at highest risk as measured by APACHE II ≥ 25 (RR: 0.90, 95% CI: 0.54, 1.49. Our economic analysis based on the totality of the available clinical evidence suggests that the cost-effectiveness of drotrecogin alfa is uncertain ( Conclusion The evidence supporting the clinical and economic attractiveness of drotrecogin alfa is not conclusive and further research appears to be indicated.

Study of the transgalactosylation activity of ß-galactosidase from a new strain Kluyveromyces lactis 3

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ILIA ILIEV

2012-01-01

Full Text Available Beta-galactosidase (EC.3.2.1.23 is an important enzyme industrially used for the hydrolysis of lactose from milk and milk whey for several applications. Lately, the importance of this enzyme was enhanced by its galactosyltransferase activity, which is responsible for synthesis of transgalactosylated oligosaccharides that act as prebiotics with several beneficial effects on the consumers. ß-Galactosidase production by Kluyveromyces lactis 3 was studied in shake flask culture. The highest enzymatic activity was obtained at 10-th hour of the fermentation. The optimum temperature for transferase activity was 50°C. When incubated with 30% lactose in 50 mM phosphate buffer (pH 6.0 the enzyme can synthesize up to 41% galacto-oligosaccharides (GalOS. β-Galactosidase from strain Kluyveromyces lactis 3 produces mainly oligosaccharides with degree of polymerization (DP 6 at 40°C and with DP 3 at 50°C.

Efeitos do levosimendan sobre TNF-alfa, PNB e MMP-1 em pacientes com insuficiência cardíaca com anemia

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Mutlu Büyüklü

2012-07-01

Full Text Available FUNDAMENTO: O levosimendan é conhecido pelo seu efeito bilateral de fortalecimento contração das miofibrilas sem aumentar a demanda de oxigênio no miocárdio. A anemia é uma deterioração que causa aumento da dosagem de fármacos em pacientes com insuficiência cardíaca. OBJETIVO: No presente estudo comparamos a eficácia do tratamento com levosimendan em pacientes com insuficiência cardíaca descompensada com ou sem anemia. MÉTODOS: Foram incluídos no estudo 23 pacientes anêmicos com insuficiência cardíaca classe 3 ou 4, segundo a New York Heart Association (NYHA e fração de ejeção abaixo de 35%. Outros 23 pacientes com o mesmo diagnóstico cardíaco, mas sem anemia, serviu como grupo controle. Ao tratamento da insuficiência cardíaca tradicional desses pacientes foi acrescido um tratamento de 24 horas de levosimendan. Amostras foram tomadas para dosar os níveis séricos do fator de necrose tumoral alfa sérico (TNF-alfa, peptídeo natriurético cerebral aminoterminal (NT-proPNB e metaloproteinase da matriz 1 (MMP-1, antes e após a administração. RESULTADOS: Não houve diferença significativa entre os níveis séricos de TNF-alfa e MMP-1, antes e depois do tratamento (p > 0,05. Embora o nível de NT-proBNP tenha diminuído em ambos os grupos após o tratamento, não foi estatisticamente significativo (p = 0,531 e p = 0,913 para os grupos de anemia e de controle, respectivamente. Uma restauração significativa da capacidade funcional foi observada em ambos os grupos avaliados, de acordo com a NYHA (p < 0,001 e p = 0,001 para os grupos de anemia e controle, respectivamente. CONCLUSÃO: O tratamento com levosimendan apresenta efeitos semelhantes em pacientes com insuficiência cardíaca, com anemia e sem anemia. No entanto, o efeito precoce desse tratamento sobre os níveis de TNF-alfa, NT-proPNB e MMP-1 não é evidente. Ele oferece uma melhora significativa na capacidade funcional, sem a influência da anemia.

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.

Production of α-Galactosidase by Aspergillus oryzae through solid-state fermentation and its application in soymilk Galactooligosaccharide hydrolysis

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Shankar Kapnoor

2010-02-01

Full Text Available α-Galactosidase was produced by Aspergillus oryzae on red gram plant waste-wheat bran based media in solid-state fermentation (SSF. Optimum temperature for α-galactosidase production was 35 0C and upto 4 cm of bed height of substrate had no inhibitory effect on enzyme production. Hydrolysis of galactooligosaccharides in soymilk was carried out by α-galactosidase. Optimum temperature and pH for the hydrolysis of raffinose and stachyose of soymilk were 55(0C and 5.2-6.2, respectively. The enzymatic treatment for 3 h completely removed the raffinose oligosaccharides in soymilk. Crude extract also showed considerable amount of invertase activity.

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.

Crystallization and preliminary X-ray crystallographic analysis of β-galactosidase from Kluyveromyces lactis

International Nuclear Information System (INIS)

Pereira-Rodríguez, Ángel; Fernández-Leiro, Rafael; González Siso, M. Isabel; Cerdán, M. Esperanza; Becerra, Manuel; Sanz-Aparicio, Julia

2010-01-01

β-Galactosidase from K. lactis has been expressed in S. cerevisiae, purified by affinity chromatography and crystallized in its native form. β-Galactosidase from Kluyveromyces lactis catalyses the hydrolysis of the β-galactosidic linkage in lactose. Owing to its many industrial applications, the biotechnological potential of this enzyme is substantial. This protein has been expressed in yeast and purified for crystallization trials. However, optimization of the best crystallization conditions yielded crystals with poor diffraction quality that precluded further structural studies. Finally, the crystal quality was improved using the streak-seeding technique and a complete diffraction data set was collected at 2.8 Å resolution

Synthesis of oligosaccharides derived from lactulose (OsLu using soluble and immobilized Aspergillus oryzae b-galactosidase

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ALEJANDRA eCARDELLE COBAS

2016-03-01

Full Text Available b-galactosidase from Aspergillus oryzae offers a high yield for the synthesis of oligosaccharides derived from lactulose (OsLu by transgalactosylation. Oligosaccharides with degree of polymerization (DP ≥ 3 have shown to possess higher in vitro bifidogenic effect than di- and tetrasaccharides. Thus, in this work, an optimization of reaction conditions affecting the specific selectivity of A. oryzae b-galactosidase for synthesis of OsLu has been carried out to enhance OsLu with DP ≥ 3 production. Assays with b-galactosidase immobilized onto a glutaraldehyde-agarose support were also carried out with the aim of making the process cost-effective and industrially viable. Optimal conditions with both soluble and immobilized enzyme for the synthesis of OsLu with DP ≥ 3 were 50 °C, pH 6.5, 450 g/L of lactulose and 8 U/mL of enzyme, reaching yields of ca. 50% (w/v of total OsLu and ca. 20% (w/v of OsLu-3, being 6′-galactosyl-lactulose the major one, after a short reaction time. Selective formation of disaccharides, however, was favored at 60 °C, pH 4.5, 450 g/L of lactulose and 8 U/mL of enzyme. Immobilization increased the enzymatic stability to temperature changes and allowed to reuse the enzyme. We can conclude that the use, under determined optimal conditions, of the A. oryzae b-galactosidase immobilized on a support of glutaraldehyde-agarose constitutes an efficient and cost-effective alternative to the use of soluble b-galactosidases for the synthesis of prebiotic OsLu mixtures.

Alpha absolute power: motor learning of practical pistol shooting Potência absoluta de alfa: aprendizagem motora do tiro prático de pistola

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Clayton Amaral Domingues

2008-06-01

Full Text Available The present study aimed at investigating changes in behavior (shooting precision and electrophysiological variables (absolute alpha power during the motor learning of practical pistol shooting. The sample was composed of 23 healthy subjects, right-handed, male, between 18 and 20 years of age. The task consisted of four learning blocks. A One-way ANOVA with repeated measures and a post hoc analysis were employed to observe modifications on behavioral and electrophysiological measures (pO presente estudo teve por objetivo investigar alterações nas variáveis comportamentais (precisão do tiro e eletrofisiológicas (potência absoluta de alfa durante o aprendizado motor do tiro prático de pistola. A amostra constituiu-se de 20 sujeitos saudáveis, destros, sexo masculino, faixa etária entre 18 e 20 anos. A tarefa consistiu de quatro blocos de aprendizagem. A análise estatística das variáveis comportamentais e eletrofisiológicas foram realizadas por meio de uma ANOVA one-way e uma análise post hoc (p<0,05. Os resultados demonstraram diferenças significativas entre os blocos em função do aprendizado motor, bem como uma sensível melhora na precisão do tiro de ambos os blocos. Foi observada uma diminuição na potência de alfa em todos os eletrodos analisados durante a execução da tarefa, quando comparados aos blocos de linha de base e controle da aprendizagem. Os achados sugerem que a potência em alfa diminui devido o aprendizado motor quando sujeitos estão engajados na execução de uma tarefa motora complexa.

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

CARBOXYLATION OF SILVER NANOPARTICLES FOR THE IMMOBILIZATION OF β-GALACTOSIDASE AND ITS EFFICACY IN GALACTO-OLIGOSACCHARIDES PRODUCTION

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Shakeel Ahmed Ansari

2015-03-01

Full Text Available The present study investigated the carboxylation of silver nanoparticles (AgNPs by 1:3 nitric acid-sulfuric acid mixtures for immobilizing Aspergillus oryzae β-galactosidase. Carboxylated AgNPs retained 93% enzyme upon immobilization and the enzyme did not leach out appreciably from the modified nanosupport in the presence of 100 mmol L-1 NaCl. Atomic force micrograph revealed the binding of β-galactosidase on the modified AgNPs. The optimal pH for soluble and carboxylated AgNPs adsorbed β-galactosidase (IβG was observed at pH 4.5 while the optimal operating temperature was broadened from 50 ºC to 60 ºC for IβG. Michaelis constant, Km was increased two and a half fold for IβG while Vmax decreases slightly as compared to soluble enzyme. β-galactosidase immobilized on surface functionalized AgNPs retained 70% biocatalytic activity even at 4% galactose concentration as compared to enzyme in solution. Our study showed that IβG produces greater amount of galacto-oligosaccharides at higher temperatures (50 ºC and 60 ºC from 0.1 mol L-1 lactose solution at pH 4.5 as compared to previous reports.

In Patients with an α-Galactosidase A Variant, Small Nerve Fibre Assessment Cannot Confirm a Diagnosis of Fabry Disease

NARCIS (Netherlands)

van der Tol, Linda; Verhamme, Camiel; van Schaik, Ivo N.; van der Kooi, Anneke J.; Hollak, Carla E. M.; Biegstraaten, Marieke

2016-01-01

Fabry disease (FD) is an X-linked lysosomal storage disorder caused by an α-galactosidase A enzyme deficiency due to pathogenic variants in the α-galactosidase A gene (GLA). An increasing number of individuals with a GLA variant, but without characteristic FD features, are identified. A definite

New Synthesis of .alfa.,.omega.-Diiodoalkynes and Capped Iodobutadiynes

Czech Academy of Sciences Publication Activity Database

Hlavatý, Jaromír; Kavan, Ladislav; Štícha, M.

- (2002), s. 705-706 ISSN 1472-7781 R&D Projects: GA ČR GA203/00/0634 Institutional research plan: CEZ:AV0Z4040901 Keywords : .alfa.,. omega .-diiodoalkynes * capped iodobutadiynes Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 2.181, year: 2002

Indução do padrão citoplasmático em forma de "bastões e anéis" através do tratamento da hepatite C: relato de caso

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Mariano Felisberto

2015-04-01

Full Text Available Paciente do sexo feminino, queixando-se de astenia e dor em hipogastro, foi admitida no pronto-socorro do Hospital Universitário do Oeste do Paraná (HUOP. Durante a anamnese relatou tratamento de infecção crônica pelo vírus da hepatite C (VHC com inteferon peguilado e ribavirina. Dentre os exames laboratoriais solicitados, a pesquisa de autoanticorpos contra antígenos celulares (PAAC-HEp-2, conhecido tradicionalmente como fator antinúcleo (FAN, apresentou fluorescência em forma de bastões e/ou anéis no citoplasma das células. Esse padrão é caracterizado por bastões de 3-10 µm e anéis com 2-5 µm de diâmetro espalhados através do citoplasma da célula. Portanto, esse novo padrão tem sido designado como "bastões e anéis" (traduzido do inglês: Rods and Rings, RR. O alvo antigênico dessa reação foi identificado como inosina-5'-monofosfato desidrogenase tipo 2 (IMPDH2 que é uma enzima chave na síntese de nucleotídeos púricos. A enzima IMPDH2 agregada ou modificada em forma de RR nos pacientes tratados com ribavirina pode tornar-se antigênica e induzir uma resposta autoimune. É possível que o interferon alfa estimule a ocorrência de reatividade anti-RR aparentemente induzida pela ribavirina. Até o momento não se sabe por que o padrão RR em células HEp-2 ocorrem apenas em uma fração de pacientes portadores do VHC. Os dados apresentados em trabalhos anteriores possibilitam afirmar que esses anticorpos associados ao padrão RR estão fortemente relacionados com o tratamento da hepatite C. Além disso, pode-se afirmar que a ocorrência de reatividade anti-RR é promovida pela terapia combinada com interferon alfa e ribavirina.

Pathophysiology of hypophosphatasia and the potential role of asfotase alfa

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Orimo H

2016-05-01

Full Text Available Hideo Orimo Division of Metabolism and Nutrition, Department of Biochemistry and Molecular Biology, Nippon Medical School, Tokyo, Japan Abstract: Hypophosphatasia (HPP is an inherited systemic bone disease that is characterized by bone hypomineralization. HPP is classified into six forms according to the age of onset and severity as perinatal (lethal, perinatal benign, infantile, childhood, adult, and odontohypophosphatasia. The causative gene of the disease is the ALPL gene that encodes tissue-nonspecific alkaline phosphatase (TNAP. TNAP is expressed ubiquitously, and its physiological role is apparent in bone mineralization. A defect in bone mineralization can manifest in several ways, including rickets or osteomalacia in HPP patients. Patients with severe forms suffer from respiratory failure because of hypoplastic chest, which is the main cause of death. They sometimes present with seizures due to a defect in vitamin B6 metabolism resulting from the lack of alkaline phosphatase activity in neuronal cells, which is also lethal. Patients with a mild form of the disease exhibit rickets or osteomalacia and a functional defect of exercise. Odontohypophosphatasia shows only dental manifestations. To date, 302 mutations in the ALPL gene have been reported, mainly single-nucleotide substitutions, and the relationships between phenotype and genotype have been partially elucidated. An established treatment for HPP was not available until the recent development of enzyme replacement therapy. The first successful enzyme replacement therapy in model mice using a modified human TNAP protein (asfotase alfa was reported in 2008, and subsequently success in patients with severe form of the disease was reported in 2012. In 2015, asfotase alfa was approved in Japan in July, followed by in the EU and Canada in August, and then by the US Food and Drug Administration in the USA in October. It is expected that therapy with asfotase alfa will drastically change

Alfa-Omega: o pensamento crítico em livro

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Sandra Reimão

Full Text Available ResumoEste artigo apresenta a síntese de uma pesquisa que, por meio de fontes bibliográficas, análise documental e entrevistas, buscou traçar a história da Editora Alfa-Omega e compreender seu espaço e sua especificidade na complexa indústria do livro no Brasil. Fundada em São Paulo, em 1973, por Fernando e Claudete Mangarielo, a editora Alfa-Omega vem desde então “publicando o pensamento crítico brasileiro”. Em seus quatro primeiros anos de atividade, de 1973 a 1976, a editora publicou 44 títulos, sendo 16 deles livros de autoria de docentes da USP, época em que o meio universitário era um importante setor de oposição ao regime. A Editora, atualmente (2014, conta com mais de 200 títulos em catálogo. Nos últimos 14 anos, a Alfa-Omega, que na década de 1990 conheceu um grande rebaixamento de vendas, investe fortemente na internet e nos recursos digitais como ferramentas de produção e comercialização de livros. A editora mantém inalterados os princípios de sua linha editorial há mais de 40 anos. Conclui-se observando a dificuldade de manutenção de editoras de porte médio segmentadas no campo político da esquerda e a importância dessas editoras na promoção e manutenção da bibliodiversidade.

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.

Adsorption of β-galactosidase of Alicyclobacillus acidocaldarius on wild type and mutants spores of Bacillus subtilis

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Sirec Teja

2012-08-01

Full Text Available Abstract Background The Bacillus subtilis spore has long been used as a surface display system with potential applications in a variety of fields ranging from mucosal vaccine delivery, bioremediation and biocatalyst development. More recently, a non-recombinant approach of spore display has been proposed and heterologous proteins adsorbed on the spore surface. We used the well-characterized β-galactosidase from the thermoacidophilic bacterium Alicyclobacillus acidocaldarius as a model to study enzyme adsorption, to analyze whether and how spore-adsorption affects the properties of the enzyme and to improve the efficiency of the process. Results We report that purified β-galactosidase molecules were adsorbed to purified spores of a wild type strain of B. subtilis retaining ca. 50% of their enzymatic activity. Optimal pH and temperature of the enzyme were not altered by the presence of the spore, that protected the adsorbed β-galactosidase from exposure to acidic pH conditions. A collection of mutant strains of B. subtilis lacking a single or several spore coat proteins was compared to the isogenic parental strain for the adsorption efficiency. Mutants with an altered outermost spore layer (crust were able to adsorb 60-80% of the enzyme, while mutants with a severely altered or totally lacking outer coat adsorbed 100% of the β-galactosidase molecules present in the adsorption reaction. Conclusion Our results indicate that the spore surface structures, the crust and the outer coat layer, have an negative effect on the adhesion of the β-galactosidase. Electrostatic forces, previously suggested as main determinants of spore adsorption, do not seem to play an essential role in the spore-β-galactosidase interaction. The analysis of mutants with altered spore surface has shown that the process of spore adsorption can be improved and has suggested that such improvement has to be based on a better understanding of the spore surface structure

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

b-GALACTOSIDASE IMMOBILIZATION ON CONTROLLED PORE SILICA

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H. C. Trevisan

1997-12-01

Full Text Available The immobilization of b -galactosidase from Kluyveromyces fragilis on controlled pore silica was investigated. Immobilization was performed on amino silica activated with glutaraldehyde and the product was applied to the hydrolysis of lactose of whey. The behaviors of the soluble and immobilized enzyme were compared by using whey and a lactose solution as the substrate. With the aim of optimizing the method, parameters such as the amount of glutaraldehyde and the size of the particles were evaluated by comparing activities and stabilities on batch and continuously fluidized bed reactors

Peginterferon Lambda-1a/Ribavirin with Daclatasvir or Peginterferon Alfa-2a/Ribavirin with Telaprevir for Chronic Hepatitis C Genotype 1b.

Science.gov (United States)

Flisiak, Robert; Kawazoe, Seiji; Znoyko, Olga; Assy, Nimer; Gadano, Adrian; Kao, Jia-Horng; Lee, Kwan-Sik; Zwirtes, Ricardo; Portsmouth, Simon; Dong, Yuping; Xu, Dong; Kumada, Hiromitsu; Srinivasan, Subasree

2016-11-01

The study objective was to compare the efficacy and safety of peginterferon lambda-1a combined with ribavirin/daclatasvir (Lambda/RBV/DCV), versus peginterferon alfa-2a combined with ribavirin/telaprevir (Alfa/RBV/TVR), in patients chronically infected with hepatitis C virus (HCV), genotype 1b. This was a prospective, randomized, open-label, phase 3 study (NCT01718158) in adults (aged ≥18 years) who were treatment naïve or prior relapsers to peginterferon alfa/ribavirin therapy. The primary endpoint was sustained virologic response at post-treatment follow-up week 12 (SVR12). Patients were randomized in a 2:1 ratio to receive 24 weeks of Lambda/RBV/DCV or response-guided 24 or 48 weeks of Alfa/RBV/TVR. Overall, 440 patients were treated (294 with Lambda/RBV/DCV; 146 with Alfa/RBV/TVR). The proportion of patients achieving SVR12 was 88.8% in the Lambda/RBV/DCV arm and 70.5% in the Alfa/RBV/TVR arm (difference between arms: 18.3%; 95% confidence interval: 9.9-25.7; P Lambda/RBV/DCV group had fewer rash-related adverse events (AEs), cytopenic abnormalities, flu-like symptoms, serious AEs, and discontinuations due to AEs, but more liver abnormalities than those in the Alfa/RBV/TVR group. In conclusion, treatment with Lambda/RBV/DCV led to higher SVR12 rates and a more favorable safety profile than Alfa/RBV/TVR in patients with chronic HCV, genotype 1b infection.

Discriminating Foot-and-Mouth Disease Virus-Infected and Vaccinated Animals by Use of β-Galactosidase Allosteric Biosensors▿ †

Science.gov (United States)

Sánchez-Aparicio, M. Teresa; Rosas, María Flora; Ferraz, Rosa Maria; Delgui, Laura; Veloso, Juan J.; Blanco, Esther; Villaverde, Antonio; Sobrino, Francisco

2009-01-01

Recombinant β-galactosidases accommodating one or two different peptides from the foot-and-mouth disease virus (FMDV) nonstructural protein 3B per enzyme monomer showed a drastic enzymatic activity reduction, which mainly affected proteins with double insertions. Recombinant β-galactosidases were enzymatically reactivated by 3B-specific murine monoclonal and rabbit polyclonal antibodies. Interestingly, these recombinant β-galactosidases, particularly those including one copy of each of the two 3B sequences, were efficiently reactivated by sera from infected pigs. We found reaction conditions that allowed differentiation between sera of FMDV-infected pigs, cattle, and sheep and those of naïve and conventionally vaccinated animals. These FMDV infection-specific biosensors can provide an effective and versatile alternative for the serological distinction of FMDV-infected animals. PMID:19553549

Possibilities of uranium recovering as by product of copper lixiviation solution in Salobo 3-Alfa-Carajas

International Nuclear Information System (INIS)

Hohn, H.; Ferraioli, R.N.M.

1983-05-01

Geochemical and geophysics regional surveys performed by Nuclebras in 'Carajas, Para' State, show several favourable areas for uranium prospecting, including the 'Salobo 3-Alfa' copper mine. Some studies for increasing the Knowledge of uranium quantity and proportion and for accompanying the pilot plant installation, aiming the uranium extraction during the cycel of ore treatment in 'Salobo 3-Alfa' mine are described [pt

The catalytic properties and stability of β-galactosidases from fungi

Science.gov (United States)

Pilipenko, O. S.; Atyaksheva, L. F.; Poltorak, O. M.; Chukhrai, E. S.

2008-12-01

The catalytic activity of β-galactosidases from fungi Penicillium canescens and Aspergillus oryzae is maximum in a weakly acidic medium and does not depend on the presence of magnesium cations in the reaction medium. The enzyme from Aspergillus oryzae fungi is more active, and that from Penicillium canescens is stabler. One of stability indications is the presence of an induction period in the kinetic curves of thermal inactivation. This period disappears at 54°C for the enzyme from Aspergillus oryzae and at 59°C for the enzyme from Penicillium canescens. The temperature dependences of the effective rate constants for the inactivation of the tetrameric enzyme from Penicillium canescens show that the main reason for enzyme inactivation is the dissociation of oligomeric forms below 66°C ( E act = 85 kJ/mol) and enzyme denaturation at higher temperatures ( E act = 480 kJ/mol). The dissociation stage is absent for monomeric β-galactosidase from Aspergillus oryzae fungi, and the activation energy of inactivation is 450 kJ/mol over the whole temperature range studied (53-60°C).

Controle genético das células-tronco humanas cultivadas Genetic control of cultivated human stem cells

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Spencer L. M. Payão

2009-05-01

Full Text Available As células-tronco apresentam uma alta capacidade de autorregeneração, assim como, um potencial de diferenciação em uma variedade de tipos celulares. Estas células podem ser classificadas como embrionárias e adultas. Apesar de apresentar propriedades de células-tronco, as mesenquimais apresentam um certo grau de dificuldade no estabelecimento das culturas, podendo induzir a perda da expressão da enzima responsável pela imortalização ou enzima telomerase. A enzima telomerase é considerada um relógio biológico, um indicador que a senescência celular irá se instalar inevitavelmente. A questão mais atual e intrigante dos pesquisadores é se o suposto potencial de divisão, por um determinado período de tempo, das células-tronco cultivadas poderia levar ao acúmulo de alterações genéticas e epigenéticas, resultando em um processo neoplásico. Daí a importância do papel da citogenética humana no controle e monitoramento das células-tronco cultivadas que serão utilizadas na terapia em seres humanos. Alterações cromossômicas estruturais, tais como deleções, translocações e inversões, representam um mecanismo importante pelo qual as células cancerígenas desenvolvem-se gradualmente, uma vez que estas alterações cromossômicas podem levar a uma expressão anormal de muitos genes, podendo desencadear assim o processo neoplásico.Stem cells have a high capacity of self-regeneration, as well as a potential to differentiate into several cell types. These cells can be classified as embryonic or adult. In spite of having inherent properties of stem cells, mesenchymal cells show a certain degree of difficulty to establish cultures. This might induce a loss of the expression of the telomerase enzyme which is considered to be a biological clock or an indicator of the senescence of the cells. The most current and intriguing question for researchers is whether the presumed division potential of cultivated stem cells, over a

Inhibition of β-galactosidase and α-glucosidase synthesis in petroleum refinery effluent bacteria by phenolic compounds

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Gideon C. Okpokwasili

2011-04-01

Full Text Available Inhibition of α-glucosidase (EC 3.2.1.20 and β-galactosidase (EC 3.2.1.23 biosynthesis by phenolic compounds (phenol, 2-chlorophenol, 4-chlorophenol, 4-bromophenol and 3,5-dimethylphenol in Escherichia coli, Bacillus and Pseudomonas species isolated from petroleum refinery wastewater was assessed. At sufficient concentrations, phenols inhibited the induction of α-glucosidase and β-galactosidase. The patterns of these toxic effects can be mathematically described with logistic and sigmoid dose-response models. The median inhibitory concentrations (IC50 varied among the phenols, the bacteria and enzymes. Quantitative structure–activity relationship (QSAR models based on the logarithm of the octanol–water partition coefficient (log10Kow were developed for each bacterium. The correlation coefficients varied between 0.84and 0.99 for the enzymes. The test results indicated α-glucosidase and β-galactosidase biosynthesis as important microbial indices for evaluation of toxicity of phenolic compounds.

A blended- rather than whole-lentil meal with or without α-galactosidase mildly increases healthy adults' appetite but not their glycemic response.

Science.gov (United States)

Anguah, Katherene O-B; Wonnell, Brittany S; Campbell, Wayne W; McCabe, George P; McCrory, Megan A

2014-12-01

Disrupting the physical structure of pulses by blending them or by using a digestive supplement (α-galactosidase) to reduce intestinal discomfort could potentially negate the previously observed beneficial effects of whole pulses of lowering appetitive and glycemic responses because of more rapid digestion. We hypothesized that blended lentils, α-galactosidase, or both increase postprandial appetite and blood glucose responses vs. whole lentils. Men and women [n = 12; means ± SDs body mass index (kg/m(2)): 23.3 ± 3.1; aged 28 ± 10 y] consumed breakfast meals containing whole (W), blended (B), or no lentils [control (C)], each with 3 α-galactosidase or placebo capsules in a randomized, crossover, double-blind placebo-controlled trial. Between each test day there was a 3- to 5-d washout period. Mixed-model ANOVA showed effects of meal on postprandial appetite and glucose (P = 0.0001-0.031). The B meal resulted in higher postprandial appetite ratings than did the W meal but not the C meal for hunger, desire to eat, and prospective consumption (Δ = 0.4-0.5 points; P = 0.002-0.044). Postprandial glucose concentration was 4.5 mg/dL lower for the B meal than for the C meal (P meal. There were no main effects of α-galactosidase, but there were meal × α-galactosidase interaction effects, with a greater postprandial desire to eat and lower postprandial fullness with the B meal than with the 2 other meals in the placebo condition but not in the α-galactosidase condition. Blending lentils increased appetite (∼6%), but not glycemic response, compared with whole lentils, whereas α-galactosidase did not. Both B and W meals may be consumed (with or without an α-galactosidase supplement) with little impact on appetite, without increasing glycemic response. This trial was registered at clinicaltrials.gov as NCT02110511. © 2014 American Society for Nutrition.

Receptor-mediated endocytosis of α-galactosidase A in human podocytes in Fabry disease.

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Thaneas Prabakaran

Full Text Available Injury to the glomerular podocyte is a key mechanism in human glomerular disease and podocyte repair is an important therapeutic target. In Fabry disease, podocyte injury is caused by the intracellular accumulation of globotriaosylceramide. This study identifies in the human podocyte three endocytic receptors, mannose 6-phosphate/insulin-like growth II receptor, megalin, and sortilin and demonstrates their drug delivery capabilities for enzyme replacement therapy. Sortilin, a novel α-galactosidase A binding protein, reveals a predominant intracellular expression but also surface expression in the podocyte. The present study provides the rationale for the renal effect of treatment with α-galactosidase A and identifies potential pathways for future non-carbohydrate based drug delivery to the kidney podocyte and other potential affected organs.

A New Approach to the ALFA Trigger Simulator

CERN Document Server

Dziedzic, Bartosz

2016-01-01

The idea and principle of operation of a device which is used to test the trigger system of the ALFA detectors of the ATLAS experiment at the LHC are discussed. A new approach to the application control is presented. The application runs under control of the ArchLinuxARM operating system. Also, the drawbacks of the new solution are discussed.

The in vitro synthesis of β-galactosidase induced in a subcellular structure of Escherichia coli (1961)

International Nuclear Information System (INIS)

Nisman, B.; Kayser, A.; Demailly, J.; Genin, C.

1961-01-01

Isopropyl-thio-galactoside (IPTG), an inducer of 3-galactosidase, makes it possible to synthesise this enzyme in vitro with the subcellular structure (P 1 ). The enzyme is isolated from the bacteria Escherichia coli K 12 which are inductive but not induced. The incorporation of radioactive amino-acids, which is stimulated by the presence of an inducer, was studied during the course of the enzyme synthesis. Saccharose suppresses the induction of β-galactosidase. The presence of a specific inhibitor in the structure studied is considered. (authors) [fr

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.

Deproteinization: an integrated-solution approach to increase efficiency in β-galactosidase production using cheese whey powder (CWP solution

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Leandro Freire dos Santos

2017-08-01

Full Text Available Whey is the liquid that results from the coagulation of milk during cheese manufacture. Cheese whey is also an important environmental pollution source. The present experiment sought to compare β-galactosidase (EC 3.2.1.23 production by Aspergillus oryzae from deproteinized and un-deproteinized CWP solutions. β-galactosidase was produced by submerged fermentation in deproteinized or un-deproteinized CWP solutions. To determine the activity of the enzyme, a reaction mixture containing cell-free extract and ortho Nitrophenyl β galactoside (ONPG was used. The results indicated that β-galactosidase induction was greater when using deproteinized CWP solution compared to the un deproteinized CWP solution. These results may enable an alternative management of cheese whey, thereby decreasing its impact on the environment and producing value-added biomacromolecules.

A novel cold-active β-D-galactosidase from the Paracoccus sp. 32d - gene cloning, purification and characterization

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Wierzbicka-Woś Anna

2011-12-01

Full Text Available Abstract Background β-D-Galactosidases (EC 3.2.1.23 catalyze the hydrolysis of terminal non-reducing β-D-galactose residues in β-D-galactosides. Cold-active β-D-galactosidases have recently become a focus of attention of researchers and dairy product manufactures owing to theirs ability to: (i eliminate of lactose from refrigerated milk for people afflicted with lactose intolerance, (ii convert lactose to glucose and galactose which increase the sweetness of milk and decreases its hydroscopicity, and (iii eliminate lactose from dairy industry pollutants associated with environmental problems. Moreover, in contrast to commercially available mesophilic β-D-galactosidase from Kluyveromyces lactis the cold-active counterparts could make it possible both to reduce the risk of mesophiles contamination and save energy during the industrial process connected with lactose hydrolysis. Results A genomic DNA library was constructed from soil bacterium Paracoccus sp. 32d. Through screening of the genomic DNA library on LB agar plates supplemented with X-Gal, a novel gene encoding a cold-active β-D-galactosidase was isolated. The in silico analysis of the enzyme amino acid sequence revealed that the β-D-galactosidase Paracoccus sp. 32d is a novel member of Glycoside Hydrolase Family 2. However, owing to the lack of a BGal_small_N domain, the domain characteristic for the LacZ enzymes of the GH2 family, it was decided to call the enzyme under study 'BgaL'. The bgaL gene was cloned and expressed in Escherichia coli using the pBAD Expression System. The purified recombinant BgaL consists of two identical subunits with a combined molecular weight of about 160 kDa. The BgaL was optimally active at 40°C and pH 7.5. Moreover, BgaL was able to hydrolyze both lactose and o-nitrophenyl-β-D-galactopyranoside at 10°C with Km values of 2.94 and 1.17 mM and kcat values 43.23 and 71.81 s-1, respectively. One U of the recombinant BgaL would thus be capable

Efficacy of Alfa EEG wave biofeedback in the management of anxiety

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Pookala Bhat

2010-01-01

Full Text Available Background: Biofeedback is a technique in which people are trained to improve their health by learning to control certain internal bodily processes that normally occur involuntarily. Various studies in the past have shown usefulness of Alfa electroencephalographic (EEG biofeedback in the alleviation of anxiety symptoms. Though most of the psychiatric centers in the armed forces have this facility, not much work has been done in our setup to assess its efficacy in the management of anxiety. Hence this study was undertaken. Materials and Methods: This study was carried out in a multispecialty Command Hospital by enrolling 100 patients with psychiatric diagnosis from both inpatient and outpatient services. The anxiety level was assessed clinically and by using Hamilton Anxiety Scale and Taylor′s Manifest Anxiety Scale. One group of 50 patients was treated with Alfa EEG biofeedback sessions only, 5 times in a week for 8 weeks, along with specific pharmacotherapy. The other group was treated with appropriate dose of anxiolytics. The anxiety level was reassessed after 4 weeks and 8 weeks. Results: The response was better for mixed anxiety and depressive disorder with pharmacotherapy than with the biofeedback, but female patients showed better response with EEG biofeedback. Conclusion: In the short term, Alfa EEG biofeedback therapy is almost as efficacious as pharmacological intervention in the management of anxiety symptoms, and relatively more useful in females.

Alfa tahab TeliaSonerat Türgis üle pakkuda / Tõnis Arnover

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Arnover, Tõnis, 1952-

2005-01-01

TeliaSoneraga, kellele kuulub 37 protsenti Türgi operaatorfirma Turkcell aktsiatest, konkureerib täiendava 27 protsendise aktsiapaki ostmises Venemaa operaatorfirmaga Alfa Telecom. Lisa: Kaasosanike huvid põrkuvad. Diagramm: Konkurendid Vene MegaFonis

Production and Characterization of α-Galactosidase by a Multiple Mutant of Aspergillus niger in Solid-State Fermentation

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Muhammad Siddique Awan

2009-01-01

Full Text Available α-Galactosidase is applied in the sugar industry to enhance sugar recovery from sugar beet syrup and to improve nutritional value of the soymilk. In the present investigation, the influence of process variables on the production of this important enzyme has been explored in a newly isolated multiple mutant strain of Aspergillus niger in solid-state fermentation (SSF. Defined fermentation parameters include substrate type (pure lactose and by-products of rice and flour mills as prime substrates, nitrogen source, incubation time, initial pH of the medium and incubation temperature. Extracellular α-galactosidase reached the value of 135.4 IU/g of dry substrate (IU/g after 96 h of fermentation. Supplementation with 2 g of glucose and 3 g of corn steep liquor significantly increased the enzyme production, and maximum value of product yield (318 IU/g by the mutant strain was significantly higher than that reported by the wild type (this work, or other A. niger mutants, recombinants and yeasts reported in literature as producers of elevated levels of α-galactosidase. Among three α-galactosidases, one possessing high subunit molecular mass proteins (99 and 100 kDa has been characterized in both wild and mutant organisms. Thermal properties of the purified enzymes indicate that the mutation decreased the values of activation energy for the formation of enzyme-substrate (ES complex, enthalpy, Gibbs free energy demand for substrate binding, and transition state stabilization. A thermodynamic study of irreversible inactivation of enzymes suggests that the mutant–derived enzyme is more thermostable than the native enzyme, which is attributable to amino acids involved in active catalysis. Because of these properties, the mutant organism is a novel organism and may be exploited for bulk production of thermostable α-galactosidase for the above industrial and nutritional applications.

β-galactosidase from Aspergillus lacticoffeatus: A promising biocatalyst for the synthesis of novel prebiotics.

Science.gov (United States)

Cardoso, Beatriz B; Silvério, Sara C; Abrunhosa, Luís; Teixeira, José A; Rodrigues, Lígia R

2017-09-18

β-galactosidase (EC 3.2.1.23) are interesting enzymes able to catalyze lactose hydrolysis and transfer reactions to produce lactose-based prebiotics with potential application in the pharmaceutical and food industry. In this work, Aspergillus lacticoffeatus is described, for the first time, as an effective β-galactosidase producer. The extracellular enzyme production was evaluated in synthetic and alternative media containing cheese whey and corn steep liquor. Although β-galactosidase production occurred in all media (expect for the one composed solely by cheese whey), the highest enzymatic activity values (460U/mL) were obtained for the synthetic medium. Ochratoxin A production in synthetic medium was also evaluated and 9days of fermentation was identified as a suitable fermentation time to obtain a crude extract enzyme with mycotoxin concentration below the legal comparable value established for wine and grape juices (2ng/mL). The optimal pH and temperature for the crude extract enzyme was found in the range of 3.5-4.5 and 50-60°C, respectively. The β-galactosidase activity was reduced in the presence of Ba 2+ , Fe 2+ , Li + , K + and galactose, while additives (except for ascorbic acid) and detergents exhibited a positive effect on enzymatic activity. This enzyme was able to catalyze the synthesis of prebiotics, namely lactulose (2.5g/L) and a galacto-oligosaccharide (trisaccharide, 6.3g/L), either when whole cells or crude enzyme was used as biocatalyst. The lactulose production using fungal whole cells is herein reported for the first time. Additionally, A. lacticoffeatus was also found to produce an enzyme with fructosyltransferase activity and other prebiotics, namely fructo-oligosaccharide 1-kestose (2.4g/L). Copyright © 2017 Elsevier B.V. All rights reserved.

Carbonaceous Polymers and Nanotubes from .alfa., .omega.-Dialkali Hexatriynides

Czech Academy of Sciences Publication Activity Database

Hlavatý, Jaromír; Kavan, Ladislav; Okabe, K.; Oya, A.

2002-01-01

Roč. 40, č. 7 (2002), s. 1147-1150 ISSN 0008-6223 R&D Projects: GA ČR GA203/00/0634 Institutional research plan: CEZ:AV0Z4040901 Keywords : carbon nanotubes * chemically modified carbon * .alfa.,. omega .-dialkali hexatriynides Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.048, year: 2002

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.

Cloning, overexpression, purification, and characterization of a polyextremophilic β-galactosidase from the Antarctic haloarchaeon Halorubrum lacusprofundi

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Karan Ram

2013-01-01

Full Text Available Abstract Background Halorubrum lacusprofundi is a cold-adapted halophilic archaeon isolated from Deep Lake, a perennially cold and hypersaline lake in Antarctica. Its genome sequencing project was recently completed, providing access to many genes predicted to encode polyextremophilic enzymes active in both extremely high salinity and cold temperatures. Results Analysis of the genome sequence of H. lacusprofundi showed a gene cluster for carbohydrate utilization containing a glycoside hydrolase family 42 β-galactosidase gene, named bga. In order to study the biochemical properties of the β-galactosidase enzyme, the bga gene was PCR amplified, cloned, and expressed in the genetically tractable haloarchaeon Halobacterium sp. NRC-1 under the control of a cold shock protein (cspD2 gene promoter. The recombinant β-galactosidase protein was produced at 20-fold higher levels compared to H. lacusprofundi, purified using gel filtration and hydrophobic interaction chromatography, and identified by SDS-PAGE, LC-MS/MS, and ONPG hydrolysis activity. The purified enzyme was found to be active over a wide temperature range (−5 to 60°C with an optimum of 50°C, and 10% of its maximum activity at 4°C. The enzyme also exhibited extremely halophilic character, with maximal activity in either 4 M NaCl or KCl. The polyextremophilic β-galactosidase was also stable and active in 10–20% alcohol-aqueous solutions, containing methanol, ethanol, n-butanol, or isoamyl alcohol. Conclusion The H. lacusprofundi β-galactosidase is a polyextremophilic enzyme active in high salt concentrations and low and high temperature. The enzyme is also active in aqueous-organic mixed solvents, with potential applications in synthetic chemistry. H. lacuprofundi proteins represent a significant biotechnology resource and for developing insights into enzyme catalysis under water limiting conditions. This study provides a system for better understanding how H. lacusprofundi is

A Blended- Rather Than Whole-Lentil Meal with or without α-Galactosidase Mildly Increases Healthy Adults’ Appetite but Not Their Glycemic Response123

Science.gov (United States)

Anguah, Katherene O-B; Wonnell, Brittany S; Campbell, Wayne W; McCabe, George P; McCrory, Megan A

2014-01-01

Background: Disrupting the physical structure of pulses by blending them or by using a digestive supplement (α-galactosidase) to reduce intestinal discomfort could potentially negate the previously observed beneficial effects of whole pulses of lowering appetitive and glycemic responses because of more rapid digestion. Objective: We hypothesized that blended lentils, α-galactosidase, or both increase postprandial appetite and blood glucose responses vs. whole lentils. Methods: Men and women [n = 12; means ± SDs body mass index (kg/m2): 23.3 ± 3.1; aged 28 ± 10 y] consumed breakfast meals containing whole (W), blended (B), or no lentils [control (C)], each with 3 α-galactosidase or placebo capsules in a randomized, crossover, double-blind placebo-controlled trial. Between each test day there was a 3- to 5-d washout period. Results: Mixed-model ANOVA showed effects of meal on postprandial appetite and glucose (P = 0.0001–0.031). The B meal resulted in higher postprandial appetite ratings than did the W meal but not the C meal for hunger, desire to eat, and prospective consumption (Δ = 0.4–0.5 points; P = 0.002–0.044). Postprandial glucose concentration was 4.5 mg/dL lower for the B meal than for the C meal (P meal. There were no main effects of α-galactosidase, but there were meal × α-galactosidase interaction effects, with a greater postprandial desire to eat and lower postprandial fullness with the B meal than with the 2 other meals in the placebo condition but not in the α-galactosidase condition. Conclusions: Blending lentils increased appetite (∼6%), but not glycemic response, compared with whole lentils, whereas α-galactosidase did not. Both B and W meals may be consumed (with or without an α-galactosidase supplement) with little impact on appetite, without increasing glycemic response. This trial was registered at clinicaltrials.gov as NCT02110511. PMID:25411033

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.

Mechanism-based fluorescent labeling of beta-galactosidases. An efficient method in proteomics for glycoside hydrolases.

Science.gov (United States)

Kurogochi, Masaki; Nishimura, Shin-Ichiro; Lee, Yuan Chuan

2004-10-22

(4-N-5-Dimethylaminonaphthalene-1-sulfonyl-2-difluoromethylphenyl)-beta-d-galactopyranoside was synthesized and successfully tested on beta-galactosidases from Xanthomonas manihotis (Wong-Madden, S. T., and Landry, D. Glycobiology (1995) 5, 19-28 and Taron, C. H., Benner, J. S., Hornstra, L. J., and Guthrie, E. P. (1995) Glycobiology 5, 603-610), Escherichia coli (Jacobson, R. H., Zhang, X. J., DuBose, R. F., and Matthews, B. W. (1994) Nature 369, 761-766), and Bacillus circulans (Fujimoto, H., Miyasato, M., Ito, Y., Sasaki, T., and Ajisaka, K. (1988) Glycoconj. J. 15, 155-160) for the rapid identification of the catalytic site. Reaction of the irreversible inhibitor with enzymes proceeded to afford a fluorescence-labeled protein suitable for further high throughput characterization by using antidansyl antibody and matrix-assisted laser desorption ionization time-of-flight/time-of-flight (MALDI-TOF/TOF). Specific probing by a fluorescent aglycon greatly facilitated identification of the labeled peptide fragments from beta-galactosidases. It was demonstrated by using X. manihotis beta-galactosidase that the Arg-58 residue, which is located within a sequence of 56IPRAYWKD63, was labeled by nucleophilic attack of the guanidinyl group. This sequence including Arg-58 (Leu-46 to Tyr-194) was similar to that (Met-1 to Tyr-151) of Thermus thermophilus A4, which is the first known structure of glycoside hydrolases family 42 (Hidaka, M., Fushinobu, S., Ohtsu, N., Motoshima, H., Matsuzawa, H., Shoun, H., and Wakagi, T. (2002) J. Mol. Biol. 322, 79-91). A catalytic glutamic acid (Glu-537) of E. coli beta-galactosidase was proved to be labeled by the same procedure, suggesting that the modification site with this irreversible substrate might depend both on the nucleophilicity of the amino acids and their spatial arrangement in the individual catalytic cavity. Similarly, a Glu-259 in 257TLEE260 was selectively labeled using B. circulans beta-galactosidase, indicating that Glu

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

Directory of Open Access Journals (Sweden)

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

The pharmacokinetics of a B-domain truncated recombinant factor VIII, turoctocog alfa (NovoEight®), in patients with hemophilia A.

Science.gov (United States)

Jiménez-Yuste, V; Lejniece, S; Klamroth, R; Suzuki, T; Santagostino, E; Karim, F A; Saugstrup, T; Møss, J

2015-03-01

Turoctocog alfa (NovoEight(®)) is a human recombinant coagulation factor VIII (rFVIII) for the treatment of patients with hemophilia A. To evaluate the pharmacokinetics of turoctocog alfa in all age groups across clinical trials. Data from previously treated patients with severe hemophilia A (FVIII activity level of ≤ 1%) with no history of FVIII inhibitors, in a non-bleeding state, were included. The pharmacokinetics were assessed following a wash-out period and a subsequent single intravenous 50 IU kg(-1) dose of turoctocog alfa. Blood was sampled during a 48-h period postdose. Standard pharmacokinetic (PK) parameters were estimated on the basis of plasma FVIII activity vs. time (PK profiles) with non-compartmental methods. Furthermore, a population PK analysis was conducted. Data from 76 patients (aged 1-60 years) enrolled globally across six clinical trials were included, totaling 105 turoctocog alfa PK profiles. Single-dose PK results 3-6 months after the first dose of turoctocog alfa were comparable with the results obtained after the first dose. Similar PK characteristics were shown for different lots and strengths of the drug product. Overall, area under the plasma concentration (activity) curve from administration to infinity (AUC) and t1(/2) tended to increase with increasing age, with lower AUC and shorter t(1/2) being seen in children than in adolescents and adults. The PK profiles of turoctocog alfa and other commercially available plasma-derived FVIII and rFVIII products were similar in all age groups. The PK characteristics of turoctocog alfa have been thoroughly studied, and shown to be consistent over time, reproducible between different lots and strengths of drug product, and similar to those observed for other FVIII products. © 2014 International Society on Thrombosis and Haemostasis.

Senescence-associated β-galactosidase activity in the in vitro ovarian stromal fibroblasts

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Lilian Chuaire-Noack

2011-04-01

Full Text Available A growing biological research field is the cellular senescence, a mechanism that has been associated, under certain circumstances, withmalignant transformation. Given the high incidence of ovarian cancerand its main origin from the ovarian surface epithelium, as well asthe possibility that an epithelial-mesenchymal transition occurs, weevaluated both the in vitro growth of stromal fibroblasts from the ovarian cortex and their β-galactosidase activity at pH 6,enzyme whose expression is considered as a marker of replicativesenescence. Methods: 48 samples of ovarian cortical fibroblasts fromdonors without a history of cancer were serially cultured untilthe end of their replicative life. β-galactosidase activity at pH 6was quantified in each passage by the chemiluminiscent method. Ascontrol, we used ovarian epithelial cell cultures from the samedonors. The enzyme activity was also evaluated in fibroblastspreviously induced to senescence by exposure to hydrogen peroxide.Results: The analysis of the enzyme activity and the replicativecapacity taken together showed that the fibroblast cultures reachedthe senescent state at passages 4-5, as what happened with the control epithelial cells. Fibroblasts induced to senescence showed high variability in the values of enzymatic activity. Conclusions:The similarity between both types of cells in reaching the senescent state deserves to be taken into account in relation to theepithelialmesenchymal transition that has been proposed to explaintheir behavior in the genesis of cancer arising from ovarian surfaceepithelium. Low β-galactosidase activity values at pH 6 would suggestpossible inactivation of the response pathways to oxidative stress.

Autoantibodies to alfa-fodrin in patients with Hashimoto thyroiditis and Sjögren's syndrome: possible markers for a common secretory disorder.

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Szanto, Antonia; Csipo, Istvan; Horvath, Ildiko; Biro, Edit; Szodoray, Peter; Zeher, Margit

2008-09-01

Presence of autoantibodies to alfa-fodrin was investigated in patients with Sjögren's syndrome (n = 61), Hashimoto thyroiditis (n = 27), Sjögren's syndrome associated with Hashimoto thyroiditis (n = 31) and in healthy persons (n = 77). In each group, level of alfa-fodrin antibodies was higher than in the controls. There was no significant difference in their presence either between patients with Hashimoto thyroiditis with or without Sjögren's syndrome, or-in IgA isotype-between Sjögren's and Hashimoto thyroiditis patients. Correlation was found between the level of IgG alfa-fodrin and anti-thyroglobulin antibodies. Based on these findings, fodrin can be associated with both endocrine and exocrine glandular secretion. Antibodies to alfa-fodrin might have a role in the pathogenesis of Hashimoto thyroiditis concerning the "final common effectory pathway", secretion. Alfa-fodrin antibodies can be good markers of secretory disorders. Assessment of these autoantibodies might help the diagnosis and follow-up of patients with impaired secretory capability of not only autoimmune origin.

Kinetics of β-galactosidase Production by Lactobacillus bulgaricus During pH Controlled Batch Fermentation in Three Commercial Bulk Starter Media

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Saeed Abbasalizadeh

2015-10-01

Full Text Available The potential of bulk starter fermentation strategy for production of a cost-effective and GRAS source of β-galactosidase from a starter culture strain Lactobacillus bulgaricus was investigated. Three different media were selected and the strain, L. bulgaricus DSM 20081 was cultivated in these media under pH-controlled condition (pH = 5.6 at 43°C. The media were: bulk starter medium based on skim milk + whey, bulk starter medium based on whey, and skim milk. Growth and β-lactic acid production parameters were estimated from experimental data with the Garcia and Luedeking-Piret models, respectively. β-galactosidase production kinetics was also simulated using models based on biomass concentration and lactic acid production. Growth in the bulk starter medium based on skim milk + whey resulted in a higher rate of lactic acid production (7.35 ± 0.23  mg lactic acid ml-1 media h-1 and β-galactosidase activity (800.1± 0.7 nmol ONP ml-1 media compared to the other two media (P<0.01. Simulation of β- galactosidase production based on rate of lactic acid production resulted in very good agreement with experimental data for all three tested media. The results revealed the potential of bulk starter fermentation strategy and skim milk + whey based medium for in-house and relatively low cost production of food-grade β-galactosidase by dairy plants.

De-squeeze the beams: the TOTEM and ATLAS/ALFA experiments

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Stefania Pandolfi

2016-01-01

A special week-long proton–proton run with larger beam sizes at the interaction point is intended to probe the p-p elastic scattering regime at small angles.   Nicola Turini, deputy spokesperson for TOTEM, in front of one of the experiment’s ‘Roman Pot’ detectors in the LHC tunnel. (Photo: Maximilien Brice/CERN) Usually, the motto of the LHC is “maximum luminosity”. But for a few days per year, the LHC ignores its motto to run at very low luminosity for the forward experiments. This week, the LHC will provide the TOTEM and ATLAS/ALFA experiments with data for a broad physics programme. The TOTEM experiment at Point 5 and the ATLAS/ALFA experiment at Point 1 study the elastic scattering of protons, which are not observable in normal operation runs. In the elastic scattering process, the two protons survive their encounter intact and only change directions by exchanging momentum. To allow this special run, the operators play with the so-c...

Efeito da suplementação com vitamina E sobre a concentração de alfa-tocoferol no colostro humano Effect of vitamin E supplementation on alpha-tocopherol levels in human colostrum

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Roberto Dimenstein

2011-06-01

Full Text Available OBJETIVO: Avaliar a concentração de alfa-tocoferol no colostro humano em condições de suplementação com cápsulas de vitamina A acrescidas de vitamina E sintética. MÉTODOS: Foram recrutadas para o estudo 30 parturientes saudáveis atendidas em uma maternidade pública. Após jejum noturno, foram coletadas amostras de sangue e de colostro (2 mL das parturientes. Após a primeira coleta, as mesmas receberam suplemento na forma de uma cápsula de palmitato de retinila (200000 UI ou 60 mg acrescido de vitamina E sintética (49,4 mg de dl-alfa-tocoferol. Após 24 horas da suplementação, foi realizada nova coleta de 2 mL de colostro, também em jejum. RESULTADOS: A concentração sérica de alfa-tocoferol foi de 1 042,9 ± 319,0 μg/dL. Os teores de alfa-tocoferol no colostro antes da suplementação foram de 1155,4 ± 811,0 μg/dL, vs. 1396,3 ± 862,2 μg/dL 24 horas depois da suplementação (P > 0,05. Foi encontrada correlação entre o alfa-tocoferol do colostro na condição de jejum antes da suplementação e 24 horas após a suplementação (P = 0,001; r = 0,58, mas não entre soro e o colostro em nenhuma das condições acima citadas. CONCLUSÕES: Não houve aumento na concentração de alfa-tocoferol do colostro 24 horas após a suplementação. Isso indica que não é vantajoso suplementar a mãe com vitamina E sintética. Entretanto, os resultados também sugerem que, se o estado nutricional prévio à suplementação estiver adequado, mais tocoferol será encontrado no colostro após a suplementação. Novos estudos devem ser realizados para investigar o efeito da suplementação com a forma natural do alfa-tocoferol.OBJECTIVE: To assess the levels of alpha-tocopherol in human colostrum following supplementation with capsules containing vitamin A plus synthetic vitamin E. METHODS: Thirty healthy women about to give birth were recruited from a public maternity hospital. After overnight fasting, blood samples as well as colostrum

Measurement of the total cross section with ALFA Detector at ATLAS

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Trzebinski, M; The ATLAS collaboration

2017-01-01

The main goals of the Absolute Luminosity For ATLAS (ALFA) detector is to provide an absolute luminosity and total cross section measurement. The measurement method used, the detector alignment and the quality of the collected data are discussed.

The role of corifollitropin alfa in controlled ovarian stimulation for IVF in combination with GnRH antagonist

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Seyhan A

2011-08-01

Full Text Available Ayse Seyhan, Baris AtaDivision of Reproductive Endocrinology and Infertility, Department of Obstetrics and Gynecology, McGill University, Montreal, QC, CanadaAbstract: Corifollitropin alfa is a synthetic recombinant follicle-stimulating hormone (rFSH molecule containing a hybrid beta subunit, which provides a plasma half-life of ∼65 hours while maintaining its pharmocodynamic activity. A single injection of corifollitropin alfa can replace daily FSH injections for the first week of ovarian stimulation for in vitro fertilization. Stimulation can be continued with daily FSH injections if the need arises. To date, more than 2500 anticipated normoresponder women have participated in clinical trials with corifollitropin alfa. It is noteworthy that one-third of women did not require additional gonadotropin injections and reached human chorionic gonadotropin criterion on day 8. The optimal corifollitropin dose has been calculated to be 100 µg for women with a body weight ≤ 60 kg and 150 µg for women with a body weight >60 kg, respectively. Combination of corifollitropin with daily gonadotropin-releasing hormone antagonist injections starting on stimulation day 5 seems to yield similar or significantly higher numbers of oocytes and good quality embryos, as well as similar ongoing pregnancy rates compared with women stimulated with daily rFSH injections. Stimulation characteristics, embryology, and clinical outcomes seem consistent with repeated corifollitropin-stimulated assisted reproductive technologies cycles. Multiple pregnancy or ovarian hyperstimulation syndrome rates with corifollitropin were not increased over daily FSH regimen. The corifollitropin alfa molecule does not seem to be immunogenic and does not induce neutralizing antibody formation. Drug hypersensitivity and injection-site reactions are not increased. Incidence and nature of adverse events and serious adverse events are similar to daily FSH injections. Current trials do not

Production of a Highly Protease-Resistant Fungal α-Galactosidase in Transgenic Maize Seeds for Simplified Feed Processing.

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Yang, Wenxia; Zhang, Yuhong; Zhou, Xiaojin; Zhang, Wei; Xu, Xiaolu; Chen, Rumei; Meng, Qingchang; Yuan, Jianhua; Yang, Peilong; Yao, Bin

2015-01-01

Raffinose-family oligosaccharide (RFO) in soybeans is one of the major anti-nutritional factors for poultry and livestocks. α-Galactosidase is commonly supplemented into the animal feed to hydrolyze α-1,6-galactosidic bonds on the RFOs. To simplify the feed processing, a protease-resistant α-galactosidase encoding gene from Gibberella sp. strain F75, aga-F75, was modified by codon optimization and heterologously expressed in the embryos of transgentic maize driven by the embryo-specific promoter ZM-leg1A. The progenies were produced by backcrossing with the commercial inbred variety Zheng58. PCR, southern blot and western blot analysis confirmed the stable integration and tissue specific expression of the modified gene, aga-F75m, in seeds over four generations. The expression level of Aga-F75M reached up to 10,000 units per kilogram of maize seeds. In comparison with its counterpart produced in Pichia pastoris strain GS115, maize seed-derived Aga-F75M showed a lower temperature optimum (50 °C) and lower stability over alkaline pH range, but better thermal stability at 60 °C to 70 °C and resistance to feed pelleting inactivation (80 °C). This is the first report of producing α-galactosidase in transgenic plant. The study offers an effective and economic approach for direct utilization of α-galactosidase-producing maize without any purification or supplementation procedures in the feed processing.

Evaluating the transport layer of the ALFA framework for the Intel(®) Xeon Phi(™) Coprocessor

OpenAIRE

Santogidis, Aram; Hirstius, Andreas; Lalis, Spyros

2015-01-01

The ALFA framework supports the software development of major High Energy Physics experiments. As part of our research effort to optimize the transport layer of ALFA, we focus on profiling its data transfer performance for inter-node communication on the Intel Xeon Phi Coprocessor. In this article we present the collected performance measurements with the related analysis of the results. The optimization opportunities that are discovered, help us to formulate the future plans of enabling high...

Valorization of Tunisian alfa fibres and sumac tannins for the elaboration of biodegradable insulating panels

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Saad, Houda; Charrier, Bertrand; Ayed, Naceur; Charrier-El-Bouhtoury, Fatima

2017-10-01

Alfa leaves are important renewable raw materials in Tunisia where they are used basically in handcrafts and paper industry. Sumac is also an abundant species in Tunisia known for its high tannin content and is basically used in traditional medicine. To valorize these natural resources, we studied, for the first time, the possibility of making insulating panels based on alfa fibres and sumac tannins based adhesive. Firstly, alfa leaves were treated with an alkali solution as it is one of the standard procedures commonly used in the paper industry to extract cellulosic fibres. Mercerization effects were studied by characterizing fibres thermal properties and fibres surface morphology. Secondly, the sumac tannin based resin was formulated and characterized. Finally, the insulating panel was elaborated and characterized by determining its thermal conductivity. The thermal gravimetric analysis results show improvement in the thermal stability of fibres after alkali treatment. Environmental Scanning Electron Microscopy showed changes on treated alfa surface which could promote the fibre-matrix adhesion. The reactivity of sumac tannins to formaldehyde test (Stiasny number) showed the possible use of sumac tannins in wood adhesive formulation. Thermomechanical analysis and strength analysis of sumac tannin/hexamin based resin highlighted acceptable bonding properties. The thermal conductivity measurement showed an average value equal to 0.110 W/m K. Contribution to the topical issue "Materials for Energy harvesting, conversion and storage II (ICOME 2016)", edited by Jean-Michel Nunzi, Rachid Bennacer and Mohammed El Ganaoui

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

A Patient Friendly Corifollitropin Alfa Protocol without Routine Pituitary Suppression in Normal Responders.

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Wang, Huai-Ling; Lai, Hsing-Hua; Chuang, Tzu-Hsuan; Shih, Yu-Wei; Huang, Shih-Chieh; Lee, Meng-Ju; Chen, Shee-Uan

2016-01-01

The release of corifollitropin alfa simplifies daily injections of short-acting recombinant follicular stimulating hormone (rFSH), and its widely-used protocol involves short-acting gonadotropins supplements and a fixed GnRH antagonist regimen, largely based on follicle size. In this study, the feasibility of corifollitropin alfa without routine pituitary suppression was evaluated. A total of 288 patients were stimulated by corifollitropin alfa on cycle day 3 following with routine serum hormone monitoring and follicle scanning every other day after 5 days of initial stimulation, and a GnRH antagonist (0.25 mg) was only used prophylactically when the luteinizing hormone (LH) was ≧ 6 IU/L (over half of the definitive LH surge). The incidence of premature LH surge (≧ 10 IU/L) was 2.4% (7/288) before the timely injection of a single GnRH antagonist, and the elevated LH level was dropped down from 11.9 IU/L to 2.2 IU/L after the suppression. Two hundred fifty-one patients did not need any antagonist (87.2% [251/288]) throughout the whole stimulation. No adverse effects were observed regarding oocyte competency (fertilization rate: 78%; blastocyst formation rate: 64%). The live birth rate per OPU cycle after the first cryotransfer was 56.3% (161/286), and the cumulative live birth rate per OPU cycle after cyrotransfers was 69.6% (199/286). Of patients who did and did not receive GnRH antagonist during stimulation, no significant difference existed in the cumulative live birth rates (78.4% vs. 68.3%, p = 0.25). The results demonstrated that the routine GnRH antagonist administration is not required in the corifollitropin-alfa cycles using a flexible and hormone-depended antagonist regimen, while the clinical outcome is not compromised. This finding reveals that the use of a GnRH antagonist only occasionally may be needed.

A Patient Friendly Corifollitropin Alfa Protocol without Routine Pituitary Suppression in Normal Responders.

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Huai-Ling Wang

Full Text Available The release of corifollitropin alfa simplifies daily injections of short-acting recombinant follicular stimulating hormone (rFSH, and its widely-used protocol involves short-acting gonadotropins supplements and a fixed GnRH antagonist regimen, largely based on follicle size. In this study, the feasibility of corifollitropin alfa without routine pituitary suppression was evaluated. A total of 288 patients were stimulated by corifollitropin alfa on cycle day 3 following with routine serum hormone monitoring and follicle scanning every other day after 5 days of initial stimulation, and a GnRH antagonist (0.25 mg was only used prophylactically when the luteinizing hormone (LH was ≧ 6 IU/L (over half of the definitive LH surge. The incidence of premature LH surge (≧ 10 IU/L was 2.4% (7/288 before the timely injection of a single GnRH antagonist, and the elevated LH level was dropped down from 11.9 IU/L to 2.2 IU/L after the suppression. Two hundred fifty-one patients did not need any antagonist (87.2% [251/288] throughout the whole stimulation. No adverse effects were observed regarding oocyte competency (fertilization rate: 78%; blastocyst formation rate: 64%. The live birth rate per OPU cycle after the first cryotransfer was 56.3% (161/286, and the cumulative live birth rate per OPU cycle after cyrotransfers was 69.6% (199/286. Of patients who did and did not receive GnRH antagonist during stimulation, no significant difference existed in the cumulative live birth rates (78.4% vs. 68.3%, p = 0.25. The results demonstrated that the routine GnRH antagonist administration is not required in the corifollitropin-alfa cycles using a flexible and hormone-depended antagonist regimen, while the clinical outcome is not compromised. This finding reveals that the use of a GnRH antagonist only occasionally may be needed.

Efficacy and safety of once-weekly intravenous epoetin alfa in maintaining hemoglobin levels in hemodialysis patients.

Science.gov (United States)

Locatelli, Francesco; Villa, Giuseppe; Messa, Piergiorgio; Filippini, Armando; Cannella, Giuseppe; De Ferrari, Giacomo; Naso, Agostino; Rossi, Egidio; Formica, Marco; Lombardi, Luigi; Rotolo, Ugo; Conte, Feruccio

2008-01-01

Although an erythropoiesis-stimulating agent (ESA) is most frequently administered intravenously for treatment of anemia in patients with chronic kidney disease who are on dialysis, few studies have compared the efficacy of different intravenous (i.v.) dosing schedules. This multicenter, phase IIIb, open-label, controlled study randomized 289 stable hemodialysis patients to continue with conventional dosing of i.v. epoetin alfa or darbepoetin, or to switch to once-weekly i.v. epoetin alfa at the same cumulative weekly starting dose, to maintain hemoglobin levels at 11.0-13.0 g/dL, and within 1.0 g/dL of the baseline value. Hemoglobin levels and ESA doses were recorded every 4 weeks for 28 weeks. Hemoglobin levels fell significantly and ESA doses increased significantly between baseline and week 28 (mean of week 16-28 values) in the once-weekly epoetin alfa group, compared with the conventional treatment group (phemoglobin levels between the groups was 0.73 g/dL (greater than the threshold for therapeutic equivalence of 0.5 g/dL). The changes between groups from baseline was significant at all time points for hemoglobin levels (0.36, 0.46, 0.81, 0.87, 0.78, 0.62 and 0.49 g/dL) and from week 12 for ESA dose (718.5, 1,326.5, 1,732.0, 1,839.7 and 1,959.1 IU/week; p=0.005). Hemoglobin was maintained at the target level in 78% and 84% of patients on conventional dosing, and 67% and 64% of those on once-weekly epoetin alfa in the intention-to-treat (p=0.1) and per protocol (p=0.016) populations, respectively. This study did not show therapeutic equivalence of once-weekly i.v. epoetin alfa with conventional dosing regimens.

LONG TERM MUCOLYTIC THERAPY WITH DORNASE ALFA AT NOVOSIBIRSK CENTER FOR DIAGNOSTICS, TREATMENT, RHEABILITATION AND ADAPTATION OF CHILDREN WITH CYSTIC FIBROSIS

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N.I. Romanenko

2006-01-01

Full Text Available The center for diagnostic, treatment, rehabilitation and social adaptation of patients with cystic fibrosis was established in pulmonology department of the children's clinical hospital № 3 of Novosibirsk in 1994. Since 1995 the database of the center has registered 23 patients. the center has carried out an open prospective study of clinical efficacy and safety of dornase alfa (pulmozyme, «F. hoffmannala roche ltd.», basel in 10 patients with cystic fibrosis. The patients have received mucolytic therapy with dornase alfa for 2 years (2003–2005. They have been administered a daily inhalation of 2.5 ml of the solution (pulmozyme inhalation solution containing 2.5 of dornaza alfa. The study has shown an improvement of pulmonary function and elevation of oxygen saturation. The number of respiratory infections have dropped by 33%, the administration of intravenous antibiotics has shortened by 25%, the nutrition status has trended towards improvement. regardless of it's high price, dornasa alfa has proven its' benefits in the treatment of cystic fibrosis in patients of any age and disease severity.Key words: dornase alfa, children, cystic fibrosis, treatment.

Maintenance therapy with peginterferon alfa-2b does not prevent hepatocellular carcinoma in cirrhotic patients with chronic hepatitis C.

Science.gov (United States)

Bruix, Jordi; Poynard, Thierry; Colombo, Massimo; Schiff, Eugene; Burak, Kelly; Heathcote, Elizabeth J L; Berg, Thomas; Poo, Jorge-Luis; Mello, Carlos Brandao; Guenther, Rainer; Niederau, Claus; Terg, Ruben; Bedossa, Pierre; Boparai, Navdeep; Griffel, Louis H; Burroughs, Margaret; Brass, Clifford A; Albrecht, Janice K

2011-06-01

Several studies have reported that low doses of interferon can delay the development of hepatocellular carcinoma (HCC) and progression of chronic hepatitis C. We investigated the incidence of clinical events among participants of the Evaluation of PegIntron in Control of Hepatitis C Cirrhosis (EPIC)3 program. Data were analyzed from an open-label randomized study of patients with chronic hepatitis C who had failed to respond to interferon alfa plus ribavirin. All patients had compensated cirrhosis with no evidence of HCC. Patients received peginterferon alfa-2b (0.5 μg/kg/week; n=311) or no treatment (controls, n=315) for a maximum period of 5 years or until 98 patients had a clinical event (hepatic decompensation, HCC, death, or liver transplantation). The primary measure of efficacy was time until the first clinical event. There was no significant difference in time to first clinical event among patients who received peginterferon alfa-2b compared with controls (hazard ratio [HR], 1.452; 95% confidence interval [CI]: 0.880-2.396). There was no decrease in the development of HCC with therapy. The time to disease progression (clinical events or new or enlarged varices) was significantly longer for patients who received peginterferon alfa-2b compared with controls (HR, 1.564; 95% CI: 1.130-2.166). In a prospectively defined subanalysis of patients with baseline portal hypertension, peginterferon alfa-2b significantly increased the time to first clinical event compared with controls (P=.016). There were no new safety observations. Maintenance therapy with peginterferon alfa-2b is not warranted in all patients and does not prevent HCC. However, there is a potential clinical benefit of long-term suppressive therapy in patients with preexisting portal hypertension. Copyright © 2011 AGA Institute. Published by Elsevier Inc. All rights reserved.

Structural features of Aspergillus niger β-galactosidase define its activity against glycoside linkages.

Science.gov (United States)

Rico-Díaz, Agustín; Ramírez-Escudero, Mercedes; Vizoso-Vázquez, Ángel; Cerdán, M Esperanza; Becerra, Manuel; Sanz-Aparicio, Julia

2017-06-01

β-Galactosidases are biotechnologically interesting enzymes that catalyze the hydrolysis or transgalactosylation of β-galactosides. Among them, the Aspergillus niger β-galactosidase (AnβGal) belongs to the glycoside hydrolase family 35 (GH35) and is widely used in the industry due to its high hydrolytic activity which is used to degrade lactose. We present here its three-dimensional structure in complex with different oligosaccharides, to illustrate the structural determinants of the broad specificity of the enzyme against different glycoside linkages. Remarkably, the residues Phe264, Tyr304, and Trp806 make a dynamic hydrophobic platform that accommodates the sugar at subsite +1 suggesting a main role on the recognition of structurally different substrates. Moreover, complexes with the trisaccharides show two potential subsites +2 depending on the substrate type. This feature and the peculiar shape of its wide cavity suggest that AnβGal might accommodate branched substrates from the complex net of polysaccharides composing the plant material in its natural environment. Relevant residues were selected and mutagenesis analyses were performed to evaluate their role in the catalytic performance and the hydrolase/transferase ratio of AnβGal. Thus, we generated mutants with improved transgalactosylation activity. In particular, the variant Y304F/Y355H/N357G/W806F displays a higher level of galacto-oligosaccharides production than the Aspergillus oryzae β-galactosidase, which is the preferred enzyme in the industry owing to its high transferase activity. Our results provide new knowledge on the determinants modulating specificity and the catalytic performance of fungal GH35 β-galactosidases. In turn, this fundamental background gives novel tools for the future improvement of these enzymes, which represent an interesting target for rational design. Structural data are available in PDB database under the accession numbers 5IFP (native form), 5IHR (in complex with 6

GSI Helmholz Centre for Heavy Ion Research - ALFA: Next generation concurrent framework for ALICE and FAIR experiments

CERN Multimedia

CERN. Geneva

2015-01-01

FAIR is a new, unique international accelerator facility for the research with antiprotons and ions. It is being built at the GSI Darmstadt in Hesse, Germany. The commonalities between the ALICE and FAIR experiments and their computing requirements led to the development of a common software framework in an experiment independent way; ALFA (ALICE-FAIR framework). ALFA is designed for high quality parallel data processing and reconstruction on heterogeneous computing systems. It provides a data transport layer and the capability to coordinate multiple data processing components. ALFA is a flexible, elastic system which balances reliability and ease of development with performance by using a message based multi-processing in addition to multi-threading. The framework allows for usage of heterogeneous computing architectures by offloading (portions of code are accelerated on the device) or natively (where the full program is executed on the device ).

An enzymatic glycosylation of nucleoside analogues using beta-galactosidase from Escherichia coli

Czech Academy of Sciences Publication Activity Database

Blažek, Jiří; Jansa, Petr; Baszczyňski, Ondřej; Kaiser, Martin Maxmilian; Otmar, Miroslav; Krečmerová, Marcela; Dračínský, Martin; Holý, Antonín; Králová, B.

2012-01-01

Roč. 20, č. 9 (2012), s. 3111-3118 ISSN 0968-0896 R&D Projects: GA MŠk 1M0508 Institutional research plan: CEZ:AV0Z40550506 Keywords : glycosylation * galactosylation * beta-galactosidase * enzymatic synthesis * nucleoside * acyclic nucleoside analogues Subject RIV: CC - Organic Chemistry Impact factor: 2.903, year: 2012

Beta-galactosidase catalyzed selective galactosylation of aromatic compounds.

Science.gov (United States)

Bridiau, Nicolas; Taboubi, Selma; Marzouki, Nejib; Legoy, Marie Dominique; Maugard, Thierry

2006-01-01

A new approach to galacto-oligosaccharides and galacto-conjugates synthesis performed by the beta-galactosidase from Kluyveromyces lactis is reported. The enzymatic galactosylation of eight kinds of adsorbed aromatic primary alcohols, in particular the two drugs guaifenesin and chlorphenesin, gave the corresponding beta-D-galacto-pyranosides in yields ranging between approximately 10% and 96%. For the first time, we have showed that the adsorption of acceptor substrates onto solid supports such as silica gel influences the yield and the selectivity of galacto-conjugates synthesis. In particular, we observed that adsorption of acceptor favored the synthesis of digalactosylated compounds.

Three-dimensional immobilization of beta-galactosidase on a silicon surface.

Science.gov (United States)

Betancor, Lorena; Luckarift, Heather R; Seo, Jae H; Brand, Oliver; Spain, Jim C

2008-02-01

Many alternative strategies to immobilize and stabilize enzymes have been investigated in recent years for applications in biosensors. The entrapment of enzymes within silica-based nanospheres formed through silicification reactions provides high loading capacities for enzyme immobilization, resulting in high volumetric activity and enhanced mechanical stability. Here we report a strategy for chemically associating silica nanospheres containing entrapped enzyme to a silicon support. beta-galactosidase from E. coli was used as a model enzyme due to its versatility as a biosensor for lactose. The immobilization strategy resulted in a three-dimensional network of silica attached directly at the silicon surface, providing a significant increase in surface area and a corresponding 3.5-fold increase in enzyme loading compared to enzyme attached directly at the surface. The maximum activity recovered for a silicon square sample of 0.5 x 0.5 cm was 0.045 IU using the direct attachment of the enzyme through glutaraldehyde and 0.16 IU when using silica nanospheres. The immobilized beta-galactosidase prepared by silica deposition was stable and retained more than 80% of its initial activity after 10 days at 24 degrees C. The ability to generate three-dimensional structures with enhanced loading capacity for biosensing molecules offers the potential to substantially amplify biosensor sensitivity. (c) 2007 Wiley Periodicals, Inc.

Andexanet Alfa for Acute Major Bleeding Associated with Factor Xa Inhibitors

NARCIS (Netherlands)

Connolly, Stuart J.; Milling, Truman J.; Eikelboom, John W.; Gibson, C. Michael; Curnutte, John T.; Gold, Alex; Bronson, Michele D.; Lu, Genmin; Conley, Pamela B.; Verhamme, Peter; Schmidt, Jeannot; Middeldorp, Saskia; Cohen, Alexander T.; Beyer-Westendorf, Jan; Albaladejo, Pierre; Lopez-Sendon, Jose; Goodman, Shelly; Leeds, Janet; Wiens, Brian L.; Siegal, Deborah M.; Zotova, Elena; Meeks, Brandi; Nakamya, Juliet; Lim, W. Ting; Crowther, Mark; Connolly, S. C.; Crowther, M.; Eikelboom, J.; Gibson, M.; Milling, T. J.; Albaladejo, P.; Cohen, A.; Lopez-Sendon, J.; Schmidt, J.; Verhamme, P.; Beyer-Westendorf, J.; Wyse, D. G.; Garcia, D.; Prins, M. [=Martin H.; Nakamya, J.; Büller, H. R.; Mahaffey, K.; Alexander, J.; Demchuk, A.; Raskob, G.; Schulman, S.; Meeks, B.; Zotova, E.; Holadyk-Gris, I.; Coppens, M.

2016-01-01

Background Andexanet alfa (andexanet) is a recombinant modified human factor Xa decoy protein that has been shown to reverse the inhibition of factor Xa in healthy volunteers. Methods In this multicenter, prospective, open-label, single-group study, we evaluated 67 patients who had acute major

Evaluation of pharmacokinetics, user handling, and tolerability of peginterferon alfa-2a (40 kDa) delivered via a disposable autoinjector device

Science.gov (United States)

Varunok, Peter; Lawitz, Eric; Beavers, Kimberly L; Matusow, Gary; Leong, Ruby; Lambert, Nathalie; Bernaards, Coen; Solsky, Jonathan; Brennan, Barbara J; Wat, Cynthia; Bertasso, Anne

2011-01-01

Background Peginterferon alfa-2a (40 kDa) is currently administered using a prefilled syringe. The peginterferon alfa-2a disposable autoinjector is a new safety-engineered device designed to facilitate injection and reduce the risk of needlestick injuries. The analysis of two open-label Phase I trials evaluated the pharmacokinetics, successful administration, and tolerability of peginterferon alfa-2a when using the autoinjector. The studies were performed to support the filing and registration of the autoinjector device. Methods In trial 1, 50 healthy adult subjects received one 180 μg dose of peginterferon alfa-2a via the autoinjector. Serial blood samples were collected predose, up to 336 hours following drug administration, and at follow-up (28 ± 3 days post-dosing) for noncompartmental pharmacokinetic analysis. Trial 2 randomized 60 adult patients with chronic hepatitis C to 180 μg peginterferon alfa-2a once weekly by the autoinjector or prefilled syringe for 3 weeks followed by the alternative device (prefilled syringe or autoinjector, respectively) for 3 weeks. Patients also received ribavirin. Administration by the devices was evaluated under direct observation by a study staff member and by patient subjective assessment. Results In trial 1, following a single dose of peginterferon alfa-2a, the maximum plasma concentration was 16.1 ± 5.3 ng/mL (mean ± standard deviation), and area under the concentration time curve (0–168 hours) was 1996 ± 613 ng · hour/mL, similar to that reported using a vial/syringe or prefilled syringe. In trial 2, few patients showed handling difficulties with either device. Generally, patients were observed to be more satisfied and confident, followed instructions better, and successfully initiated injection with the autoinjector versus the prefilled syringe. Patients reported the autoinjector to be more convenient and easier to use. No pain or discomfort was experienced using the autoinjector. The autoinjector safety profile

Is dibotermin alfa a cost-effective substitute for autologous iliac crest bone graft in single level lumbar interbody spine fusion?

Science.gov (United States)

Svedbom, Axel; Paech, Daniel; Leonard, Catherine; Donnell, David; Song, Fujian; Boszcyk, Bronek; Rothenfluh, Dominique A; Lloyd, Andrew; Borgman, Benny

2015-11-01

To evaluate the cost-effectiveness of dibotermin alfa compared with autologous iliac crest bone graft (ICBG) for patients undergoing single level lumbar interbody spinal fusion in a UK hospital setting. An individual patient data (IPD) meta-analysis of six randomized controlled clinical trials and two single arm trials compared dibotermin alfa on an absorbable collagen implantation matrix (ACIM) (n = 456) and ICBG (n = 244) on resource use, re-operation rates, and SF-6D (Short form 6-dimension) health utility (total N = 700). Failure-related second surgery, operating time, post-operative hospital stay, and quality-adjusted life years (QALYs) derived from the IPD meta-analysis were included as inputs in an economic evaluation undertaken to assess the cost-effectiveness of dibotermin alfa/ACIM versus ICBG for patients undergoing single level lumbar interbody spinal fusion. A four year time horizon and the United Kingdom (UK) National Health Service (NHS) and Personal Social Services (PSS) perspective was adopted in the base case, with sensitivity analyses performed to gauge parameter uncertainty. In the base case analysis, patients treated using dibotermin alfa/ACIM (12 mg pack) accrued 0.055 incremental QALYs at an incremental cost of £ 737, compared with patients treated with ICBG. This resulted in an incremental cost-effectiveness ratio (ICER) of £ 13,523, indicating that at a willingness-to-pay threshold of £ 20,000, dibotermin alfa/ACIM is a cost-effective intervention relative to ICBG from the NHS and PSS perspective. In a UK hospital setting, dibotermin alfa/ACIM is a cost-effective substitute for ICBG for patients who require lumbar interbody arthrodesis.

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.

Pathophysiology of hypophosphatasia and the potential role of asfotase alfa.

Science.gov (United States)

Orimo, Hideo

2016-01-01

Hypophosphatasia (HPP) is an inherited systemic bone disease that is characterized by bone hypomineralization. HPP is classified into six forms according to the age of onset and severity as perinatal (lethal), perinatal benign, infantile, childhood, adult, and odontohypophosphatasia. The causative gene of the disease is the ALPL gene that encodes tissue-nonspecific alkaline phosphatase (TNAP). TNAP is expressed ubiquitously, and its physiological role is apparent in bone mineralization. A defect in bone mineralization can manifest in several ways, including rickets or osteomalacia in HPP patients. Patients with severe forms suffer from respiratory failure because of hypoplastic chest, which is the main cause of death. They sometimes present with seizures due to a defect in vitamin B6 metabolism resulting from the lack of alkaline phosphatase activity in neuronal cells, which is also lethal. Patients with a mild form of the disease exhibit rickets or osteomalacia and a functional defect of exercise. Odontohypophosphatasia shows only dental manifestations. To date, 302 mutations in the ALPL gene have been reported, mainly single-nucleotide substitutions, and the relationships between phenotype and genotype have been partially elucidated. An established treatment for HPP was not available until the recent development of enzyme replacement therapy. The first successful enzyme replacement therapy in model mice using a modified human TNAP protein (asfotase alfa) was reported in 2008, and subsequently success in patients with severe form of the disease was reported in 2012. In 2015, asfotase alfa was approved in Japan in July, followed by in the EU and Canada in August, and then by the US Food and Drug Administration in the USA in October. It is expected that therapy with asfotase alfa will drastically change treatments and prognosis of HPP.

Arecibo pulsar survey using ALFA: probing radio pulsar intermittency and transients

NARCIS (Netherlands)

Deneva, J.S.; Cordes, J.M.; McLaughlin, M.A.; Nice, D.J.; Lorimer, D.R.; Crawford, F.; Bhat, N.D.R.; Camilo, F.; Champion, D.J.; Freire, P.C.C.; Edel, S.; Kondratiev, V.I.; Hessels, J.W.T.; Jenet, F.A.; Kasian, L.; Kaspi, V.M.; Kramer, M.; Lazarus, P.; Ransom, S.M.; Stairs, I.H.; Stappers, B.W.; van Leeuwen, J.; Brazier, A.; Venkataraman, A.; Zollweg, J.A.; Bogdanov, S.

2009-01-01

We present radio transient search algorithms, results, and statistics from the ongoing Arecibo Pulsar ALFA (PALFA) survey of the Galactic plane. We have discovered seven objects through a search for isolated dispersed pulses. All of these objects are Galactic and have measured periods between 0.4

Towards a Total Cross Section Measurement with the ALFA Detector at ATLAS

CERN Document Server

Trzebiński, Maciej

2013-01-01

The main goals of the Absolute Luminosity For ATLAS (ALFA) detector is to provide an absolute luminosity and total cross section measurement. The measurement method used, the detector alignment and the quality of the collected data are discussed.

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

OpenAIRE

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...

Glycosylation of phenolic compounds by the site-mutated β-galactosidase from Lactobacillus bulgaricus L3.

Science.gov (United States)

Lu, Lili; Xu, Lijuan; Guo, Yuchuan; Zhang, Dayu; Qi, Tingting; Jin, Lan; Gu, Guofeng; Xu, Li; Xiao, Min

2015-01-01

β-Galactosidases can transfer the galactosyl from lactose or galactoside donors to various acceptors and thus are especially useful for the synthesis of important glycosides. However, these enzymes have limitations in the glycosylation of phenolic compounds that have many physiological functions. In this work, the β-galactosidase from Lactobacillus bulgaricus L3 was subjected to site-saturation mutagenesis at the W980 residue. The recombinant pET-21b plasmid carrying the enzyme gene was used as the template for mutation. The mutant plasmids were transformed into Escherichia coli cells for screening. One recombinant mutant, W980F, exhibited increased yield of glycoside when using hydroquinone as the screening acceptor. The enzyme was purified and the effects of the mutation on enzyme properties were determined in detail. It showed improved transglycosylation activity on novel phenolic acceptors besides hydroquinone. The yields of the glycosides produced from phenol, hydroquinone, and catechol were increased by 7.6% to 53.1%. Moreover, it generated 32.3% glycosides from the pyrogallol that could not be glycosylated by the wild-type enzyme. Chemical structures of these glycoside products were further determined by MS and NMR analysis. Thus, a series of novel phenolic galactosides were achieved by β-galactosidase for the first time. This was a breakthrough in the enzymatic galactosylation of the challenging phenolic compounds of great values.

Glycosylation of phenolic compounds by the site-mutated β-galactosidase from Lactobacillus bulgaricus L3.

Directory of Open Access Journals (Sweden)

Lili Lu

Full Text Available β-Galactosidases can transfer the galactosyl from lactose or galactoside donors to various acceptors and thus are especially useful for the synthesis of important glycosides. However, these enzymes have limitations in the glycosylation of phenolic compounds that have many physiological functions. In this work, the β-galactosidase from Lactobacillus bulgaricus L3 was subjected to site-saturation mutagenesis at the W980 residue. The recombinant pET-21b plasmid carrying the enzyme gene was used as the template for mutation. The mutant plasmids were transformed into Escherichia coli cells for screening. One recombinant mutant, W980F, exhibited increased yield of glycoside when using hydroquinone as the screening acceptor. The enzyme was purified and the effects of the mutation on enzyme properties were determined in detail. It showed improved transglycosylation activity on novel phenolic acceptors besides hydroquinone. The yields of the glycosides produced from phenol, hydroquinone, and catechol were increased by 7.6% to 53.1%. Moreover, it generated 32.3% glycosides from the pyrogallol that could not be glycosylated by the wild-type enzyme. Chemical structures of these glycoside products were further determined by MS and NMR analysis. Thus, a series of novel phenolic galactosides were achieved by β-galactosidase for the first time. This was a breakthrough in the enzymatic galactosylation of the challenging phenolic compounds of great values.

Lactose hydrolysis potential and thermal stability of commercial β-galactosidase in UHT and skimmed milk

Directory of Open Access Journals (Sweden)

Alessandra BOSSO

2016-03-01

Full Text Available Abstract The commercial enzyme (E.C. = 3.2.1.23 from Kluyveromyces lactis (liquid and Aspergillus oryzae(lyophilized was investigated for its hydrolysis potential in lactose substrate, UHT milk, and skimmed milk at different concentrations (0.7; 1.0 and 1.5%, pH values (5.0; 6.0; 6.5 and 7.0, and temperature (30; 35; 40 and 55 ºC. High hydrolysis rates were observed for the enzyme from K. lactis at pH 7.0 and 40 ºC, and from A. oryzae at pH 5.0 and 55 ºC. The enzyme from K. lactis showed significantly higher hydrolysis rates when compared to A. oryzae. The effect of temperature and β-galactosidase concentration on the lactose hydrolysis in UHT milk was higher than in skimmed milk, for all temperatures tested. With respect to the thermal stability, a decrease in hydrolysis rate was observed at pH 6.0 at 35 ºC for K. lactisenzyme, and at pH 6.0 at 55 ºC for the enzyme from A. oryzae. This study investigate the hydrolysis of β-galactosidase in UHT and skimmed milk. The knowledge about the characteristics of the β-galactosidase fromK. lactis and A. oryzae enables to use it most efficiently to control the enzyme concentration, temperature, and pH in many industrial processes and product formulations.

Pegylated interferons Lambda-1a and alfa-2a display different gene induction and cytokine and chemokine release profiles in whole blood, human hepatocytes and peripheral blood mononuclear cells.

Science.gov (United States)

Freeman, J; Baglino, S; Friborg, J; Kraft, Z; Gray, T; Hill, M; McPhee, F; Hillson, J; Lopez-Talavera, J C; Wind-Rotolo, M

2014-06-01

Pegylated interferon-lambda-1a (Lambda), a type III interferon (IFN) in clinical development for the treatment of chronic HCV infection, has shown comparable efficacy and an improved safety profile to a regimen based on pegylated IFN alfa-2a (alfa). To establish a mechanistic context for this improved profile, we investigated the ex vivo effects of Lambda and alfa on cytokine and chemokine release, and on expression of IFN-stimulated genes (ISGs) in primary human hepatocytes and peripheral blood mononuclear cells (PBMCs) from healthy subjects. Our findings were further compared with changes observed in blood analysed from HCV-infected patients treated with Lambda or alfa in clinical studies. mRNA transcript and protein expression of the IFN-λ-limiting receptor subunit was lower compared with IFN-α receptor subunits in all cell types. Upon stimulation, alfa and Lambda induced ISG expression in hepatocytes and PBMCs, although in PBMCs Lambda-induced ISG expression was modest. Furthermore, alfa and Lambda induced release of cytokines and chemokines from hepatocytes and PBMCs, although differences in their kinetics of induction were observed. In HCV-infected patients, alfa treatment induced ISG expression in whole blood after single and repeat dosing. Lambda treatment induced modest ISG expression after single dosing and showed no induction after repeat dosing. Alfa and Lambda treatment increased IP-10, iTAC, IL-6, MCP-1 and MIP-1β levels in serum, with alfa inducing higher levels of all mediators compared with Lambda. Overall, ex vivo and in vivo induction profiles reported in this analysis strongly correlate with clinical observations of fewer related adverse events for Lambda vs those typically associated with alfa. © 2014 John Wiley & Sons Ltd.

Análise da utilização na prática dos termos atribuídos aos fenômenos de enfermagem da CIPE-Versão Alfa

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Maria Miriam Lima da Nóbrega

2001-09-01

Full Text Available Este estudo teve como propósito a análise da utilização dos termos atribuídos aos fenômenos de enfermagem na prática das enfermeiras paraibanas. Os resultados evidenciam que todos os termos são utilizados pelas enfermeiras, com alguma freqüência. No que diz respeito à utilização dos termos, de acordo com o estrato amostral, os resultados evidenciam que as enfermeiras dos demais Municípios Paraibanos utilizam um maior número de termos, estatisticamente significativo, quando comparado com os dois outros estratos - João Pessoa e Campina Grande. A partir dos resultados do estudo, pode-se concluir, que os termos atribuídos aos fenômenos constantes na CIPE - Versão Alfa representam problemas ou situações da prática de enfermagem das enfermeiras paraibanas, corroborando a afirmação do Conselho Internacional de Enfermeiras de que estes termos são comuns às enfermeiras de todo o mundo.

O carvedilol potencializa o efeito antioxidante das vitaminas E e C na cardiopatia chagásica crônica

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Patrícia Budni

2013-10-01

Full Text Available FUNDAMENTO: A doença de Chagas continua a ser uma importante doença endêmica no país, sendo o acometimento cardíaco a sua manifestação mais grave. OBJETIVO: Verificar se o uso concomitante de carvedilol potencializará o efeito antioxidante das vitaminas E e C na atenuação do estresse oxidativo sistêmico na cardiopatia chagásica crônica. MÉTODOS: Foram estudados 42 pacientes com cardiopatia chagásica, agrupados de acordo com a classificação modificada de Los Andes, em quatro grupos: 10 pacientes no grupo IA (eletrocardiograma e ecocardiograma normais: sem envolvimento do coração, 20 pacientes do grupo IB (eletrocardiograma normal e ecocardiograma anormal: ligeiro envolvimento cardíaco, oito pacientes no grupo II (eletrocardiograma e ecocardiograma anormais, sem insuficiência cardíaca: moderado envolvimento cardíaco e quatro pacientes no grupo III (eletrocardiograma e ecocardiograma anormais com insuficiência cardíaca: grave envolvimento cardíaco. Os marcadores de estresse oxidativo foram medidos no sangue, antes e após um período de seis meses de tratamento com carvedilol e após seis meses de terapia combinada com vitaminas E e C. Os marcadores foram: atividades da superóxido dismutase, catalase, glutationa peroxidase, glutationa S-transferase e redutase, mieloperoxidase e adenosina deaminase, e os níveis de glutationa reduzida, de espécies reativas do ácido tiobarbitúrico, proteína carbonilada, vitamina E e óxido nítrico. RESULTADOS: Após o tratamento com carvedilol, todos os grupos apresentaram diminuições significativas dos níveis de proteína carbonilada e glutationa reduzida, enquanto os níveis de óxido nítrico e atividade da adenosina aumentaram significativamente apenas no grupo menos acometido (IA. Além disso, a maioria das enzimas antioxidantes mostrou atividades diminuídas nos grupos menos acometidos (IA e IB. Com a adição das vitaminas ao carvedilol houve diminuição dos danos em prote

Cooperation between two ClpB isoforms enhances the recovery of the recombinant {beta}-galactosidase from inclusion bodies

Energy Technology Data Exchange (ETDEWEB)

Guenther, Izabela [Department of Biochemistry, University of Gdansk, Wita Stwosza 59, 80-308 Gdansk (Poland); Zolkiewski, Michal [Department of Biochemistry, Kansas State University, Manhattan, KS 66506 (United States); Kedzierska-Mieszkowska, Sabina, E-mail: kedzie@biotech.ug.gda.pl [Department of Biochemistry, University of Gdansk, Wita Stwosza 59, 80-308 Gdansk (Poland)

2012-10-05

Highlights: Black-Right-Pointing-Pointer An important role of synergistic cooperation between the two ClpB isoforms. Black-Right-Pointing-Pointer Both ClpB isoforms are associated with IBs of {beta}-galactosidase. Black-Right-Pointing-Pointer ClpB is a key chaperone in IB protein release. -- Abstract: Bacterial ClpB is a molecular chaperone that solubilizes and reactivates aggregated proteins in cooperation with the DnaK chaperone system. The mechanism of protein disaggregation mediated by ClpB is linked to translocation of substrates through the central channel within the ring-hexameric structure of ClpB. Two isoforms of ClpB are produced in vivo: the full-length ClpB95 and the truncated ClpB80 (ClpB{Delta}N), which does not contain the N-terminal domain. The functional specificity of the two ClpB isoforms and the biological role of the N-terminal domain are still not fully understood. Recently, it has been demonstrated that ClpB may achieve its full potential as an aggregate-reactivating chaperone through the functional interaction and synergistic cooperation of its two isoforms. It has been found that the most efficient resolubilization and reactivation of stress-aggregated proteins occurred in the presence of both ClpB95 and ClpB80. In this work, we asked if the two ClpB isoforms functionally cooperate in the solubilization and reactivation of proteins from insoluble inclusion bodies (IBs) in Escherichia coli cells. Using the model {beta}-galactosidase fusion protein (VP1LAC), we found that solubilization and reactivation of enzymes entrapped in IBs occurred more efficiently in the presence of ClpB95 with ClpB80 than with either ClpB95 or ClpB80 alone. The two isoforms of ClpB chaperone acting together enhanced the solubility and enzymatic activity of {beta}-galactosidase sequestered into IBs. Both ClpB isoforms were associated with IBs of {beta}-galactosidase, what demonstrates their affinity to this type of aggregates. These results demonstrate a synergistic

Lactococcus lactis expressing food-grade β-galactosidase alleviates lactose intolerance symptoms in post-weaning Balb/c mice.

Science.gov (United States)

Li, Jingjie; Zhang, Wen; Wang, Chuan; Yu, Qian; Dai, Ruirui; Pei, Xiaofang

2012-12-01

The endogenous β-galactosidase expressed in intestinal microbes is demonstrated to help humans in lactose usage, and treatment associated with the promotion of beneficial microorganism in the gut is correlated with lactose tolerance. From this point, a kind of recombinant live β-galactosidase delivery system using food-grade protein expression techniques and selected probiotics as vehicle was promoted by us for the purpose of application in lactose intolerance subjects. Previously, a recombinant Lactococcus lactis MG1363 strain expressing food-grade β-galactosidase, the L. lactis MG1363/FGZW, was successfully constructed and evaluated in vitro. This study was conducted to in vivo evaluate its efficacy on alleviating lactose intolerance symptoms in post-weaning Balb/c mice, which were orally administered with 1 × 10⁶ CFU or 1 × 10⁸ CFU of L. lactis MG1363/FGZW daily for 4 weeks before lactose challenge. In comparison with naïve mice, the mice administered with L. lactis MG1363/FGZW showed significant alleviation of diarrhea symptoms in less total feces weight within 6 h post-challenge and suppressed intestinal motility after lactose challenge, although there was no significant increase of β-galactosidase activity in small intestine. The alleviation also correlated with higher species abundance, more Bifidobacterium colonization, and stronger colonization resistance in mice intestinal microflora. Therefore, this recombinant L. lactis strain effectively alleviated diarrhea symptom induced by lactose uptake in lactose intolerance model mice with the probable mechanism of promotion of lactic acid bacteria to differentiate and predominantly colonize in gut microbial community, thus making it a promising probiotic for lactose intolerance subjects.

Biochemical characterization of a novel β-galactosidase from Paenibacillus barengoltzii suitable for lactose hydrolysis and galactooligosaccharides synthesis.

Science.gov (United States)

Liu, Yu; Chen, Zhou; Jiang, Zhengqiang; Yan, Qiaojuan; Yang, Shaoqing

2017-11-01

A β-galactosidase gene (PbBGal2A) was cloned from Paenibacillus barengoltzii and expressed in Escherichia coli. The in silico analysis of the deduced amino acid sequences revealed that PbBGal2A shared the highest identity of 40% with the characterized glycoside hydrolase (GH) family 2 β-galactosidase from Actinobacillus pleuropneumoniae. The recombinant β-galactosidase (PbBGal2A) was purified with a molecular mass of 124.2kDa on SDS-PAGE. The optimal pH and temperature of PbBGal2A were determined to be pH 7.5 and 45°C, respectively. PbBGal2A was stable within pH 6.0-8.0 and up to 45°C. It completely hydrolyzed the lactose in milk and whey powder solution. In addition, PbBGal2A exhibited high transglycosylation activity and a maximum yield of 47.9% (w/w) for galactooligosaccharides (GOS) production was obtained in 8h at a lactose concentration of 350g/L. These properties make PbBGal2A an ideal candidate for commercial use in the production of lactose-free milk and GOS. Copyright © 2017. Published by Elsevier B.V.

Transgalactosylation/Hydrolysis Ratios of Various β-Galactosidases Catalyzing Alkyl-β-Galactoside Synthesis in Single-Phased Alcohol Media

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Eleonora Winkelhausen

2008-01-01

Full Text Available Three microbial galactosidases, Aspergillus oryzae, Escherichia coli and Kluyveromyces marxianus β-galactosidase, were used as catalysts for transgalactosylation synthesis of alkyl-β-galactosides in single-phased alcohol media. Their selectivity towards different alcohol nucleophiles was quantified by determining the transgalactosylation/hydrolysis ratio in the water/alcohol mixtures containing water in concentrations below the level of saturation. p-Nitrophenyl-β-galactoside was used as a glycosyl donor at a concentration of 10 mM. Both the total reaction rate (transgalactosylation+hydrolysis and the ratio between the transgalactosylation (alcoholysis and hydrolysis increased with the increase of water activity. Although the A. oryzae β-galactosidase showed relatively low total activity (3.13 μmol/(min·mg protein, it exhibited the highest selectivity towards the hexanol nucleophile among the examined enzymes (0.65. The selectivity values in all the examined cases were below one, which implies that the hydrolysis, and not the synthesis, was the dominating reaction. The total reaction rate (transgalactosylation+hydrolysis was strongly affected by the water activity, and for the specific water activity in the different alcohols, it increased in the following order: n-octanol, n-hexanol, n-butanol.

Human factors engineering and design validation for the redesigned follitropin alfa pen injection device.

Science.gov (United States)

Mahony, Mary C; Patterson, Patricia; Hayward, Brooke; North, Robert; Green, Dawne

2015-05-01

To demonstrate, using human factors engineering (HFE), that a redesigned, pre-filled, ready-to-use, pre-asembled follitropin alfa pen can be used to administer prescribed follitropin alfa doses safely and accurately. A failure modes and effects analysis identified hazards and harms potentially caused by use errors; risk-control measures were implemented to ensure acceptable device use risk management. Participants were women with infertility, their significant others, and fertility nurse (FN) professionals. Preliminary testing included 'Instructions for Use' (IFU) and pre-validation studies. Validation studies used simulated injections in a representative use environment; participants received prior training on pen use. User performance in preliminary testing led to IFU revisions and a change to outer needle cap design to mitigate needle stick potential. In the first validation study (49 users, 343 simulated injections), in the FN group, one observed critical use error resulted in a device design modification and another in an IFU change. A second validation study tested the mitigation strategies; previously reported use errors were not repeated. Through an iterative process involving a series of studies, modifications were made to the pen design and IFU. Simulated-use testing demonstrated that the redesigned pen can be used to administer follitropin alfa effectively and safely.

Preparation And Properties Of Bionanocomposite Films Reinforced With Nanocellulose Isolated From Moroccan Alfa Fibres

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Youssef Benyoussif

2015-09-01

Full Text Available Nanocellulose (NC were extracted from the Moroccan Alfa plant (Stipa tenacissima L. and characterised. These Alfa cellulosic nanoparticles were used as reinforcing phase to prepare bionanocomposite films using carboxymethyl cellulose as matrix. These films were obtained by the casting/evaporation method. The crystallinity of NC was analysed by X-ray diffraction, the dimension of NC by atomic force microscopy, molecular interactions due to incorporation of NC in carboxymethyl cellulose (CMC matrix were supported by Fourier transforms infrared (FTIR spectroscopy. The properties of the ensuing bionanocomposite films were investigated using tensile tests, water vapour permeability (WVP study and thermogravimetric analysis. With the progress of purification treatment of cellulose, the crystallinity is improved compared to the untreated fibres; this can be explained by the disappearance of the amorphous areas in cellulose chain of the plant. Consequently, the tensile modulus and tensile strength of CMC film increased by 60 and 47%, respectively, in the bionanocomposite films with 10 wt% of NC, and decrease by 8.6% for WVP with the same content of NC. The NC obtained from the Moroccan Alfa fibres can be used as a reinforcing agent for the preparation of bionanocomposites, and they have a high potential for the development of completely biodegradable food packaging materials.

Alfa-Laval plate heat exchangers for the power industries

International Nuclear Information System (INIS)

Kitae, Junnosuke; Mtsuura, Kazuyuki

1979-01-01

Within power-generating plants, the transfer and conversion of heat energy of very large quantity are carried out in the process of energy conversion, accordingly the importance of heat exchangers is very high. Heretofore, multi-tube heat exchangers have been used mostly, but Alfa-Laval group developed the heat exchanger with very high efficiency to incorporate it effectively into a power-generating plant. In this plate type heat exchanger, the heat transfer efficiency is very high, and the quantity of stagnation is small as it is compact, consequently it is suitable to the secondary cooling for power-generating plant or the heat exchange of high-priced liquid heat media such as heavy water. Originally, plate type heat exchangers were used for food and chemical industries, therefore the prevention of mixing two liquids, sanitary construction, and corrosion resistance were required. Then they were adopted in iron and steel industry, and large thermal load, large heat transfer area and corrosion resistance to sea water were required. They were adopted in a nuclear power plant for the first time in 1964. In this heat exchanger, channels are formed with corrugated metal sheets, and titanium, stainless steels, Incoloy, Hastelloy and others are used as occasion demands. The Alfa-Laval heat exchangers and their features are explained. (Kako, I.)

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.

Patient evaluation of the use of follitropin alfa in a prefilled ready-to-use injection pen in assisted reproductive technology: an observational study

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Welcker J

2010-09-01

Full Text Available Abstract Background Self-administration of recombinant human follicle-stimulating hormone (r-hFSH can be performed using injection pen devices by women undergoing assisted reproductive technology procedures. The objective of this study was to explore the use of the prefilled follitropin alfa pen in routine assisted reproductive technology procedures in Germany. Methods This prospective, observational study was conducted across 43 German IVF centres over a period of 1.75 years. Patients who had used the prefilled follitropin alfa pen in the current or a previous cycle of controlled ovarian stimulation completed a questionnaire to assess their opinions of the device. Results A total of 5328 patients were included in the study. Of these, 2888 reported that they had previous experience of daily FSH injections. Significantly more patients reported that less training was required to use the prefilled follitropin alfa pen than a syringe and lyophilized powder (1997/3081 [64.8%]; p Conclusions In this questionnaire-based survey, routine use of the prefilled follitropin alfa pen was well accepted and associated with favourable patient perceptions. Users of the pen found it easier to initially learn how to use, and subsequently use, than other injection methods. In general, the prefilled follitropin alfa pen was the preferred method for self-administration of gonadotrophins. Together with previous findings, the results here indicate a high level of patient satisfaction among users of the prefilled follitropin alfa pen for daily self-administration of r-hFSH.

A comparison of the activities of three beta-galactosidases in aqueous-organic solvent mixtures

NARCIS (Netherlands)

Yoon, JH; Mckenzie, D

2005-01-01

The hydrolytic activities of beta-galactosidases from three different sources have been determined in various 50% (v/v) organic solvent-buffer mixtures with a view to finding solvent systems of reduced water content suitable for the synthesis of glycosides and oligosaccharides. K. fragilis

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

A pilot trial on subjects with lactose and/or oligosaccharides intolerance treated with a fixed mixture of pure and enteric-coated α- and ß-galactosidase

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Di Pierro F

2015-02-01

Full Text Available Francesco Di Pierro,1 Alexander Bertuccioli,2 Eleonora Marini,3 Leandro Ivaldi4 1Velleja Research, Milan, Italy; 2Italian Association Fitness and Medicine, Fano, PU, Italy; 3Pharmextracta, Pontenure, Piacenza, Italy; 4Digestive Endoscopic Department, Ceva Hospital, Ceva, Cuneo, Italy Aim: Lactose and complex carbohydrates maldigestion, common food intolerances due to low gut content of α- and ß-galactosidase, lead to abdominal symptoms including pain, diarrhea, bloating, flatulence, and cramping. Commonly, intolerant patients are advised by physicians to avoid the offending foods (dairy foods, cereals, beans, etc. This food-limiting option, however, has possible nutritional risks. We have therefore evaluated the impact of using pure, enteric-coated α- plus ß-galactosidase on gut symptoms in intolerant subjects instead of avoidance of the offending foods. Methods: Sixteen subjects intolerant to lactose and/or complex carbohydrates were enrolled and evaluated in terms of gut symptoms with 1 uncontrolled diet, 2 diet devoid of offending foods, and 3 uncontrolled diet along with pure, enteric-coated α- and ß-galactosidase (DDM Galactosidase®. Results: Even with the uncontrolled diet, intolerant subjects treated with DDM Galactosidase® exhibited reduced gut symptoms (bloating, flatulence, diarrhea, and constipation significantly better than the control treatment as well as having a diet devoid of offending foods. Conclusion: DDM Galactosidase® is a valid and safe optional treatment to counteract lactose and complex carbohydrate intolerance in subjects who prefer not to avoid, at least partially, offending foods. Keywords: lactase, lactose intolerance, complex carbohydrate intolerance

Corifollitropin alfa followed by rFSH in a GnRH antagonist protocol for poor ovarian responder patients

DEFF Research Database (Denmark)

Polyzos, Nikolaos P; Devos, Michel; Humaidan, Peter

2013-01-01

OBJECTIVE: To identify whether women with poor ovarian response may benefit from treatment with corifollitropin alfa in a GnRH antagonist protocol. DESIGN: Retrospective pilot study. SETTING: University-based tertiary care center. PATIENT(S): Poor ovarian responders fulfilling the Bologna criteria...... developed by European Society for Human Reproduction and Embryology Consensus Group. INTERVENTION(S): Corifollitropin alfa (150 μg) followed by 300 IU rFSH in a GnRH antagonist protocol. MAIN OUTCOME MEASURE(S): Endocrinologic profile and ongoing pregnancy rates. RESULT(S): Among 43 women treated...

alfa-Deuterium kinetic isotope effects in reactions of methyllithium. Is better aggregation the cause of lower reactivity?

DEFF Research Database (Denmark)

Holm, Torkil

1996-01-01

The value of kH/kD for alfa deuterium kinetic isotope effects for the reaction of methyllithium and methylmagnesium iodid with a series of substrates are consistently ca. 10-15 % higher for the lithium reagent. This may indicate a pre-equilibrium......The value of kH/kD for alfa deuterium kinetic isotope effects for the reaction of methyllithium and methylmagnesium iodid with a series of substrates are consistently ca. 10-15 % higher for the lithium reagent. This may indicate a pre-equilibrium...

AVALIAÇÃO DAS DIETAS SIMPLIFICADAS E SEMI-SIMPLIFICADAS, COM BASE NA MISTURA DE FORRAGEIRAS, COM E SEM ENZIMAS, PARA COELHOS EM CRESCIMENTO

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Luiz Carlos Machado

2012-09-01

Full Text Available The simplified and half-simplified diets are a new technology that combines economy and welfare intestinal. The main objective of this study was to evaluate the effect of simplified and half-simplified diets, with and without exogenous enzymes, on the productive performance of growing rabbits. A Reference diet, two simplified diets and eight half-simplified diets had been utilized. The main fibrous sources used were the alfalfa hay (ALH, hay from the upper third of the cassava foliage (HUTCF and cassava leaf meal (CLM, using a completely randomized design with 11 treatments and eight repetitions. The enzymes used were the carbohydrases (alpha-galactosidase, galactomanase, xylanase and beta-glucanase and phytase. The consumption of animals fed with diets with high inclusion of CLM was lower than the other. The daily weight gain was higher in animals fed with the reference diet, half-simplified diet based on HUTCF and half-simplified diets based on the mix of ALH and CLM. A positive effect of exogenous enzymes inclusion, on the performance parameters, was not observed. Based on these results we conclude that the simplified and half-simplified diets provide decrease in productive performance. Considering the proportionate economy in the cost of feed, the half simplified based on HUTCF may be an interesting alternative.

Evaluation of pharmacokinetics, user handling, and tolerability of peginterferon alfa-2a (40 kDa delivered via a disposable autoinjector device

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Varunok P

2011-11-01

Full Text Available Peter Varunok1, Eric Lawitz2, Kimberly L Beavers3, Gary Matusow4, Ruby Leong5, Nathalie Lambert6, Coen Bernaards7, Jonathan Solsky5, Barbara J Brennan5, Cynthia Wat8, Anne Bertasso51Gastroenterology Associates, Poughkeepsie, NY, USA; 2Alamo Medical Research, San Antonio, TX, USA; 3Asheville Gastroenterology, Asheville, NC, USA; 4Gastroenterology Group, South Jersey, NJ, USA; 5Roche, Nutley, NJ, USA; 6Roche, Basel, Switzerland; 7Roche, San Francisco, CA, USA; 8Roche, Welwyn, UKBackground: Peginterferon alfa-2a (40 kDa is currently administered using a prefilled syringe. The peginterferon alfa-2a disposable autoinjector is a new safety-engineered device designed to facilitate injection and reduce the risk of needlestick injuries. The analysis of two open-label Phase I trials evaluated the pharmacokinetics, successful administration, and tolerability of peginterferon alfa-2a when using the autoinjector. The studies were performed to support the filing and registration of the autoinjector device.Methods: In trial 1, 50 healthy adult subjects received one 180 µg dose of peginterferon alfa-2a via the autoinjector. Serial blood samples were collected predose, up to 336 hours following drug administration, and at follow-up (28 ± 3 days post-dosing for noncompartmental pharmacokinetic analysis. Trial 2 randomized 60 adult patients with chronic hepatitis C to 180 µg peginterferon alfa-2a once weekly by the autoinjector or prefilled syringe for 3 weeks followed by the alternative device (prefilled syringe or autoinjector, respectively for 3 weeks. Patients also received ribavirin. Administration by the devices was evaluated under direct observation by a study staff member and by patient subjective assessment.Results: In trial 1, following a single dose of peginterferon alfa-2a, the maximum plasma concentration was 16.1 ± 5.3 ng/mL (mean ± standard deviation, and area under the concentration time curve (0–168 hours was 1996 ± 613 ng · hour

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

OpenAIRE

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...

A Randomized Study of Peginterferon Lambda-1a Compared to Peginterferon Alfa-2a in Combination with Ribavirin and Telaprevir in Patients with Genotype-1 Chronic Hepatitis C.

Science.gov (United States)

Flisiak, Robert; Shiffman, Mitchell; Arenas, Juan; Cheinquer, Hugo; Nikitin, Igor; Dong, Yuping; Rana, Khurram; Srinivasan, Subasree

2016-01-01

A randomized, double-blind, multinational, phase 3 study was conducted comparing the efficacy and safety of peginterferon lambda-1a (Lambda)/ribavirin (RBV)/telaprevir (TVR) vs. peginterferon alfa-2a (Alfa)/RBV/TVR in patients with chronic hepatitis C virus (HCV) genotype-1 (GT-1) infection. Patients (treatment-naïve or relapsers on prior Alfa/RBV treatment) were randomly assigned in a 2:1 ratio to receive Lambda/RBV/TVR or Alfa/RBV/TVR. Total duration of treatment was either 24 or 48 weeks (response-guided treatment), with TVR administered for the first 12 weeks. The primary endpoint was the proportion of patients who achieved a sustained virologic response at post treatment week 12 (SVR12), which was tested for noninferiority of Lambda/RBV/TVR. A total of 838 patients were enrolled, and 617 were treated; 411 and 206 patients received Lambda/RBV/TVR and Alfa/RBV/TVR, respectively. The majority of patients were treatment-naïve, with HCV GT-1b and a high baseline viral load (≥800,000 IU/mL). Less than 10% of patients had cirrhosis (Lambda, 7.5%; Alfa, 6.8%). Lambda/RBV/TVR did not meet the criterion for noninferiority (lower bound of the treatment difference interval was -12.3%); the SVR12 in all patients (modified intent-to-treat) was 76.2% in the Lambda arm and 82.0% in the Alfa arm. Overall, the frequency of adverse events in each arm was comparable (Lambda, 91.7%; Alfa, 97.1%). As expected based on the safety profile of the 2 interferons, there were more hepatobiliary events observed in the Lambda arm and more hematologic events in the Alfa arm. In this comparison of Lambda/RBV/TVR and Alfa/RBV/TVR in patients who were treatment-naïve or had relapsed on prior Alfa/RBV treatment, Lambda failed to demonstrate noninferiority based on SVR12 results. Treatment with Lambda/RBV/TVR was associated with a higher incidence of relapse. More patients discontinued Lambda/RBV/TVR treatment during the first 4 weeks of study treatment, mainly due to hepatobiliary

Discovery of Intermediates of lacZ β-Galactosidase Catalyzed Hydrolysis Using dDNP NMR

DEFF Research Database (Denmark)

Kjeldsen, Christian; Ardenkjær-Larsen, Jan Henrik; Duus, Jens Ø.

2018-01-01

Using dissolution dynamic nuclear polarization, the sensitivity of single scan solution state 13C NMR can be improved up to 4 orders of magnitude. In this study, the enzyme lacZ β-galactosidase from Escherichia coli was subjected to hyperpolarized substrate, and previously unknown reaction...

Discovery of Intermediates of lacZ beta-Galactosidase Catalyzed Hydrolysis Using dDNP NMR

DEFF Research Database (Denmark)

Kjeldsen, Christian; Ardenkjær-Larsen, Jan Henrik; Duus, Jens Øllgaard

2018-01-01

Using dissolution dynamic nuclear polarization, the sensitivity of single scan solution state C-13 NMR can be improved up to 4 orders of magnitude. In this study, the enzyme lacZ beta-galactosidase from Escherichia coli was subjected to hyperpolarized substrate, and previously unknown reaction...

Characterization of a novel ß-galactosidase from Bifidobacterium adolescentis DSM 20083 active towards transgalactooligosaccharides

NARCIS (Netherlands)

Laere, van K.M.J.; Abee, T.; Schols, H.A.; Beldman, G.; Voragen, A.G.J.

2000-01-01

This paper reports on the effects of both reducing and nonreducing transgalactooligosaccharides (TOS) comprising 2 to 8 residues on the growth of Bifidobacterium adolescentis DSM 20083 and on the production of a novel -galactosidase (-Gal II). In cells grown on TOS, in addition to the

Salivary gland proteins of the human malaria vector, Anopheles dirus B (Diptera: Culicidae Proteínas das glândulas salivares do Anopheles dirus B (Diptera: Culicidae, vetor da malária humana

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Narissara Jariyapan

2007-02-01

Full Text Available Salivary gland proteins of the human malaria vector, Anopheles dirus B were determined and analyzed. The amount of salivary gland proteins in mosquitoes aged between 3 - 10 days was approximately 1.08 ± 0.04 µg/female and 0.1 ± 0.05 µg/male. The salivary glands of both sexes displayed the same morphological organization as that of other anopheline mosquitoes. In females, apyrase accumulated in the distal regions, whereas alpha-glucosidase was found in the proximal region of the lateral lobes. This differential distribution of the analyzed enzymes reflects specialization of different regions for sugar and blood feeding. SDS-PAGE analysis revealed that at least seven major proteins were found in the female salivary glands, of which each morphological region contained different major proteins. Similar electrophoretic protein profiles were detected comparing unfed and blood-fed mosquitoes, suggesting that there is no specific protein induced by blood. Two-dimensional polyacrylamide gel analysis showed the most abundant salivary gland protein, with a molecular mass of approximately 35 kilodaltons and an isoelectric point of approximately 4.0. These results provide basic information that would lead to further study on the role of salivary proteins of An. dirus B in disease transmission and hematophagy.Proteínas das glândulas salivares do Anopheles dirus B (Diptera: Culicidae, vetor da malária humana foram determinadas e analisadas. A quantidade de proteínas das glândulas salivares em mosquitos com três a 10 dias de idade foi de aproximadamente 1,08 ± 0,04 µg/ fêmea e de 0,1 ± 0,05 µg/macho. As glândulas salivares de ambos os sexos mostraram organização morfológica semelhante à de outros mosquitos anofelinos. Em fêmeas, apirase acumula-se nas regiões distais, enquanto alfa-glucosidase foi encontrada na região proximal dos lóbulos laterais. Esta distribuição diferencial das enzimas analisadas reflete a especialização de

Functionalized agarose as an effective and novel matrix for immobilizing Cicer arietinum β-galactosidase and its application in lactose hydrolysis

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Rukhsana Satar

Full Text Available Abstract The present study demonstrates the immobilization of β-galactosidase from Cicer arietinum on a simple and inexpensive matrix, glutaraldehyde functionalized agarose (GFA, to suggest its potential application in hydrolyzing whey lactose in biotechnology industries. The designed matrix provided large surface area for the immobilization of β-galactosidase, apart from exhibiting greater biocatalytic activity in terms of selectivity, loading and stability. GFA retained 83% enzyme activity as a result of immobilization. Soluble and GFA bound Cicer arietinum β-galactosidase showed the same pH and temperature-optima at pH 5.0 and at 50 °C, respectively. However, immobilized enzyme exhibited a greater fraction of activity at both acidic and basic pH, and at higher temperature ranges. GFA bound enzyme lost only 20 % enzyme in the presence of 3% galactose, and retained 70 % activity even after its sixth repeated use. Immobilized enzyme showed pronounced lactose hydrolysis from whey in batch processes at 55 °C as compared to enzyme in solution.

Accurate Learning with Few Atlases (ALFA): an algorithm for MRI neonatal brain extraction and comparison with 11 publicly available methods.

Science.gov (United States)

Serag, Ahmed; Blesa, Manuel; Moore, Emma J; Pataky, Rozalia; Sparrow, Sarah A; Wilkinson, A G; Macnaught, Gillian; Semple, Scott I; Boardman, James P

2016-03-24

Accurate whole-brain segmentation, or brain extraction, of magnetic resonance imaging (MRI) is a critical first step in most neuroimage analysis pipelines. The majority of brain extraction algorithms have been developed and evaluated for adult data and their validity for neonatal brain extraction, which presents age-specific challenges for this task, has not been established. We developed a novel method for brain extraction of multi-modal neonatal brain MR images, named ALFA (Accurate Learning with Few Atlases). The method uses a new sparsity-based atlas selection strategy that requires a very limited number of atlases 'uniformly' distributed in the low-dimensional data space, combined with a machine learning based label fusion technique. The performance of the method for brain extraction from multi-modal data of 50 newborns is evaluated and compared with results obtained using eleven publicly available brain extraction methods. ALFA outperformed the eleven compared methods providing robust and accurate brain extraction results across different modalities. As ALFA can learn from partially labelled datasets, it can be used to segment large-scale datasets efficiently. ALFA could also be applied to other imaging modalities and other stages across the life course.

Mango resistance to fruit flies. II - resistance of the alfa cultivar

Energy Technology Data Exchange (ETDEWEB)

Rossetto, C.J.; Bortoletto, N., E-mail: rossetto@iac.sp.gov.b [Agencia Paulista de Tecnologia dos Agronegocios (APTA), Votuporanga, SP (Brazil). Polo Regional do Noroeste Paulista; Walder, J.M.M.; Mastrangelo, T. de A., E-mail: jmwalder@cena.usp.b [Centro de Energia Nuclear na Agricultura (CENA/USP), Piracicaba, SP (Brazil); Carvalho, C.R.L.; Castro, J.V. de, E-mail: climonta@iac.sp.gov.b, E-mail: josalba@iac.sp.gov.b [Instituto Agronomico de Campinas, SP (Brazil); Pinto, A.C. de Q. [EMBRAPA, Brasilia, DF (Brazil); Cortelazzo, A.L., E-mail: angelo@unicamp.b [Universidade Estadual de Campinas (UNICAMP), SP (Brazil). Inst. de Biologia

2006-07-01

The percentage of infested mango fruits of five selected mango varieties was evaluated during three years under field conditions. Three varieties with field resistance to fruit flies had less then 10% of fruits infested. Tommy Atkins, the susceptible commercial check, had 42,9% and the susceptible check had 98.9 % of infested fruits. The three field resistant varieties plus the susceptible commercial check, Tommy Atkins, were further tested in laboratory, under caged conditions, with artificial infestation of Anastrepha obliqua. The attempts of oviposition and the number of pupae developed from each fruit were evaluated. Under caged conditions, the cultivar Alfa maintained its field resistance and Espada Stahl and IAC 111 lost the field resistance and were as susceptible as Tommy Atkins. The attempts of oviposition were positively and highly correlated with the number of pupae developed in the fruits. Non preference for oviposition was confirmed as the main mechanism of resistance of mango fruits to fruit flies. In the absence of a more susceptible variety (no choice test) the cultivar Alfa has kept the resistance (author)

Mango resistance to fruit flies. II - resistance of the alfa cultivar

International Nuclear Information System (INIS)

Rossetto, C.J.; Bortoletto, N.; Carvalho, C.R.L.; Castro, J.V. de; Pinto, A.C. de Q.; Cortelazzo, A.L.

2006-01-01

The percentage of infested mango fruits of five selected mango varieties was evaluated during three years under field conditions. Three varieties with field resistance to fruit flies had less then 10% of fruits infested. Tommy Atkins, the susceptible commercial check, had 42,9% and the susceptible check had 98.9 % of infested fruits. The three field resistant varieties plus the susceptible commercial check, Tommy Atkins, were further tested in laboratory, under caged conditions, with artificial infestation of Anastrepha obliqua. The attempts of oviposition and the number of pupae developed from each fruit were evaluated. Under caged conditions, the cultivar Alfa maintained its field resistance and Espada Stahl and IAC 111 lost the field resistance and were as susceptible as Tommy Atkins. The attempts of oviposition were positively and highly correlated with the number of pupae developed in the fruits. Non preference for oviposition was confirmed as the main mechanism of resistance of mango fruits to fruit flies. In the absence of a more susceptible variety (no choice test) the cultivar Alfa has kept the resistance (author)

Hydrolysis of whey lactose by immobilized β-galactosidase in a bioreactor with a spirally wound membrane.

Science.gov (United States)

Vasileva, Nastya; Ivanov, Yavor; Damyanova, Stanka; Kostova, Iliana; Godjevargova, Tzonka

2016-01-01

The β-galactosidase was covalently immobilized onto a modified polypropylene membrane, using glutaraldehyde. The optimal conditions for hydrolysis of lactose (4.7%) by immobilized β-galactosidase in a batch process were determined 13.6 U enzyme activity, 40°C, pH 6.8 and 10h. The obtained degree of hydrolysis was compared with results received by a free enzyme. It was found, that the lactose hydrolysis by an immobilized enzyme was 1.6 times more effective than the lactose hydrolysis by a free enzyme. It was determined that the stability of the immobilized enzyme was 2 times higher in comparison with the stability of free enzyme. The obtained immobilized system β-galactosidase/polypropylene membrane was applied to produce glucose-galactose syrup from waste whey. The whey characteristics and the different preliminary treatments of the whey were investigated. Then the whey lactose hydrolysis in a bioreactor by an immobilized enzyme on a spirally wound membrane was performed. The optimal membrane surface and the optimal flow rate of the whey through the membrane module were determined, respectively 100 cm(2) and 1.0 mL min(-1). After 10h, the degree of lactose hydrolysis was increased to 91%. The operation stability was studied. After 20th cycle the yield of bioreactor was 69.7%. Copyright © 2015 Elsevier B.V. All rights reserved.

Mutations of alpha-galactosidase A gene in two unusual cases of Fabry disease

NARCIS (Netherlands)

Beyer, EM; Kopishinskaya, SV; Van Amstel, JKP; Tsvetkova, [No Value

1999-01-01

The mutation analysis of alpha-galactosidase A gene was carried out in two families with Fabry disease described by us earlier. In the family P. a new point mutation E341K (a G to A transition at position 10999 of the gene) was identified. The mutation causes a Glu341Lys substitution in

Application of the thermostable β-galactosidase, BgaB, from Geobacillus stearothermophilus as a versatile reporter under anaerobic and aerobic conditions

DEFF Research Database (Denmark)

Jensen, Torbjørn Ølshøj; Pogrebnyakov, Ivan; Falkenberg, Kristoffer Bach

2017-01-01

Use of thermophilic organisms has a range of advantages, but the significant lack of engineering tools limits their applications. Here we show that β-galactosidase from Geobacillus stearothermophilus (BgaB) can be applicable in a range of conditions, including different temperatures and oxygen...... and encompassed fivefold variation. The experimental pipeline allowed construction and measurement of expression levels of the library in just 4 days. This β-galactosidase provides a promising tool for engineering of aerobic, anaerobic, and thermophilic production organisms such as Geobacillus species....

Improving Properties of a Novel β-Galactosidase from Lactobacillus plantarum by Covalent Immobilization

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Rocio Benavente

2015-04-01

Full Text Available A novel β-galactosidase from Lactobacillus plantarum (LPG was over-expressed in E. coli and purified via a single chromatographic step by using lowly activated IMAC (immobilized metal for affinity chromatography supports. The pure enzyme exhibited a high hydrolytic activity of 491 IU/mL towards o-nitrophenyl β-d-galactopyranoside. This value was conserved in the presence of different divalent cations and was quite resistant to the inhibition effects of different carbohydrates. The pure multimeric enzyme was stabilized by multipoint and multisubunit covalent attachment on glyoxyl-agarose. The glyoxyl-LPG immobilized preparation was over 20-fold more stable than the soluble enzyme or the one-point CNBr-LPG immobilized preparation at 50 °C. This β-galactosidase was successfully used in the hydrolysis of lactose and lactulose and formation of different oligosaccharides was detected. High production of galacto-oligosaccharides (35% and oligosaccharides derived from lactulose (30% was found and, for the first time, a new oligosaccharide derived from lactulose, tentatively identified as 3'-galactosyl lactulose, has been described.

Effect of Carrot-Juice on Exopolisaccharides and β-D Galactosidase Activity in Yogurt

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Lilik Eka Radiati

2016-09-01

Full Text Available Carrot juice and milk  were  blended and fermented by  culture bacteria Streptococcus thermophillus  and Lactobacillus bulgaricus.  Carrot juice  affect significantly  on   lactic acid content   (1.09± 0.12% – 1.15± 0.01%,   pH value (3.80±0.06 – 4.17± 0.10,   viscosity  (133±2.30 cP–146±2.10cP,  β-carotene (0-173.19±1.02 µg/g, EPS (11.90 ±0.50 - 18.00 ±0.40 mg/100g,   β-D-galactosidase activity (2.27±0.30-192.40±0.48 µ/g yogurt, but did not affect significantly on  bacteria number (9.0± 0.5 – 9.8±0.4 log CFU/g.    Carrot juice increased the yogurt culture activity with increasing acidifying, β-catotene, EPS and β-D-galactosidase, suggesting that yogurt could be fortified with carrot juice.

A new cold-adapted β-D-galactosidase from the Antarctic Arthrobacter sp. 32c – gene cloning, overexpression, purification and properties

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Kur Józef

2009-07-01

Full Text Available Abstract Background The development of a new cold-active β-D-galactosidases and microorganisms that efficiently ferment lactose is of high biotechnological interest, particularly for lactose removal in milk and dairy products at low temperatures and for cheese whey bioremediation processes with simultaneous bio-ethanol production. Results In this article, we present a new β-D-galactosidase as a candidate to be applied in the above mentioned biotechnological processes. The gene encoding this β-D-galactosidase has been isolated from the genomic DNA library of Antarctic bacterium Arthrobacter sp. 32c, sequenced, cloned, expressed in Escherichia coli and Pichia pastoris, purified and characterized. 27 mg of β-D-galactosidase was purified from 1 L of culture with the use of an intracellular E. coli expression system. The protein was also produced extracellularly by P. pastoris in high amounts giving approximately 137 mg and 97 mg of purified enzyme from 1 L of P. pastoris culture for the AOX1 and a constitutive system, respectively. The enzyme was purified to electrophoretic homogeneity by using either one step- or a fast two step- procedure including protein precipitation and affinity chromatography. The enzyme was found to be active as a homotrimeric protein consisting of 695 amino acid residues in each monomer. Although, the maximum activity of the enzyme was determined at pH 6.5 and 50°C, 60% of the maximum activity of the enzyme was determined at 25°C and 15% of the maximum activity was detected at 0°C. Conclusion The properties of Arthrobacter sp. 32cβ-D-galactosidase suggest that this enzyme could be useful for low-cost, industrial conversion of lactose into galactose and glucose in milk products and could be an interesting alternative for the production of ethanol from lactose-based feedstock.

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

OpenAIRE

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

OpenAIRE

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...

Characterization of ß-Galactosidase Isoforms from Bacillus circulans and Their Contribution to GOS Production

NARCIS (Netherlands)

Warmerdam, A.; Paudel, E.; Wanqing, J.; Boom, R.M.; Janssen, A.E.M.

2013-01-01

A ß-galactosidase preparation from Bacillus circulans consists of four isoforms called ß-gal-A, ß-gal-B, ß-gal-C, and ß-gal-D. These isoforms differ in lactose hydrolysis and galacto-oligosaccharide (GOS) synthesis at low substrate concentrations. For this reason, using a selection of the isoforms

Pancreatitis induced by pegylated interferon alfa-2b in a patient affected by chronic hepatitis C.

Science.gov (United States)

Cecchi, Enrica; Forte, Paolo; Cini, Elisabetta; Banchelli, Grazia; Ferlito, Chiara; Mugelli, Alessandro

2004-01-01

A middle-aged man was admitted to the ED because of nausea and vomiting, abdominal distention and fainting. A blood analysis revealed high levels of serum amylase and lipase, confirming a diagnosis of acute pancreatitis. The history showed that the patient had self-administered a single dose of pegylated interferon alfa-2b and ribavirin daily for 7 days for chronic hepatitis C. The medications were stopped and his condition gradually improved. In agreement with the literature and the Naranjo algorythm result, pegylated interferon alfa-2b is associated with acute pancreatitis. Identification of a few signs and symptoms is the first 'signal' in preventing a serious drug-induced adverse event.

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.

Elosulfase alfa enzyme replacement therapy attenuates disease progression in a non-ambulatory Japanese patient with Morquio A syndrome (case report

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Misako Hiramatsu

2017-12-01

Full Text Available Enzyme replacement therapy (ERT with elosulfase alfa is the only approved therapy in Japan for patients with Morquio A syndrome, a lysosomal storage disorder inherited in an autosomal recessive fashion. The experience with ERT in severely affected, non-ambulatory patients has not been reported in previous studies. This case report describes clinical evidence for the 1-year efficacy and safety of ERT with elosulfase alfa in a severely affected, non-ambulatory, 47-year-old patient with Morquio A syndrome who needs intensive respiratory management. ERT with elosulfase alfa was well tolerated in this patient. Because of the possibility of potential hypersensitivity adverse events, special attention is needed when using ERT in patients with respiratory disorders. However, under the appropriate management of specialists, the patient in this case report showed significant respiratory improvement after starting ERT, and abdominal bloating was improved by gas evacuation. In addition, the patient was able to lift up her arms, reach behind her back, and move her legs slightly, and she recovered her grip strength. Her hearing loss improved and she could hear without a hearing aid. This report shows that ERT with elosulfase alfa can be used with appropriate respiratory care in patients with severe respiratory dysfunction.

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.

Efeito de agentes anti-hipertensivos sobre as células estreladas durante a regeneração hepática em ratos

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Ramalho Fernando S.

2000-01-01

Full Text Available O efeito de um inibidor da enzima conversora da angiotensina (lisinopril, de um antagonista do receptor da angiotensina II (losartan e da bradicinina na população de células estreladas (CE durante o fenômeno regenerativo hepático foi estudado. Ratos machos Wistar receberam lisinopril, losartan, bradicinina ou solução salina em volumes proporcionais, intraperitonealmente, antes e após hepatectomia parcial a 70% (HP. Cinco animais de cada grupo experimental e controle foram sacrificados sob anestesia com éter em 36 horas após a HP. A população de CE marcadas para alfa-actina de músculo liso foi estimada nas zonas periportal e pericentral das amostras hepáticas. A população de CE foi menor no grupo tratado com losartan, e maior nos grupos tratados com bradicinina e lisinopril que no grupo controle. Estes resultados sugerem que o losartan pode inibir, e a bradicinina e o lisinopril podem estimular a população de CE durante a regeneração hepática em ratos.

Atividade de fosfatases em gramíneas forrageiras em resposta à disponibilidade de fósforo no solo e à altura de corte das plantas Phosphatase activity in forage grasses as influenced by soil phosphorus availability and plant cutting height

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Flancer Novais Nunes

2008-10-01

Full Text Available A eficiência de utilização de um nutriente, expressa pela relação entre a biomassa produzida e a quantidade absorvida, é uma importante característica adaptativa das plantas, particularmente daquelas cultivadas em solos com baixa disponibilidade desse nutriente. Maior eficiência de utilização de P (EUP é conferida pela maior taxa de remobilização de P, que consiste na translocação desse nutriente para regiões de maior demanda metabólica na planta. Maiores taxas de remobilização têm sido associadas a maiores atividades das enzimas: fosfatase ácida (APase e ribonuclease (RNase. Neste trabalho, avaliou-se a atividade dessas enzimas em plantas de Brachiaria decumbens, pouco exigente, e de Panicum maximum cv. Tanzânia, mais exigente em P, cultivadas em solos que receberam diferentes doses desse nutriente e submetidas a diferentes alturas de corte. O ensaio foi realizado em casa de vegetação, utilizando amostra do horizonte B de um Latossolo Vermelho-Amarelo argiloso. Os tratamentos resultaram da combinação fatorial das duas gramíneas, das doses de 100, 200 e 500 mg dm-3 de P e das três alturas de corte: sem corte e corte a 15 e 30 cm a partir da superfície do solo para Brachiaria; e sem corte e corte a 20 e 40 cm para Panicum. As unidades experimentais consistiram de vasos com 10 dm³ de solo, com 10 plantas. Os resultados indicaram que não houve diferença significativa na produção de biomassa das duas gramíneas, mas a adição de P propiciou maior produção e maior teor desse nutriente na parte aérea de ambas. As plantas submetidas a cortes mais severos apresentaram menor produção e maiores teores de P na parte aérea. Ocorreu efeito negativo das doses de P na atividade da APase e da RNase nas duas forrageiras. As plantas cultivadas com a menor dose de P apresentaram maior atividade dessas enzimas e maior EUP. A atividade de ambas as fosfatases decresceu com a idade das plantas. No tratamento com o corte mais

Structural and functional analysis of lysosomal ss-galactosidase and its relation to the protective protein.

NARCIS (Netherlands)

H. Morreau (Hans)

1992-01-01

textabstractLysosomal B-galactosidase is the glycosidase, that cleaves B-linked galactosyl mmenes from a variety of natural and synthetic substrates. In normal tissues of various species this enzyme appears to associate with two other hydrolases, N-acetyl-o:-neuraminidase and the protective protein.

Entrappment of alkaline protease and {beta}-galactosidase in radiation stitched together poly-N-vinylcaprolactam.; Vklyuchenie shchelochnoj proteazy i {beta}-galaktozidazy v radiatsionno sshityj poli-N-vinilkaprolaktam.

Energy Technology Data Exchange (ETDEWEB)

Davidenko, T I; Kravchenko, I A [AN Ukrainskoj SSR, Odessa (Ukraine). Fiziko-Khimicheskij Inst.

1996-03-01

The gel formations by poly-N-vinylcaprolactam upon its {gamma}-irradiation by the 20-25 kGy dose as a results of partial polymer stitching together is shown, which is confirmed by the CD-and thermogravimetric data. By the alkaline protease and {beta}-galactosidase entrapment in poly-N- vinylcaprolactam stitched together by {gamma}-irradiation, the active preparations are obtained with 90-98 % and 30-35 % activity retained for alkaline protease and {beta}-galactosidase, respectively. The increased stability of alkaline protease at acidic pH values and higher temperature was noted, and for {beta}-galactosidase - the possibility of repeated use of the obtained preparation for lactose hydrolysis.

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.

The expression of β-galactosidase during long-term cultured goat skin fibroblasts and the effect of donor cell passage on in vitro development of nuclear transfer embryos.

Science.gov (United States)

Liu, Haijun; Peng, Hui; Liu, Fang; Ma, Qun; Zhang, Wenchang

2016-05-01

The present study aimed to detect the expression of β-galactosidase during long-term cultured goat skin fibroblasts and investigate the effects of donor goat age, sex, and cell passage on senescence and the effects of donor cell passage on in vitro development of nuclear transfer embryos. The results showed that, in the same cell passage, more β-galactosidase-positive cells were detected in cells from older donors than younger donors. Irrespective of the donor age, the number of positive cells was higher in later passages from passages 20 to 50. In the same passage from 20 to 50, the β-galactosidase-positive rate was higher in cells from 5-yr female goat than 5-yr male goat. Using fibroblasts from male goats at various passages as donor cells, reconstructed embryos had similar fusion and cleavage rates, but the blastocyst rate was higher for cells at passages 10 and 20 than passage 30. In conclusion, donor goat age and cell passage had significant effects on the β-galactosidase-positive rate; also, cells from 5-yr female goat had a higher β-galactosidase-positive rate than those from 5-yr male goat, and the donor cell passage affected the developmental potential of nuclear transfer embryos.

Structural prediction and comparative docking studies of psychrophilic β- Galactosidase with lactose, ONPG and PNPG against its counter parts of mesophilic and thermophilic enzymes.

Science.gov (United States)

Kumar, Ponnada Suresh; Pulicherla, Kk; Ghosh, Mrinmoy; Kumar, Anmol; Rao, Krs Sambasiva

2011-01-01

Enzymes from psychrophiles catalyze the reactions at low temperatures with higher specific activity. Among all the psychrophilic enzymes produced, cold active β-galactosidase from marine psychrophiles revalorizes a new arena in numerous areas at industrial level. The hydrolysis of lactose in to glucose and galactose by cold active β-galactosidase offers a new promising approach in removal of lactose from milk to overcome the problem of lactose intolerance. Herein we propose, a 3D structure of cold active β-galactosidase enzyme sourced from Pseudoalteromonas haloplanktis by using Modeler 9v8 and best model was developed having 88% of favourable region in ramachandran plot. Modelling was followed by docking studies with the help of Auto dock 4.0 against the three substrates lactose, ONPG and PNPG. In addition, comparative docking studies were also performed for the 3D model of psychrophilic β-galactosidase with mesophilic and thermophilic enzymes. Docking studies revealed that binding affinity of enzyme towards the three different substrates is more for psychrophilic enzyme when compared with mesophilic and thermophilic enzymes. It indicates that the enzyme has high specific activity at low temperature when compared with mesophilic and thermophilic enzymes.

Desenvolvimento de populações duplo-haplóides de cevada cervejeira associadas à atividade das enzimas (1-3, 1-4-β-glucanases Development of doubled-haploids populations in malting barley associated to activity of enzymes (1-3, 1-4-β-glucanases

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Janaína Endres Georg-Kraemer

2011-05-01

Full Text Available Populações duplo-haplóides apresentam especial vantagem para análises genéticas, uma vez que a informação que elas oferecem pode ser maximizada, devido ao fato que todos os locos encontram-se em homozigose. O objetivo deste trabalho foi o desenvolvimento de duas populações duplo-haplóides (DHs de cevada (Hordeum vulgare ssp. vulgare L. segregantes para a atividade das enzimas (1-3, 1-4-β-glucanases, através da técnica de cultura de anteras. Foram realizados dois cruzamentos com cultivares contrastantes para esta característica. As cultivares parentais selecionadas foram 'MN 698' e 'CEV 97047', para o desenvolvimento da população "malte verde" (MV, e 'Embrapa 127' e 'CEV 96025' para o desenvolvimento da população "malte seco" (MS. Foram cultivadas 10.734 anteras da população MS e 4.139 anteras da população MV. A população MV produziu 50% mais plântulas verdes quando comparada à população MS, refletindo a importância do genótipo na resposta à cultura de anteras e na regeneração. A maioria das plantas adultas duplo-haplóides foi obtida através da duplicação espontânea in vitro do genoma haplóide, ocorrendo em 66% das plantas da população MS e 76% das plantas da população MV. Também foram observadas, em menor frequência, plantas haplóides, triplóides e tetraplóides. Através da técnica de cultura de anteras, foram desenvolvidas 204 linhagens duplo-haplóides, das quais 72 linhagens são da população "malte seco" e 132 linhagens são da população "malte verde". Este material constitui um importante germoplasma para o melhoramento genético da cevada.Doubled haploid populations offer special advantages in genetic analyses, since the information they provide may be maximized due to the fact that all loci are homozygous. The aim of this study was to develop two barley (Hordeum vulgare ssp.vulgare L. doubled-haploid (DHs populations segregating to (1-3, 1-4-β-glucanases activity, utilizing the anther

Milhos com diferentes texturas de endosperma e adição de alfa-amilase na dieta de leitões The effect of corn kernel texture and alpha-amylase addition in performance and digestibility of diets for weaned pigs

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Vanessa Piovesan

2011-11-01

Full Text Available Dois experimentos foram realizados para avaliar a influência de milhos com diferentes texturas de endosperma e adição de alfa amilase na digestibilidade de dietas e no desempenho de leitões na fase de creche. No ensaio de digestibilidade, foram usados 12 leitões castrados (7,96±0,31kg, alojados individualmente em gaiolas de metabolismo, enquanto no desempenho utilizaram-se 75 leitões (7,87±0,67kg distribuídos em 15 gaiolas de creche. Nos dois experimentos, os tratamentos consistiram de três dietas formuladas para serem isonutritivas, mas com milhos de diferentes texturas e adição ou não de alfa amilase, conforme descrito a seguir: 1. grãos de milho semi-dentado; 2. grãos de milho duro e; 3. grãos de milho duro + alfa-amilase. Os resultados de metabolismo mostraram que os valores de energia digestível (ED e metabolizável (EM das dietas foram influenciados pelos tipos de milho, sendo o semi-dentado superior (P0,05 entre os tratamentos no consumo de ração, ganho de peso e/ou conversão alimentar. Dietas com milho duro ou semi-dentado e adição de alfa-amilase não alteram o desempenho dos leitões. Por outro lado, dietas contendo milho semi-dentado apresentam maior energia digestível que dietas com milho duro e a adição de alfa-amilase aumenta a energia de dietas com milho duro.Two experiments were carried out to evaluate the effects of diets with different corn grain textures and amylase supplementation on nutrients digestibility and nursery piglets' performance. Twelve castrated piglets (7.96±0.31kg were individually housed in metabolic crates and 75 piglets (7.87±0.67kg penned in 15 nursery pens. In both experiments the treatments consisted of three isonutritive diets: 1. semi dent corn grain; 2. flint corn grain, and 3. flint corn grain + amylase. There was effect of treatments (P0.05 the performance characteristics studied. In conclusion, diets with flint or semi dent corns and amylase supplementation did not improve

An open-label clinical trial to investigate the efficacy and safety of corifollitropin alfa combined with hCG in adult men with hypogonadotropic hypogonadism.

Science.gov (United States)

Nieschlag, Eberhard; Bouloux, Pierre-Marc G; Stegmann, Barbara J; Shankar, R Ravi; Guan, Yanfen; Tzontcheva, Anjela; McCrary Sisk, Christine; Behre, Hermann M

2017-03-07

Hypogonadotropic hypogonadism (HH) in men results in insufficient testicular function and deficiencies in testosterone and spermatogenesis. Combinations of human chorionic gonadotropin (hCG) and recombinant follicle-stimulating hormone (recFSH) have been successful in the treatment of HH. Corifollitropin alfa is a long-acting FSH-analog with demonstrated action in women seeking infertility care. The aim of this study was to investigate the efficacy and safety of corifollitropin alfa combined with hCG to increase testicular volume and induce spermatogenesis in men with HH. This was a Phase III, multi-center, open-label, single-arm trial of corifollitropin alfa in azoospermic men aged 18 to 50 years with HH. After 16 weeks of pretreatment of 23 subjects with hCG alone, 18 subjects with normalized testosterone (T) levels who remained azoospermic entered the 52-week combined treatment phase with hCG twice-weekly and 150 μg corifollitropin alfa every other week. The increase in testicular volume (primary efficacy endpoint) and induction of spermatogenesis resulting in a sperm count ≥1 × 10 6 /mL (key secondary efficacy endpoint) during 52 weeks of combined treatment were assessed. Safety was evaluated by the presence of anti-corifollitropin alfa antibodies and the occurrence of adverse events (AEs). Mean (±SD) testicular volume increased from 8.6 (±6.09) mL to 17.8 (±8.93) mL (geometric mean fold increase, 2.30 [95% CI: 2.03, 2.62]); 14 (77.8%) subjects reached a sperm count ≥1 × 10 6 /mL. No subject developed confirmed anti-corifollitropin alfa antibodies during the trial. Treatment was generally well tolerated. Corifollitropin alfa 150 μg administrated every other week combined with twice-weekly hCG for 52 weeks increased testicular volume significantly, and induced spermatogenesis in >75% of men with HH who had remained azoospermic after hCG treatment alone. ClinicalTrials.gov: NCT01709331 .

Lactose hydrolysis in aqueous two-phase system by whole-cell {beta}-galactosidase of Kluyveromyces marxianus. Semicontinuous and continuous processes

Energy Technology Data Exchange (ETDEWEB)

Tomaska, M [Slovak Technical Univ., Bratislava (Slovakia). Dept. of Biochemical Technology; Stredansky, M [Slovak Technical Univ., Bratislava (Slovakia). Dept. of Biochemical Technology; Tomaskova, A [Slovak Technical Univ., Bratislava (Slovakia). Dept. of Biochemical Technology; Sturdik, E [Slovak Technical Univ., Bratislava (Slovakia). Dept. of Biochemical Technology

1995-01-01

Semicontinuous and continuous hydrolysis of lactose in aqueous two-phase systems (polyethylene glycol 20000/ dextran 40) with whole-cell {beta}-galactosidase of K. marxianus were studied. Both phase polymers had no effect on {beta}-galactosidase activity confined in cells. Good operational stability of the biocatalyst during 55 cycles of semicontinuous process was observed without appreciable decrease in product concentration. Continuous hydrolysis of lactose was performed in the stirred bioreactor, connected with the phase separator. The satisfactory degree of hydrolysis (between 82-88%) and volumetric productivity (21.6 g/l/h) were reached during 72 hours of continuous hydrolysis of 5% (w/w) lactose. (orig.)

Ability of Lactobacillus fermentum to overcome host α-galactosidase deficiency, as evidenced by reduction of hydrogen excretion in rats consuming soya α-galacto-oligosaccharides

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Sesma Fernando

2008-01-01

Full Text Available Abstract Background Soya and its derivatives represent nutritionally high quality food products whose major drawback is their high content of α-galacto-oligosaccharides. These are not digested in the small intestine due to the natural absence of tissular α-galactosidase in mammals. The passage of these carbohydrates to the large intestine makes them available for fermentation by gas-producing bacteria leading to intestinal flatulence. The aim of the work reported here was to assess the ability of α-galactosidase-producing lactobacilli to improve the digestibility of α-galacto-oligosaccharides in situ. Results Gnotobiotic rats were orally fed with soy milk and placed in respiratory chambers designed to monitor fermentative gas excretion. The validity of the animal model was first checked using gnotobiotic rats monoassociated with a Clostridium butyricum hydrogen (H2-producing strain. Ingestion of native soy milk by these rats caused significant H2 emission while ingestion of α-galacto-oligosaccharide-free soy milk did not, thus validating the experimental system. When native soy milk was fermented using the α-galactosidase-producing Lactobacillus fermentum CRL722 strain, the resulting product failed to induce H2 emission in rats thus validating the bacterial model. When L. fermentum CRL722 was coadministered with native soy milk, a significant reduction (50 %, P = 0.019 in H2 emission was observed, showing that α-galactosidase from L. fermentum CRL722 remained active in situ, in the gastrointestinal tract of rats monoassociated with C. butyricum. In human-microbiota associated rats, L. fermentum CRL722 also induced a significant reduction of H2 emission (70 %, P = 0.004. Conclusion These results strongly suggest that L. fermentum α-galactosidase is able to partially alleviate α-galactosidase deficiency in rats. This offers interesting perspectives in various applications in which lactic acid bacteria could be used as a vector for

Feasibility of corifollitropin alfa/GnRH antagonist protocol combined with GnRH agonist triggering and freeze-all strategy in polycystic ovary syndrome patients

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Jiann-Loung Hwang

2018-06-01

Full Text Available Background/Purpose: The long-acting corifollitropin alfa is comparable to FSH in terms of pregnancy outcomes in normal responders and poor responders. Corifollitropin alfa has never been studied in polycystic ovary syndrome (PCOS patients because of concerns of excessive ovarian stimulation and ovarian hyperstimulation syndrome (OHSS. The purpose of the study was to evaluate if corifollitropin alfa can be used in PCOS patients. Methods: Forty PCOS patients who were going to undergo in vitro fertilization were enrolled in this study. A single injection of corifollitropin alfa was administered on cycle day 2 or day 3. From stimulation day 8 onwards, daily FSH was administered until the day of final oocyte maturation. Cetrorelix was administered from stimulation day 5 to prevent premature LH surge. Final oocyte maturation was triggered by: acetate. All embryos were cryopreserved and replaced in subsequent cycles. Results: All 40 patients were subjected to oocyte retrieval, and none developed moderate or severe ovarian hyperstimulation syndrome (0%, 95% CI 0–0.088. For each patient, an average of 23.4 (±7.4; 95% CI 21.0–25.7 oocytes were retrieved and a mean of 11.7 (±6.4; 95% CI 9.6–13.8 embryos were frozen. Mean serum estradiol level on the day of GnRHa triggering was 7829.9 pg/ml (±3297; 95% CI 6775–8885. The cumulated ongoing pregnancy rate after 3 frozen-thawed embryo transfers was 75.0% (95% CI 61.6%–88.4%. Conclusion: The results suggest that corifollitropin alfa/GnRH antagonist protocol can be used in PCOS patients, in combination with GnRHa triggering and embryo cryopreservation. Keywords: Corifollitropin alfa, Cryopreservation, GnRH agonist, Polycystic ovary syndrome

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.

Escherichia coli LacZ beta-galactosidase inhibition by monohydroxy acetylated glycopyranosides: Role of the acetyl groups

Czech Academy of Sciences Publication Activity Database

Brabcová, Jana; Carrasco-Lopez, C.; Bavaro, T.; Hermoso, J. A.; Palomo, J. M.

2014-01-01

Roč. 107, Sep (2014), s. 31-38 ISSN 1381-1177 Grant - others:GA AV ČR(CZ) M20055123 Institutional support: RVO:61388963 Keywords : biotransformation * galactosidase * inhibition * glycopyranosides * regioselectivity Subject RIV: CE - Biochemistry Impact factor: 2.128, year: 2014

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

ALFA detector, Background removal and analysis for elastic events

CERN Document Server

Belaloui, Nazim

2017-01-01

I worked on the ALFA project, which has the aim to measure the total cross section in PP collisions as a function of t, the momentum transfer by measuring the scattering angle of the protons. This measurement is done for all available energies; so far 7, 8 and 13 TeV. There are many analysis steps and we have focused on enhancing the signal-to-noise ratio. First of all I tried to be more familiar with ROOT, worked on understanding the code used to access to the data, plotting histograms, then cutting-off background.

Recombinant enzyme therapy for Fabry disease: Absence of editing of human alpha-galactosidase A mRNA

NARCIS (Netherlands)

Blom, Daniël; Speijer, Dave; Linthorst, Gabor E.; Donker-Koopman, Wilma G.; Strijland, Anneke; Aerts, Johannes M. F. G.

2003-01-01

For more than a decade, protein-replacement therapy has been employed successfully for the treatment of Gaucher disease. Recently, a comparable therapy has become available for the related lipid-storage disorder Fabry disease. Two differently produced recombinant alpha-galactosidase A (alpha-gal A)

High level production of β-galactosidase exhibiting excellent milk-lactose degradation ability from Aspergillus oryzae by codon and fermentation optimization.

Science.gov (United States)

Zhao, Qianqian; Liu, Fei; Hou, Zhongwen; Yuan, Chao; Zhu, Xiqiang

2014-03-01

A β-galactosidase gene from Aspergillus oryzae was engineered utilizing codon usage optimization to be constitutively and highly expressed in the Pichia pastoris SMD1168H strain in a high-cell-density fermentation. After fermentation for 96 h in a 50-L fermentor using glucose and glycerol as combined carbon sources, the recombinant enzyme in the culture supernatant had an activity of 4,239.07 U mL(-1) with o-nitrophenyl-β-D-galactopyranoside as the substrate, and produced a total of extracellular protein content of 7.267 g L(-1) in which the target protein (6.24 g L(-1)) occupied approximately 86 %. The recombinant β-galactosidase exhibited an excellent lactose hydrolysis ability. With 1,000 U of the enzyme in 100 mL milk, 92.44 % lactose was degraded within 24 h at 60 °C, and the enzyme could also accomplish the hydrolysis at low temperatures of 37, 25, and 10 °C. Thus, this engineered strain had significantly higher fermentation level of A. oryzae lactase than that before optimization and the β-galactosidase may have a good application potential in whey and milk industries.

Relationship between sol-gel conditions and enzyme stability: a case study with β-galactosidase/silica biocatalyst for whey hydrolysis.

Science.gov (United States)

Escobar, Sindy; Bernal, Claudia; Mesa, Monica

2015-01-01

The sol-gel process has been very useful for preparing active and stable biocatalysts, with the possibility of being reused. Especially those based on silica are well known. However, the study of the enzyme behavior during this process is not well understood until now and more, if the surfactant is involved in the synthesis mixture. This work is devoted to the encapsulation of β-galactosidase from Bacillus circulans in silica by sol-gel process, assisted by non-ionic Triton X-100 surfactant. The correlation between enzyme activity results for the β-galactosidase in three different environments (soluble in buffered aqueous reference solution, in the silica sol, and entrapment on the silica matrix) explains the enzyme behavior under stress conditions offered by the silica sol composition and gelation conditions. A stable β-galactosidase/silica biocatalyst is obtained using sodium silicate, which is a cheap source of silica, in the presence of non-ionic Triton X-100, which avoids the enzyme deactivation, even at 40 °C. The obtained biocatalyst is used in the whey hydrolysis for obtaining high value products from this waste. The preservation of the enzyme stability, which is one of the most important challenges on the enzyme immobilization through the silica sol-gel, is achieved in this study.

Screening and Characterization of Cold-Active β-Galactosidase Producing Psychrotrophic Enterobacter ludwigii from the Sediments of Arctic Fjord.

Science.gov (United States)

Alikkunju, Aneesa P; Sainjan, Neethu; Silvester, Reshma; Joseph, Ajith; Rahiman, Mujeeb; Antony, Ally C; Kumaran, Radhakrishnan C; Hatha, Mohamed

2016-10-01

Low-temperature-tolerant microorganisms and their cold-active enzymes could be an innovative and invaluable tool in various industrial applications. In the present study, bacterial isolates from the sediment samples of Kongsfjord, Norwegian Arctic, were screened for β-galactosidase production. Among the isolates, KS25, KS85, KS60, and KS92 have shown good potential in β-galactosidase production at 20 °C. 16SrRNA gene sequence analysis revealed the relatedness of the isolates to Enterobacter ludwigii. The optimum growth temperature of the isolate was 25 °C. The isolate exhibited good growth and enzyme production at a temperature range of 15-35 °C, pH 5-10. The isolate preferred yeast extract and lactose for the maximum growth and enzyme production at conditions of pH 7.0, temperature of 25 °C, and agitation speed of 100 rpm. The growth and enzyme production was stimulated by Mn 2+ and Mg 2+ and strongly inhibited by Zn 2+ , Ni 2+ , and Cu + . β-Galactosidases with high specific activity at low temperatures are very beneficial in food industry to compensate the nutritional problem associated with lactose intolerance. The isolate exhibited a remarkable capability to utilize clarified whey, an industrial pollutant, for good biomass and enzyme yield and hence could be well employed in whey bioremediation.

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

NUTRIENT DIGESTIBILITY AND PERFORMANCE IN BROILER CONSUMING DIETS FORMULATED WITH DIFFERENT FIBER LEVELS AND SUPLEMENTED WITH ENZYMATIC COMPLEX DIGESTIBILIDADE DOS NUTRIENTES E DESEMPENHO DE FRANGOS DE CORTE CONSUMINDO DIETAS FORMULADAS COM DIFERENTES NÍVEIS DE FIBRA E SUPLEMENTADAS COM ENZIMAS EXÓGENAS

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Bárbara Karolina Ratier da Silva

2008-12-01

, celulase, amilase, protease. No primeiro experimento (14 a 19 dias de idade dos animais, distribuíram-se aleatoriamente 190 aves em arranjo fatorial 2x2 (fibra x enzima, em gaiolas metabólicas com 9-10 aves/gaiola, cinco gaiolas por tratamento. Avaliaram-se a metabolizabilidade (MET da matéria seca (MS, a proteína bruta (PB e a energia (E, os coeficientes de digestibilidade total aparente (CDT da gordura (G e a fibra em detergente ácido (FDA,a  energia metabolizável aparente (EMA e o desempenho. Quatro repetições de cada tratamento foram mantidas para o segundo experimento. No 21º dia de idade das aves, após adaptação ao consumo das dietas com indicador, abateram-se os frangos abatidos e colheu-se conteúdo de íleo para avaliação do coeficiente de digestibilidade ileal aparente (CDI da MS, PB, E, G e peso dos cecos. O maior nível de fibra resultou em menor MET da MS, E, PB, e menor no CDT da G e FDA (P<0,05, assim como também menor CDI da MS e E (P<0,01. A suplementação enzimática reduziu o efeito da fibra sobre MET da MS, E, PB e CDT da FDA (P<0,05. Registrou-se maior peso vazio de cecos para os animais que consumiram dieta com alta fibra (P<0,01, assim como menor ganho de peso (P<0,03 e pior conversão alimentar (P<0,01. O desempenho dos animais teve maior relação com a digestibilidade ileal aparente dos nutrientes do que com a digestibilidade total aparente.

PALAVRAS-CHAVES: Cecos, desempenho, digestibilidade ileal, digestibilidade total, metabolizabilidade.

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...

Enhancement of water soluble wheat bran polyphenolic compounds using different steviol glucosides prepared by thermostable β-galactosidase

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Hee-jung Lim

2016-10-01

Full Text Available Background: Production of wheat bran (WB for human consumption is estimated to be about 90 million tons per year. WB contains an abundant source of dietary fiber, minerals, vitamins, and bioactive compounds. WB is a by-product of milling and contains an abundant source of carbohydrate (60%, protein (12%, fat (0.5%, minerals (2%, and bioactive compounds such as phenolic acids, arabinoxylans, flavonoids, caroteinoids alkylresorcinol and phytosterols. These are known for health promoting properties such as controlling glycemic index, reducing plasma cholesterol level, antioxidant, anti-inflammatory, and anticarcinogenic activities. Several terpene glycosides such as mogroside V, paenoiflorin, geniposide, rubusoside (Ru, stevioside (Ste, rebaudioside A (RebA, steviol monoside, and stevioside glucoside have been discovered to enhance the solubility of a number of pharmaceutically and medically important compounds that normally show poor solubility in water. Context and purpose of this study: In this study, in order to increase soluble extraction of polyphenol compounds of WB using Ru, the expression of β-galactosidase from Thermus thermophilus (T. thermophilus was optimized using different E. coli hosts and a different concentration of lactose inducer rather than of isopropyl-1- thio-β-D-galactopyranoside (IPTG for industrial production. Additionally, the effect of different steviol glucosides (Ru, Ste, RebA, and SG on the enhancement of polyphenol compounds extraction from wheat bran was studied. Results: β-galactosidase from T. thermophilus was used for the specific conversion of stevioside (Ste to rubusoside (Ru with 92% productivity. The enzyme was optimized to be expressed in E. coli. With 7 mM lactose, the β-galactosidase activity expressed was 34.3, 14.2, or 34.4 ± 0.5 U/mL in E. coli BL21(DE3pLysS, Rosetta(DE3pLysS, or BL21(DE3 at 37°C, and 9.8 ± 0.2, 7.0 ± 0.5, or 7.4 ± 0.2 U/mL at 28°C respectively. The expression of β-galactosidase

A Novel Process to Prepare Chitosan Macrospheres without Shrinkage and its Application to Immobilize β-Galactosidase

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Su-Fang Sun

2009-01-01

Full Text Available A new process for the preparation of chitosan macrospheres, which was simple and practicable, was suggested in this paper and various chitosans with different molecular weight were used as materials to immobilize β-galactosidase and the chitosan macrospheres with the lowest molecular weight (500 000 was selected as enzyme immobilization carrier based on the highest enzyme activity. In order to overcome the shrinkage of chitosan during drying, the wet macrospheres obtained was treated by 30% glycerol solution for 1 h before drying and the results showed that the dried chitosan macrospheres obtained could keep almost the same structure as its wet form, which was very important for chitosan as enzyme carrier in industry. Finally, β-galactosidase from Aspergillus oryzae was immobilized on above dry chitosan macrospheres and a satisfactory result of the immobilized enzyme was obtained in enzyme activity yield, pH stability, thermal stability, operational stability, Michaelis constants Km and the maximum velocity (Vm

Synthesis of the Galactosyl Derivative of Gluconic Acid With the Transglycosylation Activity of β-Galactosidase

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Aleksandra Wojciechowska

2017-01-01

Full Text Available Bionic acids are bioactive compounds demonstrating numerous interesting properties. They are widely produced by chemical or enzymatic oxidation of disaccharides. This paper focuses on the galactosyl derivative of gluconic acid as a result of a new method of bionic acid synthesis which utilises the transglycosylation properties of β-galactosidase and introduces lactose as a substrate. Products obtained in such a process are characterised by different structures (and, potentially, properties than those resulting from traditional oxidation of disaccharides. The aim of this study is to determine the effect of selected parameters (concentration and ratio of substrates, dose of the enzyme, time, pH, presence of salts on the course of the reaction carried out with the enzymatic preparation Lactozym, containing β-galactosidase from Kluyveromyces lactis. Research has shown that increased dry matter content in the baseline solution (up to 50 %, by mass per volume and an addition of NaCl contribute to higher yield. On the other hand, reduced content of the derivative is a result of increased pH from 7.0 to 9.0 and an addition of magnesium and manganese salts. Moreover, exceeding the β-galactosidase dose over approx. 35 000 U per 100 g of lactose also leads to reduced yield of the process. The most favourable molar ratio of sodium gluconate to lactose is 2.225:0.675. Depending on the conditions of the synthesis, the product concentration ranged between 17.3 and 118.3 g/L of the reaction mixture, which corresponded to the mass fraction of 6.64–23.7 % of dry matter. The data obtained as a result of the present study may be useful for designing an industrial process.

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

A role for α-galactosidase in the degradation of the endosperm cell walls of lettuce seeds, cv. Grand Rapids.

Science.gov (United States)

Leung, D W; Bewley, J D

1983-04-01

Isolated endosperms of Grand Rapids lettuce (Lactuca sativa L.) seeds undergo extensive cell-wall degradation and sugars are released into the surrounding incubation medium. One sugar so released is galactose. α-Galactosidase (EC 3.2.122) is present at the same level in both dry and imbibed isolated endosperms and is responsible for the release of galactose. However, this enzyme does not act upon the native endosperm cell wall, but requires first its partial hydrolysis and the production of oligomers by the action of endo-β-mannanase (EC 3.2.1.787). Galactose is then cleaved from these oligomers, allowing their further subsequent hydrolysis by endo-β-mannanase. Thus α-galactosidase and endo-β-mannanase act cooperatively to effect the hydrolysis of the lettuce endosperm cell walls.

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...

Kinetics and Thermal Properties of Crude and Purified β-Galactosidase with Potential for the Production of Galactooligosaccharides

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Anna Rafaela Cavalcante Braga

2013-01-01

Full Text Available β-Galactosidase is an enzyme that catalyzes the hydrolysis of lactose. It has potential importance due to various applications in the food and dairy industries, involving lactose-reduced ingredients. The properties of two β-galactosidase enzymes, crude and purified, from different sources, Kluyveromyces marxianus CCT 7082 and Kluyveromyces marxianus ATCC 16045, were analyzed. The pH and temperature optima, deactivation energy, thermal stability and kinetic and thermodynamic parameters were determined, as well as the ability to hydrolyze lactose and produce galactooligosaccharides. Purification process improved the properties of the enzymes, and the results showed that purified enzymes from both strains had a higher optimum temperature, and lower values of Km, thus showing greater affinity for o-nitrophenyl-β-D-galactopiranoside than the crude enzymes. The production of galactooligosaccharides was also greater when using purified enzymes, increasing the synthesis by more than 30 % by both strains.

The influence of monovalent cations on trimeric G protein Gi1alfa activity in HEK293 cells stably expressing DOR-Gi1alfa (Cys351-Ile351) fusion protein

Czech Academy of Sciences Publication Activity Database

Vošahlíková, Miroslava; Svoboda, Petr

2011-01-01

Roč. 60, č. 3 (2011), s. 541-547 ISSN 0862-8408 R&D Projects: GA AV ČR(CZ) IAA500110606; GA MŠk(CZ) LC554; GA ČR(CZ) GD305/08/H037 Institutional research plan: CEZ:AV0Z50110509 Keywords : delta-opioid receptor (DOR) * monovalent ions * G(i)1alfa protein Subject RIV: CE - Biochemistry Impact factor: 1.555, year: 2011

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

A redesigned follitropin alfa pen injector for infertility: results of a market research study

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Abbotts C

2011-06-01

Full Text Available Carole Abbotts1, Cristiana Salgado-Braga2, Céline Audibert-Gros31Pharmaceutical Marketing Research Consultancy, London, UK; 2Fertility and Endocrinology Global Business Unit, 3Business Intelligence, Merck Serono SA, Geneva, SwitzerlandBackground: The purpose of this study was to evaluate patient-learning and nurse-teaching experiences when using a redesigned prefilled, ready-to-use follitropin alfa pen injector.Methods: Seventy-three UK women of reproductive age either administering daily treatment with self-injectable gonadotropins or about to start gonadotropin treatment for infertility (aged 24–47 years; 53 self-injection-experienced and 20 self-injection-naïve and 28 nurses from UK infertility clinics were recruited for the study. Following instruction, patients and nurses used the redesigned follitropin alfa pen to inject water into an orange and completed questionnaires to evaluate their experiences with the pen immediately after the simulated injections.Results: Most (88%, n = 64 patients found it easy to learn how to use the pen. Among injection-experienced patients, 66% (n = 35 agreed that the redesigned pen was easier to learn to use compared with their current method and 70% (n = 37 also said they would prefer its use over current devices for all injectable fertility medications. All nurses considered the redesigned pen easy to learn and believed it would be easy to teach patients how to use. Eighty-six percent (n = 24 of the nurses thought it was easy to teach patients to determine the remaining dose to be dialed and injected in a second pen if the initial dose was incomplete. Compared with other injection devices, 96% (n = 27 thought it was "much easier" to "as easy" to teach patients to use the redesigned pen. Based on ease of teaching, 68% (n = 19 of nurses would choose to teach the pen in preference to any other injection method. Almost all (93%, n = 26 nurses considered that having the same pen format for a range of

Galactosylation of steroidal saponins by β-galactosidase from Lactobacillus bulgaricus L3.

Science.gov (United States)

Zhang, Jie; Lu, Lili; Lu, Li; Zhao, Yang; Kang, Liping; Pang, Xu; Liu, Jingyuan; Jiang, Tengchuan; Xiao, Min; Ma, Baiping

2016-02-01

The galactosylation of furostanosides and spirostanosides were investigated by using β-galactosidase from Lactobacillus bulgaricus L3 as a catalyst and lactose as a sugar donor. Four novel galactosylated furostanoside products (compounds 1-4) from compound F, compound G, compound I, and compound H were obtained and their structures were identified by HR-ESI-MS, 1D and 2D NMR spectra. The result showed that galactosyl moiety was found to be added to the 6-OH of the 26-O-glucosyl in these four furostanoside substrates.

Impact of weight-based ribavirin with peginterferon alfa-2b in African Americans with hepatitis C virus genotype 1.

Science.gov (United States)

Jacobson, Ira M; Brown, Robert S; McCone, Jonathan; Black, Martin; Albert, Clive; Dragutsky, Michael S; Siddiqui, Firdous A; Hargrave, Thomas; Kwo, Paul Y; Lambiase, Louis; Galler, Greg W; Araya, Victor; Freilich, Bradley; Harvey, Joann; Griffel, Louis H; Brass, Clifford A

2007-10-01

WIN-R (Weight-based dosing of pegINterferon alfa-2b and Ribavirin) was a multicenter, randomized, open-label, investigator-initiated trial involving 236 community and academic sites in the United States, comparing response to pegylated interferon (PEG-IFN) alfa-2b plus a flat or weight-based dose of ribavirin (RBV) in treatment-naive patients with chronic hepatitis C and compensated liver disease. Patients were randomized to receive PEG-IFN alfa-2b at 1.5 microg/kg/week plus flat-dose (800 mg/day) or weight-based-dose RBV (800 mg/day for weight 85-105 kg, or 1400 mg/day for >105- or =65 kg was the primary end point. Low SVR rates have been reported among African American individuals, in whom there is a preponderance of HCV genotype 1. This subanalysis of WIN-R was conducted to evaluate the efficacy of weight-based dosing among African American individuals with genotype 1 infection enrolled in the trial. Of 362 African American patients in the primary efficacy analysis, 188 received RBV flat dosing and 174 received weight-based dosing. SVR rates were higher (21% versus 10%; P = 0.0006) and relapse rates were lower (22% versus 30%) in the weight-based-dose group than in the flat-dose group. Safety and rates of drug discontinuation were similar between the 2 groups. Weight-based dosing of RBV is more effective than flat dosing in combination with PEG-IFN alfa-2b in African American individuals with HCV genotype 1. Even with weight-based dosing, response rates in African American individuals are lower than reported in other ethnic groups.

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

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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

Investigations to determine whether viable microorganisms are required during intestinal lactose hydrolysis of fermented milk products by microbial ß-galactosidase using gnotobiotic Göttingen minipigs

OpenAIRE

Winchenbach, Andrea

2010-01-01

The most common reason worldwide for the indigestibility of milk is the lack of ß-galactosidases in the small intestine, leading to the malabsorbtion of lactose. Fermented dairy products are very often much better tolerated than raw (not fermented) milk, because of the microbial ß-galactosidases they contain. The aim of this thesis was to elucidate the question as to weather lactose hydrolysis in the small intestine requires the presence of living bacteria (with their microbial ß-galac...

Disertación endocrina de la preeclampsia y propuesta de una hipótesis Endocrine dissertation on preeclampsia and proposal of a hypothesis

Directory of Open Access Journals (Sweden)

Bernardo Agudelo Jaramillo

1992-01-01

Full Text Available

Se propone que la facies androgenizada en las embarazadas podría servir como elemento clínico para la búsqueda de la preeclampsia, con el fin de lograr un enfoque preventivo. El balance de las prostaglandinas vasodilatadoras y vasoconstrictoras es, en último término, responsable del control de la presión arterial en la gestante. Para ello se debe lograr un adecuado equilibrio en la producción de hormonas gestacionales; de ellas parece ser fundamental la dihidroprogesterona que se obtiene a través de la acción de la enzima 5-alfa reductasa. En el metabolismo de los andrógenos esta enzima cumple un papel único al transformar los andrógenos circulantes en la hormona trófica, la dihidrotestosterona. La hipótesis propone que la utilización androgénica de la enzima 5-alfa reductasa durante la gestación alteraría la producción de dihidroprogesterona con lo cual crearía un desequilibrio entre las diferentes hormonas y, finalmente, entre las prostaglandinas.

The hypothesis that an androgenized facies In a pregnant woman could serve as a clinical clue for the search of preeclampsia, making It possible to apply a preventive approach is proposed. The balance between dilator and constrictor prostaglandins controls blood pressure during pregnancy; to keep such a balance an adequate equilibrium in the production of hormones, particularly dihydroprogesterone (DHP, is necessary. DHP is obtained through the action of 5-alpha reductase, which in androgen metabolism has the unique role of transforming circulating androgens into the trophic hormone dihydrotestosterone. The hypothesis proposes that androgenic utilization of 5- alpha reductase during pregnancy could alter the production of DHP I generating a lack of equilibrium among the various hormones, prostaglandins included.

Comparative Studies of Two-Dimensional Electrophoresis on Galactosidase Relating to Bombyx Lectin Activity

OpenAIRE

加藤, 靖夫; カトウ, ヤスオ; Yasuo, Kato

2005-01-01

"Comparative two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) analysis on the haemolymph of the domesticated silkworm, Bombyx mori and Fraction II obtained by gel filtration from the haemolymph of B. mori was performed using the 2-D mini-slab system (Atto Co.) (the first method of 2-D PAGE) and the Mini-PROTEAN mini tube gel 2-D PAGE system (Bio-Rad Laboratories, Inc.) (the second method). Moreover, two-dimensionnal electrophoresis analysis on standard β-galactosidase, grade III ...

Fluorouracil continuous infusion plus alfa interferon plus oral folinic acid in advanced colorectal cancer

NARCIS (Netherlands)

Punt, C. J.; de Mulder, P. H.; Burghouts, J. T.; Wagener, D. J.

1992-01-01

Several reports on fluorouracil (5-FU) and alfa interferon (IFN-alpha) combination therapy in patients with advanced colorectal cancer have been published. In our study high-dose continuous infusion 5-FU (60 mg/kg per 48 hours), oral folinic acid (FA) (8 x 90 mg during 5-FU), and IFN-alpha three

Cloning, expression and structural stability of a cold-adapted ß-Galactosidase from Rahnella sp.R3

Science.gov (United States)

A novel gene was isolated for the first time from a psychrophilic gram-negative bacterium Rahnella sp.R3. It encoded a cold-adapted ß-galactosidase (R-ß-Gal). Recombinant R-ß-Gal was expressed in Escherichia coli BL21 (DE3), purified, and characterized. R-ß-Gal belongs to the glycosyl hydrolase fami...

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

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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

Correction of acid beta-galactosidase deficiency in GM1 gangliosidosis human fibroblasts by retrovirus vector-mediated gene transfer: higher efficiency of release and cross-correction by the murine enzyme.

Science.gov (United States)

Sena-Esteves, M; Camp, S M; Alroy, J; Breakefield, X O; Kaye, E M

2000-03-20

Mutations in the lysosomal acid beta-galactosidase (EC 3.2.1.23) underlie two different disorders: GM1 gangliosidosis, which involves the nervous system and visceral organs to varying extents, and Morquio's syndrome type B (Morquio B disease), which is a skeletal-connective tissue disease without any CNS symptoms. This article shows that transduction of human GM1 gangliosidosis fibroblasts with retrovirus vectors encoding the human acid beta-galactosidase cDNA leads to complete correction of the enzymatic deficiency. The newly synthesized enzyme is correctly processed and targeted to the lysosomes in transduced cells. Cross-correction experiments using retrovirus-modified cells as enzyme donors showed, however, that the human enzyme is transferred at low efficiencies. Experiments using a different retrovirus vector carrying the human cDNA confirmed this observation. Transduction of human GM1 fibroblasts and mouse NIH 3T3 cells with a retrovirus vector encoding the mouse beta-galactosidase cDNA resulted in high levels of enzymatic activity. Furthermore, the mouse enzyme was found to be transferred to human cells at high efficiency. Enzyme activity measurements in medium conditioned by genetically modified cells suggest that the human beta-galactosidase enzyme is less efficiently released to the extracellular space than its mouse counterpart. This study suggests that lysosomal enzymes, contrary to the generalized perception in the field of gene therapy, may differ significantly in their properties and provides insights for design of future gene therapy interventions in acid beta-galactosidase deficiency.

Stroke in patients with type 2 diabetes mellitus, chronic kidney disease, and anemia treated with Darbepoetin Alfa

DEFF Research Database (Denmark)

Skali, Hicham; Parving, Hans-Henrik; Parfrey, Patrick S

2011-01-01

More strokes were observed in the Trial to Reduce Cardiovascular Events With Aranesp Therapy (TREAT) among patients assigned to darbepoetin alfa. We sought to identify baseline characteristics and postrandomization factors that might explain this association....

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. ...

Fast radio burst discovered in the Arecibo pulsar ALFA survey

International Nuclear Information System (INIS)

Spitler, L. G.; Freire, P. C. C.; Lazarus, P.; Lee, K. J.; Cordes, J. M.; Chatterjee, S.; Wharton, R. S.; Brazier, A.; Hessels, J. W. T.; Lorimer, D. R.; McLaughlin, M. A.; Crawford, F.; Deneva, J. S.; Kaspi, V. M.; Karako-Argaman, C.; Allen, B.; Bogdanov, S.; Camilo, F.; Jenet, F. A.; Knispel, B.

2014-01-01

Recent work has exploited pulsar survey data to identify temporally isolated, millisecond-duration radio bursts with large dispersion measures (DMs). These bursts have been interpreted as arising from a population of extragalactic sources, in which case they would provide unprecedented opportunities for probing the intergalactic medium; they may also be linked to new source classes. Until now, however, all so-called fast radio bursts (FRBs) have been detected with the Parkes radio telescope and its 13-beam receiver, casting some concern about the astrophysical nature of these signals. Here we present FRB 121102, the first FRB discovery from a geographic location other than Parkes. FRB 121102 was found in the Galactic anti-center region in the 1.4 GHz Pulsar Arecibo L-band Feed Array (ALFA) survey with the Arecibo Observatory with a DM = 557.4 ± 2.0 pc cm –3 , pulse width of 3.0 ± 0.5 ms, and no evidence of interstellar scattering. The observed delay of the signal arrival time with frequency agrees precisely with the expectation of dispersion through an ionized medium. Despite its low Galactic latitude (b = –0.°2), the burst has three times the maximum Galactic DM expected along this particular line of sight, suggesting an extragalactic origin. A peculiar aspect of the signal is an inverted spectrum; we interpret this as a consequence of being detected in a sidelobe of the ALFA receiver. FRB 121102's brightness, duration, and the inferred event rate are all consistent with the properties of the previously detected Parkes bursts.

Fast radio burst discovered in the Arecibo pulsar ALFA survey

Energy Technology Data Exchange (ETDEWEB)

Spitler, L. G.; Freire, P. C. C.; Lazarus, P.; Lee, K. J. [Max-Planck-Institut für Radioastronomie, D-53121 Bonn (Germany); Cordes, J. M.; Chatterjee, S.; Wharton, R. S.; Brazier, A. [Department of Astronomy and Space Sciences, Cornell University, Ithaca, NY 14853 (United States); Hessels, J. W. T. [ASTRON, Netherlands Institute for Radio Astronomy, Postbus 2, 7990 AA, Dwingeloo (Netherlands); Lorimer, D. R.; McLaughlin, M. A. [Department of Physics and Astronomy, West Virginia University, Morgantown, WV 26506 (United States); Crawford, F. [Department of Physics and Astronomy, Franklin and Marshall College, Lancaster, PA 17604-3003 (United States); Deneva, J. S. [Naval Research Laboratory, 4555 Overlook Ave SW, Washington, DC 20375 (United States); Kaspi, V. M.; Karako-Argaman, C. [Department of Physics, McGill University, Montreal, QC H3A 2T8 (Canada); Allen, B. [Physics Department, University of Wisconsin-Milwaukee, Milwaukee, WI 53211 (United States); Bogdanov, S.; Camilo, F. [Columbia Astrophysics Laboratory, Columbia University, New York, NY 10027 (United States); Jenet, F. A. [Center for Gravitational Wave Astronomy, University of Texas at Brownsville, Brownsville, TX 78520 (United States); Knispel, B., E-mail: lspitler@mpifr-bonn.mpg.de [Leibniz Universität, Hannover, D-30167 Hannover (Germany); and others

2014-08-01

Recent work has exploited pulsar survey data to identify temporally isolated, millisecond-duration radio bursts with large dispersion measures (DMs). These bursts have been interpreted as arising from a population of extragalactic sources, in which case they would provide unprecedented opportunities for probing the intergalactic medium; they may also be linked to new source classes. Until now, however, all so-called fast radio bursts (FRBs) have been detected with the Parkes radio telescope and its 13-beam receiver, casting some concern about the astrophysical nature of these signals. Here we present FRB 121102, the first FRB discovery from a geographic location other than Parkes. FRB 121102 was found in the Galactic anti-center region in the 1.4 GHz Pulsar Arecibo L-band Feed Array (ALFA) survey with the Arecibo Observatory with a DM = 557.4 ± 2.0 pc cm{sup –3}, pulse width of 3.0 ± 0.5 ms, and no evidence of interstellar scattering. The observed delay of the signal arrival time with frequency agrees precisely with the expectation of dispersion through an ionized medium. Despite its low Galactic latitude (b = –0.°2), the burst has three times the maximum Galactic DM expected along this particular line of sight, suggesting an extragalactic origin. A peculiar aspect of the signal is an inverted spectrum; we interpret this as a consequence of being detected in a sidelobe of the ALFA receiver. FRB 121102's brightness, duration, and the inferred event rate are all consistent with the properties of the previously detected Parkes bursts.

Ca(OH2 action on TNF-alpha and NO release in macrophage culture stimulated by Pseudomonas aeruginosa LPS = Ação do Ca(OH2 sobre a produção de TNF-alfa e NO de cultura de macrófagos estimulada por LPS de Pseudomonas aeruginosa

Directory of Open Access Journals (Sweden)

Queiroz, Celso Emanoel de Souza

2008-01-01

Full Text Available Objetivo: A ação do hidróxido de cálcio [Ca(OH2] com o sistema imune e o mecanismo de neutralização das bactérias e seus subprodutos ainda não foi completamente esclarecida. Neste estudo foi avaliada a capacidade do Ca(OH2 em neutralizar o lipopolissacarídeo (LPS de Pseudomonas aeruginosa, utilizando-se duas metodologias: liberação de Óxido Nítrico (NO e Fator de Necrose Tumoral Alfa (TNF-alfa em cultura de macrófagos peritoneais de camundongos. Metodologia: No ensaio do NO, as células peritoneais foram expostas a uma solução de LPS (25mg/mL e 50mg/mL; e à suspensão de LPS/Ca(OH2 em duas concentrações (50mg/ 25mg e 25mg/25mg. Após 8 horas de incubação, foi utilizado reagente de Griess, e a liberação de NO foi quantificada. No ensaio do TNF-alfa, a solução de LPS foi usada na concentração de 25mg/mL e o LPS/Ca(OH2 a 25mg/25mg. Após 24 horas, as células foram fixadas e coradas com cristal violeta, e os valores de absorbância foram obtidos. Os resultados foram expressos em micromols. Todos os testes foram realizados em triplicata. Resultados: A presença de Ca(OH2 nas duas concentrações avaliadas reduziu significativamente a liberação de NO e TNF-alfa. Conclusão: Pode-se concluir que o LPS bacteriano representa um forte estímulo para liberação destas citocinas, mas o hidróxido de cálcio foi capaz de neutralizar este efeito

Rat hepatocyte invasion by Listeria monocytogenes and analysis of TNF-alpha role in apoptosis Invasão de hepatócitos de rato por Listeria monocytogenes e análise do papel do TNF-alfa na apoptose

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Sânia Alves dos Santos

2005-04-01

do hospedeiro e manipulação do citoesqueleto, resultando em morte celular. As interações entre células do hospedeiro e bactérias podem resultar em produção de citocinas como o Fator de Necrose Tumoral alfa (TNF-alfa. Hepatócitos têm potencial de produzir citocinas pro-inflamatórias como TNF-alfa, quando invadidos por bactérias. No presente trabalho demonstramos o comportamento dos hepatócitos invadidos por L. monocytogenes pela análise microscópica, determinação da produção de TNF-alfa por bioensaio e análise da apoptose pela técnica TUNEL. A presença da bactéria, na razão que variou de 5 a 50.000 bactérias por célula, induziu ruptura das monocamadas celulares. Observamos presença de bactérias internalizadas na 1ª hora de incubação por microscopia eletrônica. Os níveis de TNF-alfa aumentaram da 1ª hora de incubação até a 6ª hora, variando de 0 a 3749 pg/mL. Nas 7ª e 8ª horas de incubação, ocorreram quedas não significativas dos níveis de TNF-alfa, indicando possível saturação dos receptores celulares. A quantidade de TNF-alfa produzido por hepatócitos foi dependente do tempo de incubação, assim como da proporção entre bactérias e células. A taxa de apoptose aumentou diretamente com o tempo de incubação (1 h a 8 + 24 h, variando de 0 a 43%, assim como com a razão bactérias : células. Estes resultados mostram a habilidade de L. monocytogenes não-hemolítica em invadir os hepatócitos, e as principais conseqüências deste fenômeno são: liberação de TNF-alfa e indução de apoptose. Assim, podemos especular que hepatócitos usam apoptose induzida por TNF-alfa para liberar bactérias de seu interior, facilitando a destruição destas pelo sistema imune.

Separação e purificação de galacto-oligossacarídeos por nanofiltração

OpenAIRE

Mariano Michelon

2013-01-01

Resumo: Galacto-oligossacarídeos (GOS) são oligossacarídeos não digeríveis que apresentam propriedades prebióticas, pois auxiliam na proliferação de bifidobactérias no cólon. Os GOS podem ser obtidos pela reação de transgalactosilação da galactose catalisada pela enzima ?-galactosidase a partir de substratos ricos em lactose. Neste processo, as misturas de açúcares obtidas após a síntese contêm além dos GOS, mono- e dissacarídeos residuais. Portanto, uma alternativa interessante para aumentar...

Overexpression and characterization of a novel transgalactosylic and hydrolytic β-galactosidase from a human isolate Bifidobacterium breve B24.

Science.gov (United States)

Yi, Sung Hun; Alli, Inteaz; Park, Kwan Hwa; Lee, Byonghoon

2011-10-01

After the complete gene of a β-galactosidase from human isolate Bifidobacterium breve B24 was isolated by PCR and overexpressed in E. coli, the recombinant β-galactosidase was purified to homogeneity and characterized for the glycoside transferase (GT) and glycoside hydrolase (GH) activities on lactose. One complete ORF encoding 691 amino acids (2,076 bp) was the structural gene, LacA (galA) of the β-gal gene. The recombinant enzyme shown by activity staining and gel-filtration chromatography was composed of a homodimer of 75 kDa with a total molecular mass of 150 kDa. The K(m) value for lactose (95.58 mM) was 52.5-fold higher than the corresponding K(m) values for the synthetic substrate ONPG (1.82 mM). This enzyme with the optimum of pH 7.0 and 45°C could synthesize approximately 42.00% of GOS from 1M of lactose. About 97.00% of lactose in milk was also quickly hydrolyzed by this enzyme (50 units) at 45°C for 5h to produce 46.30% of glucose, 46.60% of galactose and 7.10% of GOS. The results suggest that this recombinant β-galactosidase derived from a human isolate B. breve B24 may be suitable for both the hydrolysis and synthesis of galacto-oligosaccharides (GOS) in milk and lactose processing. Copyright © 2011 Elsevier B.V. All rights reserved.

Reaction kinetics and galactooligosaccharide product profiles of the β-galactosidases from Bacillus circulans, Kluyveromyces lactis and Aspergillus oryzae

NARCIS (Netherlands)

Yin, Huifang; Bultema, Jelle B; Dijkhuizen, Lubbert; van Leeuwen, Sander S

2017-01-01

β-Galactosidase enzymes are used in the dairy industry to convert lactose into galactooligosaccharides (GOS) that are added to infant formula to mimic the molecular sizes and prebiotic functions of human milk oligosaccharides. Here we report a detailed analysis of the clearly different GOS profiles

Blood group does not correlate with disease severity in patients with Fabry disease (alpha-galactosidase A deficiency)

NARCIS (Netherlands)

Linthorst, Gabor E.; Folman, Claudia C.; Aerts, Johannes M. F. G.; Hollak, Carla E. M.

2003-01-01

Blood groups B and P1 are substrates for the lysosomal enzyme alpha-galactosidase A. Therefore, patients with alpha-Gal A deficiency and blood groups B or P1 may exhibit more severe disease. In 48 Fabry patients distribution of blood group was not different from that in the Dutch population. 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.

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.

Identification of a Bacillus subtilis secretion mutant using a ß-galactosidase screening procedure

DEFF Research Database (Denmark)

Jacobs, Myra F.; Andersen, Jens Bo; Borchert, Torben V.

1995-01-01

High-level synthesis of exportable beta-galactosidase (LacZ) fusion proteins in Bacillus subtilis results in a lethal phenotype, and has been suggested as a tool for the selection of secretion mutants. We tested a plasmid-based, inducible lacZ fusion gene system for this purpose, but frequent...... mutations in cis, which reduced expression of the fusion gene, forced abandonment of the induction-selection strategy. Instead, after modification of the indicator plasmid, a screening procedure for increased basal LacZ activity levels was adopted. This led to the identification of a conditional B. subtilis...

Identification of a Bacillus subtilis secretion mutant using a beta-galactosidase screening procedure

DEFF Research Database (Denmark)

Jacobs, M F; Borchert, T V; Kontinen, V P

1995-01-01

High-level synthesis of exportable beta-galactosidase (LacZ) fusion proteins in Bacillus subtilis results in a lethal phenotype, and has been suggested as a tool for the selection of secretion mutants. We tested a plasmid-based, inducible lacZ fusion gene system for this purpose, but frequent...... mutations in cis, which reduced expression of the fusion gene, forced abandonment of the induction-selection strategy. Instead, after modification of the indicator plasmid, a screening procedure for increased basal LacZ activity levels was adopted. This led to the identification of a conditional B. subtilis...

FTIR spectroscopy and X-ray powder diffraction characterization of microcrystalline cellulose obtained from alfa fibers

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Trache D.

2013-07-01

Full Text Available Many cereal straws have been used as raw materials for the preparation of microcrystalline cellulose (MCC. These raw materials were gradually replaced with wood products; nevertheless about 10% of the world overall pulp production is obtained from non-wood raw material. The main interest in pulp made from straw is that it provides excellent fibres for different industries with special properties, and that it is the major available source of fibrous raw material in some geographical areas. The aim of the present work was to characterize microcrystalline cellulose prepared from alfa fibers using the hydrolysis process. The products obtained are characterized with FTIR spectroscopy and X-ray powder diffraction. As a result, FTIR spectroscopy is an appropriate technique for studying changes occurred by any chemical treatment. The spectrum of alfa grass stems shows the presence of lignin and hemicelluloses. However, the cellulose spectrum indicates that the extraction of lignin and hemicellulose was effective. The X-ray analysis indicates that the microcrystalline cellulose is more crystalline than the source material.

Immunogenicity of glycans on biotherapeutic drugs produced in plant expression systems—The taliglucerase alfa story

Science.gov (United States)

Rup, Bonita; Alon, Sari; Amit-Cohen, Bat-Chen; Brill Almon, Einat; Chertkoff, Raul; Rudd, Pauline M.

2017-01-01

Plants are a promising alternative for the production of biotherapeutics. Manufacturing in-planta adds plant specific glycans. To understand immunogenic potential of these glycans, we developed a validated method to detect plant specific glycan antibodies in human serum. Using this assay, low prevalence of pre-existing anti-plant glycan antibodies was found in healthy humans (13.5%) and in glucocerebrosidase-deficient Gaucher disease (GD) patients (5%). A low incidence (9% in naïve patient and none in treatment experienced patients) of induced anti-plant glycan antibodies was observed in GD patients after up to 30 months replacement therapy treatment with taliglucerase alfa, a version of human glucocerebrosidase produced in plant cells. Detailed evaluation of clinical safety and efficacy endpoints indicated that anti-plant glycan antibodies did not affect the safety or efficacy of taliglucerase alfa in patients. This study shows the benefit of using large scale human trials to evaluate the immunogenicity risk of plant derived glycans, and indicates no apparent risk related to anti-plant glycan antibodies. PMID:29088235

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

OpenAIRE

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...

A randomised, double-blind, placebo-controlled, crossover study to assess the efficacy and safety of three dosing schedules of agalsidase alfa enzyme replacement therapy for Fabry disease.

Science.gov (United States)

Hughes, D A; Deegan, P B; Milligan, A; Wright, N; Butler, L H; Jacobs, A; Mehta, A B

2013-07-01

Anecdotal reports suggest that the currently approved dosing interval of agalsidase alfa (0.2 mg/kg/2 weeks) for Fabry disease treatment is too long. This randomised, double-blind, placebo-controlled, crossover study investigated three altered dosing intervals. 18 Fabry patients received three agalsidase alfa dosing schedules, each for four weeks (A: 0.2 mg/kg∗2 weeks, B: 0.1 mg/kg/week, C: 0.2 mg/kg/week). Health state, pain levels, sweat volume and latency and plasma and urinary globotriaosylceramide levels were recorded throughout the study. No significant differences were found among the schedules for the primary efficacy outcome of self-assessed health state, or for pain scores. A trend toward increased sweat volume on QSART testing, and reduced urine globotriaosylceramide concentration were seen with treatment schedule C. Agalsidase alfa was safe and well tolerated with all schedules. In conclusion, the primary analyses did not find weekly infusions of agalsidase alfa to be statistically better than the approved dosing schedule however the data indicates that further studies with more patients over a longer period are required to more accurately determine the optimum dose and schedule. Copyright © 2013 Elsevier Inc. All rights reserved.

The G209A mutation in the alpha-synuclein gene in Brazilian families with Parkinson's disease Mutação G209A no gene da alfa-sinucleína em famílias brasileiras com doença de Parkinson

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Hélio A.G. Teive

2001-09-01

Full Text Available A missense G209A mutation of the alpha-synuclein gene was recently described in a large Contursi kindred with Parkinson's disease (PD. The objective of this study is to determine if the mutation G209A of the alpha-synuclein gene was present in 10 Brazilian families with PD. PD patients were recruited from movement disorders clinics of Brazil. A family history with two or more affected in relatives was the inclusion criterion for this study. The alpha-synuclein G209A mutation assay was made using polymerase chain reaction and the restriction enzyme Tsp45I. Ten patients from 10 unrelated families were studied. The mean age of PD onset was 42.7 years old. We did not find the G209A mutation in our 10 families with PD. Our results suggest that alpha-synuclein mutation G209A is uncommon in Brazilian PD families.Recentemente foi detectada mutação missense G209A no gene da alfa-sinucleína em uma grande família com doença de Parkinson (DP de Contursi, Itália. Este estudo tem o objetivo de determinar se a mutação G209A está presente em 10 famílias brasileiras com DP. Pacientes com DP foram recrutados em clínicas de distúrbio do movimento no Brasil. O critério de inclusão no estudo foi à presença de dois ou mais familiares acometidos pela DP. A mutação G209A do gene da alfa-sinucleína foi pesquisada usando a técnica de reação em cadeia de polimerase e a enzima de restrição Tsp45I. Foram estudados 10 pacientes de famílias não-relacionadas. A idade média do início dos sintomas da DP foi 42,7 anos. Não encontramos a mutação estudada neste grupo de pacientes. Nossos resultados sugerem que a mutação G209A é incomum em famílias brasileiras com DP.

Applying neutral drift to the directed molecular evolution of a β-glucuronidase into a β-galactosidase: Two different evolutionary pathways lead to the same variant

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Hale Jennifer R

2011-05-01

Full Text Available Abstract Background Directed protein evolution has been used to modify protein activity and research has been carried out to enhance the production of high quality mutant libraries. Many theoretical approaches suggest that allowing a population to undergo neutral selection may be valuable in directed evolution experiments. Findings Here we report on an investigation into the value of neutral selection in a classical model system for directed evolution, the conversion of the E. coli β-glucuronidase to a β-galactosidase activity. We find that neutral selection, i.e. selection for retaining glucuronidase activity, can efficiently identify the majority of sites of mutation that have been identified as beneficial for galactosidase activity in previous experiments. Each variant demonstrating increased galactosidase activity identified by our neutral drift experiments contained a mutation at one of four sites, T509, S557, N566 or W529. All of these sites have previously been identified using direct selection for beta galactosidase activity. Conclusions Our results are consistent with others that show that a neutral selection approach can be effective in selecting improved variants. However, we interpret our results to show that neutral selection is, in this case, not a more efficient approach than conventional directed evolution approaches. However, the neutral approach is likely to be beneficial when the resulting library can be screened for a range of related activities. More detailed statistical studies to resolve the apparent differences between this system and others are likely to be a fruitful avenue for future research.

Antiviral activity of telaprevir (VX-950) and peginterferon alfa-2a in patients with hepatitis C

NARCIS (Netherlands)

Forestier, Nicole; Reesink, Hendrik W.; Weegink, Christine J.; McNair, Lindsay; Kieffer, Tara L.; Chu, Hui-May; Purdy, Susan; Jansen, Peter L. M.; Zeuzem, Stefan

2007-01-01

Telaprevir (VX-950), an inhibitor of the hepatitis C virus (HCV) NS3/4A protease, substantially decreased plasma HCV RNA levels in a prior clinical study. The present study evaluated viral kinetics and safety during dosing with telaprevir alone and in combination with peginterferon alfa-2a for 14

Using ALFA for high throughput, distributed data transmission in the ALICE O2 system

Science.gov (United States)

Wegrzynek, A.; ALICE Collaboration

2017-10-01

ALICE (A Large Ion Collider Experiment) is a heavy-ion detector designed to study the physics of strongly interacting matter (the Quark-Gluon Plasma at the CERN LHC (Large Hadron Collider). ALICE has been successfully collecting physics data in Run 2 since spring 2015. In parallel, preparations for a major upgrade of the computing system, called O2 (Online-Offline), scheduled for the Long Shutdown 2 in 2019-2020, are being made. One of the major requirements of the system is the capacity to transport data between so-called FLPs (First Level Processors), equipped with readout cards, and the EPNs (Event Processing Node), performing data aggregation, frame building and partial reconstruction. It is foreseen to have 268 FLPs dispatching data to 1500 EPNs with an average output of 20 Gb/s each. In overall, the O2 processing system will operate at terabits per second of throughput while handling millions of concurrent connections. The ALFA framework will standardize and handle software related tasks such as readout, data transport, frame building, calibration, online reconstruction and more in the upgraded computing system. ALFA supports two data transport libraries: ZeroMQ and nanomsg. This paper discusses the efficiency of ALFA in terms of high throughput data transport. The tests were performed with multiple FLPs pushing data to multiple EPNs. The transfer was done using push-pull communication patterns and two socket configurations: bind, connect. The set of benchmarks was prepared to get the most performant results on each hardware setup. The paper presents the measurement process and final results - data throughput combined with computing resources usage as a function of block size. The high number of nodes and connections in the final set up may cause race conditions that can lead to uneven load balancing and poor scalability. The performed tests allow us to validate whether the traffic is distributed evenly over all receivers. It also measures the behaviour of

The influence of a polymorphism in the gene encoding angiotensin converting enzyme (ACE on treatment outcomes in late-onset Pompe patients receiving alglucosidase alfa

Directory of Open Access Journals (Sweden)

Rena C. Baek

2016-09-01

Full Text Available Correlations between angiotensin-converting enzyme (ACE genotype (I/I, I/D, D/D, disease severity at baseline and response to enzyme replacement therapy (ERT were assessed in the Pompe disease Late-Onset Treatment Study (LOTS. No correlations were observed between ACE genotype and disease severity at baseline. However, D/D patients appeared to have a reduced response to alglucosidase alfa treatment than I/I or I/D patients, suggesting that ACE polymorphisms may influence the response to alglucosidase alfa treatment and warrants further investigation.

easyDAS: Automatic creation of DAS servers

Directory of Open Access Journals (Sweden)

Jimenez Rafael C

2011-01-01

Full Text Available Abstract Background The Distributed Annotation System (DAS has proven to be a successful way to publish and share biological data. Although there are more than 750 active registered servers from around 50 organizations, setting up a DAS server comprises a fair amount of work, making it difficult for many research groups to share their biological annotations. Given the clear advantage that the generalized sharing of relevant biological data is for the research community it would be desirable to facilitate the sharing process. Results Here we present easyDAS, a web-based system enabling anyone to publish biological annotations with just some clicks. The system, available at http://www.ebi.ac.uk/panda-srv/easydas is capable of reading different standard data file formats, process the data and create a new publicly available DAS source in a completely automated way. The created sources are hosted on the EBI systems and can take advantage of its high storage capacity and network connection, freeing the data provider from any network management work. easyDAS is an open source project under the GNU LGPL license. Conclusions easyDAS is an automated DAS source creation system which can help many researchers in sharing their biological data, potentially increasing the amount of relevant biological data available to the scientific community.

Corifollitropin alfa compared to daily rFSH or HP-HMG in GnRH antagonist controlled ovarian stimulation protocol for patients undergoing assisted reproduction.

Science.gov (United States)

Souza, Priscila Morais Galvão; Carvalho, Bruno Ramalho de; Nakagawa, Hitomi Miura; Rassi, Thalita Reis Esselin; Barbosa, Antônio César Paes; Silva, Adelino Amaral

2017-06-01

This study aimed to compare the outcomes of controlled ovarian stimulation (COS) with corifollitropin alfa versus daily recombinant follicle-stimulating hormone (rRFSH) or highly purified human menopausal gonadotropin (HP-HMG) in patients undergoing in vitro fertilization (IVF) cycles based on gonadotropin-releasing hormone (GnRH) antagonist protocols. The primary endpoints were total number of oocytes and mature oocytes. This retrospective study looked into 132 controlled ovarian stimulation cycles from IVF or oocyte cryopreservation performed in a private human reproduction center between January 1 and December 31, 2014. Enrollment criteria: women aged 0.05). There were no significant differences in fertilization (76.9% vs. 76.8%, p=1.0), biochemical pregnancy (66.7% vs. 47.2%, p=0.1561) or embryo implantation rates (68.7% vs. 50%, p=0.2588) between the groups using corifollitropin alfa and rFSH or HMG, respectively. Corifollitropin alfa seems to be as effective as rFSH or HP-HMG when used in the first seven days of ovulation induction for patients undergoing assisted reproduction in GnRH antagonist protocols.

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

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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.

Fungal Alfa Amilaz ve Deaktif Kuru Maya Preparatları Kullanılarak Hızlandırılmış Ekmek Yapımı

OpenAIRE

Bilgiçli, Nermin; Türker, Selman; Elgün, Adem

2015-01-01

The aim of this research is the determination of optimum fermentation time giving the best bread quality with fungal alfa amylase and a reducing agent-deactive dry yeast preparetes at different flour types in bread making. In the experiment two different enzyme praperates (Fungal Alfa Amylase and Reducing Agent-Deactive dry yeast) were added at three different types of flour (Type 550, 650 and 850) and prepared bread dough. The dough was rested during three different bulk fermentation time (1...

Continuous Packed Bed Reactor with Immobilized β-Galactosidase for Production of Galactooligosaccharides (GOS

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Barbara Rodriguez-Colinas

2016-11-01

Full Text Available The β-galactosidase from Bacillus circulans was covalently attached to aldehyde-activated (glyoxal agarose beads and assayed for the continuous production of galactooligosaccharides (GOS in a packed-bed reactor (PBR. The immobilization was fast (1 h and the activity of the resulting biocatalyst was 97.4 U/g measured with o-nitrophenyl-β-d-galactopyranoside (ONPG. The biocatalyst showed excellent operational stability in 14 successive 20 min reaction cycles at 45 °C in a batch reactor. A continuous process for GOS synthesis was operated for 213 h at 0.2 mL/min and 45 °C using 100 g/L of lactose as a feed solution. The efficiency of the PBR slightly decreased with time; however, the maximum GOS concentration (24.2 g/L was obtained after 48 h of operation, which corresponded to 48.6% lactose conversion and thus to maximum transgalactosylation activity. HPAEC-PAD analysis showed that the two major GOS were the trisaccharide Gal-β(1→4-Gal-β(1→4-Glc and the tetrasaccharide Gal-β(1→4-Gal-β(1→4-Gal-β(1→4-Glc. The PBR was also assessed in the production of GOS from milk as a feed solution. The stability of the bioreactor was satisfactory during the first 8 h of operation; after that, a decrease in the flow rate was observed, probably due to partial clogging of the column. This work represents a step forward in the continuous production of GOS employing fixed-bed reactors with immobilized β-galactosidases.

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...

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 ...

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

Coexpression of β-D-galactosidase and L-arabinose isomerase in the production of D-tagatose: a functional sweetener.

Science.gov (United States)

Zhan, Yijing; Xu, Zheng; Li, Sha; Liu, Xiaoliu; Xu, Lu; Feng, Xiaohai; Xu, Hong

2014-03-19

The functional sweetener, d-tagatose, is commonly transformed from galactose by l-arabinose isomerase. To make use of a much cheaper starting material, lactose, hydrolization, and isomerization are required to take place collaboratively. Therefore, a single-step method involving β-d-galactosidase was explored for d-tagatose production. The two vital genes, β-d-galactosidase gene (lacZ) and l-arabinose isomerase mutant gene (araA') were extracted separately from Escherichia coli strains and incorporated into E. coli simultaneously. This gave us E. coli-ZY, a recombinant producing strain capable of coexpressing the two key enzymes. The resulted cells exhibited maximum d-tagatose producing activity at 34 °C and pH 6.5 and in the presence of borate, 10 mM Fe(2+), and 1 mM Mn(2+). Further monitoring showed that the recombinant cells could hydrolyze more than 95% lactose and convert 43% d-galactose into d-tagatose. This research has verified the feasibility of single-step d-tagatose fermentation, thereby laying down the foundation for industrial usage of lactose.

Reaction kinetics and galactooligosaccharide product profiles of the β-galactosidases from Bacillus circulans, Kluyveromyces lactis and Aspergillus oryzae.

Science.gov (United States)

Yin, Huifang; Bultema, Jelle B; Dijkhuizen, Lubbert; van Leeuwen, Sander S

2017-06-15

β-Galactosidase enzymes are used in the dairy industry to convert lactose into galactooligosaccharides (GOS) that are added to infant formula to mimic the molecular sizes and prebiotic functions of human milk oligosaccharides. Here we report a detailed analysis of the clearly different GOS profiles of the commercial β-galactosidases from Bacillus circulans, Kluyveromyces lactis and Aspergillus oryzae. Also the GOS yields of these enzymes differed, varying from 48.3% (B. circulans) to 34.9% (K. lactis), and 19.5% (A. oryzae). Their incubation with lactose plus the monosaccharides Gal or Glc resulted in altered GOS profiles. Experiments with 13 C 6 labelled Gal and Glc showed that both monosaccharides act as acceptor substrates in the transgalactosylation reactions. The data shows that the lactose isomers β-d-Galp-(1→2)-d-Glcp, β-d-Galp-(1→3)-d-Glcp and β-d-Galp-(1→6)-d-Glcp are formed from acceptor reactions with free Glc and not by rearrangement of Glc in the active site. Copyright © 2017 Elsevier Ltd. All rights reserved.

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%.

Actividad biológica de la prostaglandina F2 alfa y del metil carboprost en el fundus gástrico aislado de rata

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Roberto Rojas Martínez

1997-08-01

Full Text Available Se comparó la potencia contráctil de una muestra del éster metílico de 15 metil prostaglandina F2 alfa (metil carboprost (I, producida por la Industria Farmacéutica Cubana, con una muestra comercial de prostaglandina F2 alfa (II, de la firma Chinoin, mediante el ensayo de los "tres puntos". También se comparó la duración de la contracción inducida por dichas muestras utilizando un análisis de regresión lineal de la relajación observada en función del tiempo, luego de suprimir el contacto entre la droga y el órgano efector. Se utilizó el fundus gástrico aislado de rata como órgano efector. La actividad contráctil se registró mediante transductores fuerza-desplazamiento. La muestra I presentó una potencia relativa de 164,0 % con respecto a la II, con límites de confianza de 111,2 y 247,8 % (p=0,95; N=7, y mostró un efecto más prolongado en relación con la II. En las tiras contraídas por la muestra I debió transcurrir 6 min y 46 s, luego del lavado, para que la contracción se redujera a la mitad de la respuesta máxima observada (N=7, en tanto que en las tiras contraídas por la II fue de 1 min y 48 s (N= 8.The contractil potency of a sample of methylic ester of 15 methyl prostaglandin F2 alpha (methyl carboprost (I produced by the Cuban Pharmaceutical Industry, was compared to a commercial sample of prostaglandin F2 alpha (II from a Chinoin firm through the "three point" assay. The duration of the contraction induced by such samples was also compared using the analysis of linear regression of the relaxation observed in function of time after the supression of contact between the drug and the effector organ. The gastric fundus isolated from a rat was used as the effector organ. The contractil activity was recorded by strength-displacement transducers. Sample I showed a relative potency of 164.0 % with respect to sample II, with confidence limits of 111.2 and 247.8 % (P = 0.95; N=7 and it also showed a more prolonged

Daclatasvir plus peginterferon alfa and ribavirin for treatment-naive chronic hepatitis C genotype 1 or 4 infection

DEFF Research Database (Denmark)

Hézode, Christophe; Hirschfield, Gideon M; Ghesquiere, Wayne

2015-01-01

OBJECTIVE: To evaluate the safety and efficacy of daclatasvir, an HCV NS5A inhibitor with pangenotypic activity, administered with peginterferon-alfa-2a/ribavirin. DESIGN: In this Phase 2b double-blind, placebo-controlled study, treatment-naive adults with HCV genotype 1 (N=365) or 4 (N=30...

Survival of freeze-dried microcapsules of α-galactosidase producing probiotics in a soy bar matrix.

Science.gov (United States)

Chen, Mo; Mustapha, Azlin

2012-05-01

Soy oligosaccharides, mainly α-galactosides, are prevalently present in soy protein products, and can result in unfavorable digestive effects when consumed. The aim of this research was to investigate the efficiency of α-galactoside reduction by probiotic bacterial hydrolysis and if such bacteria could be maintained in a high number in a soy protein product in a microencapsulated and freeze-dried form. The probiotic bacterium, Lactobacillus acidophilus LA-2, when induced by raffinose, exhibited a high level of α-galactosidase activity at 5.0 U/mg. To preserve probiotics with high viability, cells were microencapsulated and freeze-dried. Optimization of microencapsulation presented that a combination of κ-carrageenan and inulin at a proportion of 1.9:0.1 (w:w) as capsule wall materials, significantly retained the viability of the probiotics through freeze-drying (P ≤ 0.05). Scanning electron microscopic images confirmed that the morphology of the microcapsules was well preserved after freeze-drying. Upon incorporation into soy protein bars, the freeze-dried microcapsules of L. acidophilus LA-2 remained in high numbers throughout 14 weeks of storage at 4 °C. Results of this work with the support of other studies on microencapsulation benefits indicate a promising use of freeze-dried α-galactosidase positive microencapsulated probiotics in a soy food. Copyright © 2011 Elsevier Ltd. All rights reserved.

Utilization of whey powder as substrate for low-cost preparation of β-galactosidase as main product, and ethanol as by-product, by a litre-scale integrated process.

Science.gov (United States)

You, Shengping; Chang, Hongxing; Yin, Qingdian; Qi, Wei; Wang, Mengfan; Su, Rongxin; He, Zhimin

2017-12-01

Whey powder, a by-product of dairy industry, is an attractive raw material for value-added products. In this study, utilization of whey powder as substrate for low-cost preparation of β-galactosidase as main product and ethanol as by-product were investigated by a litre-scale integrated strategy, encompassing fermentation, isolation, permeabilization and spray drying. Firstly, through development of low-cost industrial culture and fed-batch strategies by Kluyveromyces lactis, 119.30U/mL β-galactosidase activity and 16.96mg/mL by-product ethanol were achieved. Afterward, an up-dated mathematic model for the recycling permeabilization was established successfully and 30.4g cells sediment isolated from 5L fermentation broth were permeabilized completely by distilled ethanol from broth supernatant. Then β-galactosidase product with 5.15U/mg from protection of gum acacia by spray drying was obtained. Furthermore, by-product ethanol with 31.08% (v/v) was achieved after permeabilization. Therefore, the integrated strategy using whey powder as substrate is a feasible candidate for industrial-scale implementation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of the medium composition on formation of amylase by Bacillus sp

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Eliana de Oliveira. Santos

2003-01-01

Full Text Available Studies on the alpha -amylase synthesis was carried out with a moderately thermophilic, facultatively anaerobic Bacillus sp, isolated from soil samples. The cells were cultivated in a complex medium containing soluble starch or maltose as carbon source. The levels of the alpha -amylaseactivity detected in culture supernatants varied greatly with the type of carbon source used. Maltose, soluble starch and citrate stimulated alpha -amylaseformation. Addition of exogenous glucose repressed formation of alpha -amylase, demonstrating that a classical glucose effect was operative in this organism. The concentration of yeast extract was found to be important factor in the alpha -amylase synthesis bythe isolate.The activity of the enzyme increased between 2 and 5 g/L yeast extract concentration and then fell very rapidly beyond this point. The best concentration of peptone to alpha-amylase formation was found to be around 10g/L.Estudos sobre a síntese de alfa -amilase foram realizados com uma bactéria termofílica moderada e facultativa anaeróbica, isolada de amostras de solo. As células foram cultivadas em um meio complexo contendo amido solúvel ou maltose como fonte de carbono. Os níveis da atividade de alfa -amilase detectados no sobrenadante da cultura variaram grandemente com o tipo da fonte de carbono utilizada. Amido solúvel, maltose e citrato estimularam a formação de alfa -amilase. A adição de glicose as culturas reprimiu a formação da alfa -amilase, demonstrando que o clássico efeito glicose foi operativo neste organismo. A concentração de extrato de levedura foi um fator importante na formação de alfa -amilase pelo isolado. A atividade da enzima aumentou entre concentrações de 2 a 5 g/L e então caiu muito rapidamente em torno deste ponto. A melhor concentração de peptona para a formação da alfa -amilase foi em torno de 10 g/L.

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

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

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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.

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 �

SPECIFIC DISORDERS OF THE RESPIRATORY SYSTEM IN CYSTIC FIBROSIS. CLINICAL EFFICACY OF THERAPY WITH DORNASE ALFA IN CHILDREN

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T.V. Simanova

2010-01-01

Full Text Available The article is devoted to specific disorders of the respiratory system in cystic fibrosis. 64 patients with cystic fibrosis (CF aged 2 months to 32 years and residing in the Udmurtian Republic were studied. Epidemiological and genetic specifics of this disease in the mentioned region of the RF were examined. Clinical, X-ray, functional and microbiological studies of the CF patients’ respiratory system were conducted. It was found that genotype delF508 and chronic infection Pseudomonas aeruginosa, Staphylococcus aureus cause severe structural changes to the bronchopulmonary system more often. The obtained data suggest the advisability of identifying the groups of CF patients at the highest risk of severe respiratory system disorders in order to optimise therapeutic efforts. The article provides indicators of clinical efficacy of a dornase alfa therapy in CF children.Key words: cystic fibrosis, genotype, delF508 mutation, respiratory organs, pseudomonas aeruginosa infection, staphylococcal infection, respiratory function, mucolytic function, dornase alfa. (Pediatric Pharmacology. – 2010; 7(6:44-48

Simultaneous knock-down of six β-galactosidase genes in petunia petals prevents loss of pectic galactan but decreases petal strength.

Science.gov (United States)

O'Donoghue, Erin M; Somerfield, Sheryl D; Deroles, Simon C; Sutherland, Paul W; Hallett, Ian C; Erridge, Zoë A; Brummell, David A; Hunter, Donald A

2017-04-01

Galactose (Gal) is incorporated into cell wall polysaccharides as flowers open, but then is lost because of β-galactosidase activity as flowers mature and wilt. The significance of this for flower physiology resides in the role of galactan-containing polysaccharides in the cell wall, which is still largely unresolved. To investigate this, transcript accumulation of six cell wall-associated β-galactosidases was simultaneously knocked down in 'Mitchell' petunia (Petunia axillaris x (P. axillaris x P. hybrida)) flower petals. The multi-PhBGAL RNAi construct targeted three bud- and three senescence-associated β-galactosidase genes. The petals of the most down-regulated line (GA19) were significantly disrupted in galactose turnover during flower opening, and at the onset of senescence had retained 86% of their galactose compared with 20% in the controls. The Gal content of Na 2 CO 3 -soluble cell wall extracts and the highly insoluble polysaccharides associated with cellulose were particularly affected. Immunodetection with the antibody LM5 showed that much of the cell wall Gal in GA19 was retained as galactan, presumably the side-chains of rhamnogalacturonan-I. The flowers of GA19, despite having retained substantially more galactan, were no different from controls in their internal cell arrangement, dimensions, weight or timing of opening and senescence. However, the GA19 petals had less petal integrity (as judged by force required to cause petal fracture) after opening and showed a greater decline in this integrity with time than controls, raising the possibility that galactan loss is a mechanism for helping to maintain petal tissue cohesion after flower opening. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Unidades fototérmicas e temperatura-base inferior de frutos de Mangueira Alfa, na Baixada Cuiabana Photothermal units and lower base temperature for alfa mango fruit on the Baixada Cuiabana

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Marcelo Paes de Barros

2010-06-01

Full Text Available O objetivo deste trabalho foi determinar a temperatura-base inferior (Tb a partir da unidade fototérmica (UF para o cultivo experimental de manga Alfa sob condições de cerrado. Foram utilizados dados diários da temperatura do ar disponibilizados pela Estação Agrometeorológica Padre Ricardo Remetter, localizada em Santo Antônio do Leverger-MT (15,8° S e 56,1° W, 140 m, e avaliação sensorial do estádio de maturação dos frutos de manga. A Tb foi determinada pelo método da menor variabilidade das unidades fototérmicas (UF acumuladas do período da floração à colheita dos frutos, variabilidade avaliada pelo coeficiente de variação (cv simulado para diferentes valores da Tb. De julho a novembro de 2007, em três plantas de um pomar demonstrativo irrigado, foram identificados 82 frutos para o acompanhamento do crescimento e maturação. Entre esses frutos, em meados de dezembro de 2007, foi possível identificar 13 frutos que atingiram a maturidade fisiológica, após um período médio de observação de 112 dias. Para exigência fototérmica de 1.878.166,1 UF, encontrou-se Tb de 10 °C, valor consistente com os apresentados na literatura para a cultivar de manga, o que comprova a eficiência do método que combina a ação da temperatura e do fotoperíodo sobre a maturação dos frutos e que confere um caráter mais racional que o método tradicional da soma térmica.In this study was to determine the lower base temperature (Tb for Alfa mango fruit, using the photothermal units (PTU method in cerrado conditions. The meteorological data for the study were acquired from "Padre Ricardo Remetter" meteorological station located at Santo Antonio do Leverger, MT, Brazil (15,8° S and 56,1° W, 140 m, and the fruit data was acquired by sensorial evaluation of mango fruit maturation state. The Tb was determined by less variability of the coefficient of variation (cv method applied to the PTU sum, from flowering to harvest, simulated for

Forward Detectors in ATLAS: ALFA, ZDC and LUCID

CERN Document Server

Fabbri, L; The ATLAS collaboration

2009-01-01

In order to determine the experimental cross sections for the observed physics processes, an estimation of the absolute luminosity is needed. In fact a careful study of “well known” processes will be one of the first steps of the LHC experiments as it can provide possible signatures of new physics which consist in deviations with respect to the Standard Model (SM) predictions. The methodologies for luminosity monitoring and total cross section estimation at the LHC will be reviewed in this talk along with the dedicated detectors of the ATLAS experiment. ATLAS will make extensive usage of the detectors in the forward region each one with a different task: LUCID (LUminosity measurement using Cherenkov Integrating Detector) is a system of 40 (2 x 20) Cherenkov tubes, surrounding the beam pipe at about 17 m from the interaction region. It will be able to monitor the collision-by-collision luminosity by detecting and counting the number of charged particles coming from the impact point. ALFA (Absolute Luminosi...

Efeito da suplementação enzimática em rações à base de milho/farelo de soja sobre o desempenho de poedeiras comerciais Effect of the enzyme supplementation of corn/soybean meal based diets on the performance of commercial laying hens

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Ednardo Rodrigues Freitas

2000-08-01

Full Text Available O experimento foi realizado com o objetivo de estudar o efeito do uso de um complexo enzimático comercial (alfa-amilase, xilanase e protease específico para dietas à base de milho e farelo de soja sobre o desempenho de poedeiras comerciais. Cento e vinte e oito poedeiras da linhagem Hy-Line, com 68 semanas de idade e no segundo ciclo de postura, foram alojadas em um delineamento de blocos ao acaso com quatro repetições de quatro aves por bloco, totalizando 32 aves por tratamento. Duas dietas isoprotéicas, porém com diferentes níveis de EM, foram formuladas à base de milho, farelo de soja, farelo de trigo e suplementadas ou não com enzimas. Os tratamentos foram: T1 - dieta contendo 2850 kcal EM/kg; T2 - dieta contendo 2850 kcal EM/kg, com 0,1% enzima; T3 - dieta contendo 2750 kcal EM/kg; e T4 - dieta contendo 2750 kcal EM/kg, com 0,1% enzima. O experimento teve duração de 112 dias, divididos em quatro períodos de 28 dias, e as variáveis estudadas foram: porcentagem de postura (ave/dia, consumo de ração (g/ave/dia, peso dos ovos (g, conversão alimentar (kg de ração/kg de ovo, ganho de peso das poedeiras (g e custo com alimentação (R$. A análise de variância dos dados demonstrou que não existiu efeito dos tratamentos sobre as variáveis estudadas. A suplementação enzimática das dietas nos níveis estudados não afetou o desempenho das poedeiras.This experiment was conducted to study the effect of the inclusion of a commercial enzyme complex (alpha-amylase, xylanase and proteases especific for corn and soybean meal based diets, on laying hens performance. One hundred and twenty eight Hy-line White Leghorns hens with 68 weeks of age, in the 2nd cycle of production were assigned to a completely randomized block design with four replications of four birds per block, totalizing 32 birds per treatment. Two isonitrogenous diets with different levels of metabolizable energy (ME were formulated based on corn, soybean meal, wheat

Experimental investigation of the mechanical properties of Alfas stone

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Konstas N. Kaklis

2017-04-01

Full Text Available This paper focuses on the experimental investigation of the mechanical properties of the Alfas natural building stone. Two series of uniaxial compression tests and indirect tensile tests (Brazilian tests were performed in order to determine the uniaxial compressive strength and the indirect tensile strength respectively. Different sets of cylindrical specimens and circular discs were prepared by varying their geometry in order to examine the size effect on the respective strength values. Also, the size effect was investigated with respect to the calculated intact rock modulus and Poisson’s ratio. All specimens were prepared by following the ISRM suggested methods and the load was applied using a stiff 1600 kN MTS hydraulic testing machine and a 500 kN load cell. Strain was measured using biaxial 0/90 stacked rosettes appropriately attached on each specimen.

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

β-Galactosidase activity of commercial lactase samples in raw and pasteurized milk at refrigerated temperatures.

Science.gov (United States)

Horner, T W; Dunn, M L; Eggett, D L; Ogden, L V

2011-07-01

Many consumers are unable to enjoy the benefits of milk due to lactose intolerance. Lactose-free milk is available but at about 2 times the cost of regular milk or greater, it may be difficult for consumers to afford. The high cost of lactose-free milk is due in part to the added cost of the lactose hydrolysis process. Hydrolysis at refrigerated temperatures, possibly in the bulk tank or package, could increase the flexibility of the process and potentially reduce the cost. A rapid β-galactosidase assay was used to determine the relative activity of commercially available lactase samples at different temperatures. Four enzymes exhibited low-temperature activity and were added to refrigerated raw and pasteurized milk at various concentrations and allowed to react for various lengths of time. The degree of lactose hydrolysis by each of the enzymes as a function of time and enzyme concentration was determined by HPLC. The 2 most active enzymes, as determined by the β-galactosidase assay, hydrolyzed over 98% of the lactose in 24h at 2°C using the supplier's recommended dosage. The other 2 enzymes hydrolyzed over 95% of the lactose in 24h at twice the supplier's recommended dosage at 2°C. Results were consistent in all milk types tested. The results show that it is feasible to hydrolyze lactose during refrigerated storage of milk using currently available enzymes. Copyright © 2011 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Desempenho, composição da cama, das tíbias, do fígado e das excretas de frangos de corte alimentados com rações contendo fitase e baixos níveis de fósforo Performance, litter composition, tibia, liver and excreta of broilers fed diets containing reduced levels of phosphorus and phytase enzyme

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Antonio Carlos de Laurentiz

2009-10-01

Full Text Available Um experimento foi realizado para avaliar o desempenho e a composição da cama, das tíbias, do fígado e das excretas de frangos de corte em diversas fases utilizando-se diferentes níveis de fósforo disponível (Pd e da enzima fitase. Foram utilizados 1.200 pintos machos de 1 dia de idade, da marca comercial Cobb, distribuídos em delineamento inteiramente casualizado, em esquema fatorial 4 × 3, composto de quatro níveis de fósforo disponível e três níveis de fitase: 0, 500 e 1.000 ftu/kg de ração, cada um com quatro repetições de 25 aves. Os níveis de fósforo disponível foram obtidos considerando quatro disponibilidades do fósforo total (Pt do milho e do farelo de soja (34, 56, 78 e 100% de disponibilidade. Foram avaliados os índices de desempenho no final de cada fase de criação e aos 42 dias de idade. Os parâmetros de desempenho foram afetados pela redução dos níveis de fósforo disponível das rações e a redução desse elemento para níveis inferiores a 19% piorou o desempenho das aves, enquanto a utilização de fitase no nível de Pd reduzido em 39% melhorou o desempenho. Nas fases de criação, a redução do teor de fósforo disponível com a utilização da enzima fitase influencia o desempenho e as características da cama e dos ossos.An experiment was conducted to evaluate the performance and composition of litter, tibia, liver and excreta of broiler chicks at different phases using different levels of available phosphorous (Pd and the enzyme phytase. One thousand and two hundred one-day-Cobb-male chicks were used, distributed in a randomized complete design, using a 4 × 3 factorial arrangement, consisting of four available phosphorus levels and three phytase levels - 0, 500 and 1,000 FTU phytase/kg of diet with four replications of 25 birds each. The levels of available phosphorus were obtained considering four different availabilities of total phosphorus (Pt, corn and soybean meal (34, 56, 78 and 100

Kinetics and Regulation Studies of the Production of β-Galactosidase from Kluyveromyces marxianus Grown on Different Substrates

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Samia Khan

2003-01-01

Full Text Available Lactose-intolerance is manifested in 50 % of the world’s population. This can be remediated by removing lactose from the diet or converting it into glucose and galactose with β-galactosidase (EC 3.2.1.23. In this work, batch production of this enzyme in the presence of lactose, galactose, cellobiose, xylose, arabinose, sucrose and glucose was investigated using Kluyveromyces marxianus in shake flask culture studies. Substrate type and temperature were the independent variables that directly regulated the specific growth and β-galactosidase production rates. Lactose (2 % supported the maximum specific product yield (YP/X, followed by galactose, sucrose, cellobiose, xylose, arabinose and glucose. Its synthesis was regulated by an induction and a growth-dependent repression mechanism. The optimum temperature for the production was found to be 35–37 °C. The highest volumetric productivity of enzyme (80.0 IU/L/h occurred on lactose-corn steep liquor medium. This was significantly higher than the calculated values reported in the literature. Thermodynamic studies revealed that the cells provided a defence mechanism against thermal inactivation. The enzyme was stable at 60 °C and pH=5.0–7.0, and it may find application in commercial lactose hydrolysis.

The Arecibo Legacy Fast ALFA Survey : X. The HI Mass Function and Omega_HI From the 40% ALFALFA Survey

NARCIS (Netherlands)

Martin, Ann M.; Papastergis, Emmanouil; Giovanelli, Riccardo; Haynes, Martha P.; Springob, Christopher M.; Stierwalt, Sabrina

2010-01-01

The Arecibo Legacy Fast ALFA (ALFALFA) survey has completed source extraction for 40% of its total sky area, resulting in the largest sample of HI-selected galaxies to date. We measure the HI mass function from a sample of 10,119 galaxies with 6.2

DAHANCA 10 – Effect of darbepoetin alfa and radiotherapy in the treatment of squamous cell carcinoma of the head and neck. A multicenter, open-label, randomized, phase 3 trial by the Danish head and neck cancer group

DEFF Research Database (Denmark)

Overgaard, Jens; Hoff, Camilla Molich; Hansen, Hanne Sand

2018-01-01

Purpose: To evaluate if correction of low hemoglobin (Hb) levels by means of darbepoetin alfa improves the outcomes of radiotherapy in patients with squamous cell carcinoma of the head and neck (HNSCC). Patients and methods: Patients eligible for primary radiotherapy and who had Hb values below 14.......0 g/dl were randomized to receive accelerated fractionated radiotherapy with or without darbepoetin alfa. Patients also received the hypoxic radiosensitizer nimorazole. Darbepoetin alfa was given weekly during radiotherapy or until the Hb value exceeded 15.5 g/dl. Results: Following a planned interim...... distributed according to the stratification parameters (gender, T and N staging, tumor site). Treatment with darbepoetin alfa increased the Hb level to the planned value in 81% of the patients. The compliance was good without excess serious adverse events. The results showed a poorer outcome with a 5-year...

Deficiência de alfa-1 antitripsina: diagnóstico e tratamento Alpha-1 antitrypsin deficiency: diagnosis and treatment

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Aquiles A Camelier

2008-07-01

Full Text Available A deficiência de alfa-1 antitripsina é um distúrbio genético de descoberta recente e que ocorre com freqüência comparável à da fibrose cística. Resulta de diferentes mutações no gene SERPINA1 e tem diversas implicações clínicas. A alfa-1 antitripsina é produzida principalmente no fígado e atua como uma antiprotease. Tem como principal função inativar a elastase neutrofílica, impedindo a ocorrência de dano tecidual. A mutação mais freqüentemente relacionada à doença clínica é o alelo Z, que determina polimerização e acúmulo dentro dos hepatócitos. O acúmulo e a conseqüente redução dos níveis séricos de alfa-1 antitripsina determinam, respectivamente, doença hepática e pulmonar, sendo que esta se manifesta principalmente sob a forma de enfisema de aparecimento precoce, habitualmente com predomínio basal. O diagnóstico envolve a detecção de níveis séricos reduzidos de alfa-1 antitripsina e a confirmação fenotípica. Além do tratamento usual para doença pulmonar obstrutiva crônica, existe atualmente uma terapia específica com infusão de concentrados de alfa-1 antitripsina. Essa terapia de reposição, aparentemente segura, ainda não teve a eficácia clínica definitivamente comprovada, e o custo-efetividade também é um tema controverso e ainda pouco abordado. Apesar da sua importância, não existem dados epidemiológicos brasileiros a respeito da prevalência da doença ou da freqüência de ocorrência dos alelos deficientes. O subdiagnóstico também tem sido uma importante limitação tanto para o estudo da doença quanto para o tratamento adequado dos pacientes. Espera-se que a criação do Registro Internacional de Alfa-1 venha a resolver essas e outras importantes questões.Alpha-1 antitrypsin deficiency is a recently identified genetic disease that occurs almost as frequently as cystic fibrosis. It is caused by various mutations in the SERPINA1 gene, and has numerous clinical

Anti-α-galactosidase A antibody response to agalsidase beta treatment

DEFF Research Database (Denmark)

Wilcox, William R; Linthorst, Gabor E; Germain, Dominique P

2012-01-01

Agalsidase beta, a form of recombinant human α-galactosidase A (αGAL), is approved for use as enzyme replacement therapy (ERT) for Fabry disease. An immunogenic response against a therapeutic protein could potentially impact its efficacy or safety. The development of anti-αGAL IgG antibodies...... was evaluated in 571 men and 251 women from the Fabry Registry who were treated with agalsidase beta. Most men developed antibodies (416 of 571, 73%), whereas most women did not (31 of 251, 12%). Women were also significantly more likely to tolerize than men; whereas 18 of 31 women tolerized (58%, 95%CI: 52......%-64%), only 47 of 416 men tolerized during the observation period (11%, 95% CI: 8%-15%). Patients who eventually tolerized had lower median peak anti-αGAL IgG antibody titers than patients who remained seropositive at their most recent assessment (400 versus 3200 in men, 200 versus 400 in women, respectively...

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.

Interferentes eritrocitários e ambientais na anemia falciforme

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Naoum Paulo C.

2000-01-01

Full Text Available A anemia falciforme tem um desenvolvimento clínico extremamente variável que se caracteriza principalmente por diferentes graus de intensidade da anemia hemolítica. As razões dessa variabilidade são parcialmente conhecidas na expressão fenotípica da doença. Apesar de ter um mesmo defeito genético, a anemia falciforme pode estar associada com níveis diferentes de Hb Fetal e interações com talassemia alfa que atuam como modeladores genéticos da doença. Entretanto, outros defeitos genéticos dos eritrócitos, com destaques para a deficiência de G-6PD, a esferocitose e as deficiências de enzimas anti-oxidantes (SOD, GPx e catalase certamente interferem no curso clínico da doença. Os diferentes haplótipos da Hb S denominados por Banto, Benin, Senegal, Camarões e Asiático, tem sido apontados também como possíveis causas da heterogeneidade fenotípica da anemia falciforme. Toda essa diversidade que caracteriza a anemia falciforme está, em parte, relacionada à sua origem multicêntrica e que envolvem populações com diferentes anormalidades genéticas de proteínas e enzimas eritrocitárias. Por outro lado, além desses fatores caracterizados como interferentes eritrocitários, há os interferentes do meio ambiente em que está inserido o doente com anemia falciforme. Entre os interferentes ambientais destacam-se as situações sociais, econômicas e culturais do doente, e que tem influência no curso de sua doença. Diante desse quadro complexo e interativo, o presente artigo mostra a influência de certos interferentes eritrocitários e ambientais na anemia falciforme. Ao finalizar o artigo é proposto um protocolo de monitoramento laboratorial das síndromes falcêmicas, com destaque para a anemia falciforme.

An Open-label, Single-dose, Pharmacokinetic Study of Factor VIII Activity After Administration of Moroctocog Alfa (AF-CC) in Male Chinese Patients With Hemophilia A.

Science.gov (United States)

Liu, Hongzhong; Wu, Runhui; Hu, Pei; Sun, Feifei; Xu, Lihong; Liang, Yali; Nepal, Sunil; Qu, Peng Roger; Huard, Francois; Korth-Bradley, Joan M

2017-07-01

Hemophilia A represents up to 80% of all hemophilia cases in China. In patients with this condition, bleeding can be prevented and controlled by administering clotting factor VIII (FVIII). Since their initial availability, recombinant FVIII products have undergone several iterations to enhance their safety. Moroctocog alfa albumin-free cell culture (AF-CC) is among the third generation of recombinant FVIII products and received regulatory approval in China in August 2012. The present study characterizes the single-dose pharmacokinetic parameters of FVIII activity (FVIII:C) after administration of moroctocog alfa (AF-CC) in male Chinese patients with hemophilia A. This multicenter, open-label, single-dose study enrolled 13 male Chinese patients diagnosed with severe hemophilia A (FVIII:C hemophilia A. The pharmacokinetic profile in older patients was similar to that previously reported with recombinant FVIII products in studies with a predominantly white population; younger patients had reduced exposure to FVIII:C. The single doses of moroctocog alfa (AF-CC) were well tolerated; 2 cases of transient, low-titer FVIII inhibitor development were observed. ClinicalTrials.gov identifier: NCT02461992. Copyright © 2017 Elsevier HS Journals, Inc. All rights reserved.

Peginterferon alfa-2a and ribavirin in Latino and non-Latino whites with hepatitis C.

Science.gov (United States)

Rodriguez-Torres, Maribel; Jeffers, Lennox J; Sheikh, Muhammad Y; Rossaro, Lorenzo; Ankoma-Sey, Victor; Hamzeh, Fayez M; Martin, Paul

2009-01-15

Race has been shown to be a factor in the response to therapy for hepatitis C virus (HCV) infection, and limited data suggest that ethnic group may be as well; however, Latinos and other ethnic subpopulations have been underrepresented in clinical trials. We evaluated the effect of Latino ethnic background on the response to treatment with peginterferon alfa-2a and ribavirin in patients infected with HCV genotype 1 who had not been treated previously. In a multicenter, open-label, nonrandomized, prospective study, 269 Latino and 300 non-Latino whites with HCV infection received peginterferon alfa-2a, at a dose of 180 microg per week, and ribavirin, at a dose of 1000 or 1200 mg per day, for 48 weeks, and were followed through 72 weeks. The primary end point was a sustained virologic response. We enrolled Latinos whose parents and grandparents spoke Spanish as their primary language; nonwhite Latinos were excluded. Baseline characteristics were similar in the Latino and non-Latino groups, although higher proportions of Latino patients were 40 years of age or younger, had a body-mass index (BMI, the weight in kilograms divided by the square of the height in meters) of more than 27 or more than 30, and had cirrhosis. The rate of sustained virologic response was higher among non-Latino whites than among Latinos (49% vs. 34%, PLatino whites at week 4 (P=0.045) and throughout the treatment period (PLatino or non-Latino background was an independent predictor of the rate of sustained virologic response in an analysis adjusted for baseline differences in BMI, cirrhosis, and other characteristics. Adherence to treatment did not differ significantly between the two groups. The numbers of patients with adverse events and dose modifications were similar in the two groups, but fewer Latino patients discontinued therapy because of adverse events. Treatment with peginterferon alfa-2a and ribavirin for 48 weeks resulted in rates of sustained virologic response among patients

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

OpenAIRE

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...

Prevention of Bacterial Contamination of a Silica Matrix Containing Entrapped β-Galactosidase through the Action of Covalently Bound Lysozymes

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Heng Li

2017-02-01

Full Text Available β-galactosidase was successfully encapsulated within an amino-functionalised silica matrix using a “fish-in-net” approach and molecular imprinting technique followed by covalent binding of lysozyme via a glutaraldehyde-based method. Transmission electron microscopy (TEM, X-ray diffraction (XRD, scanning electron microscopy (SEM, and Fourier transform infrared (FTIR spectroscopy were used to characterise the silica matrix hosting the two enzymes. Both encapsulated β-galactosidase and bound lysozyme exhibited high enzymatic activities and outstanding operational stability in model reactions. Moreover, enzyme activities of the co-immobilised enzymes did not obviously change relative to enzymes immobilised separately. In antibacterial tests, bound lysozyme exhibited 95.5% and 89.6% growth inhibition of Staphylococcus aureus ATCC (American type culture collection 653 and Escherichia coli ATCC 1122, respectively. In milk treated with co-immobilised enzymes, favourable results were obtained regarding reduction of cell viability and high lactose hydrolysis rate. In addition, when both co-immobilised enzymes were employed to treat milk, high operational and storage stabilities were observed. The results demonstrate that the use of co-immobilised enzymes holds promise as an industrial strategy for producing low lactose milk to benefit people with lactose intolerance.

A Fungal α-Galactosidase from Tricholoma matsutake with Broad Substrate Specificity and Good Hydrolytic Activity on Raffinose Family Oligosaccharides.

Science.gov (United States)

Geng, Xueran; Tian, Guoting; Zhao, Yongchang; Zhao, Liyan; Wang, Hexiang; Ng, Tzi Bun

2015-07-24

An acidic α-galactosidase designated as TMG was purified from the fruiting bodies The purification protocol entailed ion exchange chromatography on Q-Sepharose and of Tricholoma matsutake with 136-fold purification and a specific activity of 909 units/mg. Mono-Q and fast protein liquid chromatography on Superdex 75. TMG is a monomeric protein exhibiting a molecular mass of 47 kDa in SDS-PAGE and gel filtration. The purified enzyme was identified by LC-MS/MS and three inner amino acid sequences were obtained. The optimum pH and temperature for TMG with pNPGal as substrate were pH 4.5 and 55 °C, respectively. The α-galactosidase activity was strongly inhibited by K+, Ca2+, Cd2+, Hg2+, Ag+ and Zn2+ ions. The enzyme activity was inhibited by the chemical modification agent N-bromosuccinimide (NBS), indicating the importance of tryptophan residue(s) at or near the active site. Besides hydrolyzing pNPGal, TMG also efficaciously catalyzed the degradation of natural substrates such as stachyose, raffinose, and melibiose. Thus TMG can be exploited commercially for improving the nutritional value of soy milk by degradation of indigestible oligosaccharides.

A Fungal α-Galactosidase from Tricholoma matsutake with Broad Substrate Specificity and Good Hydrolytic Activity on Raffinose Family Oligosaccharides

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Xueran Geng

2015-07-01

Full Text Available An acidic α-galactosidase designated as TMG was purified from the fruiting bodies The purification protocol entailed ion exchange chromatography on Q-Sepharose and of Tricholoma matsutake with 136-fold purification and a specific activity of 909 units/mg. Mono-Q and fast protein liquid chromatography on Superdex 75. TMG is a monomeric protein exhibiting a molecular mass of 47 kDa in SDS-PAGE and gel filtration. The purified enzyme was identified by LC-MS/MS and three inner amino acid sequences were obtained. The optimum pH and temperature for TMG with pNPGal as substrate were pH 4.5 and 55 °C, respectively. The α-galactosidase activity was strongly inhibited by K+, Ca2+, Cd2+, Hg2+, Ag+ and Zn2+ ions. The enzyme activity was inhibited by the chemical modification agent N-bromosuccinimide (NBS, indicating the importance of tryptophan residue(s at or near the active site. Besides hydrolyzing pNPGal, TMG also efficaciously catalyzed the degradation of natural substrates such as stachyose, raffinose, and melibiose. Thus TMG can be exploited commercially for improving the nutritional value of soy milk by degradation of indigestible oligosaccharides.

Tratamento de hemangioma gigante com interferon alfa: relato de dois casos Treatment of giant hemangioma with interferon-alpha: report of two cases

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Ana Julia Balau

2007-12-01

Full Text Available O objetivo do trabalho é descrever o uso de interferon alfa no tratamento de pacientes com hemangioma gigante. Os autores relatam e analisam dois casos de hemangioma gigante em tratamento com interferon alfa. IBS, 3 anos, em acompanhamento no Ambulatório de Hematologia desde um ano de idade com quadro de lesão angiomatosa em praticamente toda hemiface direita, acompanhada de sangramentos gengivais importantes. Após a realização de exames complementares (Angiorressonância magnética e feito o diagnóstico de hemangioma gigante em face, foi iniciado tratamento com prednisona e, posteriormente, associação com interferon alfa e observada importante melhora do quadro, resultando na diminuição dos episódios de sangramento e no tamanho do tumor. C.N.P., 12 anos, apresentando nódulo em região lateral de joelho esquerdo há 2 anos, com aumento progressivo do tamanho e dor local. Fez uso de prednisona e, sem melhora do quadro, introduzido interferon alfa com regressão importante do tamanho do tumor. O tratamento com interferon alfa deve ser considerado no tratamento de hemangiomas, pois apresenta bons resultados em relação à diminuição do tamanho do tumor e, conseqüentemente, reduz as intercorrências clínicas associadas à sua presença, principalmente os sangramentos.The aim of this study is to describe the treatment using interferon-alpha of giant hemangiomas in children. The authors report two cases of children presenting with giant hemangiomas treated using interferon-alpha and analyze the results. IBS, 3 years-old, has been followed up in Famema Hemathology Service since she was 1 year-old with a tumor on the face and persistent bleeding. After clinical and radiologic evaluations and suggested the diagnosis of giant hemangioma, she started treatment with interferon-alpha. A great clinical improvement was observed a reducing of the number of episodes of bleedings and a decrease in of the tumor size. CNP, 12 years-old, came to

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

Interferon alfa for chronic hepatitis B infection: increased efficacy of prolonged treatment. The European Concerted Action on Viral Hepatitis (EUROHEP)

NARCIS (Netherlands)

Janssen, H. L.; Gerken, G.; Carreño, V.; Marcellin, P.; Naoumov, N. V.; Craxi, A.; Ring-Larsen, H.; Kitis, G.; van Hattum, J.; de Vries, R. A.; Michielsen, P. P.; ten Kate, F. J.; Hop, W. C.; Heijtink, R. A.; Honkoop, P.; Schalm, S. W.

1999-01-01

Interferon alfa (IFN-alpha) is the primary treatment for chronic hepatitis B. The standard duration of IFN-alpha therapy is considered 16 weeks; however, the optimal treatment length is still poorly defined. We evaluated the efficacy and acceptability of prolonged IFN-alpha treatment in patients

Nebulised dornase alfa versus placebo or hypertonic saline in adult critically ill patients

DEFF Research Database (Denmark)

Claudius, Casper; Perner, Anders; Møller, Morten Hylander

2015-01-01

a systematic review with meta-analysis and trial sequential analysis (TSA) using the Cochrane Collaboration methodology. Eligible trials were randomised clinical trials comparing nebulised dornase alfa with placebo, no prophylaxis, or hypertonic saline. The predefined outcome measures were all-cause mortality...... of the primary estimate was assessed by TSA. RESULTS: Two trials (n = 63) were included; both were judged to have high risk of bias. There was no statistically significant difference in mortality (random effects model RR (95 % CI) 0.73 (0.09-5.77); P = 0.24; I (2) = 30 %). TSA could not be conducted because less...

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...

Perfil bioquímico do soro de frangos de corte alimentados com dieta suplementada com alfa-amilase de Cryptococcus flavus e Aspergillus niger HM2003 Biochemichal serum profile of broilers fed diets suplemented with alfa-amylase from Cryptococcus flavus and Aspergillus niger HM2003

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Cibele Silva Minafra

2010-12-01

Full Text Available Avaliou-se o perfil bioquímico do soro de frangos de corte alimentados com a enzima α-amilase produzida por dois microrganismos. Produziram-se dois extratos, um com a-amilase obtida a partir de Cryptococcus flavus em meio de levedura comercial e outro com Aspergillus niger HM2003 em meio de proteína de soja e amido comercial, com atividade de 9,58 U/mL e 10,0 U/mL, respectivamente. Utilizaram-se 360 pintos de corte Cobb 500 de 1 dia de idade e com 49,72 ± 0,68 g de peso vivo inicial. As aves foram alojadas em baterias e foram criadas até os 21 dias de idade. Foram utilizados três dietas, cada uma com cinco repetições de 12 aves, em delineamento inteiramente casualizado. A primeira dieta (basal foi formulada sem adição de enzima e as outras duas receberam a suplementação de a-amilase produzida por cultivo de Cryptococcus flavus e Aspergillus niger HM2003. Dietas à base de milho e soja foram formuladas em duas fases: pré-inicial (1-7 dias e inicial (8-21 dias. Na fase pré-inicial, foram observados os seguintes valores médios para cálcio (6,90 e 5,99 mg/dL, proteína plasmática (2,0 e 2,50 g/dL e fosfatase alcalina (979,98 e 974,66 UI/L, respectivamente para Cryptococcus flavus e Aspergillus niger HM2003. A dieta acrescida de a-amilase obtida a partir de Aspergillus niger HM2003 determinou maior concentração sérica de fósforo. Na fase inicial, os resultados significativos relacionaram-se a potássio quando avaliadas dietas com adição de a-amilase pelas duas fontes. A incorporação das enzimas testadas não proporciona alterações metabólicas ou toxicidade nos animais.It was evaluated the biochemical serum profile of broilers fed rations supplemented with α-amylase produced by two microorganisms. Two extracts were produced, one was produced with a-amylase obtained from Cryptococcus flavus in a commercial yeast-based medium and the other with Aspergillus niger HM2003 produced in soybean protein and commercial starch medium

Bakterisel Alfa-Amilaz Önemi, Özellikleri ve Üretim Teknolojisi

OpenAIRE

Esener, A. Aydın

2015-01-01

Enzimler biyolojik katalizörlerdir. Çeşitli mikroorganizmalar, çevrelerini yaşamları için elverişli bir hale getirebilme çabası ile bazı enzimleri bünyelerinde sentez edip hücre duvarından dışarıya salgılamaktadırlar. Alfa-amilaz da bu tür bir enzim olup, özellikle nişasta gibi doğal kaynaklı ham maddelerin parçalanmasında kullanılmaktadır. Endüstride geniş tüketim olanakları bulan bu enzimin, modern yöntemlerle üretilmesi, özellikle fermente olabilir tarım artığı maddelerin bol olduğu Türkiy...

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.

Nucleotide sequence of the melA gene, coding for alpha-galactosidase in Escherichia coli K-12.

OpenAIRE

Liljeström, P L; Liljeström, P

1987-01-01

Melibiose uptake and hydrolysis in E.coli is performed by the MelB and MelA proteins, respectively. We report the cloning and sequencing of the melA gene. The nucleotide sequence data showed that melA codes for a 450 amino acid long protein with a molecular weight of 50.6 kd. The sequence data also supported the assumption that the mel locus forms an operon with melA in proximal position. A comparison of MelA with alpha-galactosidase proteins from yeast and human origin showed that these prot...

Alfa hemolisina induce un aumento de calcio eritrocitario: estudio por FLIM (Fluorescence Lifetime Imaging Microscopy)

OpenAIRE

Sánchez, Susana; Bakás, Laura; Gratton, Enrico; Herlax, Vanesa Silvana

2010-01-01

Alfa hemolisina (HlyA), miembro representativo de la familia de toxinas RTX (Repeat in toxin), es una toxina proteica que ciertas cepas patógenas de E.coli secretan específicamente al medio. Está demostrado que la toxina es un factor importante de virulencia en enfermedades extraintestinales. Además, se ha determinado que el desarrollo de falla renal aguda aumenta significativamente la mortalidad en pacientes con shock séptico producido por bacterias Gram negativas, en los que distintos facto...

Analysis of bone marrow stromal cell transferred bacterial {beta}-galactosidase gene by PIXE

Energy Technology Data Exchange (ETDEWEB)

Kumakawa, Toshiro [Tokyo Metropolitan Geriatric Hospital, Tokyo (Japan). Dept. of Blood Transfusion and Hematology; Hibino, Hitoshi; Tani, Kenzaburo; Asano, Shigetaka; Futatugawa, Shouji; Sera, Kouichiro

1997-12-31

PIXE, Particle Induced X-ray Emission, is a powerful, multi-elemental analysis method which has many distinguishing features and has been used in varies research fields. Recently the method of applying baby cyclotrons for nuclear medicine to PIXE has been developed. This enables us to study biomedical phenomena from the physical point of view. Mouse bone marrow stromal cells were transferred bacterial {beta}-galactosidase gene (LacZ gene) by murine retroviral vectors. Analysis of the bone marrow stromal cells with the LacZ gene by PIXE revealed remarkable changes of intracellular trace elements compared with the normal control cells. These results indicate that gene transfer by retroviral vectors may bring about a dynamic change of intracellular circumstances of the target cell. (author)

Evaluation of treatment response to enzyme replacement therapy with Velaglucerase alfa in patients with Gaucher disease using whole-body magnetic resonance imaging.

Science.gov (United States)

Laudemann, K; Moos, L; Mengel, E; Lollert, A; Hoffmann, C; Brixius-Huth, M; Wagner, D; Düber, C; Staatz, G

2016-03-01

This was a retrospective data analysis to evaluate the treatment response to enzyme replacement therapy (ERT) with Velaglucerase alfa using whole-body magnetic resonance imaging (MRI). A baseline and follow-up MRI were performed on 18 Gaucher Type 1 patients at an interval of 11.6 months. The MRI score systems determined the Bone-Marrow-Burden (BMB) score, the Düsseldorf-Gaucher score (DGS), and the Vertebra-Disc-Ratio (VDR). The Severity Score Index Type 1 (GD-DS3) was also assessed. The baseline MRI medians were: BMB, 7.00; DGS, 3.00; and VDR: 1.70; while, the follow-up MRI medians were: BMB, 7.00; DGS, 3.00; and VDR: 1.73. The baseline GD-DS3 median was 2.40 (BMB excl.: 0.50) and the follow-up median was 2.00 (BMB excl.: 0.50). There was weak statistical significance with the Wilcoxon signed-rank test for the DGS (p=0.034) and GD-DS3 (p=0.047) between both MRIs. Velaglucerase alfa therapy is a effective long-term treatment for Gaucher Type 1 patients who are newly diagnosed or switching therapies. Measurements with whole-body MRI and an objective scoring system were reliable tools for detecting early stage bone marrow activity. Further research is needed to evaluate the "Booster-Effect" of Velaglucerase alfa therapy in Gaucher skeletal disease. Copyright © 2015 Elsevier Inc. All rights reserved.

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.

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

Peginterferon alfa-2b and weight-based or flat-dose ribavirin in chronic hepatitis C patients: a randomized trial.

Science.gov (United States)

Jacobson, Ira M; Brown, Robert S; Freilich, Bradley; Afdhal, Nezam; Kwo, Paul Y; Santoro, John; Becker, Scott; Wakil, Adil E; Pound, David; Godofsky, Eliot; Strauss, Robert; Bernstein, David; Flamm, Steven; Pauly, Mary Pat; Mukhopadhyay, Pabak; Griffel, Louis H; Brass, Clifford A

2007-10-01

This prospective, multicenter, community-based and academic-based, open-label, investigator-initiated, U.S. study evaluated efficacy and safety of pegylated interferon (PEG-IFN) alfa-2b plus a flat or weight-based dose of ribavirin (RBV) in adults with chronic hepatitis C. Patients (n = 5027) were randomly assigned to receive PEG-IFN alfa-2b 1.5 microg/kg/week plus flat-dose (800 mg/day) or weight-based (800-1400 mg/day) RBV for 48 weeks (patients with genotype 1, 4, 5, or 6) and for 24 or 48 weeks (genotype 2/3 patients). Primary end point was sustained virologic response (undetectable [<125 IU/mL] serum hepatitis C virus RNA at 24-week follow-up). Sustained virologic response, but not end-of-treatment, rates were significantly higher with weight-based than with flat-dose RBV (44.2% versus 40.5%; P = 0.008). Sustained virologic response rates by intention-to-treat analysis were 34.0% and 28.9%, respectively, in genotype 1 patients (P = 0.005) and 31.2% and 26.7%, respectively, in genotype 1 patients with high baseline viral load (P = 0.056). In genotype 2/3 patients, rates were not significantly different (61.8% and 59.5%, respectively) regardless of treatment duration. Besides greater hemoglobin reductions with weight-based RBV, safety profiles were similar across RBV dosing groups, including the 1400-mg/day group. PEG-IFN alfa-2b plus weight-based RBV is more effective than flat-dose RBV, particularly in genotype 1 patients, providing equivalent efficacy across all weight groups. RBV 1400 mg/day is appropriate for patients 105 to 125 kg. For genotype 2/3 patients, 24 weeks of treatment with flat-dose RBV is adequate; no evidence of additional benefit of extending treatment to 48 weeks was demonstrated.

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

β-Galactosidase treatment is a common first-stage modification of the three major subtypes of Gc protein to GcMAF.

Science.gov (United States)

Uto, Yoshihiro; Yamamoto, Syota; Mukai, Hirotaka; Ishiyama, Noriko; Takeuchi, Ryota; Nakagawa, Yoshinori; Hirota, Keiji; Terada, Hiroshi; Onizuka, Shinya; Hori, Hitoshi

2012-06-01

The 1f1f subtype of the group-specific component (Gc) protein is converted into Gc protein-derived macrophage-activating factor (GcMAF) by enzymatic processing with β-galactosidase and sialidase. We previously demonstrated that preGc(1f1f)MAF, a full Gc(1f1f) protein otherwise lacking a galactosyl moiety, can be converted to GcMAF by treatment with mouse peritoneal fluid. Here, we investigated the effects of the β-galactosidase-treated 1s1s and 22 subtypes of Gc protein (preGc(1s1s)MAF and preGc₂₂MAF) on the phagocytic activation of mouse peritoneal macrophages. We demonstrated the presence of Gal-GalNAc disaccharide sugar structures in the Gc(1s1s) protein by western blotting using peanut agglutinin and Helix pomatia agglutinin lectin. We also found that preGc(1s1s)MAF and preGc₂₂MAF significantly enhanced the phagocytic activity of mouse peritoneal macrophages in the presence and absence of mouse peritoneal fluid. We demonstrate that preGc(1s1s)MAF and preGc₂₂MAF proteins can be used as effective macrophage activators.

Kalyan Das

Indian Academy of Sciences (India)

Home; Journals; Resonance – Journal of Science Education. Kalyan Das. Articles written in Resonance – Journal of Science Education. Volume 5 Issue 12 December 2000 pp 76-76 Book Review. Linear Algebra and Linear Models · Kalyan Das · More Details Fulltext PDF ...

EFFECTIVENESS AND SAFETY OF VELAGLUCERASE ALFA IN TREATMENT OF GAUCHER DISEASE TYPE 1 (ACCORDING TO INTERNATIONAL STUDIES

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O. S. Gundobina

2014-01-01

Full Text Available The article is dedicated to modern approaches to treatment of Gaucher’s disease. The authors list the primary aspects of the disease and present data on the origin and introduction of pathogenetic enzyme replacement therapy to clinical practice. Review of international clinical studies demonstrates high effectiveness and safety of long-term enzyme replacement therapy with velaglucerase alfa in patients with confirmed type I Gaucher’s disease. 

Prospective exploratory muscle biopsy, imaging, and functional assessment in patients with late-onset Pompe disease treated with alglucosidase alfa: The EMBASSY Study

NARCIS (Netherlands)

A.T. van der Ploeg (Ans); Carlier, P.G. (Pierre G.); Carlier, R.-Y. (Robert-Yves); Kissel, J.T. (John T.); B. Schoser; Wenninger, S. (Stephan); Pestronk, A. (Alan); Barohn, R.J. (Richard J.); Dimachkie, M.M. (Mazen M.); Goker-Alpan, O. (Ozlem); Mozaffar, T. (Tahseen); Pena, L.D.M. (Loren D.M.); Simmons, Z. (Zachary); V. Straub; Guglieri, M. (Michela); Young, P. (Peter); Boentert, M. (Matthias); Baudin, P.-Y. (Pierre-Yves); S.C.A. Wens (Stephan); Shafi, R. (Raheel); Bjartmar, C. (Carl); B. Thurberg

2016-01-01

textabstractBackground Late-onset Pompe disease is characterized by progressive skeletal myopathy followed by respiratory muscle weakness, typically leading to loss of ambulation and respiratory failure. In this population, enzyme replacement therapy (ERT) with alglucosidase alfa has been shown to

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...

Structural and biochemical characterization of novel bacterial α-galactosidases belonging to glycoside hydrolase family 31.

Science.gov (United States)

Miyazaki, Takatsugu; Ishizaki, Yuichi; Ichikawa, Megumi; Nishikawa, Atsushi; Tonozuka, Takashi

2015-07-01

Glycoside hydrolase family 31 (GH31) proteins have been reportedly identified as exo-α-glycosidases with activity for α-glucosides and α-xylosides. We focused on a GH31 subfamily, which contains proteins with low sequence identity (Pedobacter heparinus and Pedobacter saltans. The enzymes unexpectedly exhibited α-galactosidase activity, but were not active on α-glucosides and α-xylosides. The crystal structures of one of the enzymes, PsGal31A, in unliganded form and in complexes with D-galactose or L-fucose and the catalytic nucleophile mutant in unliganded form and in complex with p-nitrophenyl-α-D-galactopyranoside, were determined at 1.85-2.30 Å (1 Å=0.1 nm) resolution. The overall structure of PsGal31A contains four domains and the catalytic domain adopts a (β/α)8-barrel fold that resembles the structures of other GH31 enzymes. Two catalytic aspartic acid residues are structurally conserved in the enzymes, whereas most residues forming the active site differ from those of GH31 α-glucosidases and α-xylosidases. PsGal31A forms a dimer via a unique loop that is not conserved in other reported GH31 enzymes; this loop is involved in its aglycone specificity and in binding L-fucose. Considering potential genes for α-L-fucosidases and carbohydrate-related proteins within the vicinity of Pedobacter Gal31, the identified Gal31 enzymes are likely to function in a novel sugar degradation system. This is the first report of α-galactosidases which belong to GH31 family. © 2015 Authors; published by Portland Press Limited.

Effectiveness of interferon alfa on incidence of hepatocellular carcinoma and decompensation in cirrhosis type C. European Concerted Action on Viral Hepatitis (EUROHEP)

DEFF Research Database (Denmark)

Fattovich, G; Giustina, G; Degos, F

1997-01-01

The role of interferon alfa treatment in improving morbidity endpoints in patients with chronic hepatitis C infection is currently under debate. The aim of this study was to evaluate the effectiveness of interferon in preventing hepatocellular carcinoma and decompensation in cirrhosis type C....

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

Teores de retinol, beta-caroteno e alfa-tocoferol em leites bovinos comercializados na cidade de São Paulo Amounts of retinol, beta-carotene and alpha-tocopherol in cow milk comercialized in the city of São Paulo

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Rute BIANCHINI

1999-12-01

Full Text Available Os teores de retinol, beta-caroteno e alfa-tocoferol foram determinados por cromatografia líquida de alta eficiência em leites em pó, pasteurizados e esterilizados, comercializados na Cidade de São Paulo. Após a saponificação e extração, os compostos foram determinados simultaneamente utilizando-se coluna de sílica, fase móvel constituída por hexano:isopropanol (99:1 e fluxo de 2,0mL/min. O retinol e o beta-caroteno foram determinados no detector UV/visível e o alfa-tocoferol no detector de fluorescência, ligado em série com o anterior. Os valores de vitamina A dos leites foram calculados com e sem a consideração do beta-caroteno. A maior contribuição deste nutriente no valor de vitamina A esteve entre os leites em pó, cerca de 17% em uma das marcas. Os altos teores das vitamina A e E encontrados em alguns leites, indicam que os mesmos provavelmente receberam adição destas vitaminas, não trazendo, entretanto, tal informação no rótulo. A análise de vitaminas nestes produtos indica a necessidade de maior controle de qualidade dos mesmos.The amount of retinol, beta-carotene, alpha -tocopherol in powder, pasteurized and sterilized milk, comercialized in the city of São Paulo, were analyzed by high performance liquid chromatography. After saponification and extraction, compounds were determined simultaneously through a normal-phase column, mobile phase composed by hexan:2-propanol (99:1 and 2 mL/min flow. The retinol and beta-carotene were analysed by a UV/visible detector and the alpha-tocopherol by a fluorescence detector, both linked in series. The milk vitamin A values were calculated with and without beta-carotene. The major contribution of beta-carotene in the vitamin A value was in powder milks, around 17% in one of the brands. The high amounts of vitamin A and E found in some milks indicate that they probably were enriched with these vitamins but nothing is mentioned about this in their labels. The analysis of

Novel dense CO2 technique for beta-galactosidase immobilization in polystyrene microchannels.

Science.gov (United States)

Leclair Ellis, Jeffrey; Tomasko, David L; Dehghani, Fariba

2008-03-01

In this study we design new fabrication techniques and demonstrate the potential of using dense CO2 for facilitating crucial steps in the fabrication of polymeric lab-on-a-chip microdevices by embedding biomolecules at temperatures well below the polymer's glass transition temperature (T(g)). These new techniques are environmentally friendly and done without the use of a clean room. Carbon dioxide at 40 degrees C and between 4.48 and 6.89 MPa was used to immobilize the biologically active molecule, beta-galactosidase (beta-gal), on the surface of polystyrene microchannels. To our knowledge, this is the first time dense CO2 has been used to directly immobilize an enzyme in a microchannel. beta-gal activity was maintained and shown via a fluorescent reaction product, after enzyme immobilization and microchannel capping by the designed fabrication steps at 40 degrees C and pressures up to 6.89 MPa.

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

S Das

Indian Academy of Sciences (India)

Home; Journals; Bulletin of Materials Science. S Das. Articles written in Bulletin of Materials Science. Volume 25 Issue 6 November 2002 pp 557-560. 3-D mapping with ellipsometrically determined physical thickness/refractive index of spin coated sol–gel silica layer · S Das P Pal S Roy S Chakraboarty P K Biswas.

Food-grade host/vector expression system for Lactobacillus casei based on complementation of plasmid-associated phospho-beta-galactosidase gene lacG.

Science.gov (United States)

Takala, T M; Saris, P E J; Tynkkynen, S S H

2003-01-01

A new food-grade host/vector system for Lactobacillus casei based on lactose selection was constructed. The wild-type non-starter host Lb. casei strain E utilizes lactose via a plasmid-encoded phosphotransferase system. For food-grade cloning, a stable lactose-deficient mutant was constructed by deleting a 141-bp fragment from the phospho-beta-galactosidase gene lacG via gene replacement. The deletion resulted in an inactive phospho-beta-galactosidase enzyme with an internal in-frame deletion of 47 amino acids. A complementation plasmid was constructed containing a replicon from Lactococcus lactis, the lacG gene from Lb. casei, and the constitutive promoter of pepR for lacG expression from Lb. rhamnosus. The expression of the lacG gene from the resulting food-grade plasmid pLEB600 restored the ability of the lactose-negative mutant strain to grow on lactose to the wild-type level. The vector pLEB600 was used for expression of the proline iminopeptidase gene pepI from Lb. helveticus in Lb. casei. The results show that the food-grade expression system reported in this paper can be used for expression of foreign genes in Lb. casei.

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

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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.

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

Limb-girdle muscular dystrophy: an immunohistochemical diagnostic approach Distrofias musculares de cinturas: uma abordagem diagnóstica imuno-histoquímica

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Enio Alberto Comerlato

2005-06-01

Full Text Available The limb-girdle muscle dystrophy (LGMD represents a heterogeneous group of muscular diseases with dominant and recessive inheritance, individualized by gene mutation. A group of 56 patients, 32 males and 24 females, with suggestive LGMD diagnosis were submitted to clinical evaluation, serum muscle enzymes, electromyography, muscle biopsy, and the immunoidentification (ID of sarcoglycans (SG alpha, beta, gamma and delta, dysferlin and western blot for calpain-3. All the patients had normal ID for dystrophin (rod domain, carboxyl and amine terminal. The alpha-SG was normal in 42 patients, beta-SG in 28, beta-SG in 45, delta-SG in 32, dysferlin in 37 and calpain-3 in 9. There was a reduction in the alpha-SG in 7 patients, beta-SG in 4, gamma-SG in 2, and delta-SG in 8. There was deficiency of alpha-SG in 7 patients, beta-SG in 6, gamma-SG in 9, delta-SG in 5, dysferlin in 8, and calpain-3 in 5. The patients were grouped according the ID as sarcoglycans deficiency 18 cases, dysferlin deficiency 8 cases and calpain-3 deficiency 5 cases. Only the sarcoglycans deficiency group showed calf hypertrophy. The dysferlin deficiency group was more frequent in females and the onset was later than sarcoglycan and calpain-3 deficiency groups. The calpain-3 deficiency group occurred only in males and showed an earlier onset and weaker muscular strength.As distrofias musculares de cinturas (DMC representam grupo heterogêneo de doenças musculares com heranças autossômicas dominante ou recessivas, caracterizadas geneticamente por mutações gênicas específicas. Cinqüenta e seis pacientes, 32 masculinos e 24 femininos, com diagnóstico sugestivo de DMC, foram submetidos a avaliação clínica, dosagem séricas das enzimas musculares, eletromiografia, biópsia muscular e imunoidentificação (ID das proteínas sarcoglicanas (SG alfa, beta, gama e delta, disferlina e calpaína-3. A ID da distrofina (domínio rod e terminais carboxila e amino era normal em todos

Measurement of elastic proton-proton scattering at $\\sqrt{s} = 7$ TeV with the ALFA sub-detector of ATLAS at the LHC

CERN Document Server

Kreutzfeldt, Kristof; Stenzel, Hasko

The ATLAS experiment with the ALFA sub-detector, provides a unique opportunity to measure elastic proton--proton scattering at the LHC at a centre-of-mass energy of $\\sqrt{s} = 7$ TeV, that has never been reached before. The ALFA detector is a tracking detector housed in Roman Pots, which makes it possible to measure elastically scattered protons down to very small scattering angles. From the proton tracks, measured during a LHC fill with special $\\beta^{*} = 90$ m beam optics, the differential elastic cross-section as a function of the four-momentum transfer squared $t$ is determined, and the total hadronic cross-section $\\sigma_\\text{tot}$, the nuclear slope parameter $B$ and further derived quantities are extracted by utilizing the optical theorem. The total hadronic cross-section is a fundamental parameter of strong interaction depending on the centre-of-mass energy. It has been measured for more than 50 years at different energies and accelerators, where a rise with energy was observed. A newly developed...

MENYOAL KESENJANGAN ANTARA DAS SOLLEN ISLAM DENGAN DAS SEIN PRAKSIS KEHIDUPAN KAUM MUSLIMIN

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Yusuf Suyono

2016-03-01

Full Text Available Abstract:This paper embarks from the question why the valuable Islamic ethics cannot be ethos grounded in the nation-state Muslim majority country-including in Indonesia? Phenomena such as the majlis taklim, majlis dhikr, interest pilgrimage exceeds the quota, the Islamic banking activity is equally excited, is real. However, it is not enough. Muslims should master the science, economics, and the strategic role of national politics. Islamic ethics is Dassollen, the Muslims condition is DasSein. ProphetMuḥammad has abled to unite Das sein andDassollenin his life, because Islam hasbecomehis bloodso that he is a mirror and store front of Islampar excellence. Muslims, as his follower, not been able todo like him. Al-Amir ArsalanSyākib, Muḥammad ‘Abduh, MohammadIqbal, Muḥammadal-Ghazālī, Ḥassan Ḥanafihavetried to formulatehow tobridge the gapbetween Das sollenandDasSein forMuslims. Theyhave adeep concern about thewide gapbetweenDasSeinpraxis in life of Muslims with DassollenIslamicteachings in slogan ya’lu walā yu’la ‘alaih. Whileatthe same timetheyseehowthe berufethos of Calvinismcouldencouragethe ethos ofmoderncapitalismto its adherentsin Western Europe, a Zen Buddhistethoscouldpushthe Japaneseintothe Asiantigers, andspirit Confucius encouragethe Korean peopleintothe Asiandragon. Abstrak:Tulisan ini berangkat dari pertanyaan mengapa etika Islam yang adiluhung itu tidak bisa membumi menjadi etos bangsa di negara-negara yang mayoritas penduduknya Muslim–termasuk di Indonesia. Fenomena seperti majlis taklim, majlis zikir, minat menunaikan ibadah haji melebihi kuota, aktivitas perbankan syariah tak kalah bersemangat, adalah nyata. Namun, itu tidak cukup. Umat Islam seharusnya lebih dari itu dalam penguasaan ilmu pengetahuan, ekonomi, dan peran strategis politik kebangsaan. Etika Islam itulah Das Sollen, keadaan kaum Muslimin itulah Das Sein. Muhammad Rasulullah telah mampu menyatukan Das Sein dan Das Sollen dalam hidupnya. Hal

Wykorzystanie metod analitycznych do wyznaczania wielowymiarowych alfa-stabilnych rozkładów prawdopodobieństwa

OpenAIRE

Łażewski, Marek; Zator, Krzysztof

2004-01-01

In this paper we study the relationship between multivariate a-stable probability distributions and their spectral measure. Analytical method based on nonabelian harmonic analysis is used to express the spectral measure using spherical harmonics on the sphere. Alfa-stabilne rozkłady prawdopodobieństwa, będące uogólnieniem rozkładów normalnych, mają ważną cechę, nie posiadają skończonej wariancji, która utrudnia stosowanie klasycznego formalizmu matematyczno-statystycznego do wyznac...

Effect of darbepoetin alfa on physical function in patients undergoing surgery for colorectal cancer

DEFF Research Database (Denmark)

Nørager, C B; Jensen, M B; Madsen, M R

2007-01-01

OBJECTIVE: To study whether perioperative treatment with darbepoetin alfa (DA) improves physical performance following colorectal cancer surgery. METHODS: Patients admitted for planned colorectal cancer surgery were randomized to receive either weekly placebo or DA 300 or 150 microg depending...... on the hemoglobin (Hb) concentration. Patients were assessed 10 days before, as well as 7 and 30 days after surgery for work capacity, postural sway, muscle strength, fatigue and quality of life (QoL). The primary outcome measure were the changes in patients' physical performance from preoperative to postoperative...... differences between the 2 groups on days 7 or 30 for fatigue, postural sway and QoL. DA treatment significantly (p

Oral sucrosomial iron versus intravenous iron in anemic cancer patients without iron deficiency receiving darbepoetin alfa: a pilot study.

Science.gov (United States)

Mafodda, Antonino; Giuffrida, D; Prestifilippo, A; Azzarello, D; Giannicola, R; Mare, M; Maisano, R

2017-09-01

Erythropoiesis-stimulating agents (ESAs) are often used in treatment of patients with chemotherapy-induced anemia. Many studies have demonstrated an improved hemoglobin (Hb) response when ESA is combined with intravenous iron supplementation and a higher effectiveness of intravenous iron over traditional oral iron formulations. A new formulation of oral sucrosomial iron featuring an increased bioavailability compared to traditional oral formulations has recently become available and could provide a valid alternative to those by intravenous (IV) route. Our study evaluated the performance of sucrosomial iron versus intravenous iron in increasing hemoglobin in anemic cancer patients receiving chemotherapy and darbepoetin alfa, as well as safety, need of transfusion, and quality of life (QoL). The present study considered a cohort of 64 patients with chemotherapy-related anemia (Hb >8 g/dL iron deficiency, scheduled to receive chemotherapy and darbepoetin. All patients received darbepoetin alfa 500 mcg once every 3 weeks and were randomly assigned to receive 8 weeks of IV ferric gluconate 125 mg weekly or oral sucrosomial iron 30 mg daily. The primary endpoint was to demonstrate the performance of oral sucrosomial iron in improving Hb response, compared to intravenous iron. The Hb response was defined as the Hb increase ≥2 g/dL from baseline or the attainment Hb ≥ 12 g/dL. There was no difference in the Hb response rate between the two treatment arms. Seventy one percent of patients treated with IV iron achieved an erythropoietic response, compared to 70% of patients treated with oral iron. By conventional criteria, this difference is considered to be not statistically significant. There were also no differences in the proportion of patients requiring red blood cell transfusions and changes in QoL. Sucrosomial oral iron was better tolerated. In cancer patients with chemotherapy-related anemia receiving darbepoetin alfa, sucrosomial oral iron provides

Interferon alfa-2b, colchicine, and benzathine penicillin versus colchicine and benzathine penicillin in Behçet's disease: a randomised trial.

Science.gov (United States)

Demiroglu, H; Ozcebe, O I; Barista, I; Dündar, S; Eldem, B

2000-02-19

Sight-threatening eye involvement is a serious complication of Behçet's disease. Extraocular complications such as arthritis, vascular occlusive disorders, mucocutaneous lesions, and central-nervous-system disease may lead to morbidity and even death. We designed a prospective study in newly diagnosed patients without previous eye disease to assess whether prevention of eye involvement and extraocular manifestations, and preservation of visual acuity are possible with combination treatments with and without interferon alfa-2b. Patients were randomly assigned 3 million units interferon alfa-2b subcutaneously every other day for the first 6 months plus 1.5 mg colchicine orally daily and 1.2 million units benzathine penicillin intramuscularly every 3 weeks (n=67), or colchicine and benzathine penicillin alone (n=68). The primary endpoint was visual-acuity loss. Analysis was by intention to treat. Significantly fewer patients who were treated with interferon had eye involvement than did patients who did not receive interferon (eight vs 27, relative risk 0.21 [95% CI 0.09-0.50], p<0.001). Ocular attack rate was 0.2 (SD 0.62) per year with interferon therapy and 1.02 (1.13) without interferon therapy (p=0.0001). Visual-acuity loss was significantly lower among patients treated with interferon than in those without interferon (two vs 13, relative risk 0.13 [95% CI 0.03-0.60], p=0.003). Arthritis episodes, vascular events, and mucocutaneous lesions were also less frequent in patients treated with interferon than in those not receiving interferon. No serious side-effects were reported. Therapy with interferon alfa-2b, colchicine, and benzathine penicillin seems to be an effective regimen in Behçet's disease for the prevention of recurrent eye attacks and extraocular complications, and for the protection of vision.

DAS performance analysis

International Nuclear Information System (INIS)

Bates, G.; Bodine, S.; Carroll, T.; Keller, M.

1984-02-01

This report begins with an overview of the Data Acquisition System (DAS), which supports several of PPPL's experimental devices. Performance measurements which were taken on DAS and the tools used to make them are then described

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

Directory of Open Access Journals (Sweden)

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

Coste-efectividad de drotrecogina alfa (activada en el tratamiento de la sepsis grave en España Cost-effectiveness of drotrecogin alpha [activated] in the treatment of severe sepsis in Spain

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José A Sacristán

2004-02-01

Full Text Available Introducción: El ensayo clínico PROWESS ha demostrado que el tratamiento con drotrecogina alfa (activada en pacientes con sepsis grave se asocia a una reducción del riesgo absoluto de muerte en comparación con el tratamiento estándar. El objetivo del estudio fue evaluar el coste-efectividad de drotrecogina alfa (activada, frente a tratamiento estándar, en el tratamiento de la sepsis grave en España. Pacientes y métodos: Se elaboró un modelo de análisis de decisión en el que se compararon los costes hasta el alta hospitalaria y la eficacia a largo plazo de drotrecogina alfa (activada frente a tratamiento estándar de pacientes con sepsis grave en España, desde la perspectiva del financiador de los servicios sanitarios. La mayoría de la información para realizar el modelo se obtuvo del ensayo clínico PROWESS. El análisis basal fue doble: a para todos los pacientes incluidos en el ensayo clínico PROWESS, y b para los pacientes con 2 o más fallos orgánicos. Las variables principales de valoración clínica fueron la reducción de la mortalidad hospitalaria y los años de vida ganados (AVG. El coste-efectividad se expresó como coste por AVG. Se realizó un análisis de sensibilidad utilizando tasas de descuento del 3 y el 5% para los AVG y modificando los patrones de atención sanitaria, los costes de la unidad de cuidados intensivos, la esperanza de vida según la comorbilidad inicial y la eficacia terapéutica de drotrecogina alfa (activada. Resultados: El tratamiento con drotrecogina alfa (activada se asoció a un descenso del riesgo absoluto de mortalidad hospitalaria del 6,0% (p = 0,005 cuando se incluyeron todos los pacientes del estudio PROWESS, y una reducción del 7,3% (p = 0,005 cuando sólo se incluyeron los pacientes con 2 o más fracasos orgánicos. El coste-efectividad de drotrecogina alfa (activada fue de 13.550 euros por AVG respecto a placebo cuando se analizó el total de pacientes, y de 9.800 euros por AVG en el

The alpha-galactosidase type A gene aglA from Aspergillus niger encodes a fully functional alpha-N-acetylgalactosaminidase

Czech Academy of Sciences Publication Activity Database

Kulik, Natallia; Weignerová, L.; Filipi, Tomáš; Pompach, Petr; Novák, Petr; Mrázek, H.; Slámová, Kristýna; Bezouška, K.; Křen, Vladimír; Ettrich, Rüdiger

2010-01-01

Roč. 20, č. 11 (2010), s. 1410-1419 ISSN 0959-6658 R&D Projects: GA MŠk(CZ) LC06010; GA ČR GAP207/10/1934; GA ČR GAP207/10/1040; GA ČR GA303/09/0477; GA ČR GAP207/10/0321 Institutional research plan: CEZ:AV0Z60870520; CEZ:AV0Z50200510; CEZ:AV0Z10750506 Keywords : α-N-acetylgalactosaminidase * α-galactosidase * Aspergillus niger * substrate binding * molecular modeling Subject RIV: CE - Biochemistry Impact factor: 3.791, year: 2010

Enzymatic hydrolysis of pretreated Alfa fibers (Stipa tenacissima) using β-d-glucosidase and xylanase of Talaromyces thermophilus from solid-state fermentation.

Science.gov (United States)

Mallek-Fakhfakh, Hanen; Fakhfakh, Jawhar; Walha, Kamel; Hassairi, Hajer; Gargouri, Ali; Belghith, Hafedh

2017-10-01

This work aims at realizing an optimal hydrolysis of pretreated Alfa fibers (Stipa tenacissima) through the use of enzymes produced from Talaromyces thermophilus AX4, namely β-d-glucosidase and xylanase, by a solid state fermentation process of an agro-industrial waste (wheat bran supplemented with lactose). The carbon source was firstly selected and the optimal values of three other parameters were determined: substrate loading (10g), moisture content (85%) and production time (10days); which led to an optimized enzymatic juice. The outcome was then supplemented with cellulases of T. reesei and used to optimize the enzymatic saccharification of alkali-pretreated Alfa fibers (PAF). The maximum saccharification yield of 83.23% was achieved under optimized conditions (substrate concentration 3.7% (w/v), time 144h and enzyme loading of 0.8 FPU, 15U CMCase, 60U β-d-glucosidase and 125U xylanase).The structural modification of PAF due to enzymatic saccharification was supported by the changes of morphologic and chemical composition observed through macroscopic representation, FTIR and X-Ray analysis. Copyright © 2017 Elsevier B.V. All rights reserved.

Epoetin alfa 40000 U once weekly and intravenous iron supply in solid tumor patients: early increase of hemoglobin level during chemotherapy

International Nuclear Information System (INIS)

Lalle, M.; Antimi, M.; Pistillucci, G.; D'Aprile, M.

2005-01-01

The objective of this observational study was the early evaluation of the impact, a week after the first administration of epoetin alfa 40000 U once weekly and i.v. dose of 62.5 mg sodium ferric gluconate for seven days in improving hemoglobin levels in cancer patients affected by mild/moderate or severe anemia during chemotherapy. Twenty patients affected by solid tumors who received epoetin alfa 40000 U once weekly and daily i.v. sodium ferric gluconate for one week were evaluated: 90% of the patients showed hemoglobin increase, with a median level of hemoglobin increase of 0.73 g/L from baseline, and 50% of them showing a hemoglobin increase > 1 gr/L. The treatment was well tolerated and no adverse event was observed. The early increase of hemoglobin level from baseline is interesting and suggestive for the possibility of achieving an adequate hemoglobin level with a short-term treatment. It is still necessary to further explore the real need of iron supplementation to maintain adequate erythropoiesis prior and during epoetin therapy

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

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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

In vivo expression of a Cicer arietinum B-galactosidase in potato tubers leads to a reduction of the Galactan side-chains in cell wall pectin

NARCIS (Netherlands)

Martin, I.; Dopico, B.; Munoz, F.J.; Esteban, R.; Oomen, R.J.F.J.; Driouich, A.; Vincken, J.P.; Visser, R.G.F.; Labrador, E.

2005-01-01

We report the generation of Solanum tuberosum transformants expressing Cicer arietinum ßIII-Gal. ßIII-Gal is a ß-galactosidase able to degrade cell wall pectins during cell wall loosening that occurs prior to cell elongation. cDNA corresponding to the gene encoding this protein was identified among

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.

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

Directory of Open Access Journals (Sweden)

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

Artvin, Rize ve Trabzon’daki Doğal Kaynak Sularında Toplam Alfa, Toplam Beta Aktivite Değerlerinin Belirlenmesi ve Jeoistatistiksel Olarak Modellenmesi

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Yaşar Kobya

2014-11-01

Full Text Available Trabzon, Rize ve Artvin yaklaşık 18000km2 alan üzerine kurulmuş ve yaklaşık 2 milyon insanın yaşadığı Türkiye’nin en kuzey doğusunda bulunan üç ildir. Bu üç ilde toplam 20 tane içme suyu kaynağından örnekler alındı ve bunların toplam alfa, toplam beta aktivite değerleri Berthold LB770 marka alfa, beta sayım cihazı kullanılarak belirlendi. Toplam alfa aktivite değerleri 1.7-114.2 Bq/L ve toplam beta aktivite değerleri ise 2.9-90.8 Bq/L aralıklarında ölçüldü. Bulunan bu değerler dünya sağlık örgütü (WHO’nün limit değerleri ve yapılmış diğer benzer çalışmalarla karşılaştırıldı. Elde edilen veriler için klasik istatistik hesaplamalar yapıldı. Ayrıca jeoistatistik analiz yöntemi kullanılarak eş-doz eğrileri oluşturuldu ve harita üzerinde gösterilerek bölge için bir risk haritası oluşturuldu.

The frequency of β-globin gene haplotypes, α-thalassemia and genetic polymorphisms of methylenetetrahydrofolate reductase, factor V Leiden and prothrombin genes in children with sickle cell disease in Rio de Janeiro, Brazil Frequência dos haplótipos da globina beta, da talassemia alfa e dos polimorfismos genéticos dos genes da metilenotetrahidrofolato redutase, do fator V Leiden e da protrombina em crianças com doença falciforme no Rio de Janeiro, Brasil

Directory of Open Access Journals (Sweden)

Isaac L. Silva Filho

2010-02-01

Full Text Available A freqüência dos haplótipos beta S e beta C do gene da globina e a prevalência de talassemia alfa e de mutações nos genes da metilenotetrahidrofolato redutase (MTHFR-C677T, do fator V de Leiden e da protrombina (G20210A foi estudada em crianças com doença falciforme do Rio de Janeiro. O haplótipo Bantu foi o mais freqüente (65,9%, 21,2% das crianças (18% heterozigotas e 3% homozigotas apresentam talassemia com mutação alfa 3.7kb, ao contrário da mutação alfa 4.2kb que não foi encontrada. Os alelos 677CT e 677TT da MTHFR foram observados em 20,2% e 4,8%, respectivamente. Os haplótipos Camarões, Árabe-Indiano e Senegal não foram detectados na amostra estudada, bem como mutações no gene do fator V de Leiden e da protrombina. Somente o haplótipo beta C CI foi observado. Esse é o primeiro estudo realizado em uma amostra proveniente do Programa de Triagem Neonatal para Hemoglobinopatias do estado do Rio de Janeiro. Apesar do Rio de Janeiro ser a segunda maior cidade brasileira e seus habitantes expressarem o elevado grau de miscigenação ocorrida no país, nossos resultados ainda coincidem com os registros históricos dos fluxos migratórios do gene beta S para o Brasil, bem como refletem a forte influência de indivíduos de origem africana na população do Rio de Janeiro.

Peginterferon alpha-2a is associated with higher sustained virological response than peginterferon alfa-2b in chronic hepatitis C: systematic review of randomized trials

DEFF Research Database (Denmark)

Awad, Tahany; Thorlund, Kristian; Hauser, Goran

2010-01-01

) is most effective. We performed a systematic review of head-to-head randomized trials to assess the benefits and harms of the two treatments. We searched the Cochrane Central Register of Controlled Trials, MEDLINE, EMBASE, and LILACS through July 2009. Using standardized forms, two reviewers independently...... extracted data from each eligible trial report. We statistically combined data using a random effects meta-analysis according to the intention-to-treat principle. We identified 12 randomized clinical trials, including 5,008 patients, that compared peginterferon alpha-2a plus ribavirin versus peginterferon...... alfa-2b plus ribavirin. Overall, peginterferon alpha-2a significantly increased the number of patients who achieved a sustained virological response (SVR) versus peginterferon alfa-2b (47% versus 41%; risk ratio 1.11, 95% confidence interval 1.04-1.19; P = 0.004 [eight trials]). Subgroup analyses...

Melibiose permease and alpha-galactosidase of Escherichia coli: Identification by selective labeling using a T7 RNA polymerase/promoter expression system

International Nuclear Information System (INIS)

Pourcher, T.; Bassilana, M.; Sarkar, H.K.; Kaback, H.R.; Leblanc, G.

1990-01-01

Identification and selective labeling of the melibiose permease and alpha-galactosidase in Escherichia coli, which are encoded by the melB and melA genes, respectively, have been accomplished by selectively labeling the two gene products with a T7 RNA polymerase expression system. Following generation of a novel EcoRI restriction site in the intergenic sequence between the two genes of the mel operon by oligonucleotide-directed, site-specific mutagenesis, melA and melB were separately inserted into plasmid pT7-6 of the T7 expression system. Expression of melB was markedly enhanced by placing a strong, synthetic ribosome binding site at an optimal distance upstream from the initiation codon of melB. Expression of cloned gene products was characterized functionally and by performing autoradiographic analysis on total cell, inner membrane, and cytoplasmic proteins from cells pulse labeled with (35S)methionine in the presence of rifampicin and resolved by sodium dodecyl sulfate/polyacrylamide gel electrophoresis. The results first confirm that alpha-galactosidase is a cytoplasmic protein with an Mr of 50K; in contrast, the membrane-bound melibiose permease is identified as a protein with an apparent Mr of 39K, a value significantly higher than that of 30K previously suggested

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

Directory of Open Access Journals (Sweden)

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

Avaliação do desempenho organizacional da empresa familiar Indústrias Alfa S.A.* com base no modelo de gestão estratégica balanced scorecard Organizational performance of a family business evaluated by the strategic performance balanced scorecard

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Christine Schneider

2006-03-01

Full Text Available O balanced scorecard é um moderno modelo estratégico de desempenho organizacional que visa traduzir a missão, visão e estratégia em objetivos e medidas, organizados sob a ótica financeira, dos clientes, dos processos internos, do aprendizado e do crescimento. O modelo disponibiliza estrutura e linguagem para comunicar a estratégia da empresa e o alcance das metas no longo prazo. A Indústrias Alfa S.A. é uma empresa familiar de segunda geração, que tem por visão estratégica "Ser a escolha preferencial de soluções hidráulicas da América Latina nos setores residencial, industrial, agrícola e predial", orientada pelo macroobjetivo de oferecer soluções e serviços em motobombas hidráulicas de modo a satisfazer as necessidades dos clientes. Partindo desses princípios, o artigo avalia o desempenho organizacional da empresa familiar Indústrias Alfa S.A. no primeiro quadrimestre do ano de 2005, com base no modelo de controle estratégico balanced scorecard.The balanced scorecard is a modern strategic model of organizational performance that seeks to translate the company mission, vision and strategy into objectives and measures organized to focus on - finance, customers, internal procedures, learning and growth. This model provides structure and language to communicate business strategy and directs efforts, abilities and specific knowledge to achieve long term objectives. The strategic vision of this second generation family business is "to be the answer for hydraulic pump needs in the Latin American residential, industrial, agricultural and building management markets" oriented by an overall objective of offering solutions and services to satisfy customer needs. Based upon these considerations, performance of this family business was evaluated during the first four months of 2005 using the strategic performance balanced scorecard.

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

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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

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

A Phase 3, multicenter, open-label, switchover trial to assess the safety and efficacy of taliglucerase alfa, a plant cell-expressed recombinant human glucocerebrosidase, in adult and pediatric patients with Gaucher disease previously treated with imiglucerase.

Science.gov (United States)

Pastores, Gregory M; Petakov, Milan; Giraldo, Pilar; Rosenbaum, Hanna; Szer, Jeffrey; Deegan, Patrick B; Amato, Dominick J; Mengel, Eugen; Tan, Ee Shien; Chertkoff, Raul; Brill-Almon, Einat; Zimran, Ari

2014-12-01

Taliglucerase alfa is a β-glucosidase enzyme replacement therapy (ERT) approved in the US and other countries for the treatment of Gaucher disease (GD) in adults and is approved in pediatric and adult patients in Australia and Canada. It is the first approved plant cell-expressed recombinant human protein. A Phase 3, multicenter, open-label, 9-month study assessed safety and efficacy of switching to taliglucerase alfa in adult and pediatric patients with GD treated with imiglucerase for at least the previous 2years. Patients with stable disease were offered taliglucerase alfa treatment using the same dose (9-60U/kg body weight) and regimen of administration (every 2weeks) as imiglucerase. This report summarizes results from 26 adult and 5 pediatric patients who participated in the trial. Disease parameters (spleen and liver volumes, hemoglobin concentration, platelet count, and biomarker levels) remained stable through 9months of treatment in adults and children following the switch from imiglucerase. All treatment-related adverse events were mild or moderate in severity and transient in nature. Exploratory parameters of linear growth and development showed positive outcomes in pediatric patients. These findings provide evidence of the efficacy and safety profile of taliglucerase alfa as an ERT for GD in patients previously treated with imiglucerase. This trial was registered at www.clinicaltrials.gov as # NCT00712348. Copyright © 2014 Elsevier Inc. All rights reserved.

MiDAS

DEFF Research Database (Denmark)

McIlroy, Simon Jon; Saunders, Aaron Marc; Albertsen, Mads

2015-01-01

The Microbial Database for Activated Sludge (MiDAS) field guide is a freely available online resource linking the identity of abundant and process critical microorganisms in activated sludge wastewater treatment systems to available data related to their functional importance. Phenotypic properties...... of some of these genera are described, but most are known only from sequence data. The MiDAS taxonomy is a manual curation of the SILVA taxonomy that proposes a name for all genus-level taxa observed to be abundant by large-scale 16 S rRNA gene amplicon sequencing of full-scale activated sludge...... communities. The taxonomy can be used to classify unknown sequences, and the online MiDAS field guide links the identity to the available information about their morphology, diversity, physiology and distribution. The use of a common taxonomy across the field will provide a solid foundation for the study...

Diffusion Kurtosis Imaging Detects Microstructural Alterations in Brain of alfa-Synuclein Overexpressing Transgenic Mouse Model of Parkinson’s Disease: A Pilot Study

Czech Academy of Sciences Publication Activity Database

Khainar, A.; Latta, P.; Dražanová, Eva; Rudá-Kučerová, J.; Szabó, N.; Arab, A.; Hutter-Paier, B.; Havas, D.; Windisch, M.; Šulcová, A.; Starčuk jr., Zenon; Rektorová, I.

2015-01-01

Roč. 28, č. 4 (2015), 281-289 ISSN 1029-8428 R&D Projects: GA MŠk(CZ) LO1212; GA MŠk ED0017/01/01 Institutional support: RVO:68081731 Keywords : diffusion kurtosis imaging * alfa-Synuclein * TNWT-61 * Parkinson’s disease * Transgenic mice * TBSS Subject RIV: BH - Optics, Masers, Lasers Impact factor: 3.140, year: 2015

Intervalos de Confianza para el coeficiente alfa de Cronbach: aportes a la investigación pediátrica

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Tomás Caycho-Rodríguez

2017-06-01

Full Text Available En la actualidad, es cada vez más común encontrar estudios que brinden evidencias de la confiabilidad de las puntuaciones obtenidas, por instrumentos de medida utilizadas en investigaciones en ciencias de la salud. Este es el caso de la valiosa contribución de Aguilar-Navarro et al, quienes adaptaron la Escala de Actitudes hacia la Alimentación Infantil de Iowa (IIFAS en México, reportando la confiabilidad a través del alfa de Cronbach.

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.

P K Das

Indian Academy of Sciences (India)

Home; Journals; Bulletin of Materials Science. P K Das. Articles written in Bulletin of Materials Science. Volume 23 Issue 4 August 2000 pp 249-253 Nitride Ceramics. Optimization of time–temperature schedule for nitridation of silicon compact on the basis of silicon and nitrogen reaction kinetics · J Rakshit P K Das.

Alpha fucosidase and beta galactosidase in serum of a Lyme disease patients as a possible marker of accelerated senescence — a preliminary study

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Anna Wasiluk

2012-07-01

Full Text Available Lyme disease (LD is the most prevalent tick-borne disease in Europe. LD is caused by the spirochete Borrelia burgdorferi. LD is a chronic disease which can attack a number of organs: skin, heart, brain, joints. Chronic, low-grade inflammation involves general production of pro-inflammatory cytokines and inflammatory markers and is a typical feature of aging. So far, the best method of diagnosing LD is a time-consuming and expensive two-stage serological method. The aim of our study was to evaluate the activity of two lysosomal exoglycosidases: α-fucosidase (FUC and β-galactosidase (GAL in the serum of patients with Lyme disease, as potential markers of LD. Due to the increasing number of patients with Lyme disease and a number of false results, new ways to diagnose this disease are still being sought. As elevated level of β-galactosidase is a manifestation of residual lysosomal activity in senescent cells, the increase in its activity in serum during chronic Lyme disease might be a marker of a potentially accelerated senescence process. The study was performed on serum taken from cubital veins of 15 patients with Lyme disease and eight healthy subjects (control group. FUC and GAL activity was measured by the method of Chatterjee et al. as modified by Zwierz et al. In the serum of patients with Lyme disease, GAL activity significantly increased (p = 0.029, and the activity of FUC had a tendency to increase (p = 0.153, compared to the control group. A significant increase in GAL activity in the serum of patients with Lyme disease indicates an increased catabolism of glycoconjugates (glycoproteins, glycolipids, proteoglycans and could be helpful in the diagnosis of Lyme disease, although this requires confirmation in a larger group of patients. As GAL is the most widely used assay for detection of senescent cells, an elevated level of β-galactosidase might be a manifestation of accelerated senescence process in the course of Lyme

Glucose buffer is suitable for blood group conversion with α-N acetylgalactosaminidase and α-galactosidase.

Science.gov (United States)

Gao, Hong-Wei; Li, Su-Bo; Bao, Guo-Qiang; Zhang, Xue; Li, Hui; Wang, Ying-Li; Tan, Ying-Xia; Ji, Shou-Ping; Gong, Feng

2014-01-01

It is well known that the buffer plays a key role in the enzymatic reaction involved in blood group conversion. In previous study, we showed that a glycine buffer is suitable for A to O or B to O blood group conversion. In this study, we investigated the use of 5% glucose and other buffers for A to O or B to O blood group conversion by α-N-acetylgalactosaminidase or α-galactosidase. We compared the binding ability of α-N-acetylgalactosaminidase/α-galactosidase with red blood cells (RBC) in different reaction buffers, such as normal saline, phosphate-buffered saline (PBS), a disodium hydrogen phosphate-based buffer (PCS), and 5% commercial glucose solution. The doses of enzymes necessary for the A/B to O conversion in different reaction buffers were determined and compared. The enzymes' ability to bind to RBC was evaluated by western blotting, and routine blood typing and fluorescence activated cell sorting was used to evaluate B/A to O conversion efficiency. The A to O conversion efficiency in glucose buffer was similar to that in glycine buffer with the same dose (>0.06 mg/mL pRBC). B to O conversion efficiency in glucose buffer was also similar to that in glycine buffer with the same dose (>0.005 mg/mL pRBC). Most enzymes could bind with RBC in glycine or glucose buffer, but few enzymes could bind with RBC in PBS, PCS, or normal saline. These results indicate that 5% glucose solution provides a suitable condition for enzymolysis, especially for enzymes combining with RBC. Meanwhile, the conversion efficiency of A/B to O was similar in glucose buffer and glycine buffer. Moreover, 5% glucose solution has been used for years in venous transfusion, it is safe for humans and its cost is lower. Our results do, therefore, suggest that 5% glucose solution could become a novel suitable buffer for A/B to O blood group conversion.

Crystal structures of β-galactosidase from Penicillium sp. and its complex with galactose

International Nuclear Information System (INIS)

Rojas, A.L.; Nagem, R.A.P.; Garratt, R.C.; Polikarpov, I.; Neustroev, K.N.; Eneyskaya, E.V.; Kulminskaya, A.A.; Golubev, A.M.; Arand, M.; Adamska, M.

2004-01-01

Glycosidase belong to a group of enzymes displaying a great variety of protein folds and substrate specificities. Two critically located acidic residues make up the catalytic machinery of these enzymes, which are responsible for the cleavage of glycosidic bonds. The applications of glycosidase in textile, food, and pulp processing and in catalysts and oligosaccharide synthesis have encouraged the engineering of these proteins to improve their catalytic properties and stability. Furthermore, structural studies broaden our understanding of the catalytic mechanism and the role of glycosidase in the recognition processes of their different substrates. In this work, we describe crystallographic studies of a fungi glycosidase. The crystallographic structures of β-galactosidase from Penicillium sp. and its complex with galactose were solved at 1.90 A and 2.10 A resolution, respectively. The X-ray structure of the enzyme-galactose complex was useful in identifying the residue Glu200 as the proton donor and residue Glu 299 as the nucleophiles involved in catalysis. (author)

Crystal structures of {beta}-galactosidase from Penicillium sp. and its complex with galactose

Energy Technology Data Exchange (ETDEWEB)

Rojas, A.L.; Nagem, R.A.P.; Garratt, R.C.; Polikarpov, I. [Universidade de Sao Paulo, Sao Carlos, SP (Brazil); Neustroev, K.N.; Eneyskaya, E.V.; Kulminskaya, A.A.; Golubev, A.M. [St. Petersburg, Gatchina (Russian Federation); Arand, M.; Adamska, M. [University of Wuerzburg (Germany)

2004-07-01

Glycosidase belong to a group of enzymes displaying a great variety of protein folds and substrate specificities. Two critically located acidic residues make up the catalytic machinery of these enzymes, which are responsible for the cleavage of glycosidic bonds. The applications of glycosidase in textile, food, and pulp processing and in catalysts and oligosaccharide synthesis have encouraged the engineering of these proteins to improve their catalytic properties and stability. Furthermore, structural studies broaden our understanding of the catalytic mechanism and the role of glycosidase in the recognition processes of their different substrates. In this work, we describe crystallographic studies of a fungi glycosidase. The crystallographic structures of {beta}-galactosidase from Penicillium sp. and its complex with galactose were solved at 1.90 A and 2.10 A resolution, respectively. The X-ray structure of the enzyme-galactose complex was useful in identifying the residue Glu200 as the proton donor and residue Glu 299 as the nucleophiles involved in catalysis. (author)

P Chaitanya Das

Indian Academy of Sciences (India)

P Chaitanya Das G Srinivasa Murthy C P Gopalakrishnan P C Deshmukh · More Details Fulltext PDF. Volume 9 Issue 7 July 2004 pp 77-85 Classroom. Motion of Charged Particles in Electromagnetic Fields and Special Theory of Relativity · P Chaitanya Das G Srinivasa Murthy P C Deshmukh K Satish Kumar T A Venkatesh.

S K Das

Indian Academy of Sciences (India)

Home; Journals; Bulletin of Materials Science. S K Das. Articles written in Bulletin of Materials Science. Volume 24 Issue 4 August 2001 pp 373-378 Metals and Alloys. Evaluation of solid–liquid interface profile during continuous casting by a spline based formalism · S K Das · More Details Abstract Fulltext PDF. A numerical ...

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

Exploitation of a Library of alfa-Galactosidases for the Synthesis of Building Blocks for Glycopolymers

Czech Academy of Sciences Publication Activity Database

Casali, M.; Tarantini, L.; Riva, S.; Huňková, Zdenka; Weignerová, Lenka; Křen, Vladimír

2002-01-01

Roč. 77, č. 1 (2002), s. 105-110 ISSN 0006-3592 R&D Projects: GA ČR GA303/99/1382; GA ČR GA203/01/1018 Keywords : glycosidases * biocatalysis * enzymatic library Subject RIV: EE - Microbiology, Virology Impact factor: 2.211, year: 2002

B P Das

Indian Academy of Sciences (India)

Home; Journals; Bulletin of Materials Science. B P Das. Articles written in Bulletin of Materials Science. Volume 25 Issue 6 November 2002 pp 517-519. Structural, dielectric and electrical properties of Sm-modified Pb(SnTi)O3 ferroelectric system · B P Das R N P Choudhary P K Mahapatra · More Details Abstract Fulltext ...

A K Das

Indian Academy of Sciences (India)

Home; Journals; Bulletin of Materials Science. A K Das. Articles written in Bulletin of Materials Science. Volume 28 Issue 2 April 2005 pp 131-136 Fly Ash. Some studies on the reaction between fly ash and lime · A Basumajumdar A K Das N Bandyopadhyay S Maitra · More Details Abstract Fulltext PDF. The reaction between ...

Estimativa do vigor das sementes e das plântulas de Bixa orellana L.

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Roberta Leopoldo Ferreira

Full Text Available RESUMO A multiplicação de espécies como as da planta de urucum tem limitações em função do conhecimento limitado das características morfológicas e fisiológicas das sementes e das plântulas e da restrição de métodos para determinar a qualidade dessas sementes. Nessa pesquisa, o objetivo foi estudar a adequação do teste de envelhecimento acelerado para estimar o vigor das sementes de urucum (Bixa orellana L., relacionando os resultados desse teste com a formação das plântulas e as diferenças de genótipo dos acessos genéticos. As sementes de urucum, representadas por quatro acessos genéticos, e por três lotes, foram avaliadas pelos testes de germinação, primeira contagem da germinação, classificação do vigor das plântulas e emergência das plântulas (total e índice de velocidade. No teste de envelhecimento acelerado foram avaliados a temperatura, de 41 ºC, e os períodos, de 48; 72 e 96 horas, de exposição das sementes às umidades relativas de 100% (água e de 76% (solução saturada de NaCl. A solução saturada reduz a quantidade de água absorvida pelas sementes de urucum, expostas às condições do teste de envelhecimento acelerado, reduzindo a deterioração das sementes, favorecendo a uniformidade dos resultados e a redução da proliferação de fungos, comuns na germinação das sementes de urucum. O teste de envelhecimento acelerado, com água ou solução salina, por 72 horas ou 96 horas, é eficiente para classificar as sementes de urucum quanto à qualidade. Assim, as variações dos teores de água das sementes de urucum devem ser entre 23;6 e 28;9% (72 horas e 29;7 e 32;9% (96 horas para a utilização da água e entre 7,3 e 9,5% para a utilização da solução salina de NaCl.

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

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

A first-year dornase alfa treatment impact on clinical parameters of patients with cystic fibrosis: the Brazilian cystic fibrosis multicenter study

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Tatiana Rozov

2013-12-01

Full Text Available OBJECTIVE: To describe the clinical impact of the first year treatment with dornase alfa, according to age groups, in a cohort of Brazilian Cystic Fibrosis (CF patients. METHODS: The data on 152 eligible patients, from 16 CF reference centers, that answered the medical questionnaires and performed laboratory tests at baseline (T0, and at six (T2 and 12 (T4 months after dornase alfa initiation, were analyzed. Three age groups were assessed: six to 11, 12 to 13, and >14 years. Pulmonary tests, airway microbiology, emergency room visits, hospitalizations, emergency and routine treatments were evaluated. Student's t-test, chi-square test and analysis of variance were used when appropriated. RESULTS: Routine treatments were based on respiratory physical therapy, regular exercises, pancreatic enzymes, vitamins, bronchodilators, corticosteroids, and antibiotics. In the six months prior the study (T0 phase, hospitalizations for pulmonary exacerbations occurred in 38.0, 10.0 and 61.4% in the three age groups, respectively. After one year of intervention, there was a significant reduction in the number of emergency room visits in the six to 11 years group. There were no significant changes in forced expiratory volume in one second (VEF1, in forced vital capacity (FVC, in oxygen saturation (SpO2, and in Tiffenau index for all age groups. A significant improvement in Shwachman-Kulczychi score was observed in the older group. In the last six months of therapy, chronic or intermittent colonization by P. aeruginosa was detected in 75.0, 71.4 and 62.5% of the studied groups, respectively, while S. aureus colonization was identified in 68.6, 66.6 and 41.9% of the cases. CONCLUSIONS: The treatment with dornase alfa promoted the maintenance of pulmonary function parameters and was associated with a significant reduction of emergency room visits due to pulmonary exacerbations in the six to 11 years age group, with better clinical scores in the >14 age group, one

Clinical and genetic investigation of a Japanese family with cardiac fabry disease. Identification of a novel α-galactosidase A missense mutation (G195V).

Science.gov (United States)

Nakagawa, Naoki; Maruyama, Hiroki; Ishihara, Takayuki; Seino, Utako; Kawabe, Jun-ichi; Takahashi, Fumihiko; Kobayashi, Motoi; Yamauchi, Atsushi; Sasaki, Yukie; Sakamoto, Naka; Ota, Hisanobu; Tanabe, Yasuko; Takeuchi, Toshiharu; Takenaka, Toshihiro; Kikuchi, Kenjiro; Hasebe, Naoyuki

2011-01-01

Fabry disease is an X-linked lysosomal storage disorder caused by mutations of the α-galactosidase A gene (GLA), and the disease is a relatively prevalent cause of left ventricular hypertrophy mimicking idiopathic hypertrophic cardiomyopathy. We assessed clinically 5 patients of a three-generation family and also searched for GLA mutations in 10 family members. The proband had left ventricular hypertrophy with localized thinning in the basal posterior wall and late gadolinium enhancement (LGE) in the near-circumferential wall in cardiovascular magnetic resonance images and her sister had vasospastic angina pectoris without organic stenosis of the coronary arteries. LGE notably appeared in parallel with decreased α-galactosidase A activity and increased NT-pro BNP in our patients. We detected a new GLA missense mutation (G195V) in exon 4, resulting in a glycine-to-valine substitution. Of the 10 family members, 5 family members each were positive and negative for this mutation. These new data extend our clinical and molecular knowledge of GLA gene mutations and confirm that a novel missense mutation in the GLA gene is important not only for a precise diagnosis of heterozygous status, but also for confirming relatives who are negative for this mutation.

Epoetin alfa improves anemia and anemia-related, patient-reported outcomes in patients with breast cancer receiving myelotoxic chemotherapy: Results of a european, multicenter, randomized, controlled trial

NARCIS (Netherlands)

P. Pronzato (Paolo); E. Cortesi (Enrico); C.C.D. van der Rijt (Carin); A. Bols (Alain); J.A. Moreno-Nogueira (José); C.F. de Oliveira; P. Barrett-Lee (Peter); P.J. Ostler (Peter); R. Rosso (Ricardo)

2010-01-01

textabstractPurpose. To evaluate the effects of epoetin alfa on patient- reported outcomes (PROs) in patients with breast cancer receiving myelotoxic chemotherapy. Materials and Methods. Women with hemoglobin concentrations ≤12.0 g/dl and an Eastern Cooperative Oncology Group performance status

Measurement of the proton-proton total cross section at √(s) = 7 TeV via elastic scattering with the ALFA sub-detector of ATLAS at the LHC

Energy Technology Data Exchange (ETDEWEB)

Kreutzfeldt, Kristof; Dueren, Michael; Heinz, Christian; Stenzel, Hasko [2. Physikalisches Institut, Universitaet Giessen (Germany)

2015-07-01

The ALFA (Absolute Luminosity for ATLAS) detector is one of the ATLAS forward detectors located about 240 m away from the interaction point in the LHC tunnel. ALFA is a scintillating fibre tracking detector housed in Roman pots and designed to measure elastic proton-proton scattering up to the smallest scattering angles. In 2011 data were taken at a centre-of-mass energy of √(s) = 7 TeV during a special low intensity fill of LHC with high β* = 90 m beam optics. Results of the measurement of the differential elastic cross section dσ/dt, the total cross section σ{sub tot}, the nuclear slope B and other derived quantities are presented in this talk. In the analysis the luminosity determination, the fine tuning of beam optics parameters and a data-driven method to determine the reconstruction efficiency are emphasized.

Diagnostic dilemma: a young woman with Fabry disease symptoms, no family history, and a "sequencing cryptic" α-galactosidase a large deletion

DEFF Research Database (Denmark)

Feldt-Rasmussen, Ulla; Dobrovolny, Robert; Nazarenko, Irina

2011-01-01

Fabry disease, an X-linked lysosomal storage disorder, results from the deficient activity of a-galactosidase A (a-Gal A). In affected males, the clinical diagnosis is confirmed by the markedly decreased a-Gal A activity. However, in female heterozygotes, the a-Gal A activity can range from low t...... on enzyme replacement therapy. Thus, gene dosage analyses can detect large deletions (>50bp) in suspect heterozygotes for X-linked and autosomal dominant diseases that are "sequencing cryptic," resolving molecular diagnostic dilemmas....

Comparable pharmacokinetics and pharmacodynamics of two epoetin alfa formulations Eporon® and Eprex® following a single subcutaneous administration in healthy male volunteers

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Yoon S

2017-10-01

Full Text Available Sumin Yoon,1 Su-jin Rhee,1 Sun Ju Heo,2 Tae Young Oh,2 Seo Hyun Yoon,1 Joo-Youn Cho,1 SeungHwan Lee,1,3 Kyung-Sang Yu1,3 1Department of Clinical Pharmacology and Therapeutics, Seoul National University College of Medicine and Hospital, Seoul, 2Dong-A ST Co. Ltd., Seoul, 3Clinical Trials Center, Seoul National University Hospital, Seoul, Republic of Korea Purpose: This study aimed to assess and compare the pharmacokinetic (PK and pharmacodynamic (PD properties following a single subcutaneous injection of epoetin alfa (Eporon® with those of the comparator (Eprex® in healthy male subjects. Subjects and methods: A randomized, double-blind, two-sequence, crossover study was conducted. Subjects were randomly assigned to receive a single dose, that is, 4,000 IU, of the test or comparator epoetin alfa. After 4 weeks, all subjects received the alternative formulation. The primary PK parameters, maximum observed concentration (Cmax and area under the curve extrapolated to infinity (AUCinf, were calculated with the serum erythropoietin (EPO concentrations from blood samples collected for 144 h after dosing. The reticulocyte, hematocrit, hemoglobin and red blood cell counts were measured up to 312 h as PD markers. The primary PD parameters, maximum observed effect (Emax and area under the effect curve (AUEC, were obtained from the baseline-corrected reticulocyte count. The serum EPO concentration and the reticulocyte count were used to assess the concentration–response relationship. The tolerability and immunogenicity profiles were assessed together. Results: Forty-two subjects completed the study. The mean EPO concentration–time profiles were comparable between the two formulations. The geometric mean ratios (90% CI of the Cmax and AUCinf were 0.908 (0.843–0.978 and 1.049 (0.999–1.101, respectively, both of which were within the regulatory range of 0.80–1.25. Additionally, the PD and tolerability profiles were similar between the two

Development of solar energy education IASEE-Argentina, Arquisur and Alfa-built

International Nuclear Information System (INIS)

Schiller, S. de; Evans, J.M.

1997-01-01

Three educational experiences are presented in this paper on the development of national and regional networks to incorporate renewable energy in curricular programmes. IASEE-Argentina, established in 1990 as the National Section of IASEE, the International Association for Solar Energy Education, was designated in 1993 as the Education Working Group of ASADES, the Argentine Association of Solar Energy. It holds annual meetings to coincide with the ASADES conference. Arquisur is a regional network established by Architectural Faculties of State Universities within the Mercosur Region, covering Argentina, southern Brazil, Paraguay and Uruguay, to promote exchange and development between the 19 faculties involved. A Working Group set up by Arquisur with members from four countries developed the programme for short post-graduate courses on bioclimatic design and rational energy use in buildings, which has been given in Argentina and Uruguay. Alfa-Built, a project supported by the European Union for promoting academic exchange in the field of energy efficient building is also introduced. This paper presents the development and initial results of these educational experiences. (author)

Projeto de Campanha Promocional por meio de Aliança Estratégica

OpenAIRE

Debastiani, Solange Maria; Meireles, Belquis Oliveira; Rojo, Claudio Antonio

2014-01-01

 Este Relato Técnico descreve o processo de idealização e negociação de um projeto de campanha promocional por meio de aliança estratégica formada por três empresas, denominadas Alfa, Beta e Gama, respectivamente. O estudo detalha as motivações, os critérios para a escolha dos parceiros, o tipo da aliança estratégica adotada e a estruturação da campanha. As motivações, além do fortalecimento das marcas, foram: (a) atrair novos clientes à empresa Alfa e fidelizar os atuais; (b) aumentar a util...

Synthesis of Galactosyl Mannitol Derivative Using β-Galactosidase from Kluyveromyces lactis

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Klewicki Robert

2017-03-01

Full Text Available The purpose of the study was to identify the influence of reactive mixture concentration (23–48 g/100 mL, pH (6.5–9.0, presence of NaCl (0.05–0.25 mol/L and enzyme dose (2850–28,500 LAU/100 g of lactose on the synthesis of galactosyl mannitol derivative using β-galactosidase from Kluyveromyces lactis. The use of the enzyme dose ranging from 2850 to 11,400 LAU/100 g lactose allowed obtaining gal-mannitol at the level of 21.8% total saccharides; higher doses intensified product decomposition. An increase in the concentration of the reactive mixture had a positive impact on the quantity of gal-mannitol obtained every single time, i.e. 4.39 g were obtained from 100 mL of a 23 g/100 mL solution and over 10 g were obtained from a 48 g/100 mL solution. A relatively low increase in product quantity (by ca. 5% occurred after the pH was increased from 6.5 to 9.0. The use of NaCl rendered better results. An increase in the maximum content of gal-mannitol in the total sugar by 12.8% was observed at the concentration of 0.25 mol/L.

Expressão do Mg+2, CK, AST e LDH em equinos finalistas de provas de enduro

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Juliana V.F. Sales

2013-01-01

Full Text Available Nos últimos anos, o equino atleta vem sendo cada vez mais requerido. Dessa forma, as exigências por alto desempenho têm fomentado o interesse pelo estudo das afecções relacionadas com a fisiopatologia de diversas enfermidades dos equinos. A relação entre o íon magnésio e o exercício físico tem recebido atenção significativa visto que este íon está intimamente relacionado ao tecido muscular estriado esquelético. Além disso, dentre as principais estratégias para a detecção e acompanhamento clínico de lesões musculares, destacam-se a avaliação das atividades das enzimas creatino quinase (CK, lactato desidrogenase (LDH e aspartato aminotransferase (AST. A busca pelo estabelecimento de parâmetros que se relacionam entre si é um fator determinante na compreensão de alterações fisiológicas encontradas diante do esforço em equinos atletas. Desta forma, o presente trabalho teve como objetivo determinar como as concentrações sanguíneas do íon magnésio e as atividades enzimáticas das enzimas CK, LDH e AST comportaram-se em equinos Puro Sangue Árabe finalistas de provas de enduro de 90km e relacionar as possíveis alterações com o tipo de esforço físico desempenhado pelos animais. Foram avaliadas a atividade enzimática das enzimas CK, LDH, AST e a concentração do íon magnésio no exercício em relação ao repouso de 14 equinos clinicamente hígidos da raça Puro Sangue Árabe, sendo 9 machos e 5 fêmeas, com idades variando entre 6 a 12 anos, submetidos a treinamento para enduro e participantes de provas de 90 km. Pode-se observar que as variáveis acima mencionadas sofreram aumento com diferença estatística em relação ao repouso. O exercício físico de enduro determinou a ocorrência de alterações nas atividades enzimáticas das enzimas CK (p≤0,001, LDH (p=0,0001, AST (p=0,0007 e na concentração do íon magnésio (p=0,0004, no exercício em relação ao repouso (p≤0,05. Fato que determinou altera

Lactose hydrolysis by free and fibre-entrapped β-galactosidase from Streptococcus thermophilus

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Zhennai Yang

1993-09-01

Full Text Available To study lactose hydrolysis by β-galactosidase, this enzyme was produced from Streptococcus thermophilus strain 11F and partially purified by acetone and ammonium sulphate fractionation, and ion exchange chromatography on a Q Sepharose FF column. Lactose hydrolysis by the enzyme was affected by lactose concentrations, sugars and milk proteins. The maximum extent of lactose hydrolysis in buffer was obtained with a 15% lactose concentration. Addition of 2% of lactose, glucose, galactose or sucrose in milk inhibited the enzymatic hydrolysis. The enzyme was activated by bovine serum albumin and a combination of αs-casein and β-casein. Of the casein fractions, the principal fraction, αs-casein, was less effective than β-casein and κ-casein. The fibre entrapped enzyme had a temperature optimum of 57°C, and a pH optimum from 7.5 to at least 9.0 with O-nitrophenyl-β-D-galactopyranoside as substrate. By recycling with whey and skim milk through a jacketed glass column (1.6 cm x 30 cm loaded with fibre-entrapped enzyme at 55°C, a lactose hydrolysis of 49.5% and 47.9% was achieved in 11 h and 7 h respectively.

Influência do desenvolvimento etário e da suplementação com vitamina E (acetato de DL-alfa-tocoferol no metabolismo oxidativo dos neutrófilos de bovinos da raça Holandesa (Bos taurus

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Joselito Nunes Costa

2004-10-01

Full Text Available O presente trabalho teve por objetivo analisar a influência do desenvolvimento etário e da suplementação com acetato de DL-alfa-tocoferol sobre o metabolismo oxidativo de neutrófilos, em bovinos da raça holandesa, no período do nascimento até os 150 dias de idade. Foram utilizados 20 bezerros divididos em dois grupos de dez animais. Os animais do grupo Tratamento receberam 2000UI de acetato de DL-alfa-tocoferol, por via intramuscular, ao nascimento, aos 15, 30, 60, 90 e 120 dias de idade, sendo o outro o grupo Controle, que não recebeu qualquer suplementação. Em ambos os grupos, o metabolismo oxidativo dos neutrófilos demonstrou pouca atividade durante os primeiros 60 dias de vida, sendo indicativo da ineficiência deste importante mecanismo bactericida. Não foi observado efeito significativo da administração do acetato de DL-alfa-tocoferol sobre o metabolismo oxidativo de neutrófilos.

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.

ALGLUCOSIDASE ALFA — A NEW STAGE IN THE THERAPY OF INFANTILE POMPE DISEASE

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E. N. Basargina

2014-01-01

Full Text Available Pompe disease is a rare severe hereditary disease caused by excessive glycogen storage in organs and target tissues due to the acid α-glucosidase gene mutation. Infantile and adult Pompe disease is characterized by involvement of cardiovascular, respiratory and muscular systems in the pathological process. The only specific method of treating Pompe disease is enzyme replacement therapy (intravenous administration of recombinant human acid glucosidase, the effectiveness whereof depends on the time the therapy started. Since such a therapy was introduced into practice, Pompe disease mortality decreased by 79%. 6 children with infantile Pompe disease were observed and treated at the cardiovascular care unit of the Scientific Center of Children’s Health in 2011‑2014. The article presents a clinical case demonstrating capabilities of diagnosing infantile Pompe disease in Russia and effective application of alglucosidase alfa in 4-month-old child. 

Surface displaced alfa-enolase of Lactobacillus plantarum is a fibronectin binding protein

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Muscariello Lidia

2009-02-01

Full Text Available Abstract Background Lactic acid bacteria of the genus Lactobacillus and Bifidobacterium are one of the most important health promoting groups of the human intestinal microbiota. Their protective role within the gut consists in out competing invading pathogens for ecological niches and metabolic substrates. Among the features necessary to provide health benefits, commensal microorganisms must have the ability to adhere to human intestinal cells and consequently to colonize the gut. Studies on mechanisms mediating adhesion of lactobacilli to human intestinal cells showed that factors involved in the interaction vary mostly among different species and strains, mainly regarding interaction between bacterial adhesins and extracellular matrix or mucus proteins. We have investigated the adhesive properties of Lactobacillus plantarum, a member of the human microbiota of healthy individuals. Results We show the identification of a Lactobacillus plantarum LM3 cell surface protein (48 kDa, which specifically binds to human fibronectin (Fn, an extracellular matrix protein. By means of mass spectrometric analysis this protein was identified as the product of the L. plantarum enoA1 gene, coding the EnoA1 alfa-enolase. Surface localization of EnoA1 was proved by immune electron microscopy. In the mutant strain LM3-CC1, carrying the enoA1 null mutation, the 48 kDa adhesin was not anymore detectable neither by anti-enolase Western blot nor by Fn-overlay immunoblotting assay. Moreover, by an adhesion assay we show that LM3-CC1 cells bind to fibronectin-coated surfaces less efficiently than wild type cells, thus demonstrating the significance of the surface displaced EnoA1 protein for the L. plantarum LM3 adhesion to fibronectin. Conclusion Adhesion to host tissues represents a crucial early step in the colonization process of either pathogens or commensal bacteria. We demonstrated the involvement of the L. plantarum Eno A1 alfa-enolase in Fn-binding, by studying

Das Bauhaus in Bewegung - wie das Bauhausgebäude in Dessau funktioniert

OpenAIRE

Seyler Thomas

2016-01-01

Analyse und Deutung des Bauhausentwurfs - wie das Bauhaus funktioniert Ein Architekt analysiert und deutet den Bauhausentwurf Das Bauhaus in Dessau gilt als Inbegriff einer räumlich freien Anordnung. Diese lässt sich jedoch auf den Prozess einer Entfaltung zurückführen. Der Autor erklärt Schritt für Schritt die Systematik des Entwurfs aus seiner beruflichen Erfahrung.

An Alpha spectrometer for measuring radon daughter individual activity concentration; Spettrometro Alfa per la misura delle concentrazioni individuali in attivita' della progenie del radon

Energy Technology Data Exchange (ETDEWEB)

Berico, M.; Formignani, M. [ENEA, Div. Protezione dell' Uomo e degli Ecosistemi, Centro Ricerche E. Clementel, Bologna (Italy); Mariotti, F. [Bologna Univ., Bologna (Italy). Dipt. di Fisica

2001-07-01

In the frame of the program of the Institute for Radiation Protection of ENEA, related to the evaluation of dose from radon and thoron progeny, an alpha spectrometer for the continuous air monitoring (CAM type) of radon and thoron has been realized. The constructive characteristics of the device are here presented together with energy and efficiency calibration. The device allows, by means of a screen type diffusion battery and a filter, to determinate the single radioactivity of each radionuclide of the progeny selecting them in relation to their diffusive behaviour (dichotomous particle size selection). The three-count filter method has been employed to measure the concentrations of {sup 218}Po, {sup 214}Pb and {sup 214}Bi in air. Radon and thoron effective doses using a dosimetric, instead of an epidemiologic approach, will be then evaluated. [Italian] Presso l'Istituto per la Radioprotezione, nell'ambito del programma di valutazione di dose da radon e' stato progettato e realizzato uno spettrometro alfa per il monitoraggio continuo in aria (CAM) della progenie del radon e del toron. Le caratteristiche costruttive dello strumento permettono, tramite l'utilizzo di batterie a diffusione a reti, di determinare l'attivita' individuale della progenie per diverse dimensioni granulometriche in particolare per la frazione attaccata e non al particolato amosferico con un taglio granulometrico di qualche nanometro. E' stato inoltre applicato un metodo spettrometrico a tre conteggi per il calcolo delle concentrazioni individuali della progenie del radon, {sup 218}Po, {sup 214}Pb and {sup 214}Bi, effettuando un conteggio alfa di {sup 218}Po e due conteggi alfa di {sup 214}Po. Tale informazione consentira' una valutazione della dose di radon utilizzando il modello dosimetrico in alternativa a quello epidemiologico.

Enhanced sialylation and in vivo efficacy of recombinant human α-galactosidase through in vitro glycosylation

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Youngsoo Sohn

2013-03-01

Full Text Available Human α-galactosidase A (GLA has been used in enzymereplacement therapy for patients with Fabry disease. Weexpressed recombinant GLA from Chinese hamster ovary cellswith very high productivity. When compared to an approvedGLA (agalsidase beta, its size and charge were found to besmaller and more neutral. These differences resulted from thelack of terminal sialic acids playing essential roles in the serumhalf-life and proper tissue targeting. Because a simplesialylation reaction was not enough to increase the sialic acidcontent, a combined reaction using galactosyltransferase,sialyltransferase, and their sugar substrates at the same timewas developed and optimized to reduce the incubation time.The product generated by this reaction had nearly the samesize, isoelectric points, and sialic acid content as agalsidasebeta. Furthermore, it had better in vivo efficacy to degrade theaccumulated globotriaosylceramide in target organs of Fabrymice compared to an unmodified version. [BMB Reports 2013;46(3: 157-162

Onset of Type 1 Diabetes Mellitus During Pegylated-interferon Alfa and Ribavirin Therapy for Chronic Hepatitis C Virus Infection

Science.gov (United States)

Ranganathan, Raghini; Janarthanan, Krishnaveni; Rajasekaran, Senthilkumar

2012-01-01

A 16-year-old female was treated with pegylated-interferon (PEG-IFN) alfa (a)-2b and ribavirin combination therapy for chronic hepatitis C virus (HCV) infection. She attained rapid virological response. She presented with diabetic ketoacidosis after 41 weeks of therapy. Anti-glutamic acid decarboxylase antibodies and islet cell antibodies were negative. Her fasting serum C-peptide level was <0.1 ng/mL, and the treatment course was completed. This case underlines the importance of periodic plasma glucose monitoring in patients during and after PEG-IFN and ribavirin therapy. PMID:25755410

I M L Das

Indian Academy of Sciences (India)

Home; Journals; Bulletin of Materials Science. I M L Das. Articles written in Bulletin of Materials Science. Volume 33 Issue 4 August 2010 pp 383-390 Electrical Properties. Temperature dependence of electromechanical properties of PLZT /57/43 ceramics · A K Shukla V K Agrawal I M L Das Janardan Singh S L Srivastava.

Seguridad de la terapia de interferón alfa 2b recombinante más ribavirina en la hepatitis crónica C

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Yoan Antonio Sánchez Rodríguez

2011-03-01

Full Text Available INTRODUCCIÓN: la hepatitis crónica C ha adquirido rango de pandemia. El virus de la hepatitis C se ha convertido en la causa principal de hepatitis crónica, cirrosis hepática, hepatocarcinoma, y trasplante de hígado a nivel mundial. OBJETIVO: identificar los efectos adversos asociados a la terapia combinada interferón alfa 2b recombinante más ribavirina durante la evolución del tratamiento y hasta 8 semanas después de finalizado, así como los principales efectos asociados a salidas temporales o definitivas de esta terapia. MÉTODOS: estudio de farmacovigilancia cuya serie estuvo conformada por 122 pacientes con hepatitis crónica C atendidos en el Instituto de Gastroenterología desde mayo de 2001 hasta mayo de 2006. Se utilizó interferón alfa 2b recombinante (3 millones de unidades 3 veces por semana más ribavirina (1 000 o 1 200 mg diarios en dependencia del peso corporal durante 48 semanas. RESULTADOS: el 88,5 % del total de casos presentó efectos adversos; de ellos el 79,5 % correspondió al síndrome seudogripal, seguido de manifestaciones hematológicas, neuropsiquiátricas, gastrointestinales, entre otras menos frecuentes. El 6,6 % de la serie presentó salidas temporales del tratamiento por efecto adverso distinto de la anemia y 4 pacientes, salidas definitivas del estudio, tres por anemia hemolítica severa y uno con hipertiroidismo no controlable. CONCLUSIONES: la terapia combinada interferón alfa 2b recombinante más ribavirina resulta segura, donde el mayor número de casos presentó síndrome seudogripal como efecto adverso más frecuente. Las manifestaciones hematológicas asociadas a las salidas definitivas del estudio permitieron recomendar seguimiento estricto de la hemoglobina y profundizar en el diagnóstico y tratamiento de los principales efectos adversos presentes en otros sistemas y asociados a esta terapia.

Adaptação transcultural e validação do World Health Organization Health and Work Performance Questionnaire para enfermeiros brasileiros

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Monica Chiodi-Toscano de Campos

2013-12-01

Full Text Available Estudo com objetivo de adaptar transculturalmente o instrumento Health and Work Performance Questionnaire (HPQ e avaliar as propriedades psicométricas da versão brasileira para enfermeiros. O processo de adaptação seguiu o Protocolo de Tradução da Organização Mundial da Saúde. Os dados para a avaliação das propriedades psicométricas foram coletados em um hospital público e de ensino de Ribeirão Preto em 2011. As propriedades psicométricas analisadas foram: validade de face e conteúdo (grupo de especialistas, confiabilidade pelo Alfa de Cronbach e estabilidade por teste-reteste. Na avaliação das propriedades psicométricas, a consistência interna da versão adaptada do HPQ, o alfa de cronbach foi de 0,94 para a seção A e 0,86 para a seção B do instrumento. Na análise das concordâncias da estabilidade teste-reteste, as concordâncias foram positivas e estatisticamente significativas. Dessa forma, a versão adaptada do HPQ mostrou-se válida e confiável na amostra estudada.

Fabrication of low cost optical detection system for E. coli 0157 based on4-methyl umbelliferyl-β-D-Galactoside--β-galactosidase reaction

International Nuclear Information System (INIS)

Racho, Joseph Michael D.; Del Rosario, Erba V.; Dawal, Karlen C.; Laurente, Dan R.; Sucgang, Raymond J.

2008-01-01

This paper reports the fabrication of a handheld, simple to use sensor which detects contamination by deadly strains of E coli in drinking water. E. coli 0157, predominately 0157:H7, has been implicated as the causative organism of hemolytic uremic syndrome and hemorrhagic colitis. The sensor works based on irreversible color changes that occur subsequent to hydrolysis of a chromogenic substrate by a β-galactosidase enzyme of E.coli. The recognition material, 4-methyl umbelliferyl-β-D-galactoside has two components: a fluorescent dye and a ''quencher'' which is sensitive to cleavage by β-galactosidase. In the absence of E. coli, the quencher binds to the dye and prevents fluorescence, while the interaction of β-galactosidase with the quencher results to the dissociation of the complex, 7-hydroxy-4-methylcoumarin, giving off a blue fluorescence. The sensor uses an LED light source shining on a photodetector. When the colorimetric reaction has taken place, light shines through the beaker containing the sample, and onto a light-sensitive meter. A clear tube of water or other clear solution in the BLANK and has zero absorbance. A fraction of the incident light that is blocked by the sample, can be used for quantitative determination of the bacterial population in the water. The E. coli population is related to the absorbance reading, following Beer's law. The amount of incident and transmitted light is expressed in foot candle units, by a LX 104 light meter. The sensor shows outstanding response over the range of 0.8 x10 7 to 1.35 x10 8 CFUs E. coli concentration. The bacterial numbers in each test solution has been confirmed using standard plate count method. Excellent sensitivity and linearity (R 2 =0.984) has been acheived using the sensor, working with 6 replicate measurements per particular concentration. The specificity of the sensing system to E. coli, was tested by spiking other bacterial strains, (Staphyloccus aureus) during the 4th to 6th determination