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Sample records for quantitativa da caspase-3

  1. Estudo citofotométrico da expressão dos marcadores tumorais Caspase-3 e Ki-67 no adenocarcinoma gástrico Cytophotometric study of the expression of tumoral markers Caspase-3 and Ki-67 in gastric adenocarcinoma

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    Pedro Manuel Gonzales Cuellar

    2007-06-01

    Full Text Available RACIONAL: A carcinogênese gástrica é processo complexo e depende de fatores genéticos, ambientais e infecciosos. Nos últimos anos, houve grandes avanços nos campos da genética e da biologia molecular, sobre o desenvolvimento dos tumores. Os marcadores tumorais são substâncias ausentes nos tecidos normais e que podem ser identificadas em tecidos com câncer. Através de procedimentos imunoistoquímicos eles podem ser estudados. OBJETIVOS: Descrever a expressão citofotométrica do marcador tumoral Ki-67 analisando a densidade óptica e o índice de marcagem no adenocarcinoma de estômago. Descrever a expressão citofotométrica do marcador tumoral Caspase-3 analisando a densidade óptica e o índice de marcagem no adenocarcinoma de estômago. Comparar o índice de marcagem e densidade óptica dos marcadores tumorais Ki-67 e Caspase-3 no adenocarcinoma de estômago. MÉTODO: Foram selecionados, inicialmente, 58 blocos com espécime de adenocarcinoma gástrico coletados nos Serviços de Anátomo-Patologia do Hospital do Gama - Brasília (DF e Hospital Dom Orione - Araguaina (TO, e analizados no Laboratório de Citologia e Histopalogia Ltda - CITOLAB, Curitiba (PR. Foram aproveitados 31 blocos para o estudo histológico e imunoistoquímico realizado pelo sistema de análise computarizado SAMBA 4000. RESULTADOS: Das 31 lâminas estudas, 15 (48% foram marcadas pelo marcador Ki-67, 22 (71% foram marcadas pelo marcador Caspase-3 e 14 (45% marcaram com os dois marcadores. CONCLUSÕES: A expressão citofométrica do marcador Ki-67 foi observada em 15 lâminas da amostra estudada e apresentaram média do índice de marcagem de 36,85%, enquanto a densidade óptica apresentou média de 29,33 pixels. A expressão citofotométrica do marcador Caspase-3 foi observada em 22 lâminas da amostra estudada e apresentaram média do índice de marcagem de 87,71% e 60,74 pixels de média para a densidade óptica. Na comparação do índice de marcagem dos

  2. Estudo da expressão citofotométrica do marcador tumoral Caspase-3 no adenocarcinoma de cólon Citophotometric expression study of tumoral marker Caspase-3 on colon adenocarcinoma

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    João Batista Monteiro Tajra

    2007-12-01

    Full Text Available RACIONAL: O adenocarcinoma de cólon é a segunda causa mais comum de morte por câncer em homens e mulheres, sendo responsável por mais de cinco milhões de mortes por ano. No momento do diagnóstico apenas 70% dos tumores são ressecáveis, 75% são curáveis e 25% poderão ter recorrência da doença. A apoptose é uma das responsáveis pelo equilíbrio homeostático entre as células. Durante o desenvolvimento do processo de degeneração maligna celular o desequilíbrio na apoptose é considerado um dos principais marcos neoplásicos. A caspase-3 é uma das mais importantes moléculas na apoptose, sendo sua efetora principal. Sua expressão e prognóstico têm sido relatados em vários estudos e revisões com seu papel valorizado desde o surgimento do pólipo até a sua transformação maligna, com a taxa de apoptose diminuindo progressivamente. OBJETIVOS: Avaliar a expressão citofotométrica computadorizada do marcador Caspase-3 no adenocarcinoma de cólon; avaliá-lo nas fases evolutivas na classificação modificada de Dukes e comparar sua expressão nos tumores do lado direito e esquerdo do cólon. MÉTODOS: Utilizaram-se 19 casos de câncer recuperados de blocos de parafina confirmados por hematoxilina-eosina e submetidos à técnica imunoistoquímica da estreptavidina-biotina com anticorpo policlonal anti-caspase-3. Após este processo as lâminas marcadas foram submetidas à leitura pelo sistema SAMBA com o software IMUNNO 4.00. Foram analisados três índices: marcagem (Label index, heterogeneidade e densidade óptica. Utilizaram-se a marcagem individual, avaliação da expressão do marcador e grupos definidos de tumores com classificação Dukes e pelo lado do tumor. RESULTADOS: A média do índice de marcagem da caspase-3 foi de 85,24 e da densidade óptica de 39,55. Na classificação Dukes de 12 tipos B tiveram índice de marcagem de 86,20 e a densidade óptica de 37,72 e para os 7 tipos C a área de marcagem foi de 85,66 e a

  3. Avaliação da caspase-3 e Ki-67 como marcadores prognósticos nas neoplasias mamárias em cadelas

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    Ana Carolina Bernardes Terzian

    2007-04-01

    Full Text Available A apoptose e a proliferação celular possuem uma participação importante na tumorigênese, determinando o crescimento tumoral e consequentemente sua agressividade. O presente estudo teve como objetivo avaliar a ocorrência da apoptose associada a proliferação celular em neoplasias mamárias em cadelas e a elas a evolução clínica do paciente. Setenta animais foram submetidos à exérese cirúrgica do tumor, sendo este submetido ao diagnóstico histopatológico e marcação imuno-histoquímica para caspase-3 e Ki-67. Estes marcadores de apoptose e proliferação celular demonstraram grande expressão nas neoplasias malignas, principalmente nos carcinomas, considerado o mais maligno dos tumores. Estes resultados corroboram os dados da literatura e contribuem para um prognóstico tumoral criterioso que complementa a classificação tumoral existente proporcionando uma melhor e maior sobrevida devido a uma adequação do procedimento terapêutico de cada paciente.

  4. ANÁLISE DESCRITIVA QUANTITATIVA DA AGUARDENTE DE CANA DURANTE O ENVELHECIMENTO EM TONEL DE CARVALHO (Quercus alba L.

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    CARDELLO Helena Maria André Bolini

    1998-01-01

    Full Text Available Apesar da importância econômica e social da aguardente de cana brasileira, são ainda muito escassos os estudos sobre sua qualidade sensorial, porém as crescentes exigências do mercado, tem feito crescer a preocupação com a qualidade dessa bebida. A aguardente de cana é muito apreciada por seu aroma e sabor característicos, que podem ainda melhorar pelo envelhecimento em recipientes de madeira. O complexo processo que ocorre durante o envelhecimento depende além de vários fatores, do tipo de madeira empregada, do tempo de maturação e obviamente da qualidade inicial do destilado. A análise descritiva quantitativa, metodologia muito aplicada na caracterização dos atributos sensoriais de alimentos e bebidas, foi utilizada neste trabalho para estudar o perfil sensorial da aguardente de cana, durante o envelhecimento, em toneis de carvalho. Nesse sentido, foram analisadas amostras de aguardente envelhecidas durante zero, 12, 24, 36 e 48 meses em um tonel de carvalho de 200 litros e duas amostras comerciais, sendo uma delas envelhecida. Dezesseis provadores pré-selecionados através de testes triangulares e análise sequencial, geraram pelo método rede (Kelly's Repertory Grid Method, os termos descritores das aguardentes. Após a etapa de treinamento, foram selecionados 10 provadores com base em seu poder de discriminação, repetibilidade e concordância com a equipe no uso de escalas. As amostras foram então apresentadas e avaliadas pelos provadores, em cabines individuais de forma monádica, com quatro repetições. Os resultados obtidos foram submetidos à análise de variância, teste de médias de Tukey e à Análise de Componentes Principais. Os termos descritores escolhidos em consenso pelos membros da equipe sensorial foram: coloração amarela, aroma alcoólico, aroma de madeira, aroma de baunilha, doçura inicial, doçura residual, sabor alcóolico inicial, sabor alcóolico residual, sabor de madeira inicial, sabor de

  5. Expression and activation of Daphnia pulex Caspase-3 are involved in regulation of aging.

    Science.gov (United States)

    Tong, Qiaoqiong; Zhang, Mengmeng; Cao, Xiao; Xu, Shanliang; Wang, Danli; Zhao, Yunlong

    2017-11-15

    Death-mediating proteases such as Caspases have been implicated in aging. Remarkably, active Caspase-3 can trigger widespread damage and degeneration, playing a key role in causing cell death. In order to explore the relationship between Caspase-3 and aging in Daphnia pulex, we cloned and analyzed the full-length cDNA sequence of its Caspase-3 gene. Both mRNA expression and activity of D. pulex Caspase-3 increased with age. Moreover, different forms of Caspase-3 appeared with aging. The expression of casp3-L was higher and decreased with age, while that of casp3-S was weak and increased with age, consistent with the trend in Caspase-3 activity. Mhc mRNA expression declined over time and was negatively correlated with age and Caspase-3. In situ hybridization results showed that Caspase-3 mRNA was expressed in different growth and reproduction stages, and its expression levels in embryos and larva were lower than that in adult D. pulex. Western blot analysis revealed the presence of Caspase-3 in the form of zymogens with a molecular weight of ~36kDa. Overall, this study explored age-associated gene regulation to provide a basis for the molecular mechanism of D. pulex reproductive conversion. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Análise da expressão do oncogene PML-RARalfa por PCR quantitativa em pacientes com leucemia aguda promielocítica

    OpenAIRE

    2006-01-01

    O gene de fusão PML-RARα é o mais freqüente marcador molecular da Leucemia Aguda Promielocítica (LAP). Com o objetivo de comparar os métodos moleculares qualitativo e quantivo, avaliar os níveis de expressão do gene correlacionar com características biológicas foram de pacientes com LAP ao diagnostico e pós-consolidação. O RNA total foi extraído a partir de e o cDNA sintetizado por RT-PCR. O gene ABL foi utilizado como controle constitutivo e a análise quantitativa realizada por curva pa...

  7. Abordagens semiquantitativa e quantitativa na avaliação da textura do solo por espectroscopia de reflectância bidirecional no VIS-NIR-SWIR

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    Marston Héracles Domingues Franceschini

    2013-12-01

    Full Text Available O objetivo deste trabalho foi avaliar o potencial da espectroscopia de reflectância no VIS-NIR-SWIR, para a caracterização granulométrica de amostras de solos de diferentes classes texturais, e obter modelos de predição dos teores de argila, silte e areia no solo. Utilizou-se um conjunto de amostras representativas de Latossolos e Argissolo de cinco locais do Estado do Mato Grosso do Sul. Os espectros do visível e do infravermelho próximo ao infravermelho de ondas curtas (de 350 a 2.500 nm das amostras foram obtidos e analisados. Empregaram-se a análise de componentes principais (ACP, agrupamento por "fuzzy c-means", regressão logística multinomial (RLM e regressão por mínimos quadrados parciais. Espectros característicos para as diferentes classes texturais e a segregação de amostras de classes texturais e de locais de coleta com características distintas, por meio da ACP, "fuzzy c-means" e RLM, mostram o potencial semiquantitativo dos dados de reflectância no VIS-NIR-SWIR. Obteve-se quantificação satisfatória quanto à argila (R²=0,92, RPD=3,59, ao silte (R²=0,80, RPD=2,15 e à areia (R²=0,87, RPD=2,62. As técnicas de espectroscopia de reflectância podem auxiliar na determinação da textura e da variabilidade espacial do solo com metodologias semiquantitativas ou quantitativas.

  8. Software para análise quantitativa da deglutição Swallowing quantitative analysis software

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    André Augusto Spadotto

    2008-02-01

    Full Text Available OBJETIVO: Apresentar um software que permita uma análise detalhada da dinâmica da deglutição. MATERIAIS E MÉTODOS: Participaram deste estudo dez indivíduos após acidente vascular encefálico, sendo seis do gênero masculino, com idade média de 57,6 anos. Foi realizada videofluoroscopia da deglutição e as imagens foram digitalizadas em microcomputador, com posterior análise do tempo do trânsito faríngeo da deglutição, por meio de um cronômetro e do software. RESULTADOS: O tempo médio do trânsito faríngeo da deglutição apresentou-se diferente quando comparados os métodos utilizados (cronômetro e software. CONCLUSÃO: Este software é um instrumento de análise dos parâmetros tempo e velocidade da deglutição, propiciando melhor compreensão da dinâmica da deglutição, com reflexos tanto na abordagem clínica dos pacientes com disfagia como para fins de pesquisa científica.OBJECTIVE: The present paper is aimed at introducing a software to allow a detailed analysis of the swallowing dynamics. MATERIALS AND METHODS: The sample included ten (six male and four female stroke patients, with mean age of 57.6 years. Swallowing videofluoroscopy was performed and images were digitized for posterior analysis of the pharyngeal transit time with the aid of a chronometer and the software. RESULTS: Differences were observed in the average pharyngeal swallowing transit time as a result of measurements with chronometer and software. CONCLUSION: This software is a useful tool for the analysis of parameters such as swallowing time and speed, allowing a better understanding of the swallowing dynamics, both in the clinical approach of patients with oropharyngeal dysphagia and for scientific research purposes.

  9. Análise quantitativa da camada de fibras nervosas da retina de cães normais e glaucomatosos através da polarimetria a laser

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    Ana Lucia Braga Martins

    2003-01-01

    Full Text Available As lesões glaucomatosas são progressivas e irreversíveis. Estudos sobre o aparecimento e comportamento desta enfermidade vêm sendo discutidos na oftalmologia em geral. O diagnóstico precoce é de extrema importância para o tratamento e estabilização da mesma. A análise da espessura da camada de fibras nervosas retinianas ''in vivo'' foi proposta na oftalmologia humana, para estabelecer as alterações produzidas pelo glaucoma, e tem demonstrado que tais alterações podem ser detectadas até seis anos antes de alterações de campo visual e aumento de pressão intraocular. Entretanto, na Medicina Veterinária, estes dados carecem de estudo e discussão. Neste experimento foram utilizados dois grupos de cães, com olhos normais e olhos glaucomatosos, que foram submetidos à análise das fibras nervosas retinianas através do Analisador de Fibras Nervosas GDx. Os resultados estatísticos demonstraram que a camada de fibras nervosas dos olhos dos cães glaucomatosos estava mais delgada (p < 0,05 quando comparados aos olhos de cães normais, confirmando a perda de axônios das células ganglionares em olhos de cães com glaucoma.

  10. Características quantitativas da carcaça de bovinos Charolês, Caracu e cruzamentos recíprocos terminados em confinamento

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    Perotto Daniel

    2000-01-01

    Full Text Available RESUMO - Foram analisadas doze características quantitativas das carcaças de 162 machos bovinos inteiros pertencentes aos grupos Caracu (Ca, Charolês (Ch, 3/4Ca+1/4Ch, 3/4Ch+1/4Ca, 5/8Ca+3/8Ch e 5/8Ch+3/8Ca, nascidos no período de 1988 a 1993. As médias para idade e peso ao início do período de confinamento, duração do confinamento, idade e peso ao abate foram, respectivamente, 712 dias, 362 kg, 99 dias, 811 dias e 489 kg. Durante o confinamento, os animais receberam silagem de milho à vontade e uma ração concentrada (79% NDT, 17,8% PB fornecida à base de 1% PV/animal/dia. Os resultados mostraram que os grupos Ch e Ca não diferiram quanto ao peso de carcaça quente, ao rendimento de carcaça quente, às porcentagens de costilhar e gordura e ao peso da porção comestível da carcaça (PCC. O Ch superou o Ca para a área da seção transversal do músculo Longissimus dorsi (AML, o escore de conformação da carcaça (CONF e as porcentagens de serrote e músculos. Animais Ca apresentaram maior espessura de gordura de cobertura e maiores porcentagens de dianteiro (PED e ossos que os do grupo Ch. Houve resposta heterótica para PCQ, AML, CONF, PCC e PED. As duas gerações avançadas de cruzamentos alternados Ch x Ca foram superiores à média das raças paternas para PCQ, AML, CONF, PCC e PED. O cruzamento alternado Ch x Ca produz animais com características de carcaça superiores às dos Charolês e Caracu puros.

  11. A face quantitativa da linguagem: um dicionário de freqüências do português

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    Maria Tereza Camargo Biderman

    2001-02-01

    Full Text Available

    O Dicionário de Freqüências do léxico do português brasileirocontemporâneo baseou-se num corpus de língua escrita, variedade brasileirade 5 milhões de palavras (1950 a 1990. Alguns resultados quantitativos:apenas 42.212 unidades léxicas diferentes totalizaram os 5 milhões de ocorrênciasdo corpus, excluídos topónimos e antropônimos. Os dados estatísticosdo dicionário registram altíssima freqüência das palavras instrumentais(artigos, preposições, pronomes, conjunções etc. bem como de verbos auxiliarese modalizadores. O mesmo ocorre com palavras de significação muito geral, arquilexemas, altamente polissêmicos. Na vertente oposta estão as palavras de baixa freqüência sobretudo os hapax legomena, que contribuemmaciçamente para o total de 42.212 lexias registradas neste corpus. De fato,as palavras de baixa freqüência totalizam grande parte desse index verborum;caso contrário, o repertório vocabular seria muito menor. A categoria substantivo contribui com a maioria de vocábulos que ocorreram apenas uma vez no corpus, assim como os tecnicismos da linguagem científica. O vocabulário jornalístico é o mais neutro e o menos temático, constituindo uma espéciede média entre os outros gêneros de linguagem.

  12. Coxsackievirus B3-induced apoptosis and Caspase-3

    Institute of Scientific and Technical Information of China (English)

    JIAN PING YUAN; WEI ZHAO; HONG TAO WANG; KAI YU WU; TAO LI; XIAO KUI GUO; SHAN QING TONG

    2003-01-01

    Cell death can be classified into two categories: apoptosis and necrosis. Apoptotic pathway can beeither caspase-dependent or caspase-independent. Caspase-independent cytopathic effect (CPE) has beendescribed. In order to evaluate the pattern of HeLa cell death induced by Coxsackievirus B3 (CVB3)and whether apoptosis involves caspase activation, we co-cultivated HeLa cells with CVB3 and detectedthe cytopathic changes, the alteration of mRNA and protein expression of caspase-3 gene plus caspase-3activity, as well as analyzing DNA fragmentation before and after caspase-3 activity inhibition. Accordingto the results, we propose that CVB3 may induce apoptosis and necrosis in HeLa cells, the latter appearingmuch earlier. Caspase-3 is activated at the levels of both transcription and translation, and procaspase-3 isproteolytically cleaved, thus leading to the continuous increasing of both caspase-3 precursor protein and itssubunit. However, besides CPE, apoptosis induced by CVB3 is not a direct consequence of the activationof caspase-3, or caspase-3 is not the only effector molecule in apoptotic cell death, for caspase-3 inhibitorcan not decrease DNA fragmentation. Some other biochemical mechanisms may participate in the process,whose role weakens the effect of inhibiting caspase-3 activity.

  13. Análise quantitativa do tratamento da escoliose idiopática com o método klapp por meio da biofotogrametria computadorizada

    OpenAIRE

    Iunes, Denise H.; Maria B. B. Cecílio; Marina A. Dozza; Almeida,Polyanna R.

    2010-01-01

    INTRODUÇÃO: Poucos trabalhos comprovam a eficácia das técnicas fisioterapêuticas para o tratamento da escoliose. OBJETIVO: Analisar a eficácia do Método Klapp no tratamento das escolioses por meio do estudo quantitativo pela biofotogrametria computadorizada. MÉTODOS: Dezesseis indivíduos com média de idade de 15±2,61 anos, portadores de escoliose idiopática, foram tratados com o método Klapp. Para análise dos resultados do tratamento, todos foram fotografados antes e após o tratamento, seguin...

  14. Análise quantitativa das descargas epileptiformes generalizadas e da neuroimagem de pacientes com epilepsia generalizada idiopática

    OpenAIRE

    Braga, Aline Marques da Silva [UNESP

    2016-01-01

    Fundamento: Evidências experimentais de modelos animais de crises de ausência sugerem focalidades no início das descargas generalizadas. Estudos clínicos indicam que pacientes com o diagnóstico de epilepsia generalizada idiopática (EGI) exibem anormalidades focais que envolvem o circuito tálamo-cortical no eletroencefalograma (EEG) e na neuroimagem. Objetivos: Investigar a presença de características focais nas descargas generalizadas interictais usando análise quantitativa do EEG (EEGq...

  15. Increased caspase-3 immunoreactivity of erythrocytes in STZ diabetic rats.

    Science.gov (United States)

    Fırat, Uğur; Kaya, Savaş; Cim, Abdullah; Büyükbayram, Hüseyin; Gökalp, Osman; Dal, Mehmet Sinan; Tamer, Mehmet Numan

    2012-01-01

    Eryptosis is a term to define apoptosis of erythrocytes. Oxidative stress and hyperglycemia, both of which exist in the diabetic intravascular environment, can trigger eryptosis of erythrocytes. In this experimental study, it is presented that the majority of erythrocytes shows caspase-3 immunoreactivity in streptozocin- (STZ)-induced diabetic rats. Besides that, caspase-3 positive erythrocytes are aggregated and attached to vascular endothelium. In conclusion, these results may start a debate that eryptosis could have a role in the diabetic complications.

  16. Características quantitativas da carcaça de bovinos Charolês, Caracu e cruzamentos recíprocos terminados em confinamento

    OpenAIRE

    Perotto Daniel; Moletta José Luiz; Cubas Antonio Carlos

    2000-01-01

    RESUMO - Foram analisadas doze características quantitativas das carcaças de 162 machos bovinos inteiros pertencentes aos grupos Caracu (Ca), Charolês (Ch), 3/4Ca+1/4Ch, 3/4Ch+1/4Ca, 5/8Ca+3/8Ch e 5/8Ch+3/8Ca, nascidos no período de 1988 a 1993. As médias para idade e peso ao início do período de confinamento, duração do confinamento, idade e peso ao abate foram, respectivamente, 712 dias, 362 kg, 99 dias, 811 dias e 489 kg. Durante o confinamento, os animais receberam silagem de milho à vont...

  17. A short caspase-3 isoform inhibits chemotherapy-induced apoptosis by blocking apoptosome assembly.

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    Frédérique Végran

    Full Text Available Alternative splicing of caspase-3 produces a short isoform caspase-3s that antagonizes caspase-3 apoptotic activity. However, the mechanism of apoptosis inhibition by caspase-3s remains unknown. Here we show that exogenous caspase-3 sensitizes MCF-7 and HBL100 breast cancers cells to chemotherapeutic treatments such as etoposide and methotrexate whereas co-transfection with caspase-3s strongly inhibits etoposide and methotrexate-induced apoptosis underlying thus the anti-apoptotic role of caspase-3s. In caspase-3 transfected cells, lamin-A and α-fodrin were cleaved when caspase-3 was activated by etoposide or methotrexate. When caspase-3s was co-transfected, this cleavage was strongly reduced. Depletion of caspase-3 by RNA interference in HBL100 containing endogenous caspase-3s caused reduction in etoposide and methotrexate-induced apoptosis, whereas the depletion of caspase-3s sensitized cells to chemotherapy. In the presence of caspase-3s, a lack of interaction between caspase-3 and caspase-9 was observed. Immunoprecipitation assays showed that caspase-3s binds the pro-forms of caspase-3. This result suggested that the absence of interaction with caspase-9 when both variants of caspase-3 are present contribute to block the apoptosome assembly and inhibit apoptosis. These data support that caspases-3s negatively interferes with caspase-3 activation and apoptosis in breast cancer, and that it can play key roles in the modulation of response to chemotherapeutic treatments.

  18. Análise quantitativa sobre a mortalidade precoce de micro e pequenas empresas da cidade de São Paulo A quantitative analysis of the premature mortality of small companies in the city of São Paulo

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    Luis Fernando Filardi Ferreira

    2012-12-01

    Full Text Available O vigor da economia de vários países tem origem no sucesso de pequenas empresas. No Brasil, o Estado de São Paulo concentra mais da metade das empresas brasileiras de pequeno porte. Assim sendo, estudos sobre o sucesso ou o insucesso de micro e pequenas empresas têm sua relevância enaltecida. Este estudo busca apresentar uma análise quantitativa e qualitativa sobre os fatores associados à mortalidade precoce das micro e pequenas empresas. Para tanto, realizou-se uma pesquisa com os gestores de micro e pequenas empresas da cidade de São Paulo que encerraram suas atividades. Para análise quantitativa da amostra, que apresenta representatividade estatística, utilizou-se de técnicas de análise multivariada. Como principal resultado, pode-se destacar que os fatores associados à mortalidade são preponderantemente de natureza estratégica, o que contraria o senso comum que professa a insuficiência de recursos como principal motivador da falência das pequenas empresas. Tal resultado tem importância para orientar as políticas públicas, os empreendedores e as agências de fomento para a formulação de suas estratégias no sentido do desenvolvimento de competências administrativas para o sucesso das micro e pequenas empresas brasileiras.The economic vigor of many countries is due to the success of small businesses. In Brazil, Sao Paulo state concentrates more than half of Brazilian small businesses. Therefore, studies dealing with high or low performance of micro and small businesses are of great relevance. This study aims at presenting a quantitative and qualitative analysis of factors associated to premature mortality of micro and small businesses in the city of Sao Paulo. For the quantitative analysis of the sample, which is statistically significant, multivariate analysis was used. The main result of this investigation is that factors associated with mortality are mainly of strategic nature, contrary to the common belief that

  19. Análise quantitativa da deglutição orofaríngea em indivíduo gastrostomizado com Lipofuscinose Ceróide Neuronal: relato de caso

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    André Vinicius Marcondes Natel Sales

    2013-12-01

    Full Text Available A presença de disfagia orofaríngea infantil na população com afecções genéticas ainda é pouco estudada. O objetivo deste estudo foi analisar o tempo de trânsito oral total (TTOT e o tempo de trânsito faríngeo (TTF em um indivíduo com diagnóstico genético clínico de Lipofuscinose Ceróide Neuronal (LCN com disfagia orofaríngea grave. Indivíduo com LCN, 3 anos de idade, gastrostomizado há dois anos e sem via oral parcial, histórico de déficit de ganho de peso anterior a via alternativa de alimentação, porém sem complicações pulmonares. A deglutição orofaríngea foi estudada por meio de videofluoroscopia de deglutição e análise quantitativa da deglutição com uso de software específico para tal avaliação. Na análise quantitativa do TTOT e TTF constatou-se, respectivamente, 45,37 segundos (padrão de normalidade em criança é de 4 segundos e de 4,53 segundos para o TTF. Constatou-se significante aumento nos tempos de trânsito orofaríngeo neste indivíduo, sendo que a disfagia orofaríngea, parte do quadro desta criança com diagnóstico de LCN, deve ser investigada e acompanhada durante a evolução da doença. Uma avaliação da deglutição orofaríngea e acompanhamento nos indivíduos com esta condição genética deve ser realizada, considerando que essa alteração pode fazer parte do fenótipo desta condição e também pelo impacto que esse aumento nos tempos da deglutição pode ocasionar na condição nutricional e pulmonar desta população.

  20. Caspase-3 activation in 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-treated mice.

    Science.gov (United States)

    Turmel, H; Hartmann, A; Parain, K; Douhou, A; Srinivasan, A; Agid, Y; Hirsch, E C

    2001-03-01

    In 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) models of Parkinson's disease (PD), dopaminergic (DA) neurons have been shown to die by apoptosis. Moreover, recent postmortem and in vitro results have indicated that apoptotic cell death induced by 1-methyl-4-phenylpyridinium (MPP(+)) may be mediated by caspase-3. To establish whether caspase-3 activation may indeed play a role in an in vivo model of PD, we studied caspase-3 activation in C57Bl/6 mice subchronically intoxicated with MPTP. We show that caspase-3 activation peaks early, at days 1 and 2 after the end of MPTP intoxication. In contrast, pycnotic neurons persist until day 7 postintoxication, indicating that caspase-3 activation is an early and transient phenomenon in apoptotic death of DA neurons. We further demonstrate that loss of tyrosine hydroxylase (TH) immunoreactivity in this model is indeed due to cell loss rather than to loss of TH protein expression. We conclude that mice subchronically intoxicated with MPTP represent a valid PD model to study and manipulate caspase activation in vivo.

  1. Discovery of Potent, Selective and Reversible Caspase-3 Inhibitors

    Institute of Scientific and Technical Information of China (English)

    Han Yongxin; John Tam; Paul Tawa; Donald W. Nicholson; Robert J. Zamboni; André Giroux; John Colucci; Christopher I. Bayly; Daniel J. Mckay; Sophie Roy; Steve Xanthoudakis; John Vaillancourt; Dita M. Rasper

    2004-01-01

    Recent studies towards understanding the molecular mechanisms of apoptosis have revealed the importance of a group of cysteinyl aspartate specific proteases, the caspases, in the programmed cell death process (Hengartner, M.O. Nature 2000, 407, 770). Caspase-3, in particular,has been characterized as the dominant effector caspase involved in the proteolytic cleavage of a variety of protein substrates including cytoskeletal proteins, kinases and DNA repair enzymes during apoptosis (Nicholson, D. W. Cell Death Differ. 1999, 6, 1028). The development of potent and selective caspase-3 inhibitors has thus emerged as an attractive therapeutic target. In the presentation,the identification of a series of potent, selective and reversible non-peptidyl caspase-3 inhibitors containing a pyrazinone core (1) will be presented. SAR optimization at R1, R2, R3 and R4 led to the discovery of inhibitors such as 2 with excellent in vitro activities (IC50 against rh-caspase-3: 5 nM; IC50 against camptothecin induced apoptotic cell death in NT2 cells: 20 nM). Compounds such as 2 also displayed excellent in vivo activities in a number of animal models of acute injuries (see: Methot, N. et al, J. Exp. Med. 2004, 119, 199; Toulmond, S. et al, British J. Pharm. 2004, 141,689; Holtzman,D.M. et al, JBC, 2002, 277, 30128), and selected examples will be discussed during the presentation.

  2. Inner ear dysfunction in caspase-3 deficient mice

    Directory of Open Access Journals (Sweden)

    Woo Minna

    2011-10-01

    Full Text Available Abstract Background Caspase-3 is one of the most downstream enzymes activated in the apoptotic pathway. In caspase-3 deficient mice, loss of cochlear hair cells and spiral ganglion cells coincide closely with hearing loss. In contrast with the auditory system, details of the vestibular phenotype have not been characterized. Here we report the vestibular phenotype and inner ear anatomy in the caspase-3 deficient (Casp3-/- mouse strain. Results Average ABR thresholds of Casp3-/- mice were significantly elevated (P Casp3+/- mice and Casp3+/+ mice at 3 months of age. In DPOAE testing, distortion product 2F1-F2 was significantly decreased (P Casp3-/- mice, whereas Casp3+/- and Casp3+/+ mice showed normal and comparable values to each other. Casp3-/- mice were hyperactive and exhibited circling behavior when excited. In lateral canal VOR testing, Casp3-/- mice had minimal response to any of the stimuli tested, whereas Casp3+/- mice had an intermediate response compared to Casp3+/+ mice. Inner ear anatomical and histological analysis revealed gross hypomorphism of the vestibular organs, in which the main site was the anterior semicircular canal. Hair cell numbers in the anterior- and lateral crista, and utricle were significantly smaller in Casp3-/- mice whereas the Casp3+/- and Casp3+/+ mice had normal hair cell numbers. Conclusions These results indicate that caspase-3 is essential for correct functioning of the cochlea as well as normal development and function of the vestibule.

  3. Avaliação clinico-quantitativa da ação do Ciba 34.647-Ba sobre a espasticidade

    Directory of Open Access Journals (Sweden)

    Israel Roitman

    1977-12-01

    Full Text Available Utilizando a quantificação clínica da sintomatologia neurológica os autores fizeram observações sobre o Ciba 34.647-Ba e avaliaram pacientes com espasticidade e incapacidade motora devidas a lesões da medula espinal. São relatados os resultados, salientando os efeitos do medicamento sobre os principais componentes da espasticidade (hipertonia, hiperreflexia, clono e automatismos. As teorias propostas para explicar o mecanismo de ação da droga são mencionadas. São também referidas as doses utiizadas, duração da ação músculo- relaxante, tolerância e efeitos colaterais.

  4. Análise quantitativa da proliferação celular no epitélio seminífero humano

    OpenAIRE

    Graça, Maria Inês Pinho dos Santos

    2009-01-01

    Apesar de décadas de investigação básica na espermatogénese humana, pouco se sabe acerca da proliferação e diferenciação das células germinais humanas em termos quantitativos. Com este trabalho pretendeu-se estudar quantitativamente os efeitos temporais e específicos de cada estadio celular, da Hormona Foliculo Estimulante (FSH) e da testosterona (T) na capacidade proliferativa do epitélio seminífero humano normal sob condições de cultura de órgão. Para tal, fragmentos de túbul...

  5. Peripheral Neuropathy in the Twitcher Mouse Involves the Activation of Axonal Caspase 3

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    Benjamin Smith

    2011-09-01

    Full Text Available Infantile Krabbe disease results in the accumulation of lipid-raft-associated galactosylsphingosine (psychosine, demyelination, neurodegeneration and premature death. Recently, axonopathy has been depicted as a contributing factor in the progression of neurodegeneration in the Twitcher mouse, a bona fide mouse model of Krabbe disease. Analysis of the temporal-expression profile of MBP (myelin basic protein isoforms showed unexpected increases of the 14, 17 and 18.5 kDa isoforms in the sciatic nerve of 1-week-old Twitcher mice, suggesting an abnormal regulation of the myelination process during early postnatal life in this mutant. Our studies showed an elevated activation of the pro-apoptotic protease caspase 3 in sciatic nerves of 15- and 30-day-old Twitcher mice, in parallel with increasing demyelination. Interestingly, while active caspase 3 was clearly contained in peripheral axons at all ages, we found no evidence of caspase accumulation in the soma of corresponding mutant spinal cord motor neurons. Furthermore, active caspase 3 was found not only in unmyelinated axons, but also in myelinated axons of the mutant sciatic nerve. These results suggest that axonal caspase activation occurs before demyelination and following a dying-back pattern. Finally, we showed that psychosine was sufficient to activate caspase 3 in motor neuronal cells in vitro in the absence of myelinating glia. Taken together, these findings indicate that degenerating mechanisms actively and specifically mediate axonal dysfunction in Krabbe disease and support the idea that psychosine is a pathogenic sphingolipid sufficient to cause axonal defects independently of demyelination.

  6. Peripheral neuropathy in the Twitcher mouse involves the activation of axonal caspase 3

    Directory of Open Access Journals (Sweden)

    Ernesto R Bongarzone

    2011-10-01

    Full Text Available Infantile Krabbe disease results in the accumulation of lipid-raft-associated galactosylsphingosine (psychosine, demyelination, neurodegeneration and premature death. Recently, axonopathy has been depicted as a contributing factor in the progression of neurodegeneration in the Twitcher mouse, a bona fide mouse model of Krabbe disease. Analysis of the temporal-expression profile of MBP (myelin basic protein isoforms showed unexpected increases of the 14, 17 and 18.5 kDa isoforms in the sciatic nerve of 1-week-old Twitcher mice, suggesting an abnormal regulation of the myelination process during early postnatal life in this mutant. Our studies showed an elevated activation of the pro-apoptotic protease caspase 3 in sciatic nerves of 15- and 30-day-old Twitcher mice, in parallel with increasing demyelination. Interestingly, while active caspase 3 was clearly contained in peripheral axons at all ages, we found no evidence of caspase accumulation in the soma of corresponding mutant spinal cord motor neurons. Furthermore, active caspase 3 was found not only in unmyelinated axons, but also in myelinated axons of the mutant sciatic nerve. These results suggest that axonal caspase activation occurs before demyelination and following a dying-back pattern. Finally, we showed that psychosine was sufficient to activate caspase 3 in motor neuronal cells in vitro in the absence of myelinating glia. Taken together, these findings indicate that degenerating mechanisms actively and specifically mediate axonal dysfunction in Krabbe disease and support the idea that psychosine is a pathogenic sphingolipid sufficient to cause axonal defects independently of demyelination.

  7. Discurso e abordagens quantitativas

    Directory of Open Access Journals (Sweden)

    Giselle Machline de Oliveira e Silva

    2001-02-01

    Full Text Available

    Este artigo revê o comportamento de diversos fenômenos de acordo com o estatuto informacional de seus referentes: a ocorrência de artigosdefinidos, a ordem do sujeito em relação a seu predicado, a topicalização deobjetos diretos, as sentenças clivadas, a ordem de orações de causa e de condição,e os marcadores discursivos. Ele mostra que a informação é uma restriçãopoderosa para a maioria dos fenômenos que tratam da ordem, mas muitomenos relevante para a colocação de artigos definidos e a co-ocorrência comalguns marcadores discursivos.

  8. Impacto da leitura intensiva em língua inglesa no repertório lexical: uma análise quantitativa

    Directory of Open Access Journals (Sweden)

    Eduardo Batista da Silva

    2016-03-01

    Full Text Available A leitura pode servir como fonte de vocabulário para o aprendiz de língua inglesa. Nesse contexto, o objetivo geral deste trabalho é analisar o desempenho de aprendizes de língua inglesa, com o propósito de investigar se por meio da exposição continuada a textos escritos com as palavras mais comuns do inglês ocorre aquisição incidental de vocabulário. Os objetivos específicos são os seguintes: 1 conscientizar professores e pesquisadores a respeito da relevância da leitura na aula de inglês e 2 discutir aspectos quantitativos relacionados ao vocabulário. A fundamentação teórica recorre principalmente às contribuições da Linguística de Corpus e da Linguística Aplicada. Quanto à metodologia, após a seleção dos participantes, foi aplicado um teste conhecido como Vocabulary Knowledge Scale (VKS, que avalia o autorreconhecimento acerca de palavras-alvo. O VKS foi aplicado antes e depois da entrega de 20 textos. No pré-teste, 14,3% das palavras-alvo eram conhecidas por mais da metade dos sujeitos, ao passo que no pós-teste, 66,7% das palavras-alvo eram conhecidas por mais da metade deles. Os resultados apontam que as atividades baseadas na leitura visando a prática lexical propiciam um alto índice de aquisição incidental.

  9. Própolis: avaliação quantitativa da produção de própolis

    OpenAIRE

    2014-01-01

    O estudo foi realizado no posto apícola do INIAV, Tapada da Ajuda, Instituto Superior de Agronomia, tendo como objetivo a avaliação da produção de própolis, durante um período de 60 dias, em plena época de floração. Foram selecionadas e padronizadas vinte colónias de abelhas Apis melifera iberiensis instaladas em colmeias modelo “Lusitana”. Realizaram-se análises do ciclo biológico e classificaram-se as colónias, estimando-se, por observação direta, o número de abelhas da co...

  10. Avaliação quantitativa de micronúcleos na citologia esfoliativa da mucosa bucal de pacientes dependentes químicos

    OpenAIRE

    Lima, Celina Faig

    2007-01-01

    O presente trabalho avaliou quantitativamente micronúcleos (MN) na citologia esfoliativa da mucosa bucal de pacientes com dependência química. Os MN contêm material genético perdido durante a mitose em eventos clastogênicos e aneugênicos e ocorrem antes de qualquer mudança histopatológica pré-neoplásica tornar-se evidente. Foram triados 24 pacientes dependentes químicos, do CAPSAD (Centro de Apoio Psicosocial Álcool Drogas) da Prefeitura de São José dos Campos e 24 pacientes não fumantes, ou ...

  11. Influência da baixa temperatura ambiental sobre a inflamação em rã-touro gigante (Rana catesbeiana: avaliações qualitativa e quantitativa

    Directory of Open Access Journals (Sweden)

    José Luiz Catão-Dias

    1999-01-01

    Full Text Available O presente trabalho teve por objetivo investigar a modulação exercida pela temperatura ambiental sobre a cinética celular inflamatória experimentalmente induzida em Rana catesbeiana, rã-touro gigante. Para tanto, 120 espécimes pós-metamórficos foram mantidos a 6ºC e 24ºC e tratados pela transfixação do tecido muscular da coxa por fio de sutura ou injeção intramuscular de carragenina. Os resultados obtidos através de avaliações qualitativa e quantitativa do foco lesional mostraram que a baixa temperatura ambiental modula significativamente a evolução do processo inflamatório. Animais mantidos a 6ºC, em ambos os modelos, apresentaram números de células inflamatórias significativamente menores que os verificados a 24ºC, independentemente do tempo de avaliação. Por outro lado, algum fator pertencente aos mecanismos de defesa do hospedeiro não deve ter sido bloqueado pela temperatura, visto que a área de reação à injúria mostrou-se equivalente na maioria dos tempos pesquisados.

  12. Análise das caracteristicas quantitativas e qualitativas da microbacia do córrego barreiro, afluente do Rio Uberaba

    Directory of Open Access Journals (Sweden)

    Jose Luiz Rodrigues Torres

    2011-08-01

    Full Text Available A associação de métodos qualitativos e quantitativos no estudo das microbacias hidrográficas é fundamental para investigar os fatores que influenciam as diversas formas de relevo e identificação das homogeneidades na área. Com os objetivos de realizar a morfometria e avaliar a qualidade da água da microbacia do córrego do Barreiro, fez-se a caracterização morfométrica da área, que foi realizada sobre uma carta topográfica do IBGE (Folha Uberaba na escala 1:100.000, com o auxílio de curvímetros, planímetros e programa computacional AutocadMap2000i. Observou-se que a microbacia possui área total de 10,97 km², perímetro de 14,33 km, com comprimento do canal principal de 6,11 km e rede de drenagem de 3ª ordem, com padrão dendrítico; o Kc de 1,21; Ic de 0,76; e Kf de 0,23 indicam o formato alongado da microbacia, o que determina a não propensão a enchentes na área; a área tem aptidão para a pecuária; a água do córrego estava imprópria para consumo humano e animal devido estar contaminada com coliformes fecais no período avaliado.

  13. Caracterização quali-quantitativa do fitoplâncton da zona de arrebentação de uma praia amazônica Phytoplankton of the surf zone in Amazon beach

    Directory of Open Access Journals (Sweden)

    Jislene Brito Matos

    2012-12-01

    Full Text Available A estrutura da comunidade fitoplanctônica da praia da Princesa (Ilha de Maiandeua foi estudada durante um ciclo nictemeral nos meses de novembro/08, março/09, junho/09 e setembro/09, de modo a verificar os efeitos de algumas variáveis ambientais sobre esta comunidade. O microfitoplâncton da praia da Princesa esteve representado por 98 táxons, com predomínio das diatomáceas, seguidas dos dinoflagelados e cianofíceas. Nas amostras qualitativas, Coscinodiscus perforatus Ehrenberg se destacou como abundante nos meses de março (45,6% e junho (45,1%, enquanto que a análise quantitativa revelou Dimeregramma minor (Gregory Ralfs como dominante, principalmente, em novembro (82,0% e setembro (83,0%. A biomassa fitoplanctônica (clorofila-a foi significativamente mais elevada em março (U= 0,0; pThe structure of the phytoplankton community of Princesa beach (Maiandeua island was investigated during a nyctemeral cycle in the months of November/08, March/09, June/09 and September/09 to verify the effects of certain environmental variables on this community. The microphytoplankton of Princesa beach was represented by 98 taxa with the dominance of the diatoms, which were followed by the dinoflagellates and cyanophytes. The qualitative study revealed that Coscinodiscus perforatus Ehrenberg was abundant in March (45.6% and June (45.1%, whereas in the quantitative analysis Dimeregramma minor (Gregory Ralfs was dominant, mainly in November/08 (82.0% and September/09 (83.0%. Biomass (chlorophyll-a content was significantly higher in March/08 (U= 0.0; p <0.05. The average total phytoplankton density was significantly higher during the rainy period (F= 6.2; p< 0.05, mainly in June (1,223 ±110 x 10³ cell L-1. K-dominance curves showed a gradual decrease in diversity along the dry period. Non-Metric Multidimensional Scaling analysis (MDS revealed the formation of three groups, whereas Principal Component Analysis (PCA showed that salinity and turbidity were

  14. Activation of GSK-3β and Caspase-3 Occurs in Nigral Dopamine Neurons during the Development of Apoptosis Activated by a Striatal Injection of 6-Hydroxydopamine

    Science.gov (United States)

    Hernandez-Baltazar, Daniel; Mendoza-Garrido, Maria E.; Martinez-Fong, Daniel

    2013-01-01

    The 6-Hydroxydopamine (6-OHDA) rat model of Parkinson's disease is essential for a better understanding of the pathological processes underlying the human disease and for the evaluation of promising therapeutic interventions. This work evaluated whether a single striatal injection of 6-OHDA causes progressive apoptosis of dopamine (DA) neurons and activation of glycogen synthase kinase 3β (GSK-3β) and caspase-3 in the substantia nigra compacta (SNc). The loss of DA neurons was shown by three neuron markers; tyrosine hydroxylase (TH), NeuN, and β-III tubulin. Apoptosis activation was determined using Apostain and immunostaining against cleaved caspase-3 and GSK-3β pY216. We also explored the possibility that cleaved caspase-3 is produced by microglia and astrocytes. Our results showed that the 6-OHDA caused loss of nigral TH(+) cells, progressing mainly in rostrocaudal and lateromedial directions. In the neostriatum, a severe loss of TH(+) terminals occurred from day 3 after lesion. The disappearance of TH(+) cells was associated with a decrease in NeuN and β-III tubulin immunoreactivity and an increase in Apostain, cleaved caspase-3, and GSK-3β pY216 in the SNc. Apostain immunoreactivity was observed from days 3 to 21 postlesion. Increased levels of caspase-3 immunoreactivity in TH(+) cells were detected from days 1 to 15, and the levels then decreased to day 30 postlesion. The cleaved caspase-3 also collocated with microglia and astrocytes indicating its participation in glial activation. Our results suggest that caspase-3 and GSK-3β pY216 activation might participate in the DA cell death and that the active caspase-3 might also participate in the neuroinflammation caused by the striatal 6-OHDA injection. PMID:23940672

  15. Avaliação quantitativa da microinfiltração em cavidades de classe V restauradas com materiais hibridos de ionomero de vidro/resina composta

    OpenAIRE

    Claudia Salami de Magalhães

    1997-01-01

    Resumo: O objetivo deste estudo "in vitro" foi avaliar quantitativamente a microinfiltração em cavidades de classe V restauradas com materiais híbridos de ionômero de vidro/resina composta, comparando-os com um cimento de ionômero de vidro e com um sistema de resina composta/adesivo dentinário. Foram preparadas cavidades cilíndricas na superfície radicular de 105 dentes humanos extraídos, a cerca de 4 mm da junção amelo-cemeotária. Os dentes foram divididos aleatoriamente em cinco grupos, cad...

  16. Composição quali-quantitativa da produção de 'Syrah' cultivada sob estresse hídrico transiente Composition and chemical-sensorial profile of 'Syrah' cultivated under transient water stress

    Directory of Open Access Journals (Sweden)

    Antonio O. Santos

    2009-06-01

    Full Text Available A produção de vinho de qualidade requer o cultivo de uvas de padrão superior, com características químicas e físicas favoráveis, resultantes das condições micrometeorológicas locais, da variedade e do manejo do solo e da cultura. A composição quantitativa e qualitativa do rendimento da videira pode ser responsiva à variação na lâmina de irrigação e ao sistema de poda utilizado. Doravante, a interação entre os dois fatores citados e o meio físico circundante foi mostrada ser influente na qualidade do vinho. Um experimento foi conduzido com uva vinífera nas estações de crescimento de 2003 a 2005; cujo objetivo foi determinar a melhor combinação entre sistemas de poda (manual e mecânica e manejo da água, para a produção de vinho de 'Syrah' (Vitis vinifera L. cultivada em condições de microclima quente e seco. Os resultados mostraram que a composição do rendimento foi consistentemente afetada pelo sistema de poda, enquanto o melhor perfil aromático foi observado para 'Syrah', cultivada sob poda mecânica e estresse hídrico baixo a moderado.The production of a quality wine depends on the cultivation of high quality grapes with favorable chemical and physical characteristics, which result from the submitted micrometeorological regime, variety and soil-plant management. Grape yield and quality performance can be responsive to varying levels of water depth and the pruning system. The interaction between these two factors and the surrounding environment has been proved to be influential upon the wine quality. An experiment was carried out with wine grapes through the growing seasons of 2003 to 2005. The objective was to determine the best combination of pruning (manual and mechanized and irrigation practices on 'Syrah' (Vitis vinifera L. envisaging wine quality achievement, under a hot and dry microclimate. Results showed that the grape composition was consistently affected by the pruning system whereas the best aroma

  17. Análise quantitativa da desmineralização do esmalte dental submetido à ação de dentifrícios fluoretados

    OpenAIRE

    Campos, Elisângela de Jesus; Araújo,Roberto Paulo Correia de; Araújo, Maria Thereza Barral; Araújo,Danilo Barral de; Lima, Max José Pimenta; Oliveira, Nayara de Sá; Dias, Alexnaldo Correia

    2005-01-01

    p. 226-235 A cárie dental é uma doença multifatorial, caracterizada como um processo dinâmico, originado pelo desequilíbrio nos fenômenos de desmineralização e remineralização. Diversos estudos têm demonstrado que a prevalência de cárie, na maioria dos países, diminuiu nas últimas duas décadas, devido à implementação de programas de saúde pública, ao uso do flúor através da fluoretação das águas de abastecimento e o uso de dentifrícios fluoretados. O flúor contido nos dentifríc...

  18. Variabilidade espacial das características qualitativas e quantitativas da pastagem de capim Marandú em topossequência no Tocantins

    Directory of Open Access Journals (Sweden)

    A. C. Santos

    2015-12-01

    Full Text Available Objetivou-se com este trabalho avaliar a variabilidade espacial das características produtivas do pasto de Brachiária brizantha cv. Marandu em vertente, em função da fertilidade do solo. O experimento foi conduzido na fazenda da Escola de Medicina Veterinária e Zootecnia da Universidade Federal do Tocantins. Para a caracterização química do solo foi realizada amostragem em pontos da área onde a malha regular possui uma distância média de 2 m de um ponto a outro longitudinalmente e doze metros perpendicularmente, perfazendo um total de 160 pontos. A forrageira também foi coletada seguindo malha regular, porém, com distância média de quatro metros longitudinalmente e 12 m perpendicularmente, totalizando 80 amostras. As análises realizadas foram às seguintes: pH (CaCl2; (matéria orgânica, fósforo disponível (P e potássio trocável (K+; cálcio trocável (Ca2+, magnésio trocável (Mg2+ e alumínio trocável (Al3+. Os dados de forragem e de solo foram submetidos à estatística descritiva e a hipótese de normalidade dos dados que foi verificada pelo teste de Kolmogorov-Smirnov. A produção do pasto apresentou baixa dependência espacial, ajustando-se ao modelo esférico para todas as variáveis analisadas. Os coeficientes de variação foram moderados. A produção de forragem foi maior nas posições de topo e pedimento, em contrapartida os atributos químicos do solo, com exceção do pH, foram mais baixos nestas posições. Os atributos químicos do solo, também apresentaram baixa dependência espacial e, com exceção do pH, todos os atributos ajustaram-se ao modelo esférico.Spatial variability of qualitative and quantitative characteristics of Marandú grass pasture in topossequence in TocantinsAbstract: The objective of this study was evaluating the spatial variability of the productive characteristics of pasture of Brachiaria brizantha cv. Marandu in front, depending on soil fertility. The experiment was conducted at

  19. Quantitative evaluation of acidity tolerance of root nodule bacteria Avaliação quantitativa da tolerância de rizóbios à acidez

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    Luiz Antonio de Oliveira

    1999-07-01

    Full Text Available Quantification of acidity tolerance in the laboratory may be the first step in rhizobial strain selection for the Amazon region. The present method evaluated rhizobia in Petri dishes with YMA medium at pH 6.5 (control and 4.5, using scores of 1.0 (sensitive, "no visible" growth to 4.0 (tolerant, maximum growth. Growth evaluations were done at 6, 9, 12, 15 and 18 day periods. This method permits preliminary selection of root nodule bacteria from Amazonian soils with statistical precision. Among the 31 rhizobia strains initially tested, the INPA strains 048, 078, and 671 presented scores of 4.0 at both pHs after 9 days of growth. Strain analyses using a less rigorous criterion (growth scores higher than 3.0 included in this highly tolerant group the INPA strains 511, 565, 576, 632, 649, and 658, which grew on the most diluted zone (zone 4 after 9 days. Tolerant strains still must be tested for nitrogen fixation effectiveness, competitiveness for nodule sites, and soil persistence before their recommendation as inoculants.A quantificação da tolerância à acidez em testes de laboratório pode ser o primeiro passo na seleção de estirpes de rizóbios para a Amazônia. O presente método avaliou isolamentos de rizóbios em placas de Petri contendo meio YMA com pHs 6,5 (controle e 4,5, usando notas de 1,0 (sensíveis, sem crescimento visual, até 4,0 (tolerantes, máximo crescimento. As avaliações foram realizadas aos 6, 9, 12, 15 e 18 dias de crescimento. O método permite selecionar preliminarmente, rizóbios isolados de solos da Amazônia, com precisão estatística. Entre as 31 estirpes inicialmente testadas, as estirpes INPA 048, 078 e 671 apresentaram notas iguais a 4,0 em ambos os pHs testados após os 9 dias de crescimento. Ao se analisar as estirpes usando um sistema menos rigoroso (nota de crescimento acima de 3,0, foi possível incluir também neste grupo, as estirpes INPA 511, 565, 576, 632, 649 e 658, que cresceram na zona mais diluída

  20. Características quantitativas da carcaça de cordeiros de diferentes genótipos submetidos a duas dietas Quantitative traits of carcass from lambs of different genotypes submitted to two diets

    Directory of Open Access Journals (Sweden)

    Felipe Queiroga Cartaxo

    2011-10-01

    Full Text Available Objetivou-se avaliar as características quantitativas de carcaça de cordeiros de diferentes genótipos recebendo diversos níveis de energia na dieta. Foram utilizados 54 cordeiros não-castrados, sendo 18 Santa Inês (SI, 18 F1Dorper × Santa Inês (Dp × SI e 18 F1 Santa Inês × Sem Raça Definida (SI × SRD, com idade média de 150 dias e peso médio de 22,60 kg no início do experimento. As dietas continham 17% de proteína bruta e 2,40 Mcal/kg MS ou 2,90 Mcal/kg MS. Os cordeiros alimentados com a dieta contendo 2,90 Mcal/kg MS apresentaram maior peso corporal e de carcaça, área de olho-de-lombo, percentual de gordura interna, índice de musculosidade, índice de compacidade da carcaça, percentual de gordura e menor relação músculo:gordura. As carcaças desses cordeiros obtiveram também menores perdas por resfriamento e maior peso e percentual de paleta e perna. As carcaças dos cordeiros Dorper × Santa Inês foram superiores em espessura de gordura subcutânea, medida GR (gordura sobre a 12ª costela a 11 cm de distância da linha média lombo, percentual de gordura, relação músculo:osso, índice de musculosidade, peso e percentual de lombo. Os cordeiros Santa Inês apresentaram maior relação músculo:gordura. A dieta e o genótipo influenciam as características quantitativas de carcaça de cordeiros.The objective of this study was to evaluate the quantitative carcass traits of lambs from different genotypes receiving different levels of energy in the diet. Fifty-four non-castrated lambs - 18 Santa Inês (SI, 18 F1Dorper × Santa Inês (Dp × SI and 18 F1Santa Inês × No defined breed (SI × NDB - of average 150 days old and 22.6-kg BW were used in the beginning of the experiment. Diets contained 17% crude protein and 2.40 Mcal/kg DM or 2.90 Mcal/kg DM. Lambs fed the diet containing 2.90 Mcal/kg DM showed higher final live and carcass weight, loin eye area, percentage of internal fat, muscularity index, compactness of the

  1. CONSTRUCTION OF ACTIVE RECOMBINANT CASPASE-3 EUKARYOTIC EXPRESSION PLA SMID AND EFFECT OF r-CASPASE-3 ON APOPTOSIS OF PANCREATIC CARCINOMA CELLS

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Objective To construct active recombinant cas pa ses-3 gene(r-caspases-3)eukaryotic expression plasmid and observe the apoptos is inducing activity of r-caspase-3 in pancreatic carcinoma cells. Methods pcDNA3.1(+)/r-caspase-3 was constructed and pan creatic carcinoma cells(PC-Ⅱ)were transfected with the pcDNA3.1(+)/r-caspases -3 by liposomes(LipofectAMINE).The expression of r-Caspase-3 mRNA in pancreat ic carcinoma cells was detected by reverse transcription process of the polymera se chain reaction(RT-PCR), and the signs of apoptosis were examined in pancreat ic carcinoma cells by the methods of the DNA electrophoresis and flow cytometry analysis(FACS).Results The sequence inserted in pBlueSKM/r-Caspase-3 p lasmid was coincident with that of the r-caspases-3. The evaluation result of pcDNA3.1(+)/r-caspases-3 through enzyme cutting was correct. A 894bp strap was observed by RT-PCR after pancreatic carcinoma cells being transfected with the pcDNA3.1(+)/r-caspases-3 by liposomes. No strap was found in control groups. A characteristic DNA ladder was observed in pancreatic carcinoma cells DNA elect r ophoresis, and transparent hypodiploid karyotype peak was found by FACS. Conclusion The plasmid of pcDNA3.1(+)/r-Caspase-3 was c onstructed successfully, the expression of r-Caspase-3 mRMA in pancreatic carc inoma cells was confirmed by RT-PCR, and pcDNA3.1(+)/r-Caspase-3 can induce a poptosis in pancreatic carcinoma cells.

  2. Quantitative analyses of maxillary sinus using computed tomography Análise quantitativa dos seios maxilares por meio da tomografia computadorizada

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    Andréia Perella

    2003-09-01

    Full Text Available The aim of this study was to evaluate the precision and accuracy of linear measurements of maxillary sinus made in tomographic films, by comparing with 3D reconstructed images. Linear measurements of both maxillary sinus in computed tomography CT of 17 patients, with or without lesion by two calibrated examiners independently, on two occasions, with a single manual caliper. A third examiner has done the same measurements electronically in 3D-CT reconstruction. The statistical analysis was performed using ANOVA (analyses of variance. Intra-observer percentage error was little in both cases, with and without lesion; it ranged from 1.14% to 1.82%. The inter-observer error was a little higher reaching a 2.08% value. The accuracy presented a higher value. The perceptual accuracy error was higher in samples, which had lesion compared to that which had not. CT had provided adequate precision and accuracy for maxillary sinus analyses. The precision in cases with lesion was considered inferior when compared to that without lesion, but it can't affect the method efficacy.O objetivo deste trabalho foi avaliar a precisão e acurácia de medidas lineares da região de seio maxilar realizadas em filmes tomográficos, por meio de comparação com a reconstrução da imagem em 3D. Medidas lineares de ambos os seios maxilares foram realizadas em tomografias computadorizadas (2D-TC de 17 pacientes, normais e com lesão, por dois examinadores, calibrados, separadamente, duas vezes cada um, utilizando um compasso. Um terceiro observador realizou as mesmas medidas eletronicamente na reconstrução 3D-TC. ANOVA (análise de variância foi utilizado para análise estatística. A porcentagem de erro intra-examinadores foi pequena em ambos os casos, com ou sem lesão, variando de 1,14% a 1,82%. O erro inter-examinadores foi maior alcançando o valor de 2,08% .O erro percentual de acurácia nas amostras com lesão foram maiores comparando-as às que não tinham lesão. A

  3. Tau and Caspase 3 as Targets for Neuroprotection

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    Anat Idan-Feldman

    2012-01-01

    Full Text Available The peptide drug candidate NAP (davunetide has demonstrated protective effects in various in vivo and in vitro models of neurodegeneration. NAP was shown to reduce tau hyperphosphorylation as well as to prevent caspase-3 activation and cytochrome-3 release from mitochondria, both characteristic of apoptotic cell death. Recent studies suggest that caspases may play a role in tau pathology. The purpose of this study was to evaluate the effect of NAP on tau hyperphosphorylation and caspase activity in the same biological system. Our experimental setup used primary neuronal cultures subjected to oxygen-glucose deprivation (OGD, with and without NAP or caspase inhibitor. Cell viability was assessed by measuring mitochondrial activity (MTS assay, and immunoblots were used for analyzing protein level. It was shown that apoptosis was responsible for all cell death occurring following ischemia, and NAP treatment showed a concentration-dependent protection from cell death. Ischemia caused an increase in the levels of active caspase-3 and hyperphosphorylated tau, both of which were prevented by either NAP or caspase-inhibitor treatment. Our data suggest that, in this model system, caspase activation may be an upstream event to tau hyperphosphorylation, although additional studies will be required to fully elucidate the cascade of events.

  4. BDNF pro-peptide regulates dendritic spines via caspase-3.

    Science.gov (United States)

    Guo, J; Ji, Y; Ding, Y; Jiang, W; Sun, Y; Lu, B; Nagappan, G

    2016-06-16

    The precursor of brain-derived neurotrophic factor (BDNF) (proBDNF) is enzymatically cleaved, by either intracellular (furin/PC1) or extracellular proteases (tPA/plasmin/MMP), to generate mature BDNF (mBDNF) and its pro-peptide (BDNF pro-peptide). Little is known about the function of BDNF pro-peptide. We have developed an antibody that specifically detects cleaved BDNF pro-peptide, but not proBDNF or mBDNF. Neuronal depolarization elicited a marked increase in extracellular BDNF pro-peptide, suggesting activity-dependent regulation of its extracellular levels. Exposure of BDNF pro-peptide to mature hippocampal neurons in culture dramatically reduced dendritic spine density. This effect was mediated by caspase-3, as revealed by studies with pharmacological inhibitors and genetic knockdown. BDNF pro-peptide also increased the number of 'elongated' mitochondria and cytosolic cytochrome c, suggesting the involvement of mitochondrial-caspase-3 pathway. These results, along with BDNF pro-peptide effects recently reported on growth cones and long-term depression (LTD), suggest that BDNF pro-peptide is a negative regulator of neuronal structure and function.

  5. Adubação da cana-de-açúcar. II - Adubação fosfatada quantitativa Quantitative phosphate fertilizing for sugar cane

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    R. Alvarez

    1957-01-01

    Full Text Available Neste trabalho são apresentados os resultados obtidos em um ensaio de adubação de cana-de-açúcar, na Usina Tamôio, município de Araraquara, em 1952. Estudaram-se cinco níveis de fósforo, em presença de nitrogênio e potássio, utilizando-se um delineamento em blocos ao acaso com cinco tratamentos e seis repetições. Os níveis dos fertilizantes empregados, em kg/ha, foram 0, 60, 120, 180 e 240 de P2O5, 30 de nitrogênio e 45 de K(20, sob as formas, respectivamente, de superfosfáto simples, salitre do Chile e cloreto de potássio. O ensaio foi conduzido sob as condições normais de cultivo para a cultura da cana. Adaptou-se às médias de produção uma curva exponencial de Mitscherlich. Esta curva, de expressão y = 66,23 [1 - 10 - 1,032(x + 0,3978] explica com grande precisão a relação entre os níveis de P2O5 aplicados e as produções obtidas. Conhecida a curva em questão, chegou-se à conclusão de que o nível mais aconselhável é o de 120 kg de P2O5 por hectare.This paper presents the results obtained in 1952 in a fertilizing experiment on sugar cane, at the Usina Tamôio, Araraquara. Five levels of phosphorus (in the presence of nitrogen and potash using a randomized blocks design with six replicates were studied. The formula used contained always 45 kg/ha of K(20 and 30 kg/ha of N, the amount of phosphorus being variable as follows: 0, 60, 120, 180 and 240 kg/ha. Phosphorus, nitrogen and potash were applied under the form of simple superphosphate, Chilean nitrate and potash muriate, respectively. An exponential Mitscherlich curve of the form y = 66,23 [1 - 10 - 1,032(x + 0,3978] was the best fit for the results. It is concluded that for the conditions of the experiment 120 kg/ha of P2O5- is the best dose to be recommended.

  6. Dual Vulnerability of Tau to Calpains and Caspase-3 Proteolysis Under Neurotoxic and Neurodegenerative Conditions

    Directory of Open Access Journals (Sweden)

    Ming Cheng Liu

    2010-11-01

    Full Text Available Axonally specific microtubule-associated protein tau is an important component of neurofibrillary tangles found in AD (Alzheimer's disease and other tauopathy diseases such as CTE (chronic traumatic encephalopathy. Such tau aggregate is found to be hyperphosphorylated and often proteolytically fragmented. Similarly, tau is degraded following TBI (traumatic brain injury. In the present study, we examined the dual vulnerability of tau to calpain and caspase-3 under neurotoxic and neurodegenerative conditions. We first identified three novel calpain cleavage sites in rat tau (four-repeat isoform as Ser130 ↓ Lys131, Gly157 ↓ Ala158 and Arg380 ↓ Glu381. Fragment-specific antibodies to target the major calpain-mediated TauBDP-35K (35 kDa tau-breakdown product and the caspase-mediated TauBDP-45K respectively were developed. In rat cerebrocortical cultures treated with excitotoxin [NMDA (N-methyl-D-aspartate], tau is significantly degraded into multiple fragments, including a dominant signal of calpain-mediated TauBDP-35K with minimal caspase-mediated TauBDP-45K. Following apoptosis-inducing EDTA treatment, tau was truncated only to TauBDP-48K/45K-exclusively by caspase. Cultures treated with another apoptosis inducer STS (staurosporine, dual fragmentation by calpain (TauBDP-35K and caspase-3 (TauBDP-45K was observed. Tau was also fragmented in injured rat cortex following TBI in vivo to BDPs of 45-42 kDa (minor, 35 kDa and 15 kDa, followed by TauBDP-25K. Calpain-mediated TauBDP-35K-specific antibody confirmed robust signals in the injured cortex, while caspase-mediated TauBDP-45K-specific antibody only detected faint signals. Furthermore, intravenous administration of a calpain-specific inhibitor SNJ-1945 strongly suppressed the TauBDP-35K formation. Taken together, these results suggest that tau protein is dually vulnerable to calpain and caspase-3 proteolysis under different neurotoxic and injury conditions.

  7. Testicular dysgenesis syndrome and the estrogen hypothesis: a quantitative meta-analysis A síndrome da disgenesia testicular e a hipótese do estrogênio: uma meta-análise quantitativa

    Directory of Open Access Journals (Sweden)

    Olwenn Martin

    2008-10-01

    ético dietilstilbestrol (DES e SDT continuam inconclusivas. Foi feita uma meta-análise quantitativa da relação entre SDT e exposição pré-natal a agentes estrogênicos. A inclusão na análise baseou-se em critérios mecanísticos e foi explorada a plausibilidade de um modo de ação mediada pelo receptor estrogênico-α (REα. Incluíram-se oito estudos sobre a etiologia das hipospádias e/ou criptorquidismo não identificados em revisões sistemáticas anteriores. Mais quatro estudos sobre estrogênios sintéticos resultaram em uma estimativa estatisticamente significativa para câncer de testículo. Os resultados das análises dos subconjuntos apontam à existência de fontes não identificadas de heterogeneidade entre estudos ou populações estudadas.

  8. Dieldrin induces apoptosis by promoting caspase-3-dependent proteolytic cleavage of protein kinase Cdelta in dopaminergic cells: relevance to oxidative stress and dopaminergic degeneration.

    Science.gov (United States)

    Kitazawa, M; Anantharam, V; Kanthasamy, A G

    2003-01-01

    We previously reported that dieldrin, one of the potential environmental risk factors for development of Parkinson's disease, induces apoptosis in dopaminergic cells by generating oxidative stress. Here, we demonstrate that the caspase-3-dependent proteolytic activation of protein kinase Cdelta (PKCdelta) mediates as well as regulates the dieldrin-induced apoptotic cascade in dopaminergic cells. Exposure of PC12 cells to dieldrin (100-300 microM) results in the rapid release of cytochrome C, followed by the activation of caspase-9 and caspase-3 in a time- and dose-dependent manner. The superoxide dismutase mimetic Mn(III)tetrakis(4-benzoic acid)porphyrin chloride significantly attenuates dieldrin-induced cytochrome C release, indicating that reactive oxygen species may contribute to the activation of pro-apoptotic factors. Interestingly, dieldrin proteolytically cleaves native PKCdelta into a 41 kDa catalytic subunit and a 38 kDa regulatory subunit to activate the kinase. The dieldrin-induced proteolytic cleavage of PKCdelta and induction of kinase activity are completely inhibited by pretreatment with 50-100 microM concentrations of the caspase inhibitors benzyloxycarbonyl-Val-Ala-Asp-fluoromethylketone (Z-VAD-FMK) and benzyloxycarbonyl-Asp-Glu-Val-Asp-fluoromethylketone (Z-DEVD-FMK), indicating that the proteolytic activation of PKCdelta is caspase-3-dependent. Additionally, Z-VAD-FMK, Z-DEVD-FMK or the PKCdelta specific inhibitor rottlerin almost completely block dieldrin-induced DNA fragmentation. Because dieldrin dramatically increases (40-80-fold) caspase-3 activity, we examined whether proteolytically activated PKCdelta amplifies caspase-3 via positive feedback activation. The PKCdelta inhibitor rottlerin (3-20 microM) dose-dependently attenuates dieldrin-induced caspase-3 activity, suggesting positive feedback activation of caspase-3 by PKCdelta. Indeed, delivery of catalytically active recombinant PKCdelta via a protein delivery system significantly

  9. Role of Caspase 3 in neuronal apoptosis after acute brain injury

    Institute of Scientific and Technical Information of China (English)

    杨新宇; 杨树源; 张建宁; 雪亮; 胡震

    2002-01-01

    To analyze the role of Caspase 3 in neuronal apoptosis after acute brain injury. Methods: Experiments were carried out with rat diffuse brain trauma model. The neuronal DNA injury in cortex and hippocampus was observed by TUNEL stain.The mRNA and protein expressions and enzyme activation of Caspase 3 were observed by Northern blot, in situ hybridization, immunohistochemistry stain and Western blot, respectively. Special Caspase 3 enzyme inhibitor was used to observe the therapeutic effect. Results: TUNEL positive neurons appeared 2 hours after severe trauma, peaked at 1 day and lasted for 7 days.Northern blot showed that the Caspase 3 mRNA expression was increased and peaked at 1 day, about twice higher than the control. In the area of cortex and hippocampus,positive mRNA staining neurons appeared most distinct on one day. With the antibody for Caspase 3 P20 subunit, the active Caspase 3 expression peaked at 1-3 days. The electrophoresis band of PARP degradation would be seen by Western blot. Caspase 3 enzyme inhibitor could reduce apoptotic neuronal death without any effect on Caspase 3 P20 subunit expression. Conclusions: After brain trauma, Caspase 3 mRNA and protein expressions and enzyme activation are enhanced in combination with neuronal apoptosis. Special Caspase 3 enzyme inhibitor can apparently decrease the neuronal apoptosis.

  10. Mechanism of mitochondrial respiratory control in caspase-3 induced positive feed back loop in apoptosis

    Institute of Scientific and Technical Information of China (English)

    2002-01-01

    Caspase-3 plays a central role in the execution of apoptosis. Besides many substrates of caspase-3, mitochondria seem to be one of the candidate targets in the apoptotic process. We evaluated the effects of caspase-3 on the isolated mitochondria in detail, and especially focused on the mechanism involved in mitochondrial functions, which were not fully assessed till now. Our results showed that recombinant caspase-3 induced the increase of superoxide production, the dissipation of mitochondrial membrane potential and rate increasing of mitochondrial state 4 respiration. Caspases inhibitor, z-VAD-fmk can inhibit these effects of caspase-3 on mitochondria. Bcl-xL and cyclosporin A were also shown to be able to inhibit these changes. These results suggested a possible mechanism in caspase-3 induced disruption of mitochondrial membrane barrier which formed a positive feedback loop in apoptosis.

  11. Low levels of Caspase-3 predict favourable response to 5FU-based chemotherapy in advanced colorectal cancer: Caspase-3 inhibition as a therapeutic approach.

    Science.gov (United States)

    Flanagan, L; Meyer, M; Fay, J; Curry, S; Bacon, O; Duessmann, H; John, K; Boland, K C; McNamara, D A; Kay, E W; Bantel, H; Schulze-Bergkamen, H; Prehn, J H M

    2016-02-04

    Colorectal cancer (CRC) is one of the most common cancers in the Western world. 5-Fluorouracil (5FU)-based chemotherapy (CT) remains the mainstay treatment of CRC in the advanced setting, and activates executioner caspases in target cells. Executioner caspases are key proteins involved in cell disassembly during apoptosis. Activation of executioner caspases also has a role in tissue regeneration and repopulation by stimulating signal transduction and cell proliferation in neighbouring, non-apoptotic cells as reported recently. Tissue microarrays (TMAs) consisting of tumour tissue from 93 stage II and III colon cancer patients were analysed by immunohistochemistry. Surprisingly, patients with low levels of active Caspase-3 had an increased disease-free survival time. This was particularly pronounced in patients who received 5FU-based adjuvant CT. In line with this observation, lower serum levels of active Caspase-3 were found in patients with metastasised CRC who revealed stable disease or tumour regression compared with those with disease progression. The role of Caspase-3 in treatment responses was explored further in primary human tumour explant cultures from fresh patient tumour tissue. Exposure of explant cultures to 5FU-based CT increased the percentage of cells positive for active Caspase-3 and Terminal Deoxynucleotidyl Transferase dUTP Nick end Labelling (TUNEL), but also the expression of regeneration and proliferation markers β-Catenin and Ki-67, as well as cyclooxygenase-2 (COX-2). Of note, selective inhibition of Caspase-3 with Ac-DNLD-CHO, a selective, reversible inhibitor of Caspase-3, significantly reduced the expression of proliferation markers as well as COX-2. Inhibition of COX-2 with aspirin or celecoxib did not affect Caspase-3 levels but also reduced Ki-67 and β-Catenin levels, suggesting that Caspase-3 acted via COX-2 to stimulate cell proliferation and tissue regeneration. This indicates that low levels of active Caspase-3 may represent a

  12. Caspase-3和bax在视网膜母细胞瘤中的表达%Expression of caspase-3 and bax gene protein in retinoblastoma

    Institute of Scientific and Technical Information of China (English)

    孙红; 惠延年; 王立勤; 马吉献

    2003-01-01

    目的: 观察凋亡及凋亡调控基因caspase-3/bax在视网膜母细胞瘤(retinoblastoma, RB)中的表达及与凋亡的相关性. 方法: 收集35例RB标本,对其分别进行caspase-3和bax免疫组织化学染色,观察表达情况及染色强度. 结果: Caspase-3及bax在未分化型(n=15)分别有较好的表达(11/12例),caspase-3及bax在分化型(n=20)中也有较好的表达(17/18例). 正常视网膜组织中无caspase-3及bax的表达. 结论: 凋亡在RB中是存在的,caspase-3及bax在RB的发生发展中起重要作用.

  13. Avaliação quantitativa da contaminação por micro-organismos no ambiente de criação do bicho-da-seda (Bombyx mori L.) na fase de chocadeira

    OpenAIRE

    Margatho,L.F.F.; Okamoto, F; Almeida, A. M.; Santos,C.A.J.P.

    2012-01-01

    O objetivo do presente trabalho foi avaliar a contaminação bacteriana e fúngica em chocadeira antes e após procedimento de limpeza e desinfecção. Foram coletadas amostras do ar e de superfícies do piso, da parede, das folhas de amoreira, das lagartas do bicho-da-seda e da máquina picadeira de folhas. A metodologia empregada seguiu a técnica da placa de sedimentação e contato de superfície com meios de cultura específicos, Ágar Nutriente, Ágar McConkey e Ágar Sabouraud Dextrose para cresciment...

  14. Análise quantitativa do crescimento populacional da cidade de Seropédica-RJ sob a influência dos estudantes da Universidade Federal Rural do Rio de Janeiro

    Directory of Open Access Journals (Sweden)

    Welber Faustino da Silva

    2016-10-01

    Full Text Available Neste trabalho pretendeu-se analisar o quanto os estudantes da Universidade Federal Rural do Rio de Janeiro (UFRRJ, que são moradores da cidade de Seropédica, influenciam quantitativamente e economicamente no crescimento da população local, uma vez que a UFRRJ tem sua sede localizada nas proximidades da cidade de Seropédica. Para isso, foi aplicado um modelo de crescimento definido por um sistema de equações diferenciais, devido a Lotka-Volterra, que envolve a iteração entre duas espécies, neste caso a população da cidade e apopulação de estudantes, sabendo que o número de moradias desocupadas é dependente da procura pelos estudantes para aluguéis e estas moradias ocupadas influenciam também a oferta e a construção de novos empreendimentos na cidade.

  15. Serum-stabilized naked caspase-3 siRNA protects autotransplant kidneys in a porcine model.

    Science.gov (United States)

    Yang, Cheng; Zhao, Tian; Zhao, Zitong; Jia, Yichen; Li, Long; Zhang, Yufang; Song, Mangen; Rong, Ruiming; Xu, Ming; Nicholson, Michael L; Zhu, Tongyu; Yang, Bin

    2014-10-01

    The naked small interfering RNA (siRNA) of caspase-3, a key player in ischemia reperfusion injury, was effective in cold preserved and hemoreperfused kidneys, but not autotransplanted kidneys in our porcine models. Here, chemically modified serum stabilized caspase-3 siRNAs were further evaluated. The left kidney was retrieved and infused by University of Wisconsin solution with/without 0.3 mg caspase-3 or negative siRNA into the renal artery for 24-hour cold storage (CS). After an intravenous injection of 0.9 mg siRNA and right-uninephrectomy, the left kidney was autotransplanted for 2 weeks. The effectiveness of caspase-3 siRNA was confirmed by caspase-3 knockdown in the post-CS and/or post-transplant kidneys with reduced apoptosis and inflammation, while the functional caspase-3 siRNA in vivo was proved by detected caspase-3 mRNA degradation intermediates. HMGB1 protein was also decreased in the post-transplanted kidneys; correlated positively with renal IL-1β mRNA, but negatively with serum IL-10 or IL-4. The minimal off-target effects of caspase-3 siRNA were seen with favorable systemic responses. More importantly, renal function, associated with active caspase-3, HMGB1, apoptosis, inflammation, and tubulointerstitial damage, was improved by caspase-3 siRNA. Taken together, the 2-week autotransplanted kidneys were protected when caspase-3 siRNA administrated locally and systemically, which provides important evidence for future clinical trials.

  16. Expression and effect of Caspase-3 in neurons after tractive spinal cord injury in rats

    Institute of Scientific and Technical Information of China (English)

    LIU Lei; PEI Fu-xing; TANG Kang-lai; XU Jian-zhong; LI Qi-hong

    2005-01-01

    Objective: To investigate Caspase-3 expression and its role in neuronal apoptosis.Methods: The T13-L2 spinal cord of rats was injured by traction after the amplitude of P1-N1 wave, monitored by a cortical somatosensory evoked potential (CSEP) monitor, decreased to seventy percent of that before operation. Then rats were killed in 6 h, 1 d, 4 d, 7 d, 14 d and 21 d respectively after operation. Flow cytometer terminal deoxynucleotldyl transferease-mediated biotinylated deoxynuridine triphosphate nick end labeling (TUNEL), Caspase-3 activity assay and immunohistochemical method were applied to investigate Caspase-3 expression in the spinal cord tissue and to study neuronal apoptosis in rats. Results: After spinal cord injury, apoptotic cells detected by flow cytometry and TUNEL-positive cells were significantly more, and positive immunohistochemical staining of Caspase-3 and Caspase-3 activity were significantly higher in Group injury than in Groups control and laminectomy, respectively (P>0.05, P>0.01). Similar trend of changes was noticed in apoptotic cells, TUNEL-positive cells and positive immunohistochemical staining of Caspase-3, all of which reached their respective peak 7 days after operation. Caspase-3 activity reached its peak, however, 4 days postoperatively. Conclusions: Increased expression and activity of Caspase-3 protein in neurons after tractive spinal cord injury is the biochemical signal of early spinal cell apoptosis. It is of great significance for understanding the mechanism of spinal cord injury.

  17. Design and Synthesis of a Novel Peptidomimetic Inhibitor of Caspase-3

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Caspases, a family of cysteine proteases, comprise of highly homologous enzymes that play an important role in apoptotic cell death. Caspase-3 shows key functions in apoptosis, mediating apoptotic cascade from the intrinsic and extrinsic activation pathways. Therefore, caspase-3 is an attractive target for therapeutic intervention. For instance,inhibitors of caspase-3 have been described as promising cardioprotectants, neuroprotectants and antiarthritic agents.A novel peptidomimetic inhibitor of caspase-3, has been designed, which still has the properties of a reversible inhibitor, while the P1 site at the C-terminal remains, and only L-amino acid has been replaced by D-amino acid. Also presented here is the synthesis of the inhibitor and its inhibitory activity against caspase-3, which was tested by the fluorescent activity assay.

  18. CLINICOPATHOLOGICAL SIGNIFICANCE OF PTEN AND CASPASE-3 EXPRESSIONS IN BREAST CANCER

    Institute of Scientific and Technical Information of China (English)

    Xue-fei Yang; Yan Xin; Li-li Mao

    2008-01-01

    Objective To investigate the expressions of PTEN and Caspase-3 proteins in human breast carcinoma, and to evaluate their clinicopathological implications during the tumorigenesis and progression of breast cancer.Methods The expressions of PTEN and Caspase-3 proteins in 95 cases of breast cancer and 15 cases of benignbreast diseases were investigated immunohistochemically. Correlations between the expression of PTEN protein,Caspase-3 protein, and clinicopathological features of breast cancers were analyzed.Results The loss expression rate of PTEN protein in tumor tissues was significantly higher than that in benignbreast diseases (33.7% vs. 0, P 0. 05). In addition,the expression of PTEN protein had significantly positive correlation with the expression of Caspase-3 protein in breast cancer (P <0.01 ).Conclusion The combination detection of PTEN and Caspase-3 may serve as an important index to estimate the pathobiological behavior and pognosis of breast cancer.

  19. Testosterone undecanoate and depo medroxyprogesterone acetate induced azoospermia through increased expression of spermatogenic cell caspase 3

    Directory of Open Access Journals (Sweden)

    Nukman Moeloek

    2008-09-01

    Full Text Available The administration of a combination of testosterone undecanoate (TU, a long-acting androgen and depo-medroxyprogesterone acetate (DMPA were investigated in term of suppression of rat sperm concentration in vivo to azoospermia through increasing activity of spermatogenic cell caspase 3. Adult Sprague Dawley rats received TU and DMPA of 2.5 mg and 1.25 mg, respectively, a regimen known to rapidly reduce intra testicular testosterone and to produce azoospermia within 12 weeks. Caspase 3 positive sperm cells increased compared with control levels during 6 weeks post-injection and increased further through 60 weeks. Immunohistochemistry for caspase 3 revealed that spermatocytes represented the predominant caspase 3 positive germ cells. Modest immunoreactivity for caspase-3 was localized to nuclear region of the germ cells of control and treated testes. Immunohistochemistry study revealed significantly increased caspase-3 expression in nuclei of germ cells during administration of TU+DMPA to rats. Additionally, the caspase 3 content was significantly increased in germ cells during rats were administered TU+DMPA (453.90±84.88 cells/200 seminiferous tubules and caspase 3 significant increase in immunoreactivity was localized to the nuclei of spermatogonia, spermatocytes, and spermatids. Taken together, these results indicated that azoospermia due to reduced intratesticular testosterone concentration was caspase-3 activation dependent and suggested that the increase in active caspase-3 in the nucleus may be involved in the induction of decreased sperm production. (Med J Indones 2008; 17: 149-56Keywords: TU, DMPA, sperm concentration, germ cells

  20. Análise quantitativa do tratamento da escoliose idiopática com o método klapp por meio da biofotogrametria computadorizada Quantitative photogrammetric analysis of the klapp method for treating idiopathic scoliosis

    Directory of Open Access Journals (Sweden)

    Denise H. Iunes

    2010-04-01

    Full Text Available INTRODUÇÃO: Poucos trabalhos comprovam a eficácia das técnicas fisioterapêuticas para o tratamento da escoliose. OBJETIVO: Analisar a eficácia do Método Klapp no tratamento das escolioses por meio do estudo quantitativo pela biofotogrametria computadorizada. MÉTODOS: Dezesseis indivíduos com média de idade de 15±2,61 anos, portadores de escoliose idiopática, foram tratados com o método Klapp. Para análise dos resultados do tratamento, todos foram fotografados antes e após o tratamento, seguindo uma padronização fotográfica. Todas as fotografias foram analisadas quantitativamente por um mesmo experimentador, utilizando o software ALCimagem 2000. A análise estatística foi realizada, utilizando-se a o teste-t pareado com nível de significância de 5%. RESULTADOS: Os resultados apontam para a melhora após o tratamento dos ângulos agromioclaviculares (AC-p=0,00 e esternoclavicular (EC-p=0,01, que avaliam a simetria dos ombros, e para o ângulo que avalia o triângulo de Tales esquerdo, (ΔTe-p=0,02. Em termos de flexibilidade, houve melhora dos ângulos tibiotársicos (ATT-p=0,01 e coxofemoral (CF-p=0,00. Não houve modificações das curvaturas vertebrais e nem melhora no posicionamento da cabeça, apenas na curvatura lombar, avaliada pelo ângulo lordose lombar (LL-p=0,00, sofreu modificação com o tratamento. CONCLUSÃO: O método Klapp foi uma técnica terapêutica eficaz para tratar as assimetrias de tronco e a flexibilidade. Não foi eficaz para assimetrias da pelve, modificações da posição da cabeça, da lordose cervical e cifose torácica.INTRODUCTION: Few studies have proved that physical therapy techniques are efficient in the treatment of scoliosis. OBJECTIVE: To analyze the efficiency of the Klapp method for the treatment of scoliosis, through a quantitative analysis using computerized biophotogrammetry. METHODS: Sixteen participants of a mean age of 15±2.61 yrs. with idiopathic scoliosis were treated using

  1. Correlação entre digestibilidade in vitro da parede celular e três métodos analíticos para a avaliação quantitativa da lignina

    Directory of Open Access Journals (Sweden)

    Fukushima Romualdo Shigueo

    2001-01-01

    Full Text Available Três métodos analíticos para a estimativa do conteúdo de lignina na parede celular, em diferentes frações vegetais, de sete espécies forrageiras, foram comparados. Os métodos empregados foram: lignina em detergente ácido (LDA, lignina permanganato de potássio e lignina solúvel em brometo de acetila (LSBA. Para o método espectrofotométrico (LSBA, foi utilizado como padrão de referência a lignina extraída da planta através do próprio reagente brometo de acetila. Não houve concordância entre os três métodos, sendo que, no geral, o método da LSBA mostrou valores mais elevados que os outros dois métodos e o método da LDA, menores valores. Na maioria das amostras, a concentração de lignina foi superior para a parede celular oriunda de planta madura em relação àquela proveniente de planta jovem, assim como o teor de lignina foi maior para a fração colmo em relação à fração folha. As leguminosas tenderam a apresentar teores mais elevados de lignina pelos métodos gravimétricos que as gramíneas, independentemente do estádio de maturidade. Entretanto, essa tendência não foi observada para o método LSBA, particularmente no estádio maduro. Preparações de parede celular foram fermentadas in vitro e os dados correlacionados com os teores de lignina obtidos por intermédio dos três métodos. O método LSBA exibiu os menores coeficientes de correlação, para a maioria das amostras, sugerindo que este método deve ser melhor avaliado.

  2. Análise quantitativa do tratamento da escoliose idiopática com o método klapp por meio da biofotogrametria computadorizada Quantitative photogrammetric analysis of the klapp method for treating idiopathic scoliosis

    OpenAIRE

    Iunes, Denise H.; Maria B. B. Cecílio; Marina A. Dozza; Almeida,Polyanna R.

    2010-01-01

    INTRODUÇÃO: Poucos trabalhos comprovam a eficácia das técnicas fisioterapêuticas para o tratamento da escoliose. OBJETIVO: Analisar a eficácia do Método Klapp no tratamento das escolioses por meio do estudo quantitativo pela biofotogrametria computadorizada. MÉTODOS: Dezesseis indivíduos com média de idade de 15±2,61 anos, portadores de escoliose idiopática, foram tratados com o método Klapp. Para análise dos resultados do tratamento, todos foram fotografados antes e após o tratamento, seguin...

  3. GT1b-induced neurotoxicity is mediated by the Akt/GSK-3/tau signaling pathway but not caspase-3 in mesencephalic dopaminergic neurons

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    Sohn Sunghyang

    2010-06-01

    Full Text Available Abstract Background Gangliosides, sialic acid-containing glycosphingolipids exist in mammalian cell membranes particularly neuronal membranes. The trisialoganglioside (GT1b is one of the major brain gangliosides and acts as an endogenous regulator in the brain. We previously showed GT1b induces mesencephalic dopaminergic (DA neuronal death, both in vivo and in vitro. We further investigate the underlying mechanisms of GT1b neurotoxicity. Results Consistent with earlier findings, GT1b attenuated the DA neuron number and dopamine uptake level in mesencephalic cultures. Morphological evidence revealed GT1b-induced chromatin condensation and nuclear fragmentation as well as an increased number of TUNEL-positive cells, compared to control cultures. Interestingly, while GT1b enhanced caspase-3 activity, DEVD, a caspase-3 inhibitor, failed to rescue DA neuronal death. Immunoblot analysis revealed that GT1b inactivates Akt through dephosphorylation at both Ser473 and Thr308, subsequent dephosphorylation of GSK-3β, a substrate of Akt, and hyperphosphorylation of tau, downstream of GSK-3β. Moreover, a GSK-3β specific inhibitor, L803-mt, attenuated tau phosphorylation and rescued DA neurons from cell death in mesencephalic cultures. Conclusion Our data provide novel evidence that a Akt/GSK-3β/tau-dependent, but not caspase-3 signaling pathway plays a pivotal role in GT1b-mediated neurotoxic actions on mesencephalic DA neurons.

  4. Caspase-3 and survivin expression in pediatric neuroblastoma and their roles in apoptosis

    Institute of Scientific and Technical Information of China (English)

    王家祥; 郑树

    2004-01-01

    Background Neuroblastoma, one of the common tumors in children, possesses the feature of natural regression that might be related to apoptosis caspase-3 and survivin are believed to respectively induce and inhibit apoptosis. We investigated the expression of caspase-3 and survivin in pediatric neuroblastoma and the role that these genes played in apoptosis.Methods The expression of caspase-3 and survivin in pediatric neuroblastoma tissue samples was detected using in situ hybridization, ter mintuesal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL), and immunohistochemical staining. The role that these genes played in apoptosis was then evaluated.Results A converse correlation was observed between the expression of survivin and caspase-3. When survivin was expressed at high levels in neuroblastoma samples, caspase-3 expression was downregulated, and the apoptotic index decreased simultaneously.Conclusion There is a converse correlation between the expression of caspase-3 and the expression of survivin in neuroblastoma cells, indicating that caspase-3 might induce apoptosis, and survivin may inhibit this process.

  5. Acteoside Binds to Caspase-3 and Exerts Neuroprotection in the Rotenone Rat Model of Parkinson's Disease

    Science.gov (United States)

    Wang, Ying; He, Xiao; Zhao, Yuwu

    2016-01-01

    Parkinson’s disease (PD) is characterized by the progressive degeneration of the dopaminergic neurons in the substantia nigra (SN) region. Acteoside has displayed multiple biological functions. Its potential role against PD and the underlying signaling mechanisms are largely unknown. Here, we showed that oral administration of acteoside significantly attenuated parkinsonism symptoms in rotenone-induced PD rats. Further, acteoside inhibited rotenone-induced α-synuclein, caspase-3 upregulation and microtubule-associated protein 2 (MAP2) downregulation in PD rats. The molecular docking and molecular dynamics (MD) simulation results indicated that acteoside may directly bind to and inhibit caspase-3. Acteoside formed hydrogen bonds with at least six residues of caspase-3: ThrA177, SerA178, GlyA238, SerB339, ArgB341 and TrpB348. In addition, a pi-pi interaction was formed between acteoside and caspase-3’s HisA237, which might further stabilize the complex. MD simulation results demonstrated that the binding affinity of the caspase-3-acteoside complex was higher than that of caspase-3 and its native ligand inhibitor. Together, we show that acteoside binds to caspase-3 and exerts neuroprotection in the rotenone rat model of PD. PMID:27632381

  6. Estudo da correlação quantitativa entre estrutura e propriedade (QSPR) usando descritores topológicos para compostos carbonílicos alifáticos

    OpenAIRE

    Amboni, Renata Dias de Mello Castanho

    2001-01-01

    Tese (doutorado) - Universidade Federal de Santa Catarina, Centro de Ciências Físicas e Matemáticas. Curso de Pós-Graduação em Química Neste trabalho foi aplicada a relação quantitativa entre estrutura e atividade, empregando-se diferentes descritores moleculares para estimar o odor frutal de ésteres alifáticos. Os parâmetros estatísticos, obtidos nas equações para os ésteres, empregando-se o método de regressão linear múltipla, foram de boa qualidade. O modelo obtido teve uma alta capacid...

  7. Características quantitativas da carcaça de bovinos Charolês, Caracu e cruzamentos recíprocos terminados em confinamento Carcass traits of Charolais, Caracu and reciprocal crosses finished in feedlot

    OpenAIRE

    Daniel Perotto; José Luiz Moletta; Antonio Carlos Cubas

    2000-01-01

    RESUMO - Foram analisadas doze características quantitativas das carcaças de 162 machos bovinos inteiros pertencentes aos grupos Caracu (Ca), Charolês (Ch), 3/4Ca+1/4Ch, 3/4Ch+1/4Ca, 5/8Ca+3/8Ch e 5/8Ch+3/8Ca, nascidos no período de 1988 a 1993. As médias para idade e peso ao início do período de confinamento, duração do confinamento, idade e peso ao abate foram, respectivamente, 712 dias, 362 kg, 99 dias, 811 dias e 489 kg. Durante o confinamento, os animais receberam silagem de milho à vont...

  8. Caspase-3 activation and increased procollagen type I in irradiated hearts

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    Samara C. Ferreira-Machado

    2013-03-01

    Full Text Available The caspase-3-cleaved presence was evaluated in this study in the heart of irradiated rats, during the decline of ventricular function. Female Wistar rats were irradiated with a single dose of radiation (15 Gy delivered directly to the heart and the molecular, histological and physiological evaluations were performed at thirteen months post-irradiation. The expressions of procollagen type I, TGF-ß1 and caspase-3-cleaved were analyzed using Western blotting. Cardiac structural and functional alterations were investigated by echocardiography and electron microscopy. In the irradiated group, the levels of procollagen type I, TGF-ß1 and caspase-3-cleaved are increased. Significant histological changes (degeneration of heart tissue and collagen deposition and functional (reduced ejection fraction were observed. Data suggest that the cardiac function decline after exposure to ionizing radiation is related, in part, to increased collagen and increased caspase-3-cleaved.

  9. MSX2 overexpression inhibits gemcitabine-induced caspase-3 activity in pancreatic cancer cells

    Institute of Scientific and Technical Information of China (English)

    Shin Hamada; Kennichi Satoh; Kenji Kimura; Atsushi Kanno; Atsushi Masamune; Tooru Shimosegawa

    2005-01-01

    AIM: To evaluate the effect of MSX2 on gemcitabineinduced caspase-3 activation in pancreatic cancer cell line Panc-1.METHODS: Using V5-tagged MSX2 expression vector,stable transfectant of MSX2 was generated from Panc-1cells (Px14 cells). Cell viability under gemcitabine administration was determined by MTT assay relative to control cell line (empty-vector transfected Panc-1 cells;P-3EV cells). Hoechst staining was used for the detection of apoptotic cell. Activation of caspase-3 was assessed using Western blotting analysis and direct measurement of caspase-3 specific activities.RESULTS: MSX2 overexpression in Panc-1 cells resulted in decreased gemcitabine-induced caspase-3 activation and increased cell viability under gemcitabine treatment in Px14 cells.CONCLUSION: MSX2 exerts repressive effects on gemcitabine-induced apoptotic pathway. This novel apoptosis-regulating function of MSX2 may provide a new therapeutic target for pancreatic cancer.

  10. Glucosidase II β-subunit, a novel substrate for caspase-3-like activity in rice, plays as a molecular switch between autophagy and programmed cell death

    Science.gov (United States)

    Cui, Jing; Chen, Bing; Wang, Hongjuan; Han, Yue; Chen, Xi; Zhang, Wei

    2016-01-01

    Endoplasmic reticulum (ER) stress activates unfolded protein response (UPR) and autophagy. However, prolonged, severe stresses activate programmed cell death (PCD) in both animal and plant cells. Compared to the well-studied UPR pathway, the molecular mechanisms of ER-stress-induced PCD are less understood. Here, we report the identification of Gas2, the glucosidase II β subunit in the ER, as a potential switch between PCD and autophagy in rice. MS analysis identified Gas2, GRP94, and HSP40 protein in a purified caspase-3-like activity from heat stressed rice cell suspensions. The three corresponding genes were down-regulated under DTT-induced ER stress. Gas2 and GRP94 were localized to the ER, while HSP40 localized to the cytoplasm. Compared to wild-type, a Gas2 RNAi cell line was much sensitive to DTT treatment and had high levels of autophagy. Both caspase-3 and heat-stressed cell suspension lysate could cleave Gas2, producing a 14 kDa N-terminal fragment. Conditional expression of corresponding C-terminal fragment resulted in enhanced caspase-3-like activity in the protoplasts under heat stress. We proposed that mild ER stress causes down-regulation of Gas2 and induces autophagy, while severe stress results in Gas2 cleavage by caspase-3-like activity and the cleavage product amplifies this activity, possibly participating in the initiation of PCD. PMID:27538481

  11. Expression of Caspase-3 in Laryngeal Squamous Cell Carcinoma and its Relationship with Cell Apoptosis

    Institute of Scientific and Technical Information of China (English)

    YU Yuan-chen; ZHONG Zhao-kun; LI Ji-xia; YU Chuan-ting

    2015-01-01

    Objective:To investigate the expression of Caspase-3 in laryngeal squamous cell carcinoma (LSCC) and its relationship with cell apoptosis. Methods: The expression of Caspase-3 protein in 43 LSCC tissues and 21 vocal cord polyp tissues was detected using immunohistochemical SP method; the apoptosis of LSCC was measured by in situ end-labeling (TUNEL) assay, and the relationships between Caspase-3 expression and clinicopathological features as well as cell apoptosis in LSCC tissue were analyzed. Results:The positive rate of Caspase-3 expression in LSCC tissue was lower than in vocal cord polyp tissue dramatically, with statistical significance (51.2%vs. 85.7%,P=0.007). Caspase-3 expression in LSCC tissue was closely related to the tumor differentiated degrees, clinical staging and presence or absence of lymph node metastasis (P=0.009, 0.001, 0.018) instead of the gender, age and tumor size (P>0.05). The apoptosis index (AI) of Caspase-3 was (4.31±0.49)% in LSCC tissue, while (24.28±1.07)% in vocal cord polyp tissue. Significant difference was presented between two groups by comparison to the AI (P<0.001). Spearman correlation analysis displayed that Caspase-3 expression in LSCC tissue had a signiifcantly positive correlation with the number of positive TUNEL cells (r=0.435,P=0.000). Conclusion: Low expression of Caspase-3 protein might promote the tumorigenesis and progression by reducing the apoptosis of tumor cells, and detection to its protein can be considered as an important index for judging the differentiation, clinical staging, inifltration and metastasis of laryngeal carcinoma.

  12. Caspase-3 triggers early synaptic dysfunction in a mouse model of Alzheimer's Disease

    OpenAIRE

    D'Amelio M; Cavallucci V; Middei S; Marchetti C; Pacioni S; Ferri A; Diamantini A; De Zio D; Carrara P; Battistini L; Moreno S; Bacci A.,; Ammassari-Teule M; Marie H; Cecconi F

    2010-01-01

    Abstract Synaptic loss is the best pathological correlate of the cognitive decline in Alzheimer's Disease; yet, the molecular mechanisms underlying synaptic failure are unknown. Here we report a non-apoptotic baseline caspase-3 activity in hippocampal dendritic spines, and an enhancement of this activity at the onset of memory decline in the Tg2576-APPswe mouse model of Alzheimer's Disease. We show that, in spines, caspase-3 activates calcineurin which, in turn, triggers dephosphor...

  13. Truncation of Caspase-3 on Phosphorylated tau%Caspase-3对磷酸化tau蛋白截断作用的研究

    Institute of Scientific and Technical Information of China (English)

    段萍; 李夏春; 邓艳秋; 张蕲; 王建枝

    2005-01-01

    磷酸化tau是阿尔茨海默病(Alzheimer's disease,AD)的特征性病理改变--神经原纤维缠结(neurofibrillarytangles,NFTs)的主要组成部分.最近的研究显示:NFT存在Glu391和Asp421位点被截断的tau片段,然而,tau蛋白的磷酸化是否会影响caspase-3的切割作用尚不清楚.首先纯化重组tau蛋白,然后利用蛋白激酶A(PKA)、钙/钙调蛋白依赖性蛋白激酶Ⅱ(CaMKⅡ)和乳鼠海马组织抽提液对其磷酸化,并用caspase-3对不同磷酸化的tau蛋白进行切割,比较caspase-3对非磷酸化和不同蛋白激酶磷酸化的tau蛋白的切割特性.结果显示:除切割非磷酸化tau蛋白外,caspase-3在体外可分别切割被PKA、CaMKⅡ和乳鼠海马组织抽提液磷酸化的tau蛋白.这一结果提示:磷酸化修饰的tau蛋白仍然是caspase-3的底物.

  14. Expression of Hsp70 and Caspase-3 in rabbits after severe traumatic brain injury

    Institute of Scientific and Technical Information of China (English)

    ZHANG Jing; TAO Dai-qin; ZHAO Hui; YIN Zhi-yong

    2012-01-01

    Objective:To investigate the expression of Caspase-3 and Hsp70 in rabbits after severe traumatic brain injury (TBI) and to explore the feasibility of its application in estimation of injury time in forensic medicine.Methods:A rabbit model of heavy TBI was developed by high velocity impact on the parietal bone with an iron stick.Totally 8 healthy adult New Zealand white rabbits were randomly divided into control group (n=2) and injury group (n=6).Four hours after injury,tissue specimens from the parietal lobe,temporal lobe,occipital lobe,cerebellum and brainstem were harvested to detect the expression of Hsp70 and Caspase-3 by immunohistochemistry.Besides,the gray values of cells positive for Hsp70 and Caspase-3 were analyzed with an image analyzer.Results:Immunohistochemistry staining demonstrated a low level of Caspase-3 and Hsp70 expression in normal control group.While in injury group,both the Caspase-3and Hsp70 expression was significantly elevated (P<0.05).Positive cells gathered around the lesion focus.Occipital lobe and cerebellum had fewer positive cells while temporal and brainstem had the fewest.Conclusion:The expression of Caspase-3 and Hsp70 at an early stage following severe TBI is characteristic and can be applied to estimate the time of injury.

  15. IMMUNOHISTOCHEMICAL ANALYSIS OF CASPASE-3 ACTIVITY IN LIVER BIOPSIES OF PATIENTS WITH MONO AND MIXED INFECTIONS

    Directory of Open Access Journals (Sweden)

    I. I. Tokin

    2015-01-01

    Full Text Available Objective: to study the activity of proapoptotic signal protein caspase-3 for determination of peculiarities of apoptosis regulation under liver chronic diseases.Subjects and methods. The immunohistochemical analysis of caspase-3 activity in 5 liver biopsies of the patients with mono infection of chronic hepatitis B and 5 liver biopsies of the patients with mixed infection of tuberculosis, chronic hepatitis C and human immunodeficiency virus was fulfilled. Morphological and morphometric analysis of serial microphotographs was performed using an image analysis system (microscope Leica DM 2500, digital camera Leica DFC320 R2 and a computer.Results. The activity of caspase-3 as dark brown granularity was revealed in all tis-sue components of liver (hepatocytes, epithelium of bile ducts, endotheliocytes, Kupffer cells of sinusoids, in compositions of lymphohistiocyte infiltrations. The maximal activity was discovered in hepatocytes nuclei. The expression of caspase-3 was significantly higher in liver biopsies of the patients with mixed infection. It is typical that the immunoreactive hepatocytes had not any morphological marks of apoptosis.Conclusion. The caspase-3 expression of proapoptotic signal protein caspase-3 may serve as an early marker of liver damage including the possibilities of apoptosis development.

  16. Caspase-3 activation as a bifurcation point between plasticity and cell death

    Institute of Scientific and Technical Information of China (English)

    Shikha Snigdha; Erica D Smith; G Aleph Prieto; Carl W Cotman

    2012-01-01

    Death-mediating proteases such as caspases and caspase-3 in particular,have been implicated in neurodegenerative processes,aging and Alzheimer's disease.However,emerging evidence suggests that in addition to their classical role in cell death,caspases play a key role in modulating synaptic function.It is remarkable that active caspases-3,which can trigger widespread damage and degeneration,aggregates in structures as delicate as synapses and persists in neurons without causing acute cell death.Here,we evaluate this dichotomy,and discuss the hypothesis that caspase-3 may be a bifurcation point in cellular signaling,able to orient the neuronal response to stress down either pathological/apoptotic pathways or towards physiological cellular remodeling.We propose that temporal,spatial and other regulators of caspase activity are key determinants of the ultimate effect of caspase-3 activation in neurons.This concept has implications for differential roles of caspase-3 activation across the lifespan.Specifically,we propose that limited caspase-3 activation is critical for synaptic function in the healthy adult brain while chronic activation is involved in degenerative processes in the aging brain.

  17. Calpain and caspase-3 play required roles in immobilization-induced limb muscle atrophy.

    Science.gov (United States)

    Talbert, Erin E; Smuder, Ashley J; Min, Kisuk; Kwon, Oh Sung; Powers, Scott K

    2013-05-15

    Prolonged skeletal muscle inactivity results in a rapid decrease in fiber size, primarily due to accelerated proteolysis. Although several proteases are known to contribute to disuse muscle atrophy, the ubiquitin proteasome system is often considered the most important proteolytic system during many conditions that promote muscle wasting. Emerging evidence suggests that calpain and caspase-3 may also play key roles in inactivity-induced atrophy of respiratory muscles, but it remains unknown if these proteases are essential for disuse atrophy in limb skeletal muscles. Therefore, we tested the hypothesis that activation of both calpain and caspase-3 is required for locomotor muscle atrophy induced by hindlimb immobilization. Seven days of immobilization (i.e., limb casting) promoted significant atrophy in type I muscle fibers of the rat soleus muscle. Independent pharmacological inhibition of calpain or caspase-3 prevented this casting-induced atrophy. Interestingly, inhibition of calpain activity also prevented caspase-3 activation, and, conversely, inhibition of caspase-3 prevented calpain activation. These findings indicate that a regulatory cross talk exists between these proteases and provide the first evidence that the activation of calpain and caspase-3 is required for inactivity-induced limb muscle atrophy.

  18. Eletrencefalografia quantitativa no transtorno obsessivo-compulsivo: resultados preliminares

    Directory of Open Access Journals (Sweden)

    FONTENELLE LEONARDO

    2000-01-01

    Full Text Available Investigamos o valor preditivo de variáveis da eletrencefalografia quantitativa (EEGq sobre a resposta ao tratamento farmacológico do transtorno obsessivo-compulsivo (TOC. Foram estudados 10 pacientes com TOC, segundo critérios da DSM-IV, inicialmente não medicados. As seguintes variáveis eletrencefalográficas foram selecionadas: potência relativa bipolar na faixa alfa para os eletrodos Cz-C3, Cz-C4, T3-T5, T4-T6, T3-F7 e T4-F8, potência relativa bipolar na faixa teta para os eletrodos Cz-C3, Cz-C4, T3-T5, T4-T6, T3-F7 e T4-F8 e potência relativa monopolar na faixa teta para os eletrodos Fp1, Fp2, F7, F3, Fz, F4 e F8. Dos 10 pacientes examinados, 5 apresentaram desvios estatisticamente significativos da potência relativa bipolar na faixa alfa em regiões centrais, quando comparados com o banco normativo de dados (grupo controle. Os demais não apresentaram desvios com significado estatístico. O grupo de pacientes com alterações eletrencefalográficas se caracterizou por idade de início do TOC significativamente mais precoce e pior resposta ao tratamento com inibidores da recaptação da serotonina.

  19. Caspase 3 siRNA decreases apoptosis in cultured neuronal cells

    Institute of Scientific and Technical Information of China (English)

    Chunting Ye; Yaoxiong Huang; Xiaohong Yang; Honghui Chen

    2009-01-01

    BACKGROUND: Lentiviral vectors, a type of retroviral vector, are able to infect cells at all phases of cell cycle. They are able to express exogenous target genes in vivo over long periods of time with limited immunological reaction. OBJECTIVE: To inhibit neuronal apoptosis by blocking the apoptotic cascade reaction, gene silencing of Caspase 3, and transfection of Caspase 3 short hairpin ribonucleic acid (shRNA) into Neuro 2a cells using a lentiviral vector. DESIGN, TIME AND SETTING: An observational, genetic engineering cellular biology experiment was performed in Guangzhou Red Cross Hospital and Guangzhou institute of Traumatic Surgery between March 2007 and June 2008.MATERIALS: PLL3.7, PCMV-VSV-G, and PH'8.9△PR plasmids were provided by the CBR institute for Biomedical Research, Harvard Medical School, USA. Staurosporine was purchased from Sigma, USA. METHODS: Caspase 3 siRNA was synthesized and cloned into the PLL3.7 plasmid. The Caspase 3 shRNA-PLL3.7 lentivirus was generated in 293T cells using a calcium phosphate transfection kit. After the lentivirus was transfected into Neuro 2a cells, apoptosis was induced in both the transfected and untransfected cells by staurosporine. Cell apoptosis was assessed by flow cytometry. MAIN OUTCOME MEASURES: Caspase 3 mRNA expression was measured by RT-PCR and Caspase protein expression was assessed by Western blot. Cellular apoptosis was determined by flow cytometry using Annevin V-PE/7aad-Cy7.RESULTS: The transfection rate of caspase 3 shRNA was>98% using the ientiviral vector. RT-PCR and Western blot results demonstrated that significantly reduced Caspase 3 mRNA and protein expression in the transfected Neuro 2a. The control group exhibited 38.7% Annexin V/7aad-positive cells, which suggested apoptotic anaphase, while only 5.0% cells in the Caspase 3 gene silencing group entered apoptotic anaphase. CONCLUSION: Caspase 3 shRNA inhibited Caspase 3 expression in Neuro 2a cells and decreased drug-induced apoptosis of

  20. ROLE OF CASPASE-3 IN ACUTE LIGHT DAMAGE TO RETINA OF RATS

    Institute of Scientific and Technical Information of China (English)

    Xiao Wang; Shi-xing Hu; Wei Li; Shao-chun Lin

    2007-01-01

    Objective To investigate the role of Caspase-3 in retinal damage caused by light exposure in rats.Methods Light injury to retina was induced by persistent exposure to illumination (intensity; 30000±50 lux) of operating microscope for 30 minutes in the right eyes of Sprague-Dawley rats. The pathological changes of retina were observed under optical and electron microscopies at different time points, which were 6 hours, 1,3,7,and 15 days after the light exposure. Apoptosis of retinal cells was analyzed by flow cytometry. The activity of Caspase-3 was evaluated by using the Caspase-3 assay kit. At the same time, the expression of Caspase-3 protease was determined with Western blot analysis.Results The examination results of optical and transmission electron microscopes showed that edema of inner and outer segments of the retina, espectially the chondriosome inside the inner segment, became obvious 6 hours after the light exposure. The change was deteriorated along with the increasing time. The structures of the discoidal valve dissociated in the outer segment simultaneously. Disorderly arranged nuclei, karyopycnosis, and thinning in the outer nuclear layer were observed. The retinal pogment epithelium almost disappeared during the later stage. The staining results of Annexin-V combined with PI demonstrated that the proportion of apoptotic cells increased with time. The proportion between 7th day (82.7%) and 15th day (80.4%), however, showed no significant difference. Caspase-3 became remarkably active with the lapse of time, which increased from 0.02 at 6th hour to the peak of 9.8 at 7th day before it started to descend. The Western blot detected a expression of the active form of Caspase-3 at 7th day and 15th day.Conclusion Apoptosis of photoreceptor cells is markedly involved in the light damage and Caspase-3 protease may play an important role in the apoptotic process of the retina after light exposure in rats.

  1. Specific inhibition of caspase-3 by a competitive DARPin: molecular mimicry between native and designed inhibitors.

    Science.gov (United States)

    Schroeder, Thilo; Barandun, Jonas; Flütsch, Andreas; Briand, Christophe; Mittl, Peer R E; Grütter, Markus G

    2013-02-05

    Dysregulation of apoptosis is associated with several human diseases. The main apoptotic mediators are caspases, which propagate death signals to downstream targets. Executioner caspase-3 is responsible for the majority of cleavage events and its therapeutic potential is of high interest with to date several available active site peptide inhibitors. These molecules inhibit caspase-3, but also homologous caspases. Here, we describe caspase-3 specific inhibitors D3.4 and D3.8, which have been selected from a library of designed ankyrin repeat proteins (DARPins). The crystal structures of D3.4 and mutants thereof show how high specificity and inhibition is achieved. They also show similarities in the binding mode with that of the natural caspase inhibitor XIAP (X-linked inhibitor of apoptosis). The kinetic data reveal a competitive inhibition mechanism. D3.4 is specific for caspase-3 and does not bind the highly homologous caspase-7. D3.4 therefore is an excellent tool to define the precise role of caspase-3 in the various apoptotic pathways.

  2. Análise quantitativa das AgNORs no carcinoma adenóide cístico intra-oral através da técnica de dupla marcação PCNA/AgNOR

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    Rivero Elena Riet-Correa

    2002-01-01

    Full Text Available A análise quantitativa das AgNORs e a imunomarcação para o PCNA têm sido empregadas de forma independente na avaliação da proliferação celular de vários tumores, e, em muitos casos, têm mostrado correlação positiva. Entretanto poucos trabalhos têm avaliado, em um mesmo corte histológico, a relação entre PCNA e AgNOR. O objetivo deste trabalho foi otimizar a técnica de dupla marcação com a finalidade de se estudar simultaneamente a correlação entre PCNA e AgNOR no carcinoma adenóide cístico (CAC de glândulas salivares menores. Foram selecionados 16 casos de CAC classificados de acordo com o subtipo histológico. A análise quantitativa das AgNORs foi feita por meio de análise de imagens. As AgNORs foram contadas em cem núcleos PCNA positivos e em cem núcleos PCNA negativos. O número médio de AgNOR nos núcleos PCNA positivos foi 2,14 ± 0,77, e, nos núcleos PCNA negativos, 1,97 ± 0,79, entretanto esta diferença não se mostrou estatisticamente significante (p = 0,2537. Nosso trabalho não mostrou correlação entre o número de AgNOR e a imunomarcação para o PCNA em CAC quando estes marcadores foram demonstrados simultaneamente através da dupla marcação. Quanto à técnica, o uso do microondas melhorou a coloração da AgNOR, permitindo uma redução no tempo de incubação com a solução de prata e uma melhor individualização das AgNORs, o que facilitou os procedimentos de contagem.

  3. Correlation between neuronal injury and Caspase-3 after focal ischemia in human hippocampus

    Institute of Scientific and Technical Information of China (English)

    戚基萍; 吴爱萍; 王德生; 王立峰; 李淑霞; 徐凤琳

    2004-01-01

    Background Cerebral ischemia is a significant clinical problem, and cerebral ischemia usually causes neuron injury such as apoptosis in various brain areas, including hippocampus. Cysteinyl aspartate-specific protease (Caspases) are fundamental factors of apoptotic mechanism. Caspase-3 inhibitors show effect in attenuating brain injury after ischemia. But all the results were from animal models in research laboratories. This study aimed at investigating the correlation between the change of ischemic neuronal injury and Caspase-3 post-ischemia in human hippocampus. Methods We selected and systematized 48 post-mortem specimens from 48 patients, who died of cerebral infarction. Morphological change was firstly analyzed by observing hematoxyline/eosin-staining hippocampal sections. The expression of Caspase-3 was investigated using the methods of in situ hybridization and immunohistochemistry. Terminal deoxynucleotidyl transferase-mediated 2'-deoxyuridine 5'-triphosphate-biotin nick-end labeling (TUNEL) method was used to clarify the involvement of Caspase-3 in neuron death. The loss of MAP 2 (MAP-2) was applied to judging the damaged area and degree of neuronal injury caused by ischemia.Results In the CA1 sector of hippocampus, Caspase-3 immunostaining modestly increased at 8 hours [8.05/high-power field (hpf)], dramatically increased at 24 hours (24.85/hpf), decreased somewhat after 72 hours. Caspase-3 mRNA was detectable at 4 hours (6.75/hpf), reached a maximum at 16 hours (17.60/hpf), faded at 72 hours. TUNEL-positive cells were detectable at 24 hours (10.76/hpf), markedly increased at 48-72 hours. The loss of MAP-2 was obviously detected at 4 hours, progressed significantly between 24 and 72 hours; MAP-2 immunoreactivity was barely detectable at 72 hours. Before 72 hours, the Caspase-3 evolution was related with the upregulation of TUNEL and the loss of MAP-2. The positive correlation between Caspase-3 mRNA and TUNEL was significant at the 0.05 level (correlation

  4. An Efficient Piecewise Linear Model for Predicting Activity of Caspase-3 Inhibitors

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    Alireza Foroumadi

    2012-09-01

    Full Text Available Background and purpose of the study Multimodal distribution of descriptors makes it more difficult to fit a single global model to model the entire data set in quantitative structure activity relationship (QSAR studies.Methods:The linear (Multiple linear regression; MLR, non-linear (Artificial neural network; ANN, and an approach based on "Extended Classifier System in Function approximation" (XCSF were applied herein to model the biological activity of 658 caspase-3 inhibitors. Results:Various kinds of molecular descriptors were calculated to represent the molecular structures of the compounds. The original data set was partitioned into the training and test sets by the K-means classification method. Prediction error on the test data set indicated that the XCSF as a local model estimates caspase-3 inhibition activity, better than the global models such as MLR and ANN. The atom-centered fragment type CR2X2, electronegativity, polarizability, and atomic radius and also the lipophilicity of the molecule, were the main independent factors contributing to the caspase-3 inhibition activity. Conclusions:The results of this study may be exploited for further design of novel caspase-3 inhibitors.

  5. Effect of Bcl-2 and caspase-3 on calcium distribution in apoptosis of HL-60 cells

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Apoptosis manifests in two major execution programs downstream of the death signal: the caspase pathway and organelle dysfunction. An important antiapoptosis factor, Bcl-2 protein, contributes in caspase pathway of apoptosis. Calcium, an important intracellular signal element in cells, is also observed to have changes during apoptosis, which maybe affected by Bcl2 protein. We have previously reported that in Harringtonine (HT) induced apoptosis of HL-60 cells, there's a change of intracellular calcium distribution, moving from cytoplast especially Golgi's apparatus to nucleus and accumulating there with the highest concentration. We report here that caspase-3 becomes activated in HT-induced apoptosis of HL-60 cells, which can be inhibited by overexpression of Bcl-2 protein. No sign of apoptosis or intracellular calcium movement from Golgi's apparatus to nucleus in HL-60 cells overexpressing Bcl-2 or treated with Ac-DEVD-CHO, a specific inhibitor of caspase-3. The results indicate that activated caspase-3 can promote the movement of intracellular calcium from Golgi's apparatus to nucleus, and the process is inhibited by Ac-DEVD-CHO (inhibitor of caspas-3), and that Bcl-2 can inhibit the movement and accumulation of intracellular calcium in nucleus through its inhibition on caspase3. Calcium relocalization in apoptosis seems to be irreversible, which is different from the intracellular calcium changes caused by growth factor.

  6. Hyperosmotic Stress Induces Tau Proteolysis by Caspase-3 Activation in SH-SY5Y Cells.

    Science.gov (United States)

    Olivera-Santa Catalina, Marta; Caballero-Bermejo, Montaña; Argent, Ricardo; Alonso, Juan C; Cuenda, Ana; Lorenzo, María J; Centeno, Francisco

    2016-12-01

    Tau is a microtubule-associated protein implicated in the pathogenesis of Alzheimer's disease and other related tauopathies. In this subset of neurodegenerative disorders, Tau auto-assembles into insoluble fibrils that accumulate in neurons as paired helical filaments (PHFs), promoting cellular dysfunction and cytotoxic effects. Growing evidence suggests that abnormal post-translational regulation, mainly hyperphosphorylation and aberrant cleavage, drives Tau to this pathological state. In this work we show that sorbitol-induced hyperosmotic stress promotes Tau proteolysis in SH-SY5Y neuroblastoma cells. The appearance of cleaved Tau was preceded by the activation of μ-calpain, the proteasome system and caspase-3. Tau proteolysis was completely prevented by caspase-3 inhibition but unaffected by neither the proteasome system nor μ-calpain activity blockade. Concomitantly, hyperosmotic stress induced apoptosis in SH-SY5Y cells, which was efficiently avoided by the inhibition of caspase-3 activity. Altogether, our results provide the first evidence that Tau protein is susceptible to caspase-3 proteolysis under hyperosmotic stress and suggest a positive relationship between Tau proteolysis and apoptosis in SH-SY5Y cells. J. Cell. Biochem. 117: 2781-2790, 2016. © 2016 Wiley Periodicals, Inc. © 2016 Wiley Periodicals, Inc.

  7. Caspase-3-dependent Cell Death in B lymphocyte Caused by Pseudomonas aeruginosa Pyocyanin

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    Heni Susilowati

    2015-08-01

    Full Text Available Objective: The aim of this study was to investigate cellular responses of B lymphocyte to the exposure of pyocyanin and the role of caspase-3 in its molecular mechanism. Methods: B lymphocytes (Raji cells were cultured overnight prior to the experiments. Cell culture in five replications were then exposed to various concentrations of pyocyanin and incubated for 24 h in antibiotics-free medium. MTT assay was performed to analyze the cytotoxicity effect of pyocyanin. In separated experiments, the cells were cultured with pyocyanin and addressed for cell morphological analysis using phase contrast microscope. To study the mechanism involved in pyocyanin-induced cellular damage, immunocytochemical analysis was run for the identification of active caspase-3 protein expression. Results: The results of this study showed that cell viability was decreased in pcyocyanin-treated groups. Statistical analysis using ANOVA (p < 0.05 demonstrated significant different between groups with significant value of 0.000. Pyocyanin induced cell death on B lymphocyte in dose-dependent manner. Nuclear fragmentation was observed in pyocyanin-induced cell death; furthermore, caspase-3 was expressed clearly in cell cytoplasm after 24 h incubation. Conclusion: It is concluded that pyocyanin is capable of inducing cell death on B lymphocyte. Caspase-3 may play important role in the molecular mechanism of pyocyanin-induced cell death.DOI: 10.14693/jdi.v22i2.403

  8. Expression of Fas ligand and Caspase-3 contributes to formation of immune escape in gastric cancer

    Institute of Scientific and Technical Information of China (English)

    Hua-Chuan Zheng; Jin-llin Sun; Zheng-Li Wei; Xue-Fei Yang; Yin-Chang Zhang; Yan Xin

    2003-01-01

    AIM: To study the role of Fas ligand (FasL) and Caspase-3expression in carcinogenesis and progression of gastric cancer and molecular mechanisms of relevant immune escape.METHODS: FasL and Caspase-3 expression was studied in adjacent epithelial cells, cancer cells and lymphocytes of primary foci, and cancer cells of metastatic foci from 113 cases of gastric cancer by streptavidin-biotin-peroxidase (S-P) immunohistochemistry. Expression of both proteins in cancer cells of primary foci was compared with clinicopathological features of gastric cancer. The relationship between FasL expression in cancer cells and Caspase-3expression in cancer cells or infiltrating lymphocytes of primary foci was investigated.RESULTS: Cancer cells of primary foci expressed FasL in 53.98 % (61/113) of gastric cancers, more than their adjacent epithelial cells (34.51%, 39/113) (P=0.003, X2=8.681), while the expression of Caspase-3 in cancer cells of primary foci was detected in 32.74 % (37/113) of gastric cancers, less than in the adjacent epithelial cells (50.44 %, 57/113)(P=0.007, X2=7.286). Infiltrating lymphocytes of the primary foci showed positive immunoreactivity to Caspase-3 in 70.80 % (80/113) of gastric cancers, more than their corresponding adjacent epithelial cells (P=0.001, X2=10.635)or cancer cells of primary foci (P=0.000, X2=32.767). FasL was less expressed in cancer cells of metastases (51.16 %,22/43) than in those of the corresponding primary foci (81.58 %, 31/38) (P=0.003, X2=9.907). Conversely,Caspase-3 was more expressed in cancer cells of metastases (58.14 %, 25/43) than in those of the corresponding primary foci (34.21%, 13/38) (P=0.031, X2=4.638). FasL expression was significantly correlated with tumor size (P=0.035,rs=0.276), invasive depth (P=0.039, rs=0.195), metastasis (P=0.039, rs=0.195), differentiation (P=0.015, rs=0.228)and Lauren′s classification (P=0.038, rs=0.196), but not with age or gender of patients, growth pattern or TNM staging of gastric cancer

  9. Vitamin C Attenuates Isoflurane-Induced Caspase-3 Activation and Cognitive Impairment.

    Science.gov (United States)

    Cheng, Baiqi; Zhang, Yiying; Wang, Arthur; Dong, Yuanlin; Xie, Zhongcong

    2015-12-01

    Anesthetic isoflurane has been reported to induce caspase-3 activation. The underlying mechanism(s) and targeted intervention(s), however, remain largely to be determined. Vitamin C (VitC) inhibits oxidative stress and apoptosis. We therefore employed VitC to further determine the up-stream mechanisms and the down-stream consequences of the isoflurane-induced caspase-3 activation. H4 human neuroglioma cells overexpressed human amyloid precursor protein (H4-APP cells) and rat neuroblastoma cells were treated either with (1) 2% isoflurane or (2) with the control condition, plus saline or 400 μM VitC for 3 or 6 h. Western blot analysis and fluorescence assay were utilized at the end of the experiments to determine caspase-3 activation, levels of reactive oxygen species and ATP, and mitochondrial function. The interaction of isoflurane (1.4% for 2 h) and VitC (100 mg/kg) on cognitive function in mice was also assessed in the fear conditioning system. Here, we show for the first time that the VitC treatment attenuated the isoflurane-induced caspase-3 activation. Moreover, VitC mitigated the isoflurane-induced increases in the levels of reactive oxygen species, opening of mitochondrial permeability transition pore, reduction in mitochondrial membrane potential, and the reduction in ATP levels in the cells. Finally, VitC ameliorated the isoflurane-induced cognitive impairment in the mice. Pending confirmation from future studies, these results suggested that VitC attenuated the isoflurane-induced caspase-3 activation and cognitive impairment by inhibiting the isoflurane-induced oxidative stress, mitochondrial dysfunction, and reduction in ATP levels. These findings would promote further research into the underlying mechanisms and targeted interventions of anesthesia neurotoxicity.

  10. Paradoxical sleep deprivation changes testicular malondialdehyde and caspase-3 expression in male rats

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    Fitranto Arjadi

    2015-08-01

    Full Text Available BACKGROUND Sleep deprivation is a significant problem among adult men and is considered as a risk factor for several diseases. Paradoxical sleep deprivation (PSD induces Leydig cell apoptosis through elevation of corticosterone, with testicular malondialdehyde (MDA and Leydig cell caspase-3 expression as parameters. The aim of this study was to observe testicular MDA level and caspase-3 expression treated with paradoxical sleep deprivation (PSD, immobilization, and footshock stress and to determine the stress model with a significant effect in white male rats (Rattus norvegicus . METHODS This experimental randomized study of posttest only with control group design was conducted on 24 white male Wistar strain rats, randomly allocated into four treatment groups, i.e. control (K1 without any stress treatment, PSD (KII, immobilization (KIII, and footshock stress (KIV. Treatments were given for 25 days to produce chronic stress. Testicular MDA concentration was examined by the ELISA method while caspase-3 was examined by the TUNEL method. RESULTS Mean testicular MDA concentration with one-way ANOVA test showed differences in means between the groups (p=0.000 and post hoc Tukey-HSD test showed significant results between PSD stress group versus control, immobilization and footshock stress groups. One-way ANOVA test showed a significant difference in caspase-3 expression in at least two treatment groups (p=0.008 and post-hoc Tuckey-LSD test showed significant differences between controls and all stress groups. CONCLUSION Sleep deprivation is a type of stress inducing changes in testicular MDA concentration and caspase-3 expression in male rat testes.

  11. Rapid cytochrome c release, activation of caspases 3, 6, 7 and 8 followed by Bap31 cleavage in HeLa cells treated with photodynamic therapy.

    Science.gov (United States)

    Granville, D J; Carthy, C M; Jiang, H; Shore, G C; McManus, B M; Hunt, D W

    1998-10-16

    Photodynamic therapy (PDT) is a clinical approach that utilizes light-activated drugs for the treatment of a variety of pathologic conditions. The initiating events of PDT-induced apoptosis are poorly defined. It has been shown for other proapoptotic stimuli that the integral endoplasmic reticulum protein Bap31 is cleaved by caspases 1 and 8, but not by caspase-3. Further, a 20 kDa Bap31 cleavage fragment is generated which can induce apoptosis. In the current report, we sought to determine whether Bap31 cleavage and generation of p20 is an early event in PDT-induced apoptosis. The mitochondrial release of cytochrome c, involvement of caspases 1, 2, 3, 4, 6, 7, 8, and 10 and the status of several known caspase substrates, including Bap31, were evaluated in PDT-treated HeLa cells. Cytochrome c appeared in the cytosol immediately following light activation of the photosensitizer benzoporphyrin derivative monoacid ring A. Activation of caspases 3, 6, 7, and 8 was evident within 1-2 h post PDT. Processing of caspases 1, 2, 4, and 10 was not observed. Cleavage of Bap31 was observed at 2-3 h post PDT. The caspase-3 inhibitor DEVD-fmk blocked caspase-8 and Bap31 cleavage suggesting that caspase-8 and Bap31 processing occur downstream of caspase-3 activation in PDT-induced apoptosis. These results demonstrate that release of mitochondrial cytochrome c into the cytoplasm is a primary event following PDT, preceding caspase activation and cleavage of Bap31. To our knowledge, this is the first example of a chemotherapeutic agent inducing caspase-8 activation and demonstrates that caspase-8 activation can occur after cytochrome c release.

  12. Bone geometry, biomechanics, and mineral content of the distal radius in uremic patients with severe hyperparathyroidism; Valutazione delle proprieta' geometriche, biomeccaniche e osteodensitometriche del radio ultradistale mediante tomografia computerizzata quantitativa periferica nei pazienti uremici affetti da iperparatiroidismo grave

    Energy Technology Data Exchange (ETDEWEB)

    Di Leo, C.; Tarolo, G.L.; Bestetti, A.; Tagliabue, L.; Del Sole, A.; Aliberti, G. [Ospedale S. Paolo, Milan (Italy). Cattedra e Scuola di Specializzazione in Medicina Nucleare. Servizio di Medicina Nucleare; Ardemagni, A.; Gallieni, M.; Brancaccio, M.; Froehlich, W. [L' Accessorio nucleare (ACN). Laboratori scientifici, Cerro Maggiore, MI (Italy)

    1999-04-01

    Bone integrity and mineral status were studied with a noninvasive method in uremic patients with severe secondary hyperparathyroidism undergoing maintenance hemodialysis. [Italian] Gli autori propongono un metodo non invasivo per studiare la densita' minerale e le caratteristiche geometriche e biomeccaniche dell'osso nei pazienti uremici affetti da iperparatiroidismo secondario grave.

  13. LA ÉTICA DE LA INVESTIGACIÓN EN CIENCIAS SOCIALES EN EL CONTEXTO DE LA GLOBALIZACIÓN: DE LA INVESTIGACIÓN CUANTITATIVA A LA CUALITATIVA A ÉTICA DA PESQUISA EM CIÊNCIAS SOCIAIS NO CONTEXTO DA GLOBALIZAÇÃO: DA PESQUISA QUANTITATIVA À QUALITATIVA SOCIAL SCIENCES’ INVESTIGATION ETHICS IN THE GLOBALISATION CONTEXT: FROM QUANTITATIVE TO QUALITATIVE INVESTIGATION

    Directory of Open Access Journals (Sweden)

    Lorenzo Agar Corbinos

    2004-01-01

    Full Text Available En las ciencias sociales existe un conjunto de cambios sociales y económicos, producto de la globalización, que nos lleva a reflexionar sobre la elección de las metodologías de estudio. El desafío consiste en reducir la brecha entre lo social, estudiado a través de las metodologías cuantitativas, y la socialidad, marcada por las metodologías cualitativas.La ética surge de la cultura. Ésta nos dice sobre el inconsciente colectivo de un pueblo. La investigación debe abordarlas formas apropiadas y validadas de aprehender la realidad. He aquí la evidencia más estrecha de la trilogía: cultura/ética/investigación. Actualmente, lo que surge desde las raíces de la vida en comunidad es una ética asociada con la socialidad y por esto es importante repensar las metodologías de investigación en las ciencias socialesNas ciências sociais existe um conjunto de mudanças sociais e econômicas, produto da globalização, que nos levam a refletir sobre a escolha das metodologias de estudo. O desfio consiste em reduzir o fosso entre ö social", estudado através das metodologias quantitativas, e "a sociabilidade", marcada pelas metodologias qualitativas. A ética surge da cultura. Esta nos fala do inconsciente coletivo de um povo. A pesquisa deve abordar as formas apropriadas e validadas de apreender a realidade. Aqui está a evidência mais clara da trilogia: cultura, ética e pesquisa. Atualmente o que surge a partir da vida em comunidade é uma ética associada com a sociabilidade e por isto é importante repensar as metodologias de pesquisa em ciências sociaisAs a result of globalisation, a number of social and economic changes are taking place in social sciences that leads us to think about the choice of studying methodologies. The challenge consists in reducing the space between what is social, studied through quantitative methodologies, and what is "sociality", marked by qualitative methodologies. Ethics arises from culture, which

  14. Carga proviral do HTLV-1 e HTLV-2: um método simples através da PCR quantitativa em tempo real HTLV-1 and HTLV-2 proviral load: a simple method using quantitative real-time PCR

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    Bruna Pedroso Tamegão-Lopes

    2006-12-01

    Full Text Available Os vírus linfotrópicos de células T humanas, quando integrados ao genoma da célula hospedeira, provírus, têm como marcador de replicação seu DNA proviral. A carga proviral parece ser um importante fator no desenvolvimento de patologias associadas a estes retrovírus. Neste estudo foi desenvolvida uma metodologia para quantificação absoluta da carga proviral dos HTLV-1 e HTLV-2 através da PCR em tempo real. Cinqüenta e três amostras de doadores de sangue com teste de ELISA reagente foram submetidas à metodologia, que utilizou o sistema TaqMan® para três seqüências alvo: HTLV-1, HTLV-2 e albumina. A quantificação proviral absoluta foi determinada através da proporção relativa entre o genoma do HTLV e o genoma da célula hospedeira, levando em consideração o número de leucócitos. O método apresentado é sensível (215 cópias/mL, prático e simples para quantificação proviral, além de eficiente e adequado para confirmação e discriminação da infecção pelos tipos virais.When the human T cell lymphotropic virus (HTLV is integrated with the host cell genome (provirus, its proviral DNA is a replication marker. Proviral load appears to be an important factor in the development of diseases related to these retroviruses. In this study, a methodology for absolute quantification of the HTLV-1 and HTLV-2 proviral load using real-time PCR was developed. Fifty-three blood donor samples with positive ELISA test result were subjected to this methodology, which utilized the TaqMan® system for three target sequences: HTLV-1, HTLV-2 and albumin. The absolute proviral load was quantified using the relative ratio between the HTLV genome and the host cell genome, taking into consideration the white blood cell count. The method presented is sensitive (215 copies/ml, practical and simple for proviral quantification, and is efficient and appropriate for confirming and discriminating infections according to viral type.

  15. Activation of caspases-3, -6, and -9 during finasteride treatment of benign prostatic hyperplasia.

    Science.gov (United States)

    Bozec, Aline; Ruffion, Alain; Decaussin, Myriam; Andre, Jean; Devonec, Marian; Benahmed, Mohamed; Mauduit, Claire

    2005-01-01

    Benign prostatic hyperplasia (BPH) results from an increase in both epithelial and stromal compartments of the human prostate. Although inhibitors of 5alpha-reductase such as finasteride have been shown to reduce the size of BPH tissues by inducing apoptosis, their mechanisms of action still remain unknown. The present study supports that such a process triggered by finasteride is caspase dependent with a possible involvement of two effector caspases (caspase-3 and 6) and two initiator caspases (caspase-8 and 9). Indeed, by using tissues from patients affected by BPH and treated by finasteride (5 mg/d) for 2-3, 6-8, or 27-32 d, we observed that the 5alpha-reductase inhibitor induced apoptosis in epithelial cells (evaluated through cell number positive for terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling) as early as 2-3 d of treatment, with a maximal activity (250-fold increase, P 8 d of treatment. However, after 27-32 d of treatment, the number of apoptotic cells was reduced and was close to control. Caspases-3, -6, -8, and -9 were immunolocalized to (basal and secretory) epithelial cells and to a lesser extent to stromal cells. Activated caspase-3 immunoexpression was restricted to epithelial secretory cells, and its immunostaining intensity appeared to be higher in BPH tissues from patients treated for 2-3 or 6-8 d. Consistently, in Western blotting analyses, activated caspases-3 and -6 were detected as early as 2-3 d of treatment in BPH tissues, and their levels were increased after 6-8 d of treatment. In real time quantitative PCR experiments, caspase-3 and -6 mRNA levels were found to be unchanged after finasteride treatment. Activated caspase-8 was not detected in the different conditions tested, whereas activated caspase-9 protein levels were maximally enhanced after 2-3 d of finasteride treatment. In conclusion, we report here that finasteride treatment of BPH tissues induced a caspase-dependent apoptotic process

  16. Detecção de locos de características quantitativas com efeito da origem parental dos alelos nos cromossomos 1, 2 e 4 de suínos

    OpenAIRE

    Silva Filho,Miguel Inácio da

    2010-01-01

    Com o objetivo de detectar QTL com efeito da origem parental dos alelos, foram utilizados dados de uma população F2 de suínos, composta de 600 animais, obtidos a partir do cruzamento de machos Piau e fêmeas comerciais. Nos animais F2, foram avaliadas características de desempenho, carcaça, órgãos e vísceras, cortes de carcaça e qualidade de carne. Para a genotipagem de todos os animais, foram utilizados 16 locos de microssatélites distribuídos nos cromossomos 1, 2 e 4. Com o resultado da g...

  17. Lentiviral-mediated RNAi targeting caspase-3 inhibits apoptosis induced by serum deprivation in rat endplate chondrocytes in vitro

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    L. Ding

    2014-06-01

    Full Text Available Current studies find that degenerated cartilage endplates (CEP of vertebrae, with fewer diffusion areas, decrease nutrient supply and accelerate intervertebral disc degeneration. Many more apoptotic cells have been identified in degenerated than in normal endplates, and may be responsible for the degenerated grade. Previous findings suggest that inhibition of apoptosis is one possible approach to improve disc regeneration. It is postulated that inhibition of CEP cell apoptosis may be responsible for the regeneration of endplates. Caspase-3, involved in the execution phase of apoptosis, is a candidate for regulating the apoptotic process. In the present study, CEP cells were incubated in 1% fetal bovine serum. Activated caspases were detected to identify the apoptotic pathway, and apoptosis was quantified by flow cytometry. Lentiviral caspase-3 short hairpin RNA (shRNA was employed to study its protective effects against serum deprivation. Silencing of caspase-3 expression was quantified by reverse transcription-polymerase chain reaction and Western blots, and inhibition of apoptosis was quantified by flow cytometry. Serum deprivation increased apoptosis of rat CEP cells through activation of a caspase cascade. Lentiviral caspase-3 shRNA was successfully transduced into CEP cells, and specifically silenced endogenous caspase-3 expression. Surviving cells were protected by the downregulation of caspase-3 expression and activation. Thus, lentiviral caspase-3 shRNA-mediated RNAi successfully silenced endogenous caspase-3 expression, preventing inappropriate or premature apoptosis.

  18. Lentiviral-mediated RNAi targeting caspase-3 inhibits apoptosis induced by serum deprivation in rat endplate chondrocytes in vitro

    Energy Technology Data Exchange (ETDEWEB)

    Ding, L.; Wu, J.P. [Fudan University, Jinshan Hospital, Department of Orthopaedics, Shanghai, China, Department of Orthopaedics, Jinshan Hospital, Fudan University, Shanghai (China); Xu, G. [Fudan University, Jinshan Hospital, Center Laboratory, Shanghai, China, Center Laboratory, Jinshan Hospital, Fudan University, Shanghai (China); Zhu, B.; Zeng, Q.M.; Li, D.F.; Lu, W. [Fudan University, Jinshan Hospital, Department of Orthopaedics, Shanghai, China, Department of Orthopaedics, Jinshan Hospital, Fudan University, Shanghai (China)

    2014-05-09

    Current studies find that degenerated cartilage endplates (CEP) of vertebrae, with fewer diffusion areas, decrease nutrient supply and accelerate intervertebral disc degeneration. Many more apoptotic cells have been identified in degenerated than in normal endplates, and may be responsible for the degenerated grade. Previous findings suggest that inhibition of apoptosis is one possible approach to improve disc regeneration. It is postulated that inhibition of CEP cell apoptosis may be responsible for the regeneration of endplates. Caspase-3, involved in the execution phase of apoptosis, is a candidate for regulating the apoptotic process. In the present study, CEP cells were incubated in 1% fetal bovine serum. Activated caspases were detected to identify the apoptotic pathway, and apoptosis was quantified by flow cytometry. Lentiviral caspase-3 short hairpin RNA (shRNA) was employed to study its protective effects against serum deprivation. Silencing of caspase-3 expression was quantified by reverse transcription-polymerase chain reaction and Western blots, and inhibition of apoptosis was quantified by flow cytometry. Serum deprivation increased apoptosis of rat CEP cells through activation of a caspase cascade. Lentiviral caspase-3 shRNA was successfully transduced into CEP cells, and specifically silenced endogenous caspase-3 expression. Surviving cells were protected by the downregulation of caspase-3 expression and activation. Thus, lentiviral caspase-3 shRNA-mediated RNAi successfully silenced endogenous caspase-3 expression, preventing inappropriate or premature apoptosis.

  19. Análise quali-quantitativa da arborização na praça XV de novembro em Ribeirão Preto - SP, Brasil

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    Gustavo de Nobrega Romani

    2012-06-01

    Full Text Available A Praça XV de Novembro, implantada em meados do século XIX, tem grande valor histórico-cultural, além de se constituir em uma das principais áreas verdes do centro da cidade de Ribeirão Preto. Visando ao conhecimento detalhado da vegetação para fins de orientação do manejo e conservação dessa área, foi feito um levantamento quali-quantitativo e fitossociológico das árvores e palmeiras da praça. Foram medidas altura e Diâmetro à Altura do Peito (DAP e identificados todos os indivíduos de porte arbóreo (árvores e palmeiras presentes na Praça, em nível de espécie. A praça ocupa uma área de 15.456,00 m², onde foram amostradas 42 espécies distribuídas por 19 famílias, num total de 161 indivíduos. Apesar de o local apresentar arborização com alto índice de diversidade de espécies (Shannon-Weaver de 3,14, os exemplares necessitam de maior atenção quanto a problemas ligados à fitossanidade e podas adequadas, fazendo que resulte em espaço seguro para os frequentadores e em boa qualidade paisagística.

  20. An activity of caspase-3 and cathepsin D at the different subtypes of ischemic stroke

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    Наталія Романівна Сохор

    2015-06-01

    Full Text Available Aim of research – To define the dynamics of activity of caspase-3, cathepsin D, apoptosis of leukocytes at the different subtypes of ischemic stroke (IS in an acute period.Methods. There were examined 232 patients in an acute period of ІS: 56 (24,1%- with hemodynamic (HDS, 62 (2,.7% – with atherothrombotic (АТS, 60 (25,9% – with cardioembolic (CЕS і 54 (23,3% – with lacunar stroke (LS. There was defined the number of leukocytes at the stage of apoptosis (ANV+-cells, necrosis (PI+-cells, with an increased content of the active forms of oxygen (AFO+-cells and with lowered mitochondrial potential (Mito+-cells, activity of caspase-3 and cathepsin D.Results. It was established that at all subtypes of IS mitochondrial dysfunction, apoptosis and necrosis of leukocytes are observed on the 1st day it were presented in increase of content of  ANV+-, PI+-, АFO+- and Mito+-cells and were the mostly apparent at ATS.   The highest activity of caspase-3 on the 1st day was noticed at LS it did not correlate with a number of cells at the stage of apoptosis and probably was connected with a predominant impact of caspase-3 on endothelium and with hyperpermeability of hematoencephalic barrier. In patients with ATS an activity of cathepsin D increased during the 1st week of disease that can indicate an activation of lysosomal way of activation of apoptosis that courses parallel to an apoptosis connected with mitochondrial dysfunction.Conclusions.  The different ways of apoptotic cellular death that depends on subtype of stroke activate in an acute period of IS

  1. Ginsenoside Rg1 Attenuates Isoflurane-induced Caspase-3 Activation via Inhibiting Mitochondrial Dysfunction

    Institute of Scientific and Technical Information of China (English)

    MIAO Hui Hui; ZHEN Yu; DING Guan Nan; HONG Fang Xiao; XIE Zhong Cong; TIAN Ming

    2015-01-01

    Objective The inhalation anesthetic isoflurane has been shown to induce mitochondrial dysfunction and caspase activation, which may lead to learning and memory impairment. Ginsenoside Rg1 is reported to be neuroprotective. We therefore set out to determine whether ginsenoside Rg1 can attenuate isoflurane-induced caspase activation via inhibiting mitochondrial dysfunction. Methods We investigated the effects of ginsenoside Rg1 at concentrations of 12.5, 25, and 50 µmol/L and pretreatment times of 12 h and 24 h on isoflurane-induced caspase-3 activation in H4 naïve and stably transfected H4 human neuroglioma cells that express full-length human amyloid precursor protein (APP) (H4-APP cells). For mitochondrial dysfunction, we assessed mitochondrial permeability transition pore (mPTP) and adenosine-5’-triphosphate (ATP) levels. We employed Western blot analysis, chemiluminescence, and flowcytometry. Results Here we show that pretreatment with 50 µmol/L ginsenoside Rg1 for 12 h attenuated isoflurane-induced caspase-3 activation and mitochondrial dysfunction in H4-APP cells, while pretreatment with 25 and 50 µmol/L ginsenoside Rg1 for 24 h attenuated isoflurane-induced caspase-3 activation and mitochondrial dysfunction in both H4 naïve and H4-APP cells. Conclusion These data suggest that ginsenoside Rg1 may ameliorate isoflurane-induced caspase-3 activation by inhibiting mitochondrial dysfunction. Pending further studies, these findings might recommend the use of ginsenoside Rg1 in preventing and treating isoflurane-induced neurotoxicity.

  2. Avaliações qualitativas e quantitativas de plantas daninhas na cultura da soja submetida aos sistemas de plantio direto e convencional Quantitative and qualitative weed evaluation of soybean crop in no-tillage and conventional tillage systems

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    Eduardo de Sá Pereira

    2000-08-01

    Full Text Available Com o objetivo de avaliar os efeitos do método de preparo do solo e do controle de plantas daninhas em resteva de aveia preta (Avena strigosa, na cultura da soja (Glycine max cv. 'IAC 14', foi conduzido um experimento de campo na Fazenda Experimental Lageado - UNESP Botucatu - SP, em 1993/94. Os tratamentos de manejo do solo foram Plantio Direto e Plantio Convencional (preparado com uma aração e três gradagens. As formas de controle de plantas daninhas estudadas corresponderam a uma testemunha sem controle do mato; controle com uso exclusivo de herbicidas aplicados em préemergência (0,28 kg/ha de metribuzim + 1,29 kg/ha de orizalin; controle com uso exclusivo de herbicidas aplicados em pósemergência (0,25 kg/ha de fluazifop-p-butil + 0,25 kg/ha de fomesafen; controle com o uso conjunto dos tratamentos em pré e pós-emergência mencionados. Tanto no plantio direto quanto no convencional, utilizou-se glyphosate para a eliminação da aveia preta e das plantas daninhas antes da implantação da cultura. O delineamento experimental adotado foi de blocos ao acaso em parcelas subdivididas, com quatro repetições. Os tipos de manejo do solo foram aplicados às parcelas e os métodos de controle das plantas daninhas às sub-parcelas. Deve ser ressaltado que durante a fase inicial do ensaio, as chuvas foram escassas, limitando tanto o crescimento da cultura quanto a atuação dos herbicidas de pré-emergência e pósemergência. As avaliações baseadas na contagem do número de plantas/m2, realizadas aos 14, 28 e 35 dias após a emergência da cultura da soja, indicaram diferenças entre os métodos de preparo do solo e os métodos de controle. Entre as espécies daninhas mais freqüentes, Brachiaria plantaginea e Amaranthus viridis foram as predominantes no sistema de plantio convencional; estas espécies apresentaram pequena importância no plantio direto, no qual predominou Euphorbia heterophylla. No plantio convencional, os herbicidas de

  3. Expression of caspase-3, p53 and Bcl-2 in generalized aggressive periodontitis

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    Özdemir B Handan

    2006-06-01

    Full Text Available Abstract Background Apoptosis, or programmed cell death is a form of physiological cell death. It is increased or decreased in the presence of infection, inflammation or tissue remodelling. Previous studies suggest that apoptosis is involved in the pathogenesis of inflammatory periodontal disease. The aim of the present study was to investigate the clinical features and known indicators of apoptosis (p53, Bcl-2, Caspase-3 in patients with generalized aggressive periodontitis (GAP Methods Eight patients with GAP, who had sites with probing depths (PD > 5 mm, and 10 periodontally-healthy persons were included in the study. Clinical examinations and PD were performed, and the plaque index and gingival index were recorded. Gingival tissues biopsies were obtained from active site of each patient and from healthy individuals. The expression of caspase-3, Bcl-2, and p53 was evaluated by immunohistochemistry Results There were no significant differences between GAP and control group with respect to levels of caspase-3 and p53 expression (P > 0.05. Contrary, the frequency of grade 3 expression of Bcl-2 was higher in GAP group than the control group. Conclusion The higher frequency of Bcl-2 expression in GAP group indicates and delayed apoptosis can lead to increasing resident inflammatory cells in periodontal tissues and resulting in progressive periodontal destruction.

  4. Caspase-3-independent pathways proceeding in bystander effect of HSV-tk/GCV system

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    Lin, Juqiang; Ma, Yan; Zeng, Shaoqun; Zhang, Zhihong

    2008-02-01

    HSV-tk/GCV system, which is the virus-directed enzyme/prodrug therapy of herpes simplex virus (HSV) thymidine kinase (tk) gene / the anti-viral reagent ganciclovir (GCV), is one of the promising approaches in the rapidly growing area of gene therapy. As gene therapy of cancer such as suicide gene therapy has entered the clinic, another therapy effect which is called 'bystander effect' was reported. Bystander effect can lead to killing of non-transduced tumor cells in the immediate vicinity of GCV-treated HSV-TK-positive cells. Now the magnitude of 'bystander effect' is an essential factor for this anti-tumor approach in vivo. However, the mechanism which HSV-tk/ACV brings "bystander effect" is poorly understood. In this study, we monitor the activation of caspase-3 in HSV-tk/GCV system by a FRET probe CD3, a FRET-based indicator for activity of caspase3, which is composed of an enhanced cyan fluorescent protein, a caspase-sensitive linker, and a red fluorescent protein from Discosoma with efficient maturation property. Through application of CD3 we have visualized the activation of caspase-3 in tk gene positive human adenoid cystic carcinoma (ACC-M) cells but not in bystander effect of HSV-tk/GCV system induced by GCV. This finding provides needed information for understanding the mechanisms by which suicide gene approaches actually kill cancer cells, and may prove to be helpful for the clinical treatment of cancers.

  5. STEREOLOGIC ESTIMATION OF KI-67, CASPASE 3 AND GSTP1 POSITIVE CELLS IN PROSTATE LESIONS

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    Luis Santamaría

    2011-05-01

    Full Text Available Cell proliferation, caspase 3 and pi-form of glutathione S transferase (GSTP1 were evaluated in prostate carcinoma (PCA, proliferative inflammatory atrophy (PIA and prostate intraepithelial neoplasia (PIN. Forty biopsies were classified as: without morphological lesions (controls: CTR, PIA, PIN and PCA. Ki67, caspase3 and GSTP1 were immunostained. The following estimates were performed: Numerical densities of Ki67+ cells (NVEPKi67, of all epithelial cells (NVEPtotal and of GSTP1+ cells (NVEPGSTP1; labelling index for Ki67 (LIKi67; volume fraction to caspase 3 positive tissue (VVcaspase 3 and of GSTP1 positive tissue (VVGSTP1. ANOVA was performed to compare the groups. NVEPtotal and NVEPKi67 were increased in PIA. LIKi67 was only increased in PCA. VVcaspase 3 was decreased in PIN and PCA. VVEGSTP1 was decreased in PCA. In our results PIA lacks the characteristics of a premalignant lesion. The result may be explained by the use of unbiased quantitative methods, the inadequate definition of PIA and the scarce inflammation observed in the samples with PIA included in this study.

  6. Avaliação quantitativa da dispensação de contraceptivos de emergência na região de Curitiba, PR, Brasil, entre 2012 e 2014

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    Maria Izabel Cândido OLIVEIRA

    2015-01-01

    Full Text Available A contracepção de emergência é caracterizada pelo uso de um fármaco com administração oral contendo 1,5 mg do Levonorgestrel, um hormônio progestágeno sintético, usado nos casos onde há falha no método contraceptivo rotineiro, na relação sexual desprotegida, não planejada ou em casos de violência sexual. Esse trabalho teve como objetivo principal analisar as taxas de vendas dos Contraceptivos de Emergência (CE, entre os anos de 2012 e 2014, em duas farmácias de dispensação, localizadas no Município de Curitiba (PR, sendo uma no bairro Pinheirinho e outra no bairro Xaxim. Observou-se que o uso dos CE vem aumentando gradativamente ao longo dos anos e que na primavera e no verão, principalmente no período de carnaval, há um pico nas vendas. Em 2014, houve um aumento nas vendas durante a Copa do Mundo realizada no Brasil nos meses de junho e julho. A farmácia do bairro Pinheirinho vendeu uma quantidade maior em relação à do Xaxim, o que pode estar relacionado ao seu tamanho e ao volume geral das vendas, à idade e ao grau de instrução da população, a fatores socioeconômicos e ao Índice de Desenvolvimento Humano (IDH do bairro menor se comparado ao bairro Pinheirinho. Pode-se concluir que o aumento gradativo na venda dos CE observado ao longo dos três anos em ambas as farmácias pode estar relacionado à falta de orientação no momento da dispensação, à escolha de outro método contraceptivo, sensação de falsa imunidade contra as doenças sexualmente transmissíveis (DST, marketing e propagandas utilizadas em épocas festivas e de estímulo à atividade sexual, além de fatores fisiológicos do próprio organismo da mulher

  7. Leaf blade quantitative anatomy of sugarcane cultivars and clones Anatomia quantitativa da lâmina foliar de genótipos de cana-de-açúcar

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    E.A. Ferreira

    2007-03-01

    Full Text Available The objective of this study was to make a quantitative assess of the anatomic characteristics of leaf blade of the sugarcane cultivars RB855113, SP80-1842, SP80-1816, RB867515 and clone RB957689 presenting different sensitivity to the mixture of sodium trifloxysulfuron + ametryn herbicides. Compared to the other cultivars assessed, RB855113 cultivar, considered more sensitive to the herbicide mixture, presented relevant differences such as greater proportion of bulliform cells, greater tissue proportion in the transverse section of the leaf blade, greater stomata and trichome density on both surfaces, thinner epidermis on the adaxial surface and length of stomata on both surfaces. The external paraclinal wall of the bulliform cells was thinner than in the common epidermis cells in all the genotypes on the adaxial and abaxial surfaces. Multivariate analysis of the data on the variables considered most relevant to explain the herbicide penetration singled out the sensitive RB855113 from the other materials. Such characteristics can explain the greater penetration, and consequently, greater sensitivity of this cultivar to the sodium trifloxysulfuron + ametryn mixture.Objetivou-se neste trabalho avaliar, quantitativamente, as características anatômicas da lâmina foliar dos cultivares de cana-de-açúcar RB855113, SP80-1842, SP80-1816 e RB867515 e do clone RB957689 com diferentes sensibilidades à mistura dos herbicidas trifloxysulfuron-sodium + ametryn. O cultivar RB855113, considerado mais sensível à mistura de herbicidas, mostrou diferenças relevantes em relação aos demais cultivares avaliados, como: maior proporção de células buliformes, tecido encontrado em maior proporção na seção transversal da lâmina foliar desse cultivar, maior densidade de estômatos e de tricomas em ambas as faces, epiderme menos espessa na face adaxial e estômatos com maior comprimento de ostíolos nas duas faces. Todos os genótipos apresentaram a parede

  8. Avaliação quantitativa da movimentação parietal regional do ventrículo esquerdo na endomiocardiofibrose Quantitative assessment of left ventricular regional wall motion in endomyocardial fibrosis

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    Charles Mady

    2005-03-01

    Full Text Available OBJETIVO: Analisar a movimentação parietal regional do ventrículo esquerdo (VE em pacientes com endomiocardiofibrose (EMF. MÉTODOS: Estudados 88 pacientes, 59 do sexo feminino, com idade média de 39±13 anos (variação de 9 a 65 com evidência ecocardiográfica e angiográfica de EMF do VE. A intensidade da deposição de tecido fibroso na cineventriculografia contrastada foi classificada como discreta, moderada ou importante. A fração de ejeção global do ventrículo esquerdo (FEVE foi determinada pelo método área-comprimento por meio da ventriculografia. O movimento foi medido em 100 cordas eqüidistantes e perpendiculares à linha média desenhada no meio dos contornos diastólico e sistólico finais e normalizadas para o tamanho cardíaco. Analisaram-se cinco segmentos do VE: A - apical; AL - ântero-lateral; AB - ântero-basal; IA - ínfero-apical; IB - ínfero-basal. A anormalidade foi expressa em unidades de desvio padrão do movimento médio em uma população de referência normal, composta por 103 pacientes com VE normal, conforme dados de clínica, eletrocardiograma e padrões angiográficos. RESULTADOS: A FEVE média foi de 0,47±0,12. O envolvimento de tecido fibroso do VE foi discreto em 12 pacientes, moderado em 40 e importante em 36. As regiões com pior movimentação parietal foram A (-1,4±1,6 desvio-padrão/cordas e IA (-1,6±1,8 desvio-padrão/cordas comparadas com AB (-0,3±1,9 desvio-padrão /cordas, AL (-0,5±1,8 desvio-padrão/cordas e IB (-0,9±1,3 desvio-padrão/cordas. Não se observou relação entre a intensidade de envolvimento do tecido fibroso e a manutenção parietal regional. CONCLUSÃO : Existe alteração da movimentação parietal regional na EMF e é independente da intensidade de deposição de tecido fibroso avaliada qualitativamente. O envolvimento não uniforme do VE deve ser levado em conta no planejamento cirúrgico dessa doença.OBJECTIVE: To analyze left ventricular (LV regional wall

  9. Uterine endoplasmic reticulum stress and its unfolded protein response may regulate caspase 3 activation in the pregnant mouse uterus.

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    Arvind Suresh

    Full Text Available We have previously proposed that uterine caspase-3 may modulate uterine contractility in a gestationally regulated fashion. The objective of this study was to determine the mechanism by which uterine caspase-3 is activated and consequently controlled in the pregnant uterus across gestation. Utilizing the mouse uterus as our gestational model we examined the intrinsic and extrinsic apoptotic signaling pathways and the endoplasmic reticulum stress response as potential activators of uterine caspase-3 at the transcriptional and translational level. Our study revealed robust activation of the uterine myocyte endoplasmic reticulum stress response and its adaptive unfolded protein response during pregnancy coinciding respectively with increased uterine caspase-3 activity and its withdrawal to term. In contrast the intrinsic and extrinsic apoptotic signaling pathways remained inactive across gestation. We speculate that physiological stimuli experienced by the pregnant uterus likely potentiates the uterine myocyte endoplasmic reticulum stress response resulting in elevated caspase-3 activation, which is isolated to the pregnant mouse myometrium. However as term approaches, activation of an elevated adaptive unfolded protein response acts to limit the endoplasmic reticulum stress response inhibiting caspase-3 resulting in its decline towards term. We speculate that these events have the capacity to regulate gestational length in a caspase-3 dependent manner.

  10. Uterine endoplasmic reticulum stress and its unfolded protein response may regulate caspase 3 activation in the pregnant mouse uterus.

    Science.gov (United States)

    Suresh, Arvind; Subedi, Kalpana; Kyathanahalli, Chandrashekara; Jeyasuria, Pancharatnam; Condon, Jennifer C

    2013-01-01

    We have previously proposed that uterine caspase-3 may modulate uterine contractility in a gestationally regulated fashion. The objective of this study was to determine the mechanism by which uterine caspase-3 is activated and consequently controlled in the pregnant uterus across gestation. Utilizing the mouse uterus as our gestational model we examined the intrinsic and extrinsic apoptotic signaling pathways and the endoplasmic reticulum stress response as potential activators of uterine caspase-3 at the transcriptional and translational level. Our study revealed robust activation of the uterine myocyte endoplasmic reticulum stress response and its adaptive unfolded protein response during pregnancy coinciding respectively with increased uterine caspase-3 activity and its withdrawal to term. In contrast the intrinsic and extrinsic apoptotic signaling pathways remained inactive across gestation. We speculate that physiological stimuli experienced by the pregnant uterus likely potentiates the uterine myocyte endoplasmic reticulum stress response resulting in elevated caspase-3 activation, which is isolated to the pregnant mouse myometrium. However as term approaches, activation of an elevated adaptive unfolded protein response acts to limit the endoplasmic reticulum stress response inhibiting caspase-3 resulting in its decline towards term. We speculate that these events have the capacity to regulate gestational length in a caspase-3 dependent manner.

  11. Cytoprotection against Hypoxic and/or MPP+ Injury: Effect of δ–Opioid Receptor Activation on Caspase 3

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    Yuan Xu

    2016-08-01

    Full Text Available The pathological changes of Parkinson’s disease (PD are, at least partially, associated with the dysregulation of PTEN-induced putative kinase 1 (PINK1 and caspase 3. Since hypoxic and neurotoxic insults are underlying causes of PD, and since δ-opioid receptor (DOR is neuroprotective against hypoxic/ischemic insults, we sought to determine whether DOR activation could protect the cells from damage induced by hypoxia and/or MPP+ by regulating PINK1 and caspase 3 expressions. We exposed PC12 cells to either severe hypoxia (0.5%–1% O2 for 24–48 h or to MPP+ at different concentrations (0.5, 1, 2 mM and then detected the levels of PINK1 and cleaved caspase 3. Both hypoxia and MPP+ reduced cell viability, progressively suppressed the expression of PINK1 and increased the cleaved caspase 3. DOR activation using UFP-512, effectively protected the cells from hypoxia and/or MPP+ induced injury, reversed the reduction in PINK1 protein and significantly attenuated the increase in the cleaved caspase 3. On the other hand, the application of DOR antagonist, naltrindole, greatly decreased cell viability and increased cleaved caspase 3. These findings suggest that DOR is cytoprotective against both hypoxia and MPP+ through the regulation of PINK1 and caspase 3 pathways.

  12. Urolitíase em cães: avaliação quantitativa da composição mineral de 156 urólitos Canine urolithiasis: quantitative evaluation of mineral composition of 156 uroliths

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    Mônica Kanashiro Oyafuso

    2010-02-01

    Full Text Available O estudo teve como objetivo avaliar os casos de urolitíase canina em que a composição mineral dos urólitos foi analisada quantitativamente. Foi avaliada quantitativamente a composição mineral de 156 urólitos obtidos de cães (nefrólitos, ureterólitos, urocistólitos e uretrólitos. Desse total, 79,5% (n=124 eram simples, 18% (n=28 eram compostos e apenas 2,5% (n=4 eram mistos. A estruvita foi o tipo mineral mais frequente nos urólitos simples (47,6%; n=59, em todos os mistos (100%; n=4 e nas camadas núcleo e pedra de urólitos compostos (32,1 e 75%, respectivamente. O oxalato de cálcio foi o segundo mineral mais frequente dos urólitos simples (37,9%, n=47. Ao contrário do que é preconizado para os urólitos simples, as recomendações para o tratamento de urólitos compostos são mais complexas, tais como protocolos de tratamento de dissolução diferentes (se composto por minerais distintos e passíveis de dissolução como urato e estruvita. Além disso, a dissolução pode não ser viável, caso ocorra presença de material insolúvel envolvendo o urólito ou se este representar mais de 20% da camada. Vinte e dois urólitos compostos (78,7% apresentaram uma camada externa não passível de dissolução (oxalato de cálcio ou fosfato de cálcio; dois (7,1% apresentaram camadas externas passíveis de dissolução (estruvita ou urato, porém camadas mais internas não solúveis, o que permitiria apenas a dissolução parcial do urólito. Assim, o conhecimento da composição de todas as camadas que compõem o urólito é essencial para o entendimento da formação do cálculo e consequentemente para a indicação do tratamento adequado, assim como para prevenção de recidivas.The aim of this study was to evaluate dogs with urolithiasis in which mineral composition of calculi was quantitatively analyzed. Quantitative mineral composition was performed in 156 canine uroliths. Simple uroliths represented 79.5% (n=124 of the cases, 18

  13. Environmental neurotoxin dieldrin induces apoptosis via caspase-3-dependent proteolytic activation of protein kinase C delta (PKCdelta: Implications for neurodegeneration in Parkinson's disease

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    Kanthasamy Anumantha G

    2008-10-01

    Full Text Available Abstract Background In previous work, we investigated dieldrin cytotoxicity and signaling cell death mechanisms in dopaminergic PC12 cells. Dieldrin has been reported to be one of the environmental factors correlated with Parkinson's disease and may selectively destroy dopaminergic neurons. Methods Here we further investigated dieldrin toxicity in a dopaminergic neuronal cell model of Parkinson's disease, namely N27 cells, using biochemical, immunochemical, and flow cytometric analyses. Results In this study, dieldrin-treated N27 cells underwent a rapid and significant increase in reactive oxygen species followed by cytochrome c release into cytosol. The cytosolic cytochrome c activated caspase-dependent apoptotic pathway and the increased caspase-3 activity was observed following a 3 hr dieldrin exposure in a dose-dependent manner. Furthermore, dieldrin caused the caspase-dependent proteolytic cleavage of protein kinase C delta (PKCδ into 41 kDa catalytic and 38 kDa regulatory subunits in N27 cells as well as in brain slices. PKCδ plays a critical role in executing the apoptotic process in dieldrin-treated dopaminergic neuronal cells because pretreatment with the PKCδ inhibitor rottlerin, or transfection and over-expression of catalytically inactive PKCδK376R, significantly attenuates dieldrin-induced DNA fragmentation and chromatin condensation. Conclusion Together, we conclude that caspase-3-dependent proteolytic activation of PKCδ is a critical event in dieldrin-induced apoptotic cell death in dopaminergic neuronal cells.

  14. Environmental neurotoxin dieldrin induces apoptosis via caspase-3-dependent proteolytic activation of protein kinase C delta (PKCdelta): Implications for neurodegeneration in Parkinson's disease.

    Science.gov (United States)

    Kanthasamy, Anumantha G; Kitazawa, Masashi; Yang, Yongjie; Anantharam, Vellareddy; Kanthasamy, Arthi

    2008-10-22

    In previous work, we investigated dieldrin cytotoxicity and signaling cell death mechanisms in dopaminergic PC12 cells. Dieldrin has been reported to be one of the environmental factors correlated with Parkinson's disease and may selectively destroy dopaminergic neurons. Here we further investigated dieldrin toxicity in a dopaminergic neuronal cell model of Parkinson's disease, namely N27 cells, using biochemical, immunochemical, and flow cytometric analyses. In this study, dieldrin-treated N27 cells underwent a rapid and significant increase in reactive oxygen species followed by cytochrome c release into cytosol. The cytosolic cytochrome c activated caspase-dependent apoptotic pathway and the increased caspase-3 activity was observed following a 3 hr dieldrin exposure in a dose-dependent manner. Furthermore, dieldrin caused the caspase-dependent proteolytic cleavage of protein kinase C delta (PKCδ) into 41 kDa catalytic and 38 kDa regulatory subunits in N27 cells as well as in brain slices. PKCδ plays a critical role in executing the apoptotic process in dieldrin-treated dopaminergic neuronal cells because pretreatment with the PKCδ inhibitor rottlerin, or transfection and over-expression of catalytically inactive PKCδ(K)³⁷⁶(R), significantly attenuates dieldrin-induced DNA fragmentation and chromatin condensation. Together, we conclude that caspase-3-dependent proteolytic activation of PKCδ is a critical event in dieldrin-induced apoptotic cell death in dopaminergic neuronal cells.

  15. Análise quantitativa e molecular de hemoglobina fetal em indivíduos da população brasileira Quantitative and molecular analysis of fetal hemoglobin in individuals from the Brazilian population

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    Paula J. A. Zamaro

    2003-01-01

    Full Text Available A hemoglobina fetal - Hb F, formada por duas cadeias gama e duas cadeias alfa, é característica do período fetal do desenvolvimento, tendo sua síntese diminuída no período pós-natal. Em algumas alterações hereditárias, a Hb F permanece aumentada, como nas delta-beta talassemia, beta talassemia e persistência hereditária de Hb F (PHHF. A síntese da globina gama também pode ser estimulada por fatores externos como leucemias, transplantes de medula óssea, induções químicas, dentre outros. Através da observação de Hb F aumentada em doadores de sangue por procedimentos eletroforéticos objetivou-se avaliar a quantidade de Hb F em amostras de sangue de candidatos à doação, visando estabelecer seus limites de normalidade na população de São José do Rio Preto e região, por meio de desnaturação alcalina e cromatografia líquida de alta pressão (HPLC, comparar as metodologias aplicadas e, nos indivíduos com Hb F aumentada, realizar estudos moleculares para identificar as mutações que alteram a expressão dos genes gama. Foram analisadas 208 amostras de sangue, sendo 119 de candidatos à doação e 89 de indivíduos sem sintomas de anemia ou achados hematológicos e com Hb F aumentada como grupo comparativo. Das 119 amostras de candidatos à doação, 110 foram utilizadas para traçar o perfil de normalidade de Hb F, comparando-se as metodologias de desnaturação alcalina e HPLC, onde se obteve a média de 1,48% e de 0,6%, respectivamente. A análise estatística por regressão linear mostrou diferença significativa na comparação entre as duas metodologias aplicadas, sendo a HPLC mais precisa para a quantificação de Hb F. Foram observados nos testes de rastreamento de hemoglobinas anormais nestas 110 amostras de sangue: 16,4% de alfa talassemia, 0,9% com Hb F aumentada, 0,9% com beta talassemia e 0,9% com hemoglobina variante de cadeia delta. Os outros nove doadores de sangue apresentaram Hb F acima de 10% em

  16. Effects of Berberine on the Expression of Caspase-3 Gene in Human Cervical Cancer Hela Cell%小檗碱对宫颈癌Hela细胞Caspase-3基因表达的影响

    Institute of Scientific and Technical Information of China (English)

    张丽萍; 张志军; 程萍; 肖劲松

    2010-01-01

    日的:研究小檗碱(berberlne)对人宫颈癌Hela细胞体外增殖、凋亡及凋亡相关基因Caspase-3表达的影响.方法:WIT法测定不同浓度(0~40 μmol/L)berberine干预Hela细胞后的凋亡率;RT-PCR检测Caspase-3 mRNA表达水平.结果:berberine呈剂量-时间依赖方式抑制Hela细胞的生长(P<0.01);且随着berberine浓度增高,细胞凋亡增加,Caspase-3 mRNA表达上调(P<0.001).结论:berberine抑制人宫颈癌Hela细胞生长、诱导凋亡的机制可能与上调Caspase-3 mRNA表达有关.

  17. Caspase-3在大鼠中枢神经系统的表达研究%Study on the expression of caspase-3 in central nervous system of rat

    Institute of Scientific and Technical Information of China (English)

    陈雪梅; 杜显刚; 官鹏; 谭志巍; 王亚琴

    2005-01-01

    目的研究caspase-3在大鼠中枢神经系统的表达及其意义.方法用western-blot方法对出生1天和3个月SD大鼠的大脑皮质、中脑和小脑组织的caspase-3进行半定量测定.结果出生1天大鼠脑的caspase-3表达较高,出生3个月大鼠脑的caspase-3表达较低.结论caspase-3在中枢神经系统的发育成熟过程中对神经元的凋亡起着关键性作用.

  18. A Crohn's disease variant in Atg16l1 enhances its degradation by caspase 3

    Science.gov (United States)

    Murthy, Aditya; Li, Yun; Peng, Ivan; Reichelt, Mike; Katakam, Anand Kumar; Noubade, Rajkumar; Roose-Girma, Merone; Devoss, Jason; Diehl, Lauri; Graham, Robert R.; van Lookeren Campagne, Menno

    2014-02-01

    Crohn's disease is a debilitating inflammatory bowel disease (IBD) that can involve the entire digestive tract. A single-nucleotide polymorphism (SNP) encoding a missense variant in the autophagy gene ATG16L1 (rs2241880, Thr300Ala) is strongly associated with the incidence of Crohn's disease. Numerous studies have demonstrated the effect of ATG16L1 deletion or deficiency; however, the molecular consequences of the Thr300Ala (T300A) variant remains unknown. Here we show that amino acids 296-299 constitute a caspase cleavage motif in ATG16L1 and that the T300A variant (T316A in mice) significantly increases ATG16L1 sensitization to caspase-3-mediated processing. We observed that death-receptor activation or starvation-induced metabolic stress in human and murine macrophages increased degradation of the T300A or T316A variants of ATG16L1, respectively, resulting in diminished autophagy. Knock-in mice harbouring the T316A variant showed defective clearance of the ileal pathogen Yersinia enterocolitica and an elevated inflammatory cytokine response. In turn, deletion of the caspase-3-encoding gene, Casp3, or elimination of the caspase cleavage site by site-directed mutagenesis rescued starvation-induced autophagy and pathogen clearance, respectively. These findings demonstrate that caspase 3 activation in the presence of a common risk allele leads to accelerated degradation of ATG16L1, placing cellular stress, apoptotic stimuli and impaired autophagy in a unified pathway that predisposes to Crohn's disease.

  19. Brain caspase-3 and intestinal FABP responses in preterm and term rats submitted to birth asphyxia

    Directory of Open Access Journals (Sweden)

    R.L. Figueira

    2016-01-01

    Full Text Available Neonatal asphyxia can cause irreversible injury of multiple organs resulting in hypoxic-ischemic encephalopathy and necrotizing enterocolitis (NEC. This injury is dependent on time, severity, and gestational age, once the preterm babies need ventilator support. Our aim was to assess the different brain and intestinal effects of ischemia and reperfusion in neonate rats after birth anoxia and mechanical ventilation. Preterm and term neonates were divided into 8 subgroups (n=12/group: 1 preterm control (PTC, 2 preterm ventilated (PTV, 3 preterm asphyxiated (PTA, 4 preterm asphyxiated and ventilated (PTAV, 5 term control (TC, 6 term ventilated (TV, 7 term asphyxiated (TA, and 8 term asphyxiated and ventilated (TAV. We measured body, brain, and intestine weights and respective ratios [(BW, (BrW, (IW, (BrW/BW and (IW/BW]. Histology analysis and damage grading were performed in the brain (cortex/hippocampus and intestine (jejunum/ileum tissues, as well as immunohistochemistry analysis for caspase-3 and intestinal fatty acid-binding protein (I-FABP. IW was lower in the TA than in the other terms (P<0.05, and the IW/BW ratio was lower in the TA than in the TAV (P<0.005. PTA, PTAV and TA presented high levels of brain damage. In histological intestinal analysis, PTAV and TAV had higher scores than the other groups. Caspase-3 was higher in PTAV (cortex and TA (cortex/hippocampus (P<0.005. I-FABP was higher in PTAV (P<0.005 and TA (ileum (P<0.05. I-FABP expression was increased in PTAV subgroup (P<0.0001. Brain and intestinal responses in neonatal rats caused by neonatal asphyxia, with or without mechanical ventilation, varied with gestational age, with increased expression of caspase-3 and I-FABP biomarkers.

  20. TNF-α Contributes to Caspase-3 Independent Apoptosis in Neuroblastoma Cells: Role of NFAT

    Science.gov (United States)

    Álvarez, Susana; Blanco, Almudena; Fresno, Manuel; Muñoz-Fernández, Ma Ángeles

    2011-01-01

    There is increasing evidence that soluble factors in inflammatory central nervous system diseases not only regulate the inflammatory process but also directly influence electrophysiological membrane properties of neurons and astrocytes. In this context, the cytokine TNF-α (tumor necrosis factor-α) has complex injury promoting, as well as protective, effects on neuronal viability. Up-regulated TNF-α expression has also been found in various neurodegenerative diseases such as cerebral malaria, AIDS dementia, Alzheimer's disease, multiple sclerosis, and stroke, suggesting a potential pathogenic role of TNF-α in these diseases as well. We used the neuroblastoma cells SK-N-MC. Transcriptional activity was measured using luciferase reporter gene assays by using lipofectin. We performed cotransfection experiments of NFAT (nuclear factor of activated T cells) promoter constructed with a dominant negative version of NFAT (dn-NFAT). Cell death was performed by MTT (3-(4,5-dimethylthiazol-2-yl)5,5-diphenyltetrazolium bromide) and TUNEL assays. NFAT translocation was confirmed by Western blot. Involvement of NFAT in cell death was assessed by using VIVIT. P53, Fas-L, caspase-3, and caspase-9 expressions were carried out by Western blot. The mechanisms involved in TNF-α-induced cell death were assessed by using microarray analysis. TNF-α causes neuronal cell death in the absence of glia. TNF-α treatment results in nuclear translocation of NFAT through activation of calcineurin in a Ca2+ independent manner. We demonstrated the involvement of FasL/Fas, cytochrome c, and caspase-9 but the lack of caspase-3 activation. NB cell death was absolutely reverted in the presence of VIVIT, and partially diminished by anti-Fas treatment. These data demonstrate that TNF-α promotes FasL expression through NFAT activation in neuroblastoma cells and this event leads to increased apoptosis through independent caspase-3 activation. PMID:21298033

  1. Densitometria da vértebra dorsal, osso pleural e osso neural em tartarugas verdes hígidas por tomografia computadorizada quantitativa Densitometry of dorsal vertebrae, pleural bone and neural bone in healthy green sea turtles by quantitative computed tomography

    Directory of Open Access Journals (Sweden)

    Jannine Forattini de Oliveira

    2012-08-01

    Full Text Available A tomografia computadorizada quantitativa (TCQ é uma técnica com alta sensibilidade, aplicabilidade e precisão no diagnóstico de perda de massa óssea, entretanto a falta de valores de normalidade para animais silvestres limita a sua aplicação clínica. O presente estudo objetiva estabelecer valores médios de radiodensidade do osso trabecular das vértebras dorsais, osso pleural e osso neural de tartarugas marinhas juvenis hígidas utilizando a TCQ. Para isso, utilizou-se um grupo experimental de cinco animais que foram submetidos a exames de tomografia computadorizada helicoidal, sendo as mensurações da radiodensidade óssea realizadas ao nível da terceira, quarta e quinta vértebras dorsais. Os valores médios de radiodensidade do osso trabecular das vértebras dorsais, osso pleural e osso neural foram, respectivamente, de 287,4±39,1, 1093,74±100,4 e 1089,6±86,5 unidades Hounsfield (HU. Estudos complementares devem ser realizados em diferentes espécies e faixas etárias de quelônios para favorecer a aplicação clínica desta técnica.Quantitative computed tomography (QCT is a technique with high sensitivity, applicability and accuracy in the diagnosis of bone loss; however the lack of normal values for wild animals limits their clinical application. This study aims to establish medium values radiodensity trabecular bone of the dorsal vertebrae, bone pleural and neural bone in healthy juvenile sea turtles using the QTC. For this we used an experimental group of five animals that underwent helical computed tomography, and measurements of bone radiodensity conducted at the level of the third, fourth and fifth dorsal vertebrae. The medium values of radiodensity trabecular bone of the dorsal vertebrae, pleural bone, and neural bone were respectively 287.4±39.1, 1093.74±100.4, and 1089.6±86.5 Hounsfield units (HU. Further studies should be performed in different species and ages of turtles to encourage the clinical application of

  2. Evaluation of Bcl-2, Bcl-x and Cleaved Caspase-3 in Malignant Peripheral Nerve Sheath Tumors and Neurofibromas

    Directory of Open Access Journals (Sweden)

    KARIN S. CUNHA

    2013-11-01

    Full Text Available AIMS: To study the expression of Bcl-2, Bcl-x, as well the presence of cleaved caspase-3 in neurofibromas and malignant peripheral nerve sheath tumors. The expression of Bcl-2 and Bcl-x and the presence of cleaved caspase 3 were compared to clinicopathological features of malignant peripheral nerve sheath tumors and their impact on survival rates were also investigated. MATERIALS AND METHODS: The evaluation of Bcl-2, Bcl-x and cleaved caspase-3 was performed by immunohistochemistry using tissue microarrays in 28 malignant peripheral nerve sheath tumors and 38 neurofibromas. Immunoquantification was performed by computerized digital image analysis. CONCLUSIONS: Apoptosis is altered in neurofibromas and mainly in malignant peripheral nerve sheath tumors. High levels of cleaved caspase-3 are more common in tumors with more aggressive histological features and it is associated with lower disease free survival of patients with malignant peripheral nerve sheath tumors.

  3. Razões entre componentes da variabilidade de características quantitativas simuladas com efeitos genéticos de dominância e sobredominância Ratios between variability components of simulated quantitative traits with genetic effects of dominance and overdominance

    Directory of Open Access Journals (Sweden)

    Elizângela Emídio Cunha

    2009-10-01

    Full Text Available Foram avaliadas as razões entre componentes da variabilidade de características quantitativas simuladas a partir de genoma incorporando efeitos genéticos não-aditivos em populações de acasalamento ao acaso e de seleção fenotípica a curto prazo. Estudaram-se uma característica de baixa (h² = 0,10 e outra de alta herdabilidade (h² = 0,60 influenciadas por 600 locos bialélicos. Cinco modelos de ação gênica foram simulados, dos quais quatro incluíram dominância completa e positiva para 25, 50, 75 e 100% dos locos (D25, D50, D75 e D100, respectivamente; e um modelo incluiu sobredominância positiva para 50% dos locos. Todos os modelos incluíram efeitos aditivos dos alelos para 100% dos locos. As principais razões quantificadas foram d² (variância de dominância/variância fenotípica e d²a (variância de dominância/variância aditiva. Para as duas características, d² e d²a aumentaram de acordo com o acréscimo na variância de dominância, decorrente da inclusão crescente de locos com desvio da dominância e sob sobredominância. No mesmo modelo, ambas as razões, sobretudo d², são mais elevadas sob alta herdabilidade, o que indica que os efeitos da dominância explicam a maior parte da variabilidade total dessa característica sob seleção.Ratios were assessed between variability components of quantitative traits simulated from the genome incorporating non-additive genetic effects in random mating populations and short-term phenotypic selection. A trait of low (h² = 0.10 heritability and another of high (h² = 0.60 heritability were studied, both influenced by 600 bi-allelic loci. Five gene action models were simulated, of which four included complete and positive dominance for 25, 50, 75 and 100% of the loci (D25, D50, D75 and D100, respectively; and one model included positive overdominance for 50% of the loci. Every model included additive effects of the alleles for 100% of the loci. The main quantified ratios were

  4. Effects of vitamin C on pathology and caspase-3 activity of kidneys with subacute endosulfan toxicity.

    Science.gov (United States)

    Ozmen, O; Mor, F

    2015-01-01

    Endosulfan is an insecticide that is composed of two stereoisomers: α- and β- endosulfan in an approximate ratio of 70:30. Owing to its widespread use, poisoning of both humans and animals is possible. We examined the toxic effects of endosulfan on New Zealand white rabbit kidneys. Rabbit kidneys were examined histopathologically and caspase-3 activity was detected using immunohistochemistry. Animals were divided into four groups: Group 1 was given a sublethal dose of endosulfan in corn oil by oral gavage daily for 6 weeks, Group 2 was given endosulfan + vitamin C during the same period, Group 3 was given corn oil daily and vitamin C on alternate days, Group 4 was given only corn oil daily throughout the experiment. By the end of experimental period, the concentration of α-endosulfan was greater than the β-endosulfan concentration in the kidneys of both of endosulfan treated groups (Groups 1 and 2). Decreased accumulation of α- and β-endosulfan was observed in Group 2, possibly because of the antioxidant effect of the vitamin C. Histopathological examination revealed hemorrhages, tubule cell necrosis, glomerular infiltration, glomerulosclerosis and proteinaceous material in the tubules, and Bowman spaces in the kidneys of Group 1. Caspase-3 reaction was stronger in Group 1 than in the other groups. Apoptotic activity was most frequent in proximal tubule cells. Endosulfan is toxic to rabbit kidneys. Vitamin C treatment reduced the accumulation of endosulfan in kidneys and reduced its toxicity.

  5. Large-scale preparation of active caspase-3 in E. coli by designing its thrombin-activatable precursors

    Directory of Open Access Journals (Sweden)

    Park Sung

    2008-12-01

    Full Text Available Abstract Background Caspase-3, a principal apoptotic effector that cleaves the majority of cellular substrates, is an important medicinal target for the treatment of cancers and neurodegenerative diseases. Large amounts of the protein are required for drug discovery research. However, previous efforts to express the full-length caspase-3 gene in E. coli have been unsuccessful. Results Overproducers of thrombin-activatable full-length caspase-3 precursors were prepared by engineering the auto-activation sites of caspase-3 precursor into a sequence susceptible to thrombin hydrolysis. The engineered precursors were highly expressed as soluble proteins in E. coli and easily purified by affinity chromatography, to levels of 10–15 mg from 1 L of E. coli culture, and readily activated by thrombin digestion. Kinetic evaluation disclosed that thrombin digestion enhanced catalytic activity (kcat/KM of the precursor proteins by two orders of magnitude. Conclusion A novel method for a large-scale preparation of active caspase-3 was developed by a strategic engineering to lack auto-activation during expression with amino acid sequences susceptible to thrombin, facilitating high-level expression in E. coli. The precursor protein was easily purified and activated through specific cleavage at the engineered sites by thrombin, generating active caspase-3 in high yields.

  6. Immunoexpression of cleaved caspase-3 shows lower apoptotic area indices in lip carcinomas than in intraoral cancer

    Science.gov (United States)

    LEITE, Ana Flávia Schueler de Assumpção; BERNARDO, Vagner Gonçalves; BUEXM, Luisa Aguirre; da FONSECA, Eliene Carvalho; da SILVA, Licínio Esmeraldo; BARROSO, Danielle Resende Camisasca; LOURENÇO, Simone de Queiroz Chaves

    2016-01-01

    ABSTRACT Objective This study aimed to evaluate apoptosis by assessing cleaved caspase-3 immunoexpression in hyperplastic, potentially malignant disorder (PMD), and malignant tumors in intraoral and lower lip sites. Material and Methods A retrospective study using paraffin blocks with tissues from patients with inflammatory fibrous hyperplasia (IFH), actinic cheilitis, oral leukoplakia, lower lip and intraoral squamous cell carcinoma (SCC) was performed. The tissues were evaluated by immunohistochemical analysis with anti-cleaved caspase-3 antibody. Apoptotic area index was then correlated with lesion type. Results From 120 lesions assessed, 55 (46%) were cleaved caspase-3-positive. The SCC samples (n=40) had the highest apoptotic area indices (n=35; 87.5%). Significant differences were detected between SCCs and PMDs (p=0.0003), as well as SCCs and IFHs (p=0.001), regarding caspase-3 immunopositivity. Carcinomas of the lower lip had lower apoptotic area indices than intraoral cancer (p=0.0015). Conclusions Cleaved caspase-3 immunoexpression showed differences in oral SCCs and PMDs and demonstrated a distinct role of apoptosis in carcinogenesis of intraoral and lower lip cancer. In future, the expression of cleaved caspase-3 with other target molecules in oral cancer may be helpful in delineating the prognosis and treatment of these tumors. PMID:27556207

  7. Analysis of expression patterns of IGF-1, caspase-3 and HSP-70 in developing human tooth germs.

    Science.gov (United States)

    Kero, Darko; Kalibovic Govorko, Danijela; Medvedec Mikic, Ivana; Vukojevic, Katarina; Cigic, Livia; Saraga-Babic, Mirna

    2015-10-01

    To analyze expression patterns of IGF-1, caspase-3 and HSP-70 in human incisor and canine tooth germs during the late bud, cap and bell stages of odontogenesis. Head areas or parts of jaw containing teeth from 10 human fetuses aged between 9th and 20th developmental weeks were immunohistochemically analyzed using IGF-1, active caspase-3 and HSP-70 markers. Semi-quantitative analysis of each marker's expression pattern was also performed. During the analyzed period, IGF-1 and HSP-70 were mostly expressed in enamel organ. As development progressed, expression of IGF-1 and HSP-70 became more confined to differentiating tissues in the future cusp tip area, as well as in highly proliferating cervical loops. Few apoptotic bodies highly positive to active caspase-3 were observed in enamel organ and dental papilla from the cap stage onward. However, both enamel epithelia moderately expressed active caspase-3 throughout the investigated period. Expression patterns of IGF-1, active caspase-3 and HSP-70 imply importance of these factors for early human tooth development. IGF-1 and HSP-70 have versatile functions in control of proliferation, differentiation and anti-apoptotic protection of epithelial parts of human enamel organ. Active caspase-3 is partially involved in formation and apoptotic removal of primary enamel knot, although present findings might reflect its ability to perform other non-death functions such as differentiation of hard dental tissues secreting cells and guidance of ingrowth of proliferating cervical loops. Copyright © 2015 Elsevier Ltd. All rights reserved.

  8. Caspase-3-dependent cleavage of Akt modulates tau phosphorylation via GSK3β kinase: implications for Alzheimer's disease.

    Science.gov (United States)

    Chu, J; Lauretti, E; Praticò, D

    2017-01-31

    The pathological hallmark of Alzheimer's disease (AD) is accumulation of misfolded amyloid-β peptides and hyperphosphorylated tau protein in the brain. Increasing evidence suggests that serine-aspartyl proteases-caspases are activated in the AD brain. Previous studies identified a caspase-3 cleavage site within the amyloid-β precursor protein, and a caspase-3 cleavage of tau as the mechanisms involved in the development of Aβ and tau neuropathology, respectively. However, the potential role that caspase-3 could have on tau metabolism remains unknown. In the current studies, we provide experimental evidence that caspase-3 directly and specifically regulates tau phosphorylation, and demonstrate that this effect is mediated by the GSK3β kinase pathway via a caspase-3-dependent cleavage of the protein kinase B (also known as Akt). In addition, we confirm these results in vivo by using a transgenic mouse model of AD. Collectively, our findings demonstrate a new role for caspase-3 in the neurobiology of tau, and suggest that therapeutic strategies aimed at inhibiting this protease-dependent cleavage of Akt may prove beneficial in preventing tau hyperphosphorylation and subsequent neuropathology in AD and related tauopathies.Molecular Psychiatry advance online publication, 31 January 2017; doi:10.1038/mp.2016.214.

  9. Caspase-3 and its inhibitor Ac-DEVD-CHO in rat lens epithelial cell apoptosis induced by hydrogen in vitro

    Institute of Scientific and Technical Information of China (English)

    姚克; 王凯军; 徐雯; 孙朝晖; 申屠形超; 邱培瑾

    2003-01-01

    Objective To investigate the role of caspase-3 and its inhibitor Ac-DEVD-CHO in rat lens epithelial cell apoptosis induced by hydrogen peroxide (H2O2) in vitro.Methods Rat lenses were incubated in modified Eagle' s medium containing 2 mmol/L H2O2 to induce apoptosis in vitro. Apoptosis in lens epithelial cells was assessed by transmission electron microscopy and annexin V-propidium iodide (PI) double staining flow cytometry after 12, 24 and 48 h of incubation. The activity of caspase-3 was analyzed by western blotting.Results Observations under transmission electron microscopy revealed that 2 mmol/L H2O2 could effectively induce lens epithelial cell apoptosis in vitro. Caspase-3 activity increased during cell apoptosis and the peak measurement occurred at 24 h after treatment with H2O2. Cell apoptosis was blocked by caspase-3 inhibitor Ac-DEVD-CHO.Conclusions The activation of caspase-3 plays an important role in executing apoptosis in H2O2-treated lens epithelial cells and in the formation of cataract. The caspase-3 inhibitor Ac-DEVD-CHO may effectively prevent lens epithelial cell apoptosis caused by oxidative injury.

  10. Cooperation of bisphenol A and leptin in inhibition of caspase-3 expression and activity in OVCAR-3 ovarian cancer cells.

    Science.gov (United States)

    Ptak, Anna; Rak-Mardyła, Agnieszka; Gregoraszczuk, Ewa L

    2013-09-01

    This study was designed to investigate the effect of bisphenol A and leptin on caspase-3 expression and activity in OVCAR-3 ovarian cancer cells. Caspase-3 and survivin expression was measured at the transcript level by real-time PCR and at the protein level by Western blotting. In addition, caspase-3 activity was measured, using a fluorometric assay, upon exposure to bisphenol A (40 nM) alone, leptin (2.5 nM) alone, and the combination of both agents. 17β-estradiol (40 nM) was used as a positive control for estrogenic properties of bisphenol A. Results showed that the interaction between bisphenol A and leptin, which was similar to that observed between 17β-estradiol and leptin, led to the inhibition of caspase-3 expression and activity in OVCAR-3 cells. Surprisingly, survivin was found to not be involved in the anti-apoptotic activity of either agent. Also, results showed that leptin inhibits caspase-3 activity by acting on the signal transducers and activators of transcription 3 (STAT3) pathway, but bisphenol A and 17β-estradiol by the extracellular-signal-regulated kinases 1/2 (ERK1/2) pathway. In conclusion, the study reveals that bisphenol A and leptin interact to inhibit caspase-3 expression and activity by modulating STAT3 and ERK1/2 signaling pathways in OVCAR-3 cells.

  11. Study on Caspase-3 activity on trichloroethylene-induced human keratinocyte apoptosis%三氯乙烯诱导人角质形成细胞凋亡中Caspase-3活力的研究

    Institute of Scientific and Technical Information of China (English)

    汪立杰; 叶良平; 沈彤; 朱启星

    2009-01-01

    目的 观察三氯乙烯(TCE)诱导离体培养的人角质形成细胞(KC)Caspase-3活力变化及细胞凋亡情况,探讨TCE诱导KC凋亡的可能信号通路.方法 以不同浓度(0.125、0.250、0.500、1.000、2.000 mmol/L)TCE对离体分离培养的KC分别染毒至4、8、12、24 h;Caspase-3抑制剂(Z-DEVD-FMK)预处理组,先用100 μmol/L Z-DEVD-FMK预处理细胞1 h,然后再用2.000 mmol/L TCE染毒12 h.用分光光度法检测细胞Caspase-3活力变化,借助Annexin-V/PI双染和流式细胞仪检测细胞凋亡情况.结果 与空白对照相比,TCE染毒4 h,各TCE剂量组Caspase-3活力无明显变化(P>0.05);染毒8 h,1.000 mmol/LTCE组Caspase-3活力和2.000 mmol/LTCE组Caspase-3活力,与对照组相比差异有显著性(P0.01).结论 在TCE诱导离体培养的KC凋亡中,Caspase-3的活化可能发挥了重要的作用.

  12. Expression and significance of caspase-3 in CD34+ cord blood cells%Caspase-3在脐血CD34+造血干/祖细胞中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    马艳萍; 邹萍; 肖娟; 黄士昂

    2002-01-01

    目的:探讨脐血CD34+ 干/祖细胞在不同细胞因子支持下的体外扩增过程中Caspase-3表达及意义.方法:采用RT-PCR、Wester blot和流式细胞仪分析技术测定脐血CD34+ 细胞在体外扩增过程中的生物学特性及Caspase-3的表达.结果:Caspase-3 mRNA在新鲜分离的脐血CD34+ 细胞中低水平表达,在细胞因子支持下体外培养3 d,扩增的CD34+ 细胞中Caspase-3 mRNA和蛋白质表达上调,但在该两种细胞中仅能检测到分子量为32 000的无活性酶原形式的Caspase-3,随着体外培养时间的延长,在IL-3、IL-6和GM-CSF组合条件下,Caspase-3被激活,可检测到分子量为20 000的裂解片段.结论:虽然造血干细胞的凋亡是个复杂的过程,但在脐血CD34+ 干/祖细胞体外扩增过程中,Caspase-3参与了凋亡事件并发挥着重要的作用.

  13. The effect of aloe emodin-encapsulated nanoliposome-mediated r-caspase-3 gene transfection and photodynamic therapy on human gastric cancer cells.

    Science.gov (United States)

    Li, Kai-Ting; Duan, Qin-Qin; Chen, Qing; He, Juan-Wen; Tian, Si; Lin, Hai-Dan; Gao, Qing; Bai, Ding-Qun

    2016-02-01

    Gastric carcinoma (GC) has high incidence and mortality rates in China. Surgery and chemotherapy are the main treatments. Photodynamic therapy (PDT) has become a new treatment modality, appearing in recent experimental studies and clinical trials in various tumors. This study explores the combined effect of gene transfection with PDT on GC cells using aloe emodin (AE)-encapsulated nanoliposomes, which acted as gene carrier as well as one photosensitizer (PS). AE-encapsulated nanoliposomes (nano-AE) were prepared by reverse evaporation method. Electron microscopy and nano-ZS90 analyzer were used to detect its morphology, size, and wavelength. Western blot was used to detect the expression of the caspase-3 after transfection. MTT assay and flow cytometry were employed to determine the cytotoxic and apoptotic rates, respectively. Hoechst 33342 staining was adopted to detect the morphological changes in death gastric cancer cells. Cellular reactive oxygen species (ROS) contents were measured by DCFH-DA staining. Outcomes demonstrated that the nano-AE has good properties as gene delivery carriers as well as a PS. The group in which the recombinant plasmid of r-caspase-3 was transfected had higher protein expression of the caspase-3 than controls, meanwhile the proliferation rates of the transfected cells were inhibited by the nano-AE-mediated PDT in an energy-dependent manner. In addition, in the transfected cells, the death rate increased to 77.3% as assessed 12 h after PDT (6.4 J/cm(2) ). Hochest 33342 staining also revealed that the death rate increased significantly in the transfected group compared with other groups. Compared to control groups, the production of ROS in nano-AE PDT group had quadrupled in SGC-7901 cells as early as 1 h after PDT, while it is similar to the group of nano-AE transfection and PDT. Nano-AE-mediated r-caspase-3 gene transfection coupled with PDT could inhibit the proliferation rate and increase the apoptotic rate remarkably in human

  14. Quantitative analysis by energy dispersive X-ray fluorescence by the transmission method applied to geological samples Análise quantitativa por fluorescência de raios X com dispersão por energia pelo método da transmissão aplicada à amostras geológicas

    Directory of Open Access Journals (Sweden)

    S.M. Simabuco

    1994-08-01

    Full Text Available Three certified samples of different matrices (Soil-5, SL-1/IAEA and SARM-4/SABS were quantitatively analysed by energy dispersive X-ray fluorescence with radioisotopic excitation. The observed errors were about 10-20% for the majority of the elements and less than 10% for Fe and Zn in the Soil-5, Mn in SL-1, and Ti, Fe and Zn in SARM-4 samples. Annular radioactive sources of Fe-55 and Cd-109 were utilized for the excitation of elements while a Si(Li semiconductor detector coupled to a multichannel emulation card inserted in a microcomputer was used for the detection of the characteristic X-rays. The fundamental parameters method was used for the determination of elemental sensitivities and the irradiator or transmission method for the correction of the absorption effect of characteristic X-rays of elements on the range of atomic number 22 to 42 (Ti to Mo and excitation with Cd-109. For elements in the range of atomic number 13 to 23 (Al to V the irradiator method cannot be applied since samples are not transparent for the incident and emergent X-rays. In order to perform the absorption correction for this range of atomic number excited with Fe-55 source, another method was developed based on the experimental value of the absorption coefficients, associated with absorption edges of the elements.Foi realizada a análise quantitativa de três amostras certificadas de diferentes matrizes (Soil-5/IAEA, SL-1/IAEA e SARM-4/SABS através da fluorescência de raios X por dispersão de energia e excitação radioisotópica. Os erros observados foram em torno de 10 a 20% para a maioria dos elementos e inferiores a 10% para Fe e Zn na amostra Soil-5, Mn na amostra SL-1 e Ti, Fe e Zn na amostra SARM-4. Para a excitação dos elementos presentes foram empregadas as fontes radioativas anelares de Fe-55 e Cd-109, enquanto que na detecção dos raios X característicos utilizou-se um detector de Si(Li, acoplado a uma placa analisadora de pulsos multicanal

  15. Empenamento de frangos de corte: efeito da restrição alimentar qualitativa e quantitativa e temperatura ambiente - DOI: 10.4025/actascianimsci.v27i3.1211 Feathering of broiler chickens: effect of feed restriction and room temperature - DOI: 10.4025/actascianimsci.v27i3.1211

    Directory of Open Access Journals (Sweden)

    Alex Maiorka

    2005-03-01

    Full Text Available Estudou-se a restrição alimentar sobre o empenamento de frangos criados em diferentes temperaturas. Para restrição qualitativa, experimento 1, utilizou-se 900 pintainhos, Cobb-500, em delineamento inteiramente ao acaso, em esquema fatorial 3 x 3 (programas alimentares:Controle – 2.950 kcal EM/kg e 22% de PB; Restrição Protéica – 2.950 kcal EM/kg e 17% de PB; Restrição Energética – 2.500 kcal EM/kg e 22% de PB e temperaturas: fria, termoneutra e quente, totalizando 9 tratamentos com 5 repetições. A diminuição de energia e proteína reduziram o empenamento. Na restrição quantitativa, experimento 2, utilizou-se 1.080 pintainhos distribuídos em um delineamento ao acaso em esquema fatorial 3 (níveis de restrição: 0%, 40% e 70% x 3 (temperatura ambiente: fria, termoneutra e quente, totalizando 9 tratamentos com 5 repetições. O empenamento foi prejudicado pela restrição de 70%. O empenamento é sensível à variação protéica e energética da dieta e à temperatura ambienteThe effect of feed restriction on feathering was studied in broilers raised under different temperatures. For qualitative feed restriction trial, experiment 1, 900 Cobb-500 chicks were used in a completely randomized design in 3 x 3 factorial scheme (3 feed programs: Control – 2,950 kcal EM.kg-1 and 22% of CP; Protein Restriction – 2,950 kcal EM.kg-1 and 17% of CP; Energy Restriction – 2,500 kcal EM.kg-1 and 22% of CP; and 3 temperatures: cold, neutral and hot with 9 treatments and 5 repetitions. Energy and Protein restriction reduced feathering. In quantitative restriction, experiment 2, 1080 chicks were distributed in a completely randomized design in 3 x 3 factorial scheme (3 restriction levels: 0%, 40% and 70%; and 3 room temperatures: cold, neutral and hot with 9 treatments and 5 repetitions. Feathering was damaged by 70% of feed restriction. The feathering seems to feel considerably the variations in diet protein and energy and at room

  16. The cyclin-dependent kinase inhibitor roscovitine and the nucleoside analog sangivamycin induce apoptosis in caspase-3 deficient breast cancer cells independent of caspase mediated P-glycoprotein cleavage

    Science.gov (United States)

    Cappellini, Alessandra; Chiarini, Francesca; Ognibene, Andrea; McCubrey, James A.; Martelli, Alberto M.

    2009-01-01

    Resistance to multiple chemotherapeutic agents is a common clinical problem which can arise during cancer treatment. Drug resistance often involves overexpression of the multidrug resistance MDR1 gene, encoding P-glycoprotein (P-gp), a 170-kDa glycoprotein belonging to the ATP-binding cassette superfamily of membrane transporters. We have recently demonstrated apoptosis-induced, caspase-3-dependent P-gp cleavage in human T-lymphoblastoid CEM-R VBL100 cells. However, P-gp contain many aspartate residues which could be targeted by caspases other than caspase-3. To test whether other caspases could cleave P-gp in vivo, we investigated the fate of P-gp during roscovitine- and sangivamycin- induced apoptosis in MCF7 human breast cancer cells, as they lack functional caspase-3. MCF7 cells were stably transfected with human cDNA encoding P-gp. P-gp was cleaved in vitro by purified recombinant caspase-3, -6 and -7. However, P-gp cleavage was not detected in vivo in MCF7 cells induced to undergoing apoptosis by either roscovitine or sangivamycin, despite activation of both caspase-6 and -7. Interestingly, P-gp overexpressing MCF7 cells were more sensitive to either roscovitine or sangivamycin than wild-type cells, suggesting a novel potential therapeutic strategy against P-gp overexpressing cells. Taken together, our results support the concept that caspase-3 is the only caspase responsible for in vivo cleavage of P-gp and also highlight small molecules which could be effective in treating P-gp overexpressing cancers. PMID:19342873

  17. 口腔白斑及口腔鳞状细胞癌中Livin和Caspase-3蛋白表达研究%Expression of Livin and Caspase- 3 protein in oral leukoplakia and oral squamous cell carcinoma

    Institute of Scientific and Technical Information of China (English)

    随丽娜

    2011-01-01

    Objective Study the expressions of Livin and caspase -3 proteins in the normal oral mucosa, oral leukoplakia and oral squamous cell carcinoma. Methods Expression of Livin and caspase- 3 protein were detected by immunohistochemistry S -P methods in the normal oral mucosa,oral leukoplakia and oral squamous cell carcinoma. Results Positive rates of Livin protein in the normal oral mucosa, oral leukoplakia and oral squamous cell carcinoma were 20.00%, 33.33% and 86.49%; Positive rates of caspase-3 protein in the normal oral mucosa, oral leukoplakia and oral squamous cell carcinoma were 100%, 83.33% and 43.24%. The expression of caspase -3 protein was obviously negative related to livin protein (P < 0.05 ). Conclusions Deletion of Caspase - 3 protein and abnormal expression of survivin protein were related to OLK cancer, and early detection of them can help to identify OLK cancer.%目的 研究Livin和caspase-3蛋白在正常口腔黏膜、口腔白斑和口腔鳞状细胞癌中的表达.方法 免疫组化SP法检测正常口腔黏膜、口腔白斑和口腔鳞状细胞癌中Livin和easpase-3蛋白的表达.结果 Livin蛋白在正常口腔黏膜、口腔白斑和口腔鳞状细胞癌中的表达率为20%、33.33%和86.49%;caspase-3蛋白在正常口腔黏膜、口腔白斑和口腔鳞状细胞癌中的表达率为100%、83.33%和43.24%;caspase-3蛋白表达与iivin蛋白表达有明显的负相关关系(P<0.05).结论 Caspase-3的缺失和Livin的高表达与口腔白斑癌变的发生发展相关,两者联合检测有助于早期识别口腔白斑癌变.

  18. Imaging of caspase-3 activation by a novel FRET probe composed of CFP and DsRed

    Science.gov (United States)

    Lin, Juquiang; Zhang, Zhihong; Liu, Bifeng; Luo, Qingming

    2006-01-01

    Caspases-3 is a kind of cysteine proteases and plays an important role in cell apoptosis. It has been reported that caspase-3 activation can be real-time detected in living cells by fluorescence resonance energy transfer (FRET) between an enhanced cyan fluorescent protein and enhanced yellow fluorescent protein. However, the large spectral overlap between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) emission and the highly sensitivity to pH of YFP restricted their detecting sensitivity and reliability. CFP and red fluorescent protein (DsRed) possess superb wavelength separation of donor and acceptor emission spectra and DsRed was insensitive to pH, so the FRET probe composed of CFP and DsRed would be more suitable for imaging caspase-3 activation than the FRET probe composed of CFP and YFP. We constructed a vector that encoded CRS (caspase-3 recognition site) fused with CFP and DsRed (CFP-CRS-DsRed). In CFP-CRS-DsRed expressing tumor cells, FRET from CFP to DsRed could be detected. In the Clinical applications of cancer chemotherapy, cisplatin is one of the most broadly used drugs. It was already confirmed that caspase-3 was activated in HeLa cell treated by cisplatin. When the cells were stimulated with cisplatin, we found that the FRET efficient was remarkably decreased and then disappeared. It indicated that actived caspase-3 cleaved the CFP-CRS-DsRed fusion protein at CRS site. Thus, the FRET probe of CFP-CRS-DsRed could sensitively and reliably monitor caspase-3 activation in living cell. This probe will be highly useful for rapid-screening potential drugs that may target the apoptotic process and for imaging tumors in vivo.

  19. Erythrocyte caspase-3 activation and oxidative imbalance in erythrocytes and in plasma of type 2 diabetic patients.

    Science.gov (United States)

    Maellaro, Emilia; Leoncini, Silvia; Moretti, Daniele; Del Bello, Barbara; Tanganelli, Italo; De Felice, Claudio; Ciccoli, Lucia

    2013-08-01

    An increased oxidative stress and a decreased life span of erythrocytes (RBCs) are reported in patients with diabetes. Aim of this study was to assess in RBCs from patients with type 2 diabetes whether downstream effector mechanisms of apoptosis, such as activation of caspase-3, is operative, and whether an iron-related oxidative imbalance, occurring inside RBCs and in plasma, could be involved in caspase-3 activation. In 26 patients with type 2 diabetes and in 12 healthy subjects, oxidative stress was evaluated by means of different markers; non-protein-bound iron, methemoglobin and glutathione were determined in RBCs, and non-protein-bound iron was also determined in plasma. Erythrocyte caspase-3 activation was evaluated by an immunosorbent enzyme assay. Arterial hypertension, demographic and standard biochemical data were also evaluated. The results show, for the first time, that type 2 diabetic RBCs put into motion caspase-3 activation, which is significantly higher than in control RBCs. Such an effector mechanism of "eryptosis" was positively correlated to blood glucose levels and to the increased plasma NPBI level. Caspase-3 activation was also positively correlated to occurrence of arterial hypertension. The results suggest that an extracellular oxidative milieu can be responsible for erythrocyte caspase-3 activation in patients with type 2 diabetes. In turn, caspase-3 activation can be envisaged as a novel mechanism which, by impairing the maintenance of erythrocyte shape and function, might contribute to the shortened life span of RBCs from patients with type 2 diabetes and to hemorheological disorders observed in these patients.

  20. Quercetin accumulation by chronic administration causes the caspase-3 activation in liver and brain of mice.

    Science.gov (United States)

    Choi, Eun Jeong; Kim, Gun-Hee

    2010-01-01

    Quercetin is an excellent antioxidant that has a variety of side effects. This study investigated whether the chronic administration of quercetin in mice induces apoptosis. Mice were divided randomly into three treatment groups. Quercetin was administered orally to two of three groups at 100 and 250 mg/kg body weight (BW) for 18 days. The serum quercetin level increased in a dose-dependent manner, although the quercetin levels in the liver and brain were lower than in serum. Nevertheless, quercetin induced apoptosis in both the liver and brain, as evidenced by increased caspase-3 expression and activity. Quercetin-induced apoptosis seems to be associated with quercetin accumulation. Moreover, with quercetin accumulation, the brain was more susceptible to apoptosis than the liver. In conclusion, quercetin administration at a high dose may lead to apoptosis in the liver and brain of mouse.

  1. BAG3 protein regulates caspase-3 activation in HIV-1-infected human primary microglial cells

    Science.gov (United States)

    Rosati, Alessandra; Khalili, Kamel; Deshmane, Satish L.; Radhakrishnan, Sujatha; Pascale, Maria; Turco, M. Caterina; Marzullo, Liberato

    2015-01-01

    BAG3, a member of the BAG co-chaperones family, is expressed in several cell types subjected to stressful conditions, such as exposure to high temperature, heavy metals, drugs. Furthermore, it is constitutively expressed in some tumors. Among the biological activities of the protein, there is apoptosis downmodulation; this appears to be exerted through BAG3 interaction with the heat shock protein (Hsp) 70, that influences cell apoptosis at several levels. We recently reported that BAG3 protein was detectable in the cytoplasm of reactive astrocytes in HIV-1-associated encephalopathy biopsies. Here we report that downmodulation of BAG3 protein levels allows caspase-3 activation by HIV-1 infection in human primary microglial cells. This is the first reported evidence of a role for BAG3 in the balance of death versus survival during viral infection. PMID:18821563

  2. Spermine triggers the activation of caspase-3 in a cell-free model of apoptosis.

    Science.gov (United States)

    Stefanelli, C; Bonavita, F; Stanic', I; Pignatti, C; Flamigni, F; Guarnieri, C; Caldarera, C M

    1999-05-21

    Polyamines are ubiquitous organic cations required for cell proliferation. However, some evidence suggested that their excessive accumulation can induce apoptosis. We show here that, in a post-nuclear extract from U937 cells, the addition of spermine triggers the death program, represented by cytochrome c exit from mitochondria, the dATP-dependent processing of pro-caspase-3 and the onset of caspase activity. Spermine is more effective than spermidine, whereas putrescine has no effect. Polyamine acetylation abolishes their pro-apoptotic power. These data demonstrate a direct mechanism responsible for polyamine toxicity and also suggest that an excessive elevation of free polyamines could be involved in the transduction of a death signal.

  3. Caspase-3-mediated degradation of condensin Cap-H regulates mitotic cell death.

    Science.gov (United States)

    Lai, S-K; Wong, C-H; Lee, Y-P; Li, H-Y

    2011-06-01

    Mitotic death is a major form of cell death in cancer cells that have been treated with chemotherapeutic drugs. However, the mechanisms underlying this form of cell death is poorly understood. Here, we report that the loss of chromosome integrity is an important determinant of mitotic death. During prolonged mitotic arrest, caspase-3 is activated and it cleaves Cap-H, a subunit of condensin I. The depletion of Cap-H results in the loss of condensin I complex at the chromosomes, thus affecting the integrity of the chromosomes. Consequently, DNA fragmentation by caspase-activated DNase is facilitated, thus driving the cell towards mitotic death. By expressing a caspase-resistant form of Cap-H, mitotic death is abrogated and the cells are able to reenter interphase after a long mitotic delay. Taken together, we provide new insights into the molecular events that occur during mitotic death.

  4. Grape Seed Proanthocyanidins Induce Apoptosis through p53, Bax, and Caspase 3 Pathways

    Directory of Open Access Journals (Sweden)

    Anshu M. Roy

    2005-01-01

    Full Text Available Grape seed proanthocyanidins (GSP have been shown to inhibit skin chemical carcinogenesis and photocarcinogenesis in mice. The mechanisms responsible for the anticarcinogenic effects of GSP are not clearly understood. Here, we report that treatment of JB6 C141 cells (a well-developed cell culture model for studying tumor promotion in keratinocytes and p53+/+ fibroblasts with GSP resulted in a dose-dependent induction of apoptosis. GSP-induced (20–80 g/ml apoptosis was observed by using immunofluorescence (27–90% apoptosis and flow cytometry (18–87% apoptosis. The induction of apoptosis by GSP was p53-dependent because it occurred mainly in cells expressing wild-type p53 (p53+/+; 15–80% to a much greater extent than in p53-deficient cells (p53-/-; 6–20%. GSP-induced apoptosis in JB6 C141 cells was associated with increased expression of the tumorsuppressor protein, p53, and its phosphorylation at Ser15. The antiapoptotic proteins, Bcl-2 and Bcl-xl, were downregulated by GSP, whereas the expression of the pro-apoptotic protein, Bax, and the levels of cytochrome c release, Apaf-1, caspase-9, and cleaved caspase 3 (p19 and p17 were markedly increased in JB6 C141 cells. The downregulation of Bcl-2 and upregulation of Bax were also observed in wild-type p53 (p53+/+ fibroblasts but was not observed in their p53-deficient counterparts. These data clearly demonstrate that GSP-induced apoptosis is p53-dependent and mediated through the Bcl-2, Bax, and caspase 3 pathways.

  5. Grape Seed Proanthocyanidins Induce Apoptosis through p53, Bax, and Caspase 3 Pathways1

    Science.gov (United States)

    Roy, Anshu M; Baliga, Manjeshwar S; Elmets, Craig A; Katiyar, Santosh K

    2005-01-01

    Abstract Grape seed proanthocyanidins (GSP) have been shown to inhibit skin chemical carcinogenesis and photocarcinogenesis in mice. The mechanisms responsible for the anticarcinogenic effects of GSP are not clearly understood. Here, we report that treatment of JB6 C141 cells (a well-developed cell culture model for studying tumor promotion in keratinocytes) and p53+/+ fibroblasts with GSP resulted in a dose-dependent induction of apoptosis. GSP-induced (20–80 g/ml) apoptosis was observed by using immunofluorescence (27–90% apoptosis) and flow cytometry (18–87% apoptosis). The induction of apoptosis by GSP was p53-dependent because it occurred mainly in cells expressing wild-type p53 (p53+/+; 15–80%) to a much greater extent than in p53-deficient cells (p53-/-; 6–20%). GSP-induced apoptosis in JB6 C141 cells was associated with increased expression of the tumor-suppressor protein, p53, and its phosphorylation at Ser15. The antiapoptotic proteins, Bcl-2 and Bcl-xl, were downregulated by GSP, whereas the expression of the pro-apoptotic protein, Bax, and the levels of cytochrome c release, Apaf-1, caspase-9, and cleaved caspase 3 (p19 and p17) were markedly increased in JB6 C141 cells. The downregulation of Bcl-2 and upregulation of Bax were also observed in wild-type p53 (p53+/+) fibroblasts but was not observed in their p53-deficient counterparts. These data clearly demonstrate that GSP-induced apoptosis is p53-dependent and mediated through the Bcl-2, Bax, and caspase 3 pathways. PMID:15720815

  6. Nascent histamine induces α-synuclein and caspase-3 on human cells

    Energy Technology Data Exchange (ETDEWEB)

    Caro-Astorga, Joaquín; Fajardo, Ignacio; Ruiz-Pérez, María Victoria; Sánchez-Jiménez, Francisca; Urdiales, José Luis, E-mail: jlurdial@uma.es

    2014-09-05

    Highlights: • Nascent histamine alters cyclin expression pattern. • Nascent histamine increases expression of α-synuclein. • Nascent histamine activates caspase-3. - Abstract: Histamine (Hia) is the most multifunctional biogenic amine. It is synthetized by histidine decarboxylase (HDC) in a reduced set of mammalian cell types. Mast cells and histaminergic neurons store Hia in specialized organelles until the amine is extruded by exocytosis; however, other immune and cancer cells are able to produce but not store Hia. The intracellular effects of Hia are still not well characterized, in spite of its physiopathological relevance. Multiple functional relationships exist among Hia metabolism/signaling elements and those of other biogenic amines, including growth-related polyamines. Previously, we obtained the first insights for an inhibitory effect of newly synthetized Hia on both growth-related polyamine biosynthesis and cell cycle progression of non-fully differentiated mammalian cells. In this work, we describe progress in this line. HEK293 cells were transfected to express active and inactive versions of GFP-human HDC fusion proteins and, after cell sorting by flow cytometry, the relative expression of a large number of proteins associated with cell signaling were measured using an antibody microarray. Experimental results were analyzed in terms of protein–protein and functional interaction networks. Expression of active HDC induced a cell cycle arrest through the alteration of the levels of several proteins such as cyclin D1, cdk6, cdk7 and cyclin A. Regulation of α-synuclein and caspase-3 was also observed. The analyses provide new clues on the molecular mechanisms underlying the regulatory effects of intracellular newly synthetized Hia on cell proliferation/survival, cell trafficking and protein turnover. This information is especially interesting for emergent and orphan immune and neuroinflammatory diseases.

  7. Lens fiber connexin turnover and caspase-3-mediated cleavage are regulated alternately by phosphorylation.

    Science.gov (United States)

    Yin, Xinye; Liu, Jialu; Jiang, Jean X

    2008-05-01

    Lens connexins are phosphorylated in vivo; however, the function and regulation of the phosphorylation remain largely unknown. We have previously identified an in vivo phosphorylation site, Ser(364), at the COOH terminus of lens connexin (Cx) Cx45.6 and phosphorylation appears to regulate connexin protein turnover. To assess the specific mechanism of Ser(364) phosphorylation in Cx45.6, exogenous wild type and Ser(364) mutant Cx45.6 were expressed in primary lens cultures through retroviral infection. Cx45.6 turnover was attenuated primarily by proteasomal inhibitors and to a lesser extent by lysosomal inhibitors. Furthermore, the level of Cx45.6 protein in ubiquitin co-expressed cells was significantly reduced as compared to the cells expressing Cx45.6 alone. Moreover, overexpression of ubiquitin led to a more significant decrease in wild type Cx45.6 than Cx45.6(S364A), a mutant deficient of phosphorylation site at Ser(364), although we did not detect any difference in the levels of ubiquitination between wild type and mutant Cx45.6. Interestingly, the mutant mimicking constitutive phosphorylation, Cx45.6(S364D), partially prevented the cleavage of Cx45.6 by caspase-3. Together, our data suggest that phosphorylation of Cx45.6 at Ser(364) appears to stimulate Cx45.6 turnover primarily through proteasome pathway and this phosphorylation inhibits the cleavage of Cx45.6 by caspase-3. These findings provide further insights into regulatory mechanism of the specific phosphorylation of connexins in the lens.

  8. 脑缺血再灌注时半暗带calpain和caspase-3的相互作用

    Institute of Scientific and Technical Information of China (English)

    孙明; 赵育梅; 徐超

    2006-01-01

    calpain和caspase-3都属于半胱氨酸蛋白酶家族。calpain主要参与兴奋毒性引起的神经细胞死亡,caspase-3主要参与细胞的凋亡。一些研究提示calpain和caspase-3之间存在功能上的关联。本研究探讨了大鼠大脑中动脉阻断1h后再灌注期间半暗带内calpain和caspase-3的相互作用。在缺血前15min经脑室给予calpain抑制剂1或caspase-3抑制剂z—DEVD—CHO。在再灌注3h和23h时留取半暗带组织,采用酪蛋白酶谱法、荧光法和免疫印迹法分别测定胞浆部分μ-和m—calpain的活性、caspase-3的活性以及calpastatin(内源性calpain抑制剂)、微管相关蛋白-2(MAP-2)和血影蛋白的含量。结果和结论如下:(1)在再灌注早期(3h)和晚期(23h),半暗带内μ-和m—calpain及caspase-3的活性明显升高,MAP-2和血影蛋白的含量明显降低,calpain和caspase-3抑制剂均可明显升高MAP-2血影蛋白的含量,表明局灶性脑缺血再灌注早期和晚期半暗带内calpain及caspase-3可被同时活化,继而水解细胞骨架蛋白;(2)在再灌注早期和晚期,半暗带内calpastatin的蛋白含量明显升高,提示局灶性脑缺血再灌注可诱导calpastatin蛋白的表达;

  9. The expression and significance of Smac、 XIAP、 caspase-3 in nonasal inverted papilloma%Smac、XIAP、caspase-3在鼻腔鼻窦内翻性乳头状瘤中的表达及意义

    Institute of Scientific and Technical Information of China (English)

    杨莉晖; 单春光; 黄红梅; 许秋荣; 赵颖; 孟雅静; 张志红

    2012-01-01

    目的:研究Smac、XIAP、caspase-3在鼻腔鼻窦内翻性乳头状瘤(NIP)的发生、发展及癌变过程中的表达及意义.方法:选取正常鼻腔黏膜(NM)10例,NIP 45例.其中的NIP组根据不同的病理分级分为3组,即无不典型增生组(25例)、伴有不典型增生组(11例)、恶变组(9例),通过免疫组织化学SP法进行Smac、XIAP、caspase-3的检测.结果:Smac、Caspase-3在NIP组中的阳性表达强度弱于在NM组中的表达,在NIP不同病理分级中,阳性表达强度随病理分化程度的降低而降低,无不典型增生组及恶变组间的表达差异有统计学意义.XIAP在NM、NIP组中的阳性表达强度呈增强趋势,在NIP不同病理分级中,组织的分化程度越低,阳性表达强度越高,无不典型增生组及恶变组间的表达具有统计学意义.Smac与XIAP的表达为负相关(rs=-0.323,P<0.05),XIAP与caspase-3的表达负相关(rs=-0.408,P<0.01),Smac与caspase-3的表达为正相关(rs=0.424,P<0.01).结论:Smac、XIAP、caspase-3与NIP的发病及恶变有关.%Objective: To explore the expression and significance of second mitochondria derived activator of caspase(Smac) ,X-linked inhibitor of apoptosis protein(XIAP)and cysteine containing aspartate specific protease 3 (caspase-3)in the growth,development and carcinogenesis of the nonasal inverted papilloma(NIP). Method:lmmu-nohistochemical method was used to detect the expression of Smac,XIAP, caspase-3 in 10 cases of nasal cavity mu-cosae(NM)and 45 cases of NIP, the group of NIP including 25 cases of NIP without dysplasia, 11 cases of NIP with dysplasia, and 9 cases of NIP with malignant transformation to squamous cell carcinoma (SCO. Result: The intensity of the positive expression of Smac., Caspase-3 in NIP were lower than NM, the intensity of the positive expression decreased with the decreasing degree of histological differentiation. There was a significant difference between NIP without dysplasia and SCC. It was presented

  10. Implication of Caspase-3 as a Common Therapeutic Target for Multineurodegenerative Disorders and Its Inhibition Using Nonpeptidyl Natural Compounds

    Directory of Open Access Journals (Sweden)

    Saif Khan

    2015-01-01

    Full Text Available Caspase-3 has been identified as a key mediator of neuronal apoptosis. The present study identifies caspase-3 as a common player involved in the regulation of multineurodegenerative disorders, namely, Alzheimer’s disease (AD, Parkinson’s disease (PD, Huntington’s disease (HD, and amyotrophic lateral sclerosis (ALS. The protein interaction network prepared using STRING database provides a strong evidence of caspase-3 interactions with the metabolic cascade of the said multineurodegenerative disorders, thus characterizing it as a potential therapeutic target for multiple neurodegenerative disorders. In silico molecular docking of selected nonpeptidyl natural compounds against caspase-3 exposed potent leads against this common therapeutic target. Rosmarinic acid and curcumin proved to be the most promising ligands (leads mimicking the inhibitory action of peptidyl inhibitors with the highest Gold fitness scores 57.38 and 53.51, respectively. These results were in close agreement with the fitness score predicted using X-score, a consensus based scoring function to calculate the binding affinity. Nonpeptidyl inhibitors of caspase-3 identified in the present study expeditiously mimic the inhibitory action of the previously identified peptidyl inhibitors. Since, nonpeptidyl inhibitors are preferred drug candidates, hence, discovery of natural compounds as nonpeptidyl inhibitors is a significant transition towards feasible drug development for neurodegenerative disorders.

  11. Role of caspase-3/E-cadherin in helicobacter pylori-induced apoptosis of gastric epithelial cells.

    Science.gov (United States)

    Yang, Yongmei; Du, Jie; Liu, Fen; Wang, Xiaoyan; Li, Xiaohui; Li, Yuanjian

    2017-08-29

    This study was designed to investigate the role of caspase-3/E-cadherin in Helicobacter pylori (H. pylori) -induced gastric epithelial apoptosis in cells, animal models and clinical gastritis patients. In cultured gastric mucosal epithelial cells, gastric glandular epithelial cells and C57BL/6 mice, H. pylori infection significantly induced apoptosis of gastric epithelial cells, down-regulated full-length E-cadherin and Bcl-2 expression, and up-regulated cleaved-caspase-3, E-cadherin/carboxy-terminal fragment 3 and Bax expression. Z-DEVD-FMK, an inhibitor of caspase-3, attenuated the effect of H. pylori. E-cadherin overexpression significantly inhibited the apoptosis of GES-1 and SGC-7901 cells induced by the H. pylori. The results from clinical studies also showed down-regulation of E-cadherin, up-regulation of cleaved-caspase-3 expression and increased apoptosis in gastric tissues from gastritis patients with H. pylori infection. These results suggest that the caspase-3/E-cadherin pathway is involved in the apoptosis of gastric epithelial cells induced by H. pylori.

  12. Expression of caspase-3 and -9 relevant to cartilage destruction and chondrocyte apoptosis in human osteoarthritic cartilage.

    Directory of Open Access Journals (Sweden)

    Matsuo M

    2001-12-01

    Full Text Available To clarify the involvement of the caspase family in the pathway of NO-induced chondrocyte apoptosis, osteoarthritis (OA cartilage obtained from 8 patients undergoing total hip arthroplasty were used for histopathological study. Cartilage samples taken from non-fibrillated areas of femoral head resected during surgery for femoral neck fracture were used for comparison. DNA fragmentation of chondrocytes was detected by the nick end-labeling (TUNEL method. Apoptosis was further confirmed by transmission electron microscopy. The distributions of nitrotyrosine (NT, caspase-3, and -9 were examined immunohistochemically. The populations of apoptotic as well as NT-, caspase-3-, and -9-positive cells were quantified by counting the number of cells in the superficial, middle, and deep layers, respectively. The TUNEL-positive cells were observed primarily in superficial proliferating chondrocytes, clustering chondrocytes, and deep-layer chondrocytes of OA cartilage. Few positive cells were seen in the proliferating chondrocytes in the middle layer. Positive reactions for caspase-3 and -9 were observed in chondrocytes in similar areas. Histological OA grade showed significant correlations with the mean populations of apoptotic chondrocytes (% apoptosis over the 3 areas. The populations of NT-positive cells (% NT over the same areas also showed significant correlation with OA grade. Positivity for caspase-3 closely correlated with the OA grade, % apoptosis and %NT. It was concluded that caspase-3 and -9 could play a role in NO-induced chondrocyte apoptosis in OA cartilage.

  13. Post-ischemic administration of progesterone reduces caspase-3 activation and DNA fragmentation in the hippocampus following global cerebral ischemia.

    Science.gov (United States)

    Espinosa-García, Claudia; Vigueras-Villaseñor, Rosa María; Rojas-Castañeda, Julio César; Aguilar-Hernández, Alejandra; Monfil, Tomas; Cervantes, Miguel; Moralí, Gabriela

    2013-08-29

    Delayed death of hippocampal CA1 pyramidal neurons following global cerebral ischemia/reperfusion may be mediated, in part, by caspase-3 activation resulting in DNA fragmentation. Progesterone (P4) is known to exert neuroprotective effects in several models of brain injury. This study was designed to assess the effect of P4 on caspase-3 levels and activation, and DNA fragmentation in the hippocampus following global cerebral ischemia/reperfusion. Adult male Sprague-Dawley rats were subjected to global ischemia by the four-vessel occlusion model. P4 (8 mg/kg), or its vehicle were administered i.v. at 15 min, 2, 6, 24, 48 and 70 h of reperfusion. Remaining pyramidal neurons were assesed by the Nissl staining technique, caspase-3 levels and activation by immunohistochemistry and an in situ activity assay, and DNA fragmentation by the TUNEL method. Post-ischemic progesterone treatment significantly reduced the ischemia/reperfusion-induced increase in caspase-3 levels and activation at 72 h, and DNA fragmentation and CA1 neuronal loss at 7 days. Present results suggest the reduction of caspase-3 levels/activation, and DNA fragmentation, as a part of the neuroprotective effects of progesterone against global cerebral ischemia/reperfusion injury. Copyright © 2013 Elsevier Ireland Ltd. All rights reserved.

  14. Interference of Caspase-3 shRNA on neural cells apoptosis induced by aluminum%Caspase-3 shRNA对染铝神经细胞凋亡的干预

    Institute of Scientific and Technical Information of China (English)

    张勤丽; 教霞; 徐丽; 李娜; 牛侨

    2012-01-01

    目的 通过体外实验来研究铝的神经毒性,探讨慢病毒载体Caspase-3 RNA于扰对染铝神经细胞凋亡的干预效果.方法 原代培养神经细胞,用AlCl3·6H2O染毒,选用病毒为载体的Caspase-3 shRNA试剂感染神经细胞,荧光标记感染成功的细胞计算神经细胞的感染效率,荧光定量PCR技术检测干预后Caspase-3基因的表达情况计算慢病毒载体的Caspase-3 shRNA的抑制率.光学显微镜、荧光染色观察神经细胞的形态学变化,并检测细胞活力;AnnexinV-PI双染法检测神经细胞感染后的凋亡坏死率.结果 慢病毒载体的Caspase-3 RNA干扰试剂的感染效率大于90%,抑制率为66.47%.用慢病毒载体的Caspase-3 shRNA感染原代培养神经细胞后可见染铝神经细胞活力显著上升(P<0.01);吖啶橙(AO)-溴化乙啶(EB)双荧光染色法可以清楚地观察到染铝细胞的早期凋亡和晚期凋亡现象明显减少;Annexin V-PI双染法检测结果显示,感染后染铝神经细胞凋亡率显著降低(P<0.01).结论 馒病毒载体的Caspase-3shRNA感染细胞后,可以显著抑制Caspase-3基因的表达,神经细胞活力明显增高,凋亡细胞的数量显著降低.%Objective To study the neurotoxicity of aluminum ( Al) , and explore the intervention effect of Caspase-3 shRNA on Al-induced apoptosis. Methods Primary cultured nerve cells were exposed to AlCl3 -6H2O, Caspase-3 shRNAs were infected in the cells. Fluorescent labeled shRNAs were used to label the successfully infected cells, and infectious rates were calculated. Interference rate of Caspase-3 shRNAs on Caspase-3 gene expression was calculated with QRT-PCR method. Morphologic characteristics of Caspase-3 shRNAs treated neural cells were observed under light microscope, and fluorescent microscope stained with AO-EB staining, the alternations of apoptosis rates were analyzed by cytometry. Results The infectious rate of Caspase-3 shRNA was above 90% , the inhibition rate was 66

  15. Características quantitativas da carcaça de bubalinos de três grupos genéticos terminados em confinamento e abatidos em diferentes estádios de maturidade Quantitative carcass traits of buffaloes from three genetic groups finished in feedlot and slaughtered at different maturities

    Directory of Open Access Journals (Sweden)

    André Mendes Jorge

    2005-12-01

    Full Text Available Objetivou-se estudar as características quantitativas da carcaça de 36 bubalinos (12 Murrah 12 Jafarabadi e 12 Mediterrâneo, com idade média de 18 meses e peso vivo inicial de 330 kg, terminados em confinamento. Os 12 animais de cada grupo genético foram divididos aleatoriamente em três subgrupos de quatro animais e submetidos aos seguintes tratamentos: Maturidade 1: 400 kg PV ao abate; Maturidade 2: 450 kg PV ao abate; Maturidade 3: 500 kg PV ao abate. Durante o experimento, uma ração única, em que 50% da MS foi composta por volumoso, foi fornecida, ad libitum, para todos os animais. Após os abates pré-fixados, determinou-se o peso corporal vazio (PCVZ dos animais pelo somatório das partes integrantes do corpo. Não houve diferença entre grupos genéticos e maturidades, em relação à porcentagem dos cortes dianteiro, paleta, traseiro total e alcatra completa. O rendimento de traseiro especial foi maior nos animais abatidos aos 400 kg PV e menor naqueles com 500 kg PV, enquanto o dos animais com 450 kg PV não diferiu dos demais. O rendimento de ponta-de-agulha, por sua vez, foi maior nos animais com 500 kg PV e menor nos com 400 kg PV. Os valores observados nesses dois pesos de abate não diferiram do obtido nos animais com 450 kg. A produção de carne a partir das raças Murrah, Jafarabadi e Mediterrâneo criadas no Brasil não difere quanto aos rendimentos de carcaça, traseiro, dianteiro e dos principais cortes básicos de interesse comercial.This work was conducted to study the quantitative carcass traits of 36 buffaloes (12 Murrah - MUR, 12 Jafarabadi - JAF and 12 Mediterranean - MED averaging 18 months old and initial body weight (BW of 330 kg finished in feedlot. Twelve animals of each genetic group were randomly assigned to three sub-groups (four animals and distributed to the following treatments: Maturity 1 - slaughter weight at 400 kg of BW, Maturity 2 - slaughter weight at 450 kg of BW, or Maturity 3 - slaughter

  16. Expression and Significance of Bcl-2, Bax, Fas and Caspase-3 in Different Phases of Human Hemangioma

    Institute of Scientific and Technical Information of China (English)

    YANG Hong; DENG Chenguo; SHEN Shengguo; ZHANG Duanlian; YUYing

    2006-01-01

    The relationship between Bcl-2, Bax, Fas, caspase-3 and development of hemangioma and the molecular mechanism was investigated. By using immunohistochemical S-P method, proliferating cell nuclear antigen was detected. According to the classification of Mulliken in combination with PCNA expression, 27 cases were identified as proliferating hemangioma and 22 cases as involutive hemangioma. Five normal skin tissues around the tumor tissue served as controls. By using immunohistochemical technique, the expression of Bcl-2, Bax, Fax and Caspase-3 was detected. The cells expressing Bcl-2, Bax, Fax and cappase-3 were identified as hemangioma endothelia by immunohistochemical staining of Ⅷ factor. The average absorbance (A) and average positive area rate of Bcl-2, Bax, Fas and caspase-3 expression were measured by using HPIAS-2000 imaging analysis system. The results showed that the expression of Bcl-2 in the endothelia of proliferating hemangioma was significantly higher that in involutive degenerative hemangioma endothelia and vascular endothelia of normal skin tissue (P<0.01). The expression of Bax, Fas and Caspase-3 in the endothelia of involutive hemangioma was obviously higher than in the endothelia of proliferating hemangioma and normal skin tissue (P<0.01). The expression of BAx and Fas in endothelia of proliferating hemangioma was higher than in those of normal skin tissue (P<0.05). It was suggested that Bcl-2,Bax, Fas and caspase-3 might be involved in the development and involution of hemangioma. Bcl-2 could promote the growth of hemangioma by inhibiting apoptosis of endothelia. Bax, Fas and caspase-3 promote the switch of hemangioma from proliferation to involution by inducing the apoptosis of hemangioma endothelia.

  17. Gênese e classificação de alguns solos da bacia do Ribeirão Tijuco Preto, Município de Rio das Pedras, SP: I - Análise mineralógica quantitativa da fração argila Genesis and classification of some soils from the hydrographic basin of Tijuco Preto Stream, Rio das Pedras county, SP: I - Quantitative mineralogical analysis of clay fraction

    Directory of Open Access Journals (Sweden)

    E. H. Escobar

    1973-01-01

    Full Text Available Foi empregada a análise mineralógica quantitativa no estudo das frações argila fina e argila grossa de quatro perfis de solos. Dois perfis são originados de um siltito da formação Corumbataí, e foram coletados numa mesma encosta. O primeiro deles, um Litossolo fase folhelho-argilito (TP-12, se encontra num estádio de intemperização inferior ao Podzólico Vermelho-Amarelo variação Piracicaba (TP-13, indicado pelo seu menor conteúdo de caulinita (31,5% na fração argila grossa do B2 em relação ao perfil TP-13 (52% no B22t. Esse resultado é confirmado pelo valor do intemperismo médio do TP-12, que é inferior ao do TP-13. Fazendo parte da mesma toposseqüência. numa posição mais elevada, encontra-se um Latossolo Vermelho-Escuro orto (TP-17, proveniente de um material de origem não identificado, que contém contribuição de rochas básicas, revelado pelo teor de 14% de Fe2O3. Esse latossolo apresentou como componente principal da fração argila grossa a caulinita (74,5%, seguindo-se a alofana (14,0%, gibbsita (3,7%, mica (2,9% e vermiculita (0,3%. Esses valores representam a média do perfil. As partes mais altas da bacia são ocupadas por um Latossolo Roxo, cujo perfil TP-18 apresentou a caulinita como principal componente (70,0% da fração argila grossa, seguindo-se-lhe a alofana (20,0%, a gibbsita (9,2%, a mica (0,9% e a vermiculita (0,6%. Tendo tomado o teor de gibbsita, ou o valor do Ki, como índice de intemperismo, verificou-se que esse latossolo é menos intemperizado do que um perfil de Latossolo Roxo de Campinas, situado também na depressão periférica. O mesmo acontece em relação a 10 perfis de Latossolo Roxo do planalto ocidental de São Paulo.The four soil profiles studied occur on a toposequence. The less weathered soils, a Lithosol-argilite-shale substratum phase (TP-12 and a Red-Yellow Podzolic Soil Piracicaba variation (TP-13 are located on the lower part of the toposequence. Both soils are derived from

  18. Quantitative characteristics of carcass and meat quality of ¾ Boer + ¼ Saanen and Saanen goat kids fed diets with dry yeast Características quantitativas de carcaça e de qualidade da carne de cabritos ¾ Boer + ¼ Saanen e Saanen alimentados com rações contendo levedura seca

    Directory of Open Access Journals (Sweden)

    Hanna Sakamoto Freitas

    2011-03-01

    Full Text Available Twenty-seven carcasses of non-castrated confined goat kids, (18 ¾ Boer + ¼ Saanen and nine Saanen with slaughtered weight of 30.1 ± 0.27 kg were used to evaluate quantitative characteristics of the carcass and qualitative characteristics of the loin and shoulder. The animals were distributed in a completely randomized design in a 2 × 3 factorial arrangement (breed group × diet. The diets consisted of inclusion of dry yeast, as follows: soybean meal, soybean meal + dried yeast or dried yeast. The other ingredients of the diets were oat hay (30% DM, ground corn and mineral mixture. Hot carcass weight, cold carcass weight, weight loss by cooling, true carcass yield and carcass compactness index were not influenced by the inclusion of dry yeast nor by racial group. Saanen goats presented the highest carcass commercial yield of carcass, but they needed 15 days more to reach the slaughter weight. The cut yields were not affected, but a significant interaction for shoulder yield occurred. Dry yeast and breed group did not influence the measurements of the loin, the muscle:bone ratio, the cooking losses, the shear force and composition. There was a significant interaction between the inclusion of dry yeast in the diets and the breed group for the muscle proportion on loin, which was higher in the ¾ Boer + ¼ Saanen group. For the shoulder, inclusion of dry yeast and breed group did not alter the proportions of muscle, fat, protein and ash. However, in Saanen goats, the proportion of bone and moisture content were higher and total lipid content was lower. The ¾ Boer + ¼ Saanen groups showed higher precocity. Dry yeast is an alternative protein source that can be included in diets for goat kids.Foram utilizadas 27 carcaças de cabritos machos não-castrados, confinados, sendo 18 ¾ Boer + ¼ Saanen e nove Saanen com peso de abate de 30,1 ± 0,27 kg, para avaliar as características quantitativas da carcaça e qualitativas do lombo e da paleta. Os

  19. MDMA诱导大鼠神经元凋亡及凋亡相关因子caspase-3的表达%Neuron apoptosis induced by 3,4-methylenedioxy methamphetamine and the expression of caspase-3

    Institute of Scientific and Technical Information of China (English)

    王雪; 李静; 祝三平

    2007-01-01

    目的 探讨3,4 -亚甲基二氧基甲基苯丙胺(MDMA)对实验大鼠神经元凋亡的诱导及凋亡相关因子半胱氨酸天冬氨酸特异性蛋白酶-3(caspase-3)的表达.方法 将20只Wistar雄性大鼠随机均分为1组对照组(A)、3组MDMA实验组(B、C、D).B组予MDMA(20 mg · kg-1, ip, single),C组予MDMA(20 mg · kg-1, 8 am,8 pm,ip×2 d),D组予MDMA (20 mg · kg-1 ,8 am,8 pm,ip×4 d);A组给予等体积生理盐水.采用TUNEL法检测神经元凋亡,免疫组织化学方法检测Caspase-3的表达.结果 给予MDMA后,大鼠各相关脑区有凋亡细胞形成,Caspase-3有不同程度的表达.结论 MDMA可导致神经元的凋亡,并诱导凋亡相关因子Caspase-3的表达.

  20. Expression of Ki-67/caspase-3 cocktail double staining in prostatic adenocarcinoma%前列腺腺癌中双染鸡尾酒抗体Ki-67/caspase-3的表达及意义

    Institute of Scientific and Technical Information of China (English)

    孙平丽; 金仁顺

    2010-01-01

    目的 探讨Ki-67/caspase-3鸡尾酒抗体在前列腺腺癌(PAC)、高级别前列腺上皮内瘤(HGPIN)及良性前列腺增生(BPH)组织中的表达及意义.方法 收集40例PAC、30例HGPIN和BPH标本,按常规方法石蜡包埋,制作蜡块连续切片,进行HE染色及Ki-67/caspase-3鸡尾酒抗体双重免疫组化染色.结果 Ki-67表达阳性率及增殖指数在PAC组织中明显高于HGPIN和BPH组织,且随着Gleason分级增高其增殖指数存在递增趋势(P0.05).Ki-67和caspase-3在PAC和HGPIN组织中的表达呈负相关,而在BPH中的表达呈正相关.结论 Ki-67/caspase-3在不同病变中表达不同,鸡尾酒抗体联合检测可用于前列腺良恶性疾病的诊断、鉴别诊断及预后的判断.

  1. Significado clínico-patológico das expressões citofotométricas do Ki-67 e Caspase-3 no carcinoma de células escamosas do esôfago Clinicopathologic significance of the Ki-67 and Caspase-3 cytophotometric expressions in the esophageal squamous cell carcinomal

    Directory of Open Access Journals (Sweden)

    Gilmar Pereira Silva

    2008-06-01

    Full Text Available RACIONAL: A escolha da forma de tratamento do carcinoma de células escamosa de esôfago ainda hoje é orientada pelo estadiamento tumoral, onde as características histopatológicas do tumor são o maior determinante. Parale-lamente, desenvolvem-se estudos para entender o comportamento da biologia tumoral por método imunoistoquímico de quantificação manual, avaliando a ati-vidade proliferativa ou apoptótica do tecido em análise. As desvantagens conti-das no modo manual fizeram surgir e desenvolver método computadorizado de análise de imagem. OBJETIVOS: Verificar as expressões dos marcadores KI-67 e Caspase-3 e correlacioná-las com as características clínico-patológicas do tumor. MÉTODOS: Foram estudados 29 blocos parafinados provenientes de pa-cientes portadores de carcinoma de células escamosas de esôfago submetidos à esofagectomia e pertencentes a acervos de laboratórios de patologia. Proce-deu-se preparo das lâminas por técnica imunoistoquímica convencional. A quantificação da imunorreatividade às proteínas Ki-67 e Caspase-3 foi realizada pelo software de análise de imagem computadorizada SAMBA (Systeme d'Analyse Microphotometrique a Balayage Automatique através do índice de marcagem encontrado. RESULTADOS: Predominaram na amostra o sexo mascu-lino (82,7%; maiores de 50 anos; tumores moderadamente diferenciados (68,98%; estágio III (72,42%; lesões >3cm e localizadas no ⅓ inferior do ór-gão. Os índices médios de marcagem identificados foram de 62,05% para o Ki-67 e 86,06% para a Caspase-3, e não mostraram correlação com as caracterís-ticas clínico-patológicas como sexo, idade, estadiamento tumoral, grau de pro-fundidade da lesão e comprometimento linfonodal. Houve significante diferença de expressão do Ki-67 entre os graus histológicos (P=0,047 e correlação entre os índices dos marcadores estudados (r=0,41 e P =0,032. CONCLUSÃO: Na presente investigação as atividades das proteínas estudadas

  2. Caspase-3 expression in spinal tissue of retinoic acid induce spiua bifida fetal rat%维甲酸诱导脊柱裂胎鼠脊髓组织中Caspase-3表达情况

    Institute of Scientific and Technical Information of China (English)

    马英桓; 袁正伟

    2012-01-01

    Objective To explore caspase-3 expression in spinal tissue of retinoic acid induced spina bifida fetal rat. Methods Pregnant Wister rats with 10 days were used. Retinoic acid dissolved in olive oil (40mg /ml) were stomach fed for preparing the rat model of spina bifida malformations 135mg / kg). Control group only received olive oil. The animals were divided into 4 groups: pregnancy of 12 days, 15 days, 17 days and 20 days. Immunohistochemical method was used to detect and compare caspase-3 expression in different groups. Results The expression of caspase-3 increased at the day 15 after pregnancy, and maintained until day 20 in the spinal tissue of modeled fetal rat, which presented significant difference compared to that of control groups at the same pregnant time. At day 15, day 17 and day 20 of pregnancy, the number of caspase-3 positive cells was more in model animals than the control. Conclusions Retinoic acid induced spina bifida fetal rat demonstrates the increased caspase-3 expression in spinal tissue of fetal rats.%目的 本文旨在探讨维甲酸诱导脊柱裂胎鼠脊髓组织Caspase-3表达情况.方法 选取孕10d Wistar大鼠,实验组用溶有维甲酸(40mg/ml)的橄榄油,以135mg/kg经胃管注入给药制作脊柱裂畸形大鼠模型;对照组选取孕10 d Wistar大鼠给等量橄榄油.将实验组及对照组按照孕12、15、17和20 d分为4组.应用免疫组织化学方法比较分析Caspase-3在对照组、畸形组胎鼠脊髓组织细胞中的分布和表达情况.结果 脊柱裂大鼠脊髓神经组织中Caspase-3在15d开始增多,一直持续到20 d胚胎大鼠.其增高情况明显高于同一时间点对照组大鼠.胚胎15、17和20 d显性脊柱裂畸形鼠脊髓组织Caspase-3阳性细胞数多于对照组,荧光强度高于对照组.结论 维甲酸诱导的脊柱裂胎鼠Caspase-3表达明显高于正常发育胎鼠.

  3. 青蒿琥酯对人胚肺成纤维细胞Caspase-3表达的影响%Effect of Artesunate on the Expression of Caspase-3 in Human Embryonic Lung Fibroblasts

    Institute of Scientific and Technical Information of China (English)

    王昌明; 黎洪秀; 张孝飞

    2011-01-01

    OBJECTIVE To study the effect of artesunate on the growth of human embryonic lung fibroblast HFL-Ⅰ cells in vitro and provide experimental data for anti-fibrosis activity of artesunate. METHODS CCK-8 assay was used to determine the effect of artesunate on the growth of HFL-Ⅰ cells in vitro. Apoptosis ratio was examined by flow cytometry (FCM). The mRNA level of Caspase-3, one of apoptosis related proteins, were assessed by RT-PCR. The expression of Caspase-3 protein was detected by Western blot. RESULTS Artesunate has a significantly inhibitory effect on the proliferation of HFL-Ⅰ cells in a dose-dependent manner in vitro. Flow cytometry assay demonstrated a higher distribution in G1 phase of HFL-Ⅰ cells with artesunate. And apoptosis rate of HFL-Ⅰ cells was significantly increased in artesunate-treated group compared with control group (P<0.01). RT-PCR analysis showed the levels of Caspase-3 mRNA were significantly increased in artesunate-treated group,compared with control group. Western bloting also showed a significant enhancement of Caspase-3 protein levels in artesunate-treated group, when compared to control group. CONCLUSION Artesunate may exert marked anti-pulmonary fibrosis effect by up-regulating the mRNA and protein level of Caspase-3, which could induce the growth inhibition and apoptosis in HFL-Ⅰ cells.%目的 研究青蒿琥酯对人胚肺成纤维细胞(human embryonic lung fibroblast,HELF)系HFL-Ⅰ细胞体外生长的影响,为青蒿琥酯抗纤维化提供实验依据.方法 采用CCK-8法检测青蒿琥酯对体外培养的HFL-Ⅰ细胞生长的影响,用流式细胞术测定细胞凋亡率;RT-PCR法测定凋亡相关蛋白Caspase-3的mRNA表达水平,Western blot法分析Caspase-3蛋白的表达情况.结果 青蒿琥酯呈浓度依赖性抑制HFL-Ⅰ细胞增殖,HFL-Ⅰ细胞经青蒿琥酯作用后细胞主要停滞于G1期,凋亡率明显增加(P<0.01),Caspase-3 mRNA的表达显著高于对照组,Caspase-3蛋白的表达亦

  4. Propofol and magnesium attenuate isoflurane-induced caspase-3 activation via inhibiting mitochondrial permeability transition pore

    Directory of Open Access Journals (Sweden)

    Zhang Yiying

    2012-08-01

    Full Text Available Abstract Background The inhalation anesthetic isoflurane has been shown to open the mitochondrial permeability transition pore (mPTP and induce caspase activation and apoptosis, which may lead to learning and memory impairment. Cyclosporine A, a blocker of mPTP opening might attenuate the isoflurane-induced mPTP opening, lessening its ripple effects. Magnesium and anesthetic propofol are also mPTP blockers. We therefore set out to determine whether propofol and magnesium can attenuate the isoflurane-induced caspase activation and mPTP opening. Methods We investigated the effects of magnesium sulfate (Mg2+, propofol, and isoflurane on the opening of mPTP and caspase activation in H4 human neuroglioma cells stably transfected to express full-length human amyloid precursor protein (APP (H4 APP cells and in six day-old wild-type mice, employing Western blot analysis and flowcytometry. Results Here we show that Mg2+ and propofol attenuated the isoflurane-induced caspase-3 activation in H4-APP cells and mouse brain tissue. Moreover, Mg2+ and propofol, the blockers of mPTP opening, mitigated the isoflurane-induced mPTP opening in the H4-APP cells. Conclusion These data illustrate that Mg2+ and propofol may ameliorate the isoflurane-induced neurotoxicity by inhibiting its mitochondrial dysfunction. Pending further studies, these findings may suggest the use of Mg2+ and propofol in preventing and treating anesthesia neurotoxicity.

  5. Eupalitin induces apoptosis in prostate carcinoma cells through ROS generation and increase of caspase-3 activity.

    Science.gov (United States)

    Kaleem, Sarjeel; Siddiqui, Sahabjada; Siddiqui, Hefazat Hussain; Badruddeen; Hussain, Arshad; Arshad, Mohammad; Akhtar, Juber; Rizvi, Aleza

    2016-02-01

    Prostate cancer is the second most common malignancy in the human reproductive system. Eupalitin is one of the O-methylated flavonol-exhibited enhanced cancer chemopreventive agents. The current study highlights the structural determination of eupalitin and aims to explore the antitumor activity of eupalitin in human prostate cancer cell (PC3) and its underlying mechanism. Eupalitin structure was determined by using FTIR, (1)H NMR, and (13)C NMR. PC3 cells were treated with increasing concentrations of eupalitin, followed by analysis of the cell viability with an MTT assay. The results demonstrated that eupalitin markedly inhibited the proliferation of PC3 cells in a concentration-dependent manner. The results from fluorescent microscopic analysis of nuclear condensation and intracellular ROS generation determined that eupalitin significantly induced ROS level lead to nuclear apoptosis. Cell cycle analysis revealed that eupalitin-induced cell cycle progression as a percentage of cells in G0/G1 phase decreased whereas S phase increased. Caspase-3 immunofluorescence analysis confirms the efficacy of eupalitin-inducing apoptotic pathway and cell death. Thus, our study is helpful in understanding the mechanism underlying these effects in prostate cancer and it may provide novel molecular targets for prostate cancer therapy. © 2015 International Federation for Cell Biology.

  6. Methylglyoxal reduces mitochondrial potential and activates Bax and caspase-3 in neurons: Implications for Alzheimer's disease.

    Science.gov (United States)

    Tajes, Marta; Eraso-Pichot, Abel; Rubio-Moscardó, Fanny; Guivernau, Biuse; Bosch-Morató, Mònica; Valls-Comamala, Victòria; Muñoz, Francisco J

    2014-09-19

    Alzheimer's disease (AD) is characterized by the oxidative stress generated from amyloid β-peptide (Aβ) aggregates. It produces protein nitrotyrosination, after the reaction with nitric oxide to form peroxynitrite, being triosephosphate isomerase (TPI) one of the most affected proteins. TPI is a glycolytic enzyme that catalyzes the interconversion between glyceraldehyde 3-phosphate (GAP) and dihydroxyacetone phosphate (DHAP). Methylglyoxal (MG) is a by-product of TPI activity whose production is triggered when TPI is nitrotyrosinated. MG is harmful to cells because it glycates proteins. Here we found protein glycation when human neuroblastoma cells were treated with Aβ. Moreover glycation was also observed when neuroblastoma cells overexpressed mutated TPI where Tyr165 or Tyr209, the two tyrosines close to the catalytic center, were changed by Phe in order to mimic the effect of nitrotyrosination. The pathological relevance of these findings was studied by challenging cells with Aβ oligomers and MG. A significant decrease in mitochondrial transmembrane potential, one of the first apoptotic events, was obtained. Therefore, increasing concentrations of MG were assayed searching for MG effect in neuronal apoptosis. We found a decrease of the protective Bcl2 and an increase of the proapoptotic caspase-3 and Bax levels. Our results suggest that MG is triggering apoptosis in neurons and it would play a key role in AD neurodegeneration.

  7. PLGA-carbon nanotube conjugates for intercellular delivery of caspase-3 into osteosarcoma cells.

    Directory of Open Access Journals (Sweden)

    Qingsu Cheng

    Full Text Available Cancer has arisen to be of the most prominent health care issues across the world in recent years. Doctors have used physiological intervention as well as chemical and radioactive therapeutics to treat cancer thus far. As an alternative to current methods, gene delivery systems with high efficiency, specificity, and safety that can reduce side effects such as necrosis of tissue are under development. Although viral vectors are highly efficient, concerns have arisen from the fact that viral vectors are sourced from lethal diseases. With this in mind, rod shaped nano-materials such as carbon nanotubes (CNTs have become an attractive option for drug delivery due to the enhanced permeability and retention effect in tumors as well as the ability to penetrate the cell membrane. Here, we successfully engineered poly (lactic-co-glycolic (PLGA functionalized CNTs to reduce toxicity concerns, provide attachment sites for pro-apoptotic protein caspase-3 (CP3, and tune the temporal release profile of CP3 within bone cancer cells. Our results showed that CP3 was able to attach to functionalized CNTs, forming CNT-PLGA-CP3 conjugates. We show this conjugate can efficiently transduce cells at dosages as low as 0.05 μg/ml and suppress cell proliferation up to a week with no further treatments. These results are essential to showing the capabilities of PLGA functionalized CNTs as a non-viral vector gene delivery technique to tune cell fate.

  8. Expression of cleaved caspase-3 in renal tubular cells in Plasmodium falciparum malaria patients.

    Science.gov (United States)

    Wichapoon, Benjamas; Punsawad, Chuchard; Viriyavejakul, Parnpen

    2017-01-01

    In Plasmodium falciparum malaria, the clinical manifestation of acute kidney injury (AKI) is commonly associated with acute tubular necrosis (ATN) in the kidney tissues. Renal tubular cells often exhibit various degrees of cloudy swelling, cell degeneration, and frank necrosis. To study individual cell death, this study evaluates the degree of renal tubular necrosis in association with apoptosis in malarial kidneys. Kidney tissues from P. falciparum malaria with AKI (10 cases), and without AKI (10 cases) were evaluated for tubular pathology. Normal kidney tissues from 10 cases served as controls. Tubular necrosis was assessed quantitatively in kidney tissues infected with P. falciparum malaria, based on histopathological evaluation. In addition, the occurrence of apoptosis was investigated using cleaved caspase-3 marker. Correlation between tubular necrosis and apoptosis was analyzed. Tubular necrosis was found to be highest in P. falciparum malaria patients with AKI (36.44% ± 3.21), compared to non-AKI (15.88% ± 1.63) and control groups (2.58% ± 0.39) (all p < 0.001). In the AKI group, the distal tubules showed a significantly higher degree of tubular necrosis than the proximal tubules (p = 0.021) and collecting tubules (p = 0.033). Tubular necrosis was significantly correlated with the level of serum creatinine (r = 0.596, p = 0.006), and the occurrence of apoptosis (r = 0.681, p = 0.001). In malarial AKI, the process of apoptosis occurs in ATN. © 2016 Asian Pacific Society of Nephrology.

  9. Kaposi's sarcoma herpesvirus microRNAs target caspase 3 and regulate apoptosis.

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    Guillaume Suffert

    2011-12-01

    Full Text Available Kaposi's sarcoma herpesvirus (KSHV encodes a cluster of twelve micro (miRNAs, which are abundantly expressed during both latent and lytic infection. Previous studies reported that KSHV is able to inhibit apoptosis during latent infection; we thus tested the involvement of viral miRNAs in this process. We found that both HEK293 epithelial cells and DG75 cells stably expressing KSHV miRNAs were protected from apoptosis. Potential cellular targets that were significantly down-regulated upon KSHV miRNAs expression were identified by microarray profiling. Among them, we validated by luciferase reporter assays, quantitative PCR and western blotting caspase 3 (Casp3, a critical factor for the control of apoptosis. Using site-directed mutagenesis, we found that three KSHV miRNAs, miR-K12-1, 3 and 4-3p, were responsible for the targeting of Casp3. Specific inhibition of these miRNAs in KSHV-infected cells resulted in increased expression levels of endogenous Casp3 and enhanced apoptosis. Altogether, our results suggest that KSHV miRNAs directly participate in the previously reported inhibition of apoptosis by the virus, and are thus likely to play a role in KSHV-induced oncogenesis.

  10. Endobronchial valves in severe emphysematous patients: CT evaluation of lung fissures completeness, treatment radiological response and quantitative emphysema analysis Valvas endobrônquicas em pacientes com enfisema grave: avaliação por TC da completude das fissuras pulmonares, resposta radiológica ao tratamento e análise quantitativa do enfisema

    Directory of Open Access Journals (Sweden)

    Marcel Koenigkam-Santos

    2013-02-01

    Full Text Available OBJECTIVE: To evaluate lung fissures completeness, post-treatment radiological response and quantitative CT analysis (QCTA in a population of severe emphysematous patients submitted to endobronchial valves (EBV implantation. MATERIALS AND METHODS: Multi-detectors CT exams of 29 patients were studied, using thin-section low dose protocol without contrast. Two radiologists retrospectively reviewed all images in consensus; fissures completeness was estimated in 5% increments and post-EBV radiological response (target lobe atelectasis/volume loss was evaluated. QCTA was performed in pre and post-treatment scans using a fully automated software. RESULTS: CT response was present in 16/29 patients. In the negative CT response group, all 13 patients presented incomplete fissures, and mean oblique fissures completeness was 72.8%, against 88.3% in the other group. QCTA most significant results showed a reduced post-treatment total lung volume (LV (mean 542 ml, reduced EBV-submitted LV (700 ml and reduced emphysema volume (331.4 ml in the positive response group, which also showed improved functional tests. CONCLUSION: EBV benefit is most likely in patients who have complete interlobar fissures and develop lobar atelectasis. In patients with no radiological response we observed a higher prevalence of incomplete fissures and a greater degree of incompleteness. The fully automated QCTA detected the post-treatment alterations, especially in the treated lung analysis.OBJETIVO: Avaliar a completude das fissuras, resposta radiológica ao tratamento e análise quantitativa por TC (AQTC em população de pacientes com enfisema grave submetidos ao implante de valvas endobrônquicas (VEB. MATERIAIS E MÉTODOS: Foram estudados exames de TC multidetectores de 29 pacientes, realizados com protocolo de baixo dose, cortes finos, sem contraste. Dois radiologistas revisaram, retrospectivamente, os exames em consenso. A completude das fissuras foi estimada em escala de 5% e

  11. Study on the enzymatic activity of Caspase-3 in response to alginic acid decomposing bacteria in Laminaria japonica Aresch.(Phaeophyta)

    Institute of Scientific and Technical Information of China (English)

    Wang Gaoge; Lin Wei; Yan Xiaojun; Duan Delin

    2005-01-01

    Caspase-3 is the major factor in apoptosis triggered by various stimuli, and plays a critical role during the apoptosis process. By using CaspGLOWTM fluorescein active caspase-3 staining method, caspase-3 enzymatic activities were detected in response to alginic acid bacteria in Laminaria japonica sporophytic tissues. Results showed that caspase-3 enzymatic activities were observed at 5 min after the infection. Caspase-3 enzymatic activity increased with the infection time, and had a tendency of moving from the infection site to outside. By applying caspase-specific peptide inhibitor Z-VAD-FMK, caspase-3 activation could be effectively abolished in the infected tissues. Our results indicate that programmed cell death (PCD) may be involved in the infected Laminaria japonica sporophytic tissues, and provide the evidence that defense mechanisms in algae may have similar caspase cascade events in animals.

  12. MAPKs and Mst1/Caspase-3 pathways contribute to H2B phosphorylation during UVB-induced apoptosis

    Institute of Scientific and Technical Information of China (English)

    2010-01-01

    Apoptosis is a highly coordinated or programmed cell suicide mechanism in eukaryotes.Histone modification is associated with nuclear events in apoptotic cells.Specifically H2B phosphorylation at serine 14 (Ser14) catalyzed by Mst1 kinase has been linked to chromatin condensation during apoptosis.We report that activation of MAPKs (ERK1/2,JNK1/2 and p38) together with Mst1 and caspase-3 is required for phosphorylation of H2B (Ser14) during ultraviolet B light (UVB)-induced apoptosis.UVB can trigger activation of MAPKs and induce H2B phosphorylation at Ser14 but not acetylation in a time-dependent manner.Inhibition of ERK1/2,JNK1/2 or p38 activity blocked H2B phosphorylation (Ser14).Furthermore,caspase-3 was activated by UVB to regulate Mst1 activity,which phosphorylates H2B at Ser14,leading to chromatin condensation.Full inhibition of caspase-3 activity reduced Mst1 activation and partially inhibited H2B phosphorylation (Ser14),but ERK1/2,JNK1/2 and p38 activities were not affected.Taken together,these data revealed that H2B phosphorylation is regulated by both MAPKs and caspase-3/Mst1 pathways during UVB-induced apoptosis.

  13. Evaluation of caspase3 and 9 gene polymorphisms in gastric cancer patients in Mazandaran province: a brief report

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    Saeid Abediankenari

    2013-11-01

    Full Text Available Background: Gastric cancer is the most prevalent cancer with poor survival in gastrointestinal tract. Caspase 3 and 9 play an important role in the development and progression of cancer. Polymorphisms in the genes for these enzymes can affect gene activity and thus may influence susceptibility to gastric cancer. In this study, caspase 3 and 9 genes polymorphisms in patients with gastric cancer were examined.Methods: In a case - control study, 100 patients with gastric cancer and 100 healthy individuals were evaluated in the region rs4647601: G> T for caspase-3 and -1263 A> G gene promoter for caspase 9. DNA extraction was performed from whole blood according to manufacture protocol. RFLP-PCR method was carrying out for detection of caspase 3 and 9 genes genotype in two groups.Results: In this study, 143 men and 57 women were evaluated. All of them were selected from the same race and geographical area. The results indicated an increase of the mutant G allele in the control group, which leads to a decreasing in the incidence of gastric cancer (P caspase 9 polymorphism could be a useful marker in personal sensitivity to gastric cancer and help to cancer treatment and prevention process. It is concluded that caspase gene variation may be a diagnostic factor in the gastric cancer.

  14. Location of Caspase 3-like Protease in the Development of Sieve Element and Tracheary Element of Stem in Cucurbita moschata

    Institute of Scientific and Technical Information of China (English)

    Xia Hao; Jie Qian; Shan Xu; Xin Song; Jian Zhu

    2008-01-01

    The casepase is considered to regulate the process of programmed cell death in the development of organisms. In this study, caspase 3-like protease was detected by immunohistochemistry and immunoelectron microscopy during the development of sieve element and tracheary element of stem in Cucurbita moschata Duch. Antibody with brown color (under light microscopy) and gold particles (under transmission electron microscopy) for detecting caspase 3-like protease was mainly displayed in inner phloem, extemal phloem and xylem in the region close to procambium. From the results it was considered that caspase 3-like protease did exist in vascular elements and played different roles during the development of sieve and tracheary elements, and different types of programmed cell death might be carried out. The caspase 3-like protease mainly participated in making cytoplasmic streaming cease and in degrading P-protein bodies; however, it rarely participated in the function for signal transferring in the developmental sieve element. However, it might induce calcium accumulation for rupturing the tonoplast in the signal of PCD in the developmental tracheary element.

  15. Location of caspase 3-like protease in the development of sieve element and tracheary element of stem in Cucurbita moschata.

    Science.gov (United States)

    Hao, Xia; Qian, Jie; Xu, Shan; Song, Xin; Zhu, Jian

    2008-12-01

    The casepase is considered to regulate the process of programmed cell death in the development of organisms. In this study, caspase 3-like protease was detected by immunohistochemistry and immunoelectron microscopy during the development of sieve element and tracheary element of stem in Cucurbita moschata Duch. Antibody with brown color (under light microscopy) and gold particles (under transmission electron microscopy) for detecting caspase 3-like protease was mainly displayed in inner phloem, external phloem and xylem in the region close to procambium. From the results it was considered that caspase 3-like protease did exist in vascular elements and played different roles during the development of sieve and tracheary elements, and different types of programmed cell death might be carried out. The caspase 3-like protease mainly participated in making cytoplasmic streaming cease and in degrading P-protein bodies; however, it rarely participated in the function for signal transferring in the developmental sieve element. However, it might induce calcium accumulation for rupturing the tonoplast in the signal of PCD in the developmental tracheary element.

  16. Analysis of caspase-3 in ASTC-a-1 cells treated with mitomycin C using acceptor photobleaching techniques

    Science.gov (United States)

    Wang, Huiying; Chen, Tongsheng; Sun, Lei

    2008-02-01

    Caspase-3 is a key activated death protease, which catalyzes the specific cleavage of many cellular proteins and induces DNA cleavage eventually. In this report, cells were treated with mitomycin C (MMC) at different concentration and its activity was detected by cell counting kit (CCK-8). Based on results of CCK-8, cells were treated with 10μg/mL MMC and Hoechst 33258 has been used to observe cell apoptosis. Fluorescence resonance energy transfer (FRET) and confocal microscopy have been used to the effect of MMC on the caspase3 activation in living cells. Human lung adenocarcinoma cells (ASTC-a-1) was transfected with plasmid SCAT3 (pSCAT3)/CKAR FRET receptor. Acceptor photobleaching techniques of FRET plasmid has been used to destruct fluorophore of cells stably expressing SCAT3 reporter on a fluorescence confocal microscope. The activity of caspase3 can be analyzed by FRET dynamics of SCAT3 in living cells. Our results show that MM C can induce ASTC-a-1 cell apoptosis through activation of caspase3.

  17. Differential Effect of Intrauterine Hypoxia on Caspase 3 and DNA Fragmentation in Fetal Guinea Pig Hearts and Brains

    Science.gov (United States)

    Evans, LaShauna C.; Liu, Hongshan; Thompson, Loren P.

    2012-01-01

    The aim of this study is to quantify the effect of intrauterine hypoxia (HPX) and the role of nitric oxide (NO) on the apoptotic enzyme, caspase 3, and DNA fragmentation in fetal heart and brain. Hypoxia and NO are important regulators of apoptosis, although this has been little studied in the fetal organs. We investigated the effect of intrauterine HPX on apoptosis and the role of NO in both fetal hearts and brains. Pregnant guinea pigs were exposed to room temperature (N = 14) or 10.5% O2 (N = 12) for 14 days prior to term (term = 65 days) and administered water or l-N6-(1-iminoethyl)-lysine (LNIL), an inducible nitric oxide synthase (iNOS) inhibitor, for 10 days. Fetal hearts and brains were excised from anesthetized near-term fetuses for study. Chronic HPX decreased pro- and active caspase 3, caspase 3 activity, and DNA fragmentation levels in fetal hearts compared with normoxic controls. l-N6-(1-iminoethyl)-lysine prevented the HPX-induced decrease in caspase 3 activity but did not alter DNA fragmentation levels. In contrast, chronic HPX increased both apoptotic indices in fetal brains, which were inhibited by LNIL. Thus, the effect of HPX on apoptosis differs between fetal organs, and NO may play an important role in modulating these effects. PMID:22383778

  18. Immunoexpression of the COX-2, p53, and caspase-3 proteins in colorectal adenoma and non-neoplastic mucosa

    Science.gov (United States)

    Nogueira, Renan Brito; Pires, Andréa Rodrigues Cordovil; Soares, Thélia Maria Santos; Rodrigues, Simone Rabello de Souza; Campos, Mariane Antonieta Menino; Toloi, Giovanna Canato; Waisberg, Jaques

    2013-01-01

    ABSTRACT Objective: To analyze the immunoexpression of the COX-2, p53, and caspase-3 proteins in colorectal adenomas and non-neoplastic mucosa. Methods: 72 individuals were subjected to colonoscopy, which provided 50 samples of adenomas and 45 samples of non-neoplastic colorectal mucosa. The tissue samples were obtained via the tissue microarray technique and subjected to immunohistochemical analysis using primary anti-p53, anti-COX-2, and anti-caspase-3 antibodies. The positivity and intensity of the immunoreaction were classified. The analyzed variables were as follows: site of the adenomas in the colon, degree of dysplasia, size, and score of positivity and intensity of immunoexpression of the p-53, caspase-3, and COX-2 proteins. Results: The immunoexpression of mutated protein p53 was positive in 30 (60%) adenoma samples and negative in 20 (40%) adenoma samples. The immunoexpression of mutated protein p53 was negative in 39 (86.6%) samples and positive in 6 (13.3%) samples of the non-neoplastic colorectal mucosa (p<0.0001). Significant differences were seen between both the largest size (p=0.006) and the highest degree of dysplasia (p<0.0001) of the adenomas and the intensity of immunoexpression of mutated protein p53. The positivity and intensity of immunoexpression of COX-2 (p=0.14) and caspase-3 (p=0.23) showed no significant differences between the adenomas and the non-neoplastic colorectal mucosa. Conclusion: Mutated protein p53 was hyperexpressed in the adenomas compared with the non-neoplastic mucosa. Greater size and greater degree of dysplasia in the adenomas were associated with higher expression of mutated protein p53. The immunoexpression of COX-2 and caspase-3 in the adenomas did not exhibit a correlation with the anatomical-pathological features of the tumors and did not differ from the corresponding expression levels in the non-neoplastic mucosa. PMID:24488384

  19. Microbiota Composition, HSP70 and Caspase-3 Expression as Marker for Colorectal Cancer Patients in Aceh, Indonesia

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    Fauzi Yusuf

    2017-02-01

    Full Text Available Aim: to investigate the relationship between microbiota composition with HSP70 and Caspase-3 expressions in colon tissue as an initial study to develop the candidate for early detection of colorectal cancer for Indonesian patients. Methods: this is a cross-sectional study on 32 patients undergoing colonoscopy; 16 patients of colorectal cancer (CRC while the other 16 patients are not (colitis and internal hemorrhoid. The composition of microbiota in stool samples was examined using 16S rRNA Denaturing Gradient Gel Electrophoresis (DDGE while expression of HSP70 was examined by immunohistochemistry and Caspase-3 by using Haematoxylin-Eosin(HE staining to determine the morphological changes in colon tissue. Results: analysis of PCR-DDGE shows a different composition of microbiota between patients with CRC and non-CRC. All CRC patients showed disappearance of dominant band from Bifidobacterium groups. Histological observation based on Inter Class Correlation (ICC test from all slide showed a high scores (5.2-9.2 in CRC patients and low scores (1.7-2.4 in non-CRC patients. HSP70 expression was increased significantly in CRC patients with the highest percentage of 84%, while expression of caspase-3 decreased with the highest percentage of 21%. Statistical analysis showed that the incidence of colorectal cancer was associated with the expression of HSP 70 (p<0.001, and Caspase 3 (p<0.001. Conclusion: bifidobacterium is an important indicator for colorectal cancer patients that show disappearance of dominant band, while expression of HSP70 increased and the Caspase-3 expression decreased significantly.

  20. Expression of Cyt C and Caspase-3 of bone marrow supernatant in patients with chronic mountain sickness%慢性高原病患者骨髓CytC和Caspase-3表达及意义研究

    Institute of Scientific and Technical Information of China (English)

    戴昕; 李建平; 李文倩; 冯建明

    2014-01-01

    目的:探讨细胞凋亡在慢性高原病(CMS)发生发展过程中的作用,从而寻求新的诊断和治疗途径。方法选取 CMS 患者20例为研究对象,健康人群14例为对照组,采用固相双抗体夹心 ELISA 方法测定 CMS 组及对照组骨髓上清液 Cyt C 和 Caspase-3水平。结果 CMS 患者骨髓上清液中 Cyt C 水平[(21.43±8.92)ng/ml]水平与对照组[(10.36±3.55)ng/ml]比较差异有统计学意义(P<0.05);CMS 患者骨髓上清液中 Caspase-3水平[(15.45±4.73)ng/ml]水平与对照组[(7.14±1.93)ng/ml]比较差异有统计学意义(P<0.05);CMS 患者骨髓上清液中 Cyt C 与 Caspase-3水平呈正相关(r=0.706,P<0.01);CMS 患者骨髓上清液中 Cyt C 和 Caspase-3表达水平均与血红蛋白(Hb)浓度呈正相关(r=0.524,P<0.01;r=0.577,P<0.01),与血氧饱和度(SaO2)呈负相关(r=-0.505,P<0.05;r=-0.554,P<0.01)。结论推测细胞凋亡可能在 CMS 病理生理过程中发挥着重要作用。%Objective To investigate the apoptosis occurred during the development of function in CMS, so as to seek a new means of diagnosis and treatment. Methods The bone marrow supernatant levels of Cyt C and Caspase-3 in 20 CMS pa-tients and 14 healthy people were determined by quantitative sandwich enzyme immunoassay. Results The bone marrow su-pernatant levels of Cyt C[(21.43±8.92)ng/ml] and Caspase-3[(10.36±3.55)ng/ml] in CMS were significantly higher than those in control group [(15.45±4.73)ng/ml] and [(7.14±1.93)ng/ml] respectively. And there were positive relationship between the levels of Cyt C and Caspase-3 (r=0.706,P<0.01),and between the levels of Cyt C and Hb (r=0.524,P<0.01),and also between the levels of Caspase-3 and Hb (r=0.577,P<0.01). There were negative relationship between the levels of Cyt C and SaO2(r=-0.554, P<0.05),and between the levels of Caspase-3 and SaO2 (r=-0.505,P<0.05). Conclusion Speculate that apoptosis may play an

  1. Astragalus saponins induce apoptosis in human gastric adenocarcinoma cells via a caspase 3-dependent pathway

    Institute of Scientific and Technical Information of China (English)

    JOSHUA K S Ko; Kathy K W Auyeung

    2008-01-01

    Objective Many Asian countriea including China, Japan and Korea have very high incidence of gastric cancer, in which about 42 % cases occur in mainland China. The precise targets and underlying mechanisms are not well understood. Our previous study revealed that Astragalus saponins (AST) showed promising effects on the suppression of the growth of HT-29 human colon cancer cells and tumor xenograft by inhibiting cell proliferation and promoting apoptosis. In the present study, we investigated the anti-carcinogenic effects of AST in AGS human gastric adenocarcinoma cells and attempted to elucidate the underlying mechanisms. Methods Growth inhibition of AGS cells was determined by using the MTT viability test. Involvement of different members of the apoptotic cascade and other growth-related factors was explored by assessment of their protein expression using Western blot analysis. Distribution of cells in different phases of the cell cycle was assessed by flow eytometry. Results Our data indicate that AST induced growth-inhibition and apoptosis in AGS cells by activating caspase 3 with subsequent poly (ADP-ribose) polymerase (PARP) cleavage. Cell cycle arrest at the G2/M phase had been observed in AST-treated AGS cells. The anti-proliferative effect of AST was associated with modulation of eydin B1 and p21. We then demonstrate that AST could downregulate the expression of VEGF, of which interaction with its receptors is important for angiogenesis during tumor formation. Conclusions Our findings suggest that AST is an effective agent in gastric cancer treatment by inducing cell cycle arrest and apoptosis, of which anti-angiogenesis could be an alternative mode of action.

  2. Oridonin induces apoptosis in gastric cancer through Apaf-1, cytochrome c and caspase-3 signaling pathway

    Institute of Scientific and Technical Information of China (English)

    Ke-Wang Sun; Ying-Yu Ma; Tian-Pei Guan; Ying-Jie Xia; Chang-Ming Shao; Le-Gao Chen; Ya-Jun Ren

    2012-01-01

    AIM:To investigate the effect and mechanism of oridonin on the gastric cancer cell line HGC-27 in vitro.METHODS:The inhibitory effect of oridonin on HGC-27 cells was detected using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay.After treatment with 10 μg/mL oridonin for 24 h and 48 h,the cells were stained with acridine orange/ethidium bromide.The morphologic changes were observed under an inverted fluorescence microscope.DNA fragmentation (a hallmark of apoptosis) and lactate dehydrogenase activity were examined using DNA ladder assay and lactate dehydrogenase-release assay.After treated with oridonin (0,1.25,2.5,5 and 10 μg/mL),HGC-27cells were collected for anexin V-phycoerythrin and 7-amino-actinomycin D double staining and tested by flow cytometric analysis,and oridonin-induced apoptosis in HGC-27 cells was detected.After treatment with oridonin for 24 h,the effects of oridonin on expression of Apaf-1,Bcl-2,Bax,caspase-3 and cytochrome c were also analyzed using reverse-transcript polymerase chain reaction (RT-PCR) and Western blotting.RESULTS:Oridonin significantly inhibited the proliferation of HGC-27 cells in a dose-and time-dependent manner.The inhibition rates of HGC-27 treated with four different concentrations of oridonin for 24 h (1.25,2.5,5 and 10 μg/mL) were 1.78% ± 0.36%,4.96% ±1.59%,10.35% ± 2.76% and 41.6% ± 4.29%,respectively,which showed a significant difference (P < 0.05).The inhibition rates of HGC-27 treated with oridonin at the four concentrations for 48 h were 14.77% ± 4.21%,21.57% ± 3.75%,30.31% ± 4.91% and 61.19% ±5.81%,with a significant difference (P < 0.05).The inhibition rates of HGC-27 treated with oridonin for 72 h at the four concentrations were 25.77% ± 4.85%,31.86% ± 3.86%,48.30% ± 4.16% and 81.80% ± 6.72%,with a significant difference (P < 0.05).Cells treated with oridonin showed typical apoptotic features with acridine orange

  3. Caspase-3在小鼠肿瘤恶病质肌肉蛋白消耗中的作用%Effect of Caspase-3 in skeletal muscle protein consumption of cancer cachexia mice

    Institute of Scientific and Technical Information of China (English)

    郑曰勇; 刘红; 李聪; 王强; 唐华

    2014-01-01

    Objective To explore the expression of caspase-3 in skeletal muscle of the mice in the state of cancer, and to elucidate the relationship between Caspase-3 and apoptosis,consumption of skeletal muscle protein in cancer cachexia.Methods 48 male BALB/c mice were randomly divided into cancer cachexia group and control group (n=24).The mice in cancer cachexia group were inoculated with mouse colon 26 adenocarcinoma cells.The body weights of the mice in two groups were detected daily.Eight mice in each group were executed to test the weight of left gastrocnemius, fiber crosscut area, the expression levels of tumor necrosis factor-α(TNF-α), interleukin-6 (IL-6),Caspase-3 proteins and the apoptotic rate of gastrocnemius cells on day 8,14,and 20,respectively. Results The mice in cancer cachexia group appeared cachectic symptoms on day 14,the non-tumor body weight was decreased more than 20% of that in control group (P<0.05).Compared with control group at the same time, the mouse body weight,non-tumor body weight,the weight of left gastrocnemius and the fiber crosscut area of the mice in cancer cachexia group were obviously decreased with the prolongation of inoculation time (P<0.05 ), whereas the expression levels of TNF-α,IL-6,Caspase-3 proteins and the apoptotic rate of muscle cells were obviously increased after tumor inoculation (P<0.05).The level of Caspase-3 protein was negatively correlated with the weight of gastrocnemius and fiber crosscut area (r=-0.716,P<0.05;r=-0.694,P<0.05),and the level of Caspase-3 was positively correlated with the levels of TNF-αand IL-6 (r=0.742,P<0.05;r=0.675,P<0.05).Conclusion Caspase-3 may be a key factor in the protein comsumption of skeletal muscle in cancer cachexia.%目的:探讨半胱氨酸天冬氨酸蛋白酶3(Caspase-3)在小鼠肿瘤恶病质状态下骨骼肌中的表达,阐明Caspase-3与细胞凋亡及肿瘤恶病质骨骼肌蛋白消耗的关系。方法:小鼠结肠腺癌 Colon26(CT26)细胞接种BALB

  4. 吗啡成瘾时脑内Fas、Bcl-2和Caspase-3蛋白表达的改变%Changes of Fas, Bcl-2 and Caspase-3 protein in rat brain during morphine addiction

    Institute of Scientific and Technical Information of China (English)

    刘立伟; 王新华; 傅舒昆; 吴青华; 傅强

    2012-01-01

    目的 观察吗啡成瘾时脑细胞中凋亡相关蛋白Fas、Caspase-3和Bcl-2表达的改变.方法 将48只体质量为190~210 g的成年SD大鼠随机分为3组:吗啡依赖组、吗啡戒断组和对照组,每组16只.依据药物递增原则,依赖组和戒断组大鼠腹腔内给予吗啡13d,建立吗啡成瘾模型.戒断组大鼠在成瘾后腹腔内注射纳洛酮5 mg/kg,诱导戒断30 min.对照组大鼠在相同的治疗时间腹腔内注射生理盐水.应用免疫组织化学、蛋白印迹分析方法检测大鼠海马区Fas、Bcl-2和Caspase-3蛋白的表达.结果 与对照组比较,吗啡依赖组和戒断组大鼠海马区Fas和Caspase-3的表达增加(P<0.01),而Bcl-2的表达降低(P<0.01).结论 长期应用吗啡可通过Fas、Caspase-3表达的增加和Bcl-2表达的降低诱发脑细胞异常凋亡,这可能是阿片类药物引起神经损害的机制之一.%Objective To investigate the changes of apoptosis-related proteins Fas,Caspase-3 and Bcl-2 expression in rat brain during morphine addiction. Methods A total of 48 adult male Sprague-Dawley rats, weighing 190-210 g, were randomly divided into 3 groups (n=16): chronic morphine-dependent group, chronic morphine-abstinent group and control group. The rats in dependent group and abstinent group were chronically treated with morphine for 13 days to establish morphine dependent model. In the abstinent group, the withdrawal syndromes were induced with intraperitoneal injection of naloxone 5 mg/kg for 30 min. The control group was injected with normal saline. Immunohistochemistry and Western blotting analysis were used to examine the expression of Fas, Bcl-2 and Caspase-3 proteins. Results Compared with the control group, the other two groups had significantly increased expression of Fas and Caspase-3 (P

  5. Caspase-3活性改变对胆道梗阻大鼠中性粒细胞凋亡的影响%Altered caspase-3 activity results in delayed polymorphonuclear neutrophil apoptosis in rats with bile duct obstruction

    Institute of Scientific and Technical Information of China (English)

    邓雪松; 倪勇; 王成友; 詹勇强; 韩庆; 周尤星

    2012-01-01

    Objective To investigate the underlying mechanisms involved in the alteration of caspase - 3 activity on peripheral polymorphonuclear neutrophil( PMN ) apoptosis in rats with bile duct obstruction( BDO ). Methods 54 SD adult rats were divided into three groups at random: normal rats termed Group A, other rats underwent either sham - ligated operation or bile duct obstruction termed Group B or Group C. Subsequently, Group B and Group C were randomly separated into subgroups of day 1,3,7, and 10. Blood samples were collected , PMN apoptosis was evaluated by flow cytometry and caspase - 3 activity was detected by fluorescence staining. Results Group C displayed significantly decreased apoptosis of PMN from day l( 54. 34 ± 2. 35 ) to day 10( 36. 01 ± 2. 11 ), as well as attenuated activity of caspase - 3 on PMN from day l( 52. 33 ± 2. 35 ) to day 10( 34. 14 ± 3. 63 ), when compared to group A( 65. 53 ± 2. 25 ), ( 60. 58 ± 5. 35 ) and each subgroup B( P <0. 01 ). Conclusion BDO rats reveal attenuated activity of caspase - 3 , which take part in regulation on PMN apoptosis process. Delayed PMN apoptosis may contribute to the excessive inflammation and severe septic complications, which plays an important role in the initiation and development of obstructive jaundice.%目的 探讨半胱天冬酶-3(Caspase-3)活性的改变对胆道梗阻(BDO)大鼠外周血中性粒细胞(PMN)凋亡的影响.方法 54只SD大鼠随机分为正常组(A组)、假手术组(B组)和胆总管结扎组(C组),B、C组术后又分为1、3、7、10 d等4个时相,每个时相6只.留取血样标本分离PMN,应用流式细胞仪检测PMN凋亡率,采用荧光分光光度法检测Caspase-3活性.结果 C组PMN凋亡率从术后1 d的54.34±2.35降至10 d的36.01±2.11,低于A组(65.53±2.25)及B组相应时相,P<0.05;Caspase-3活性从术后1 d的52.33±2.35逐步下降至10 d的34.14±3.63,低于A组(60.58±5.35)及B组相应时相,(P<0.05).结论 BDO大鼠外周血Caspase-3活性降低,

  6. Expression of Caspase-3 in Dentate Gyrus of Adult Mice with Chronic Arsenic Poisoning%慢性砷中毒对小鼠齿状回caspase-3表达的影响

    Institute of Scientific and Technical Information of China (English)

    孙宝飞; 康朝胜; 余资江; 李玉飞

    2011-01-01

    目的 观察慢性砷中毒对成年小鼠齿状回神经元的形态学影响,探讨慢性砷中毒对成年小鼠脑部的神经毒性机制.方法 选取健康成年昆明小鼠80只,雌雄各半,分为对照组、高、中、低剂量砷染毒组,每组20只,高、中、低剂量砷染毒组分别以As03的1/5、1/10、1/40 LD50(9、4.5、1.1 mg/kg)灌胃染毒,对照组以蒸馏水灌胃,连续3个月.利用免疫组织化学和蛋白印迹技术观察小鼠齿状回部位神经元半胱氨酸蛋白水解酶-3(caspase-3)蛋白的表达.结果 免疫组化染色显示,与正常对照组比较,砷染毒组小鼠齿状回caspase-3阳性细胞明显增多(P<0.01),阳性反应产物平均光密度增高(P<0.01),同时蛋白印迹结果显示随砷中毒剂量的增加,小鼠齿状回caspase-3蛋白含量随之增加(P<0.01),各剂量组雌雄间各数据差异无统计学意义(P>0.05).结论 慢性砷中毒导致脑齿状回神经元细胞凋亡可能与齿状回细胞caspase-3增加有关,同时脑细胞caspase-3随砷浓度增加而增高.%Objective To investigate the effects of chronic arsenic exposure at different doses on dentate gyrus neurons in adult mice. Methods Eighty healthy adult Kunming mice, 20-22 g, were randomly divided into four groups: normal control group, low-dose group, moderate dose group and high dose group, 20 in each (10 males and 10 females in each group), each group was fed respectively with distilled water, 1/5 LD50, 1/10 LD50 and 1/40 LD50 As203 for 3 consecutive months, and adjusting the dose according to their weight changes. The content of arsenic in brain was determined. The expression of caspase-3 in dentate gyrus neurons was detected by western blotting and immunohistochemistry and analyzed by morphology methods. Results Compared with normal control group, groups of arsenic poisoning had been the main changes: caspase-3 immunohistochemical staining positive cells increased significantly (P0.05). Conclusion The obvious up

  7. TRAIL, DR5 and caspase 3-dependent apoptosis in vessels of diseased human temporomandibular joint disc. An immunohistochemical study

    Directory of Open Access Journals (Sweden)

    C. Loreto

    2010-09-01

    Full Text Available To evaluate the apoptosis involvement in the angiogenesis as a self-limiting process in patients with temporomandibular joint (TMJ degenerated disc vessels, we assessed, by immunohistochemistry, the detection of TRAIL, its death receptor DR5 and caspase 3. TRAIL, its death receptor DR5 and caspase 3 expression were studied by immunohistochemistry in 15 TMJ discs displaced without reduction and in 4 unaffected discs. These apoptosis molecules were detected in the intima and media layers of newly formed vessels affected discs. In conclusion, vessels apoptosis activation in TMJ disc with ID could be regarded as a self-limiting process that try to leads to vessel regression; in this way an inhibition of angiogenic vessels may prove a key strategy in limiting pathological angiogenesis, by cutting off blood supply to tumors, or by reducing harmful inflammation.

  8. The calpain, caspase 12, caspase 3 cascade leading to apoptosis is altered in F508del-CFTR expressing cells.

    Directory of Open Access Journals (Sweden)

    Mathieu Kerbiriou

    Full Text Available In cystic fibrosis (CF, the most frequent mutant variant of the cystic fibrosis transmembrane conductance regulator (CFTR, F508del-CFTR protein, is misfolded and retained in the endoplasmic reticulum (ER. We previously showed that the unfolded protein response (UPR may be triggered in CF. Since prolonged UPR activation leads to apoptosis via the calcium-calpain-caspase-12-caspase-3 cascade and because apoptosis is altered in CF, our aim was to compare the ER stress-induced apoptosis pathway between wild type (Wt and F508del-CFTR expressing cells. Here we show that the calcium-calpain-caspase-12-caspase-3 cascade is altered in F508del-CFTR expressing cells. We propose that this alteration is involved in the altered apoptosis triggering observed in CF.

  9. In Vitro and Ex Vivo Evaluation of Smart Infra-Red Fluorescent Caspase-3 Probes for Molecular Imaging of Cardiovascular Apoptosis

    Directory of Open Access Journals (Sweden)

    Manuelle Debunne

    2011-01-01

    Procedure. We developed quenched near-infrared probes which become fluorescent upon cleavage by caspase-3, the key regulatory enzyme of apoptosis. Results. Probes were shown to be selectively cleaved by recombinant caspase-3. Apoptosis of cultured endothelial cells was associated with an increased fluorescent signal for the cleaved probes, which colocalized with caspase-3 and was reduced by the addition of a caspase-3 inhibitor. Flow cytometry demonstrated a similar profile between the cleaved probes and annexin V. Ex vivo experiments showed that sections of hearts obtained from mice treated with the proapoptotic drug doxorubicin displayed an increase in the fluorescent signal for the cleaved probes, which was reduced by a caspase-3 inhibitor. Conclusion. We demonstrated the capacity of these novel probes to detect apoptosis by optical imaging in vitro and ex vivo.

  10. 完全睡眠剥夺对大鼠caspase-3表达的影响%Effect of total sleep deprivation on expression of caspase-3 in cortex and hippocampus

    Institute of Scientific and Technical Information of China (English)

    先雄斌; 刘军祥; 先德海; 郑宇杰

    2008-01-01

    目的:探讨完全睡眠剥夺对大鼠caspase-3的影响及其导致记忆、认知等功能障碍的机制.方法:取Sprague-Dawley大鼠35只,随机分为正常对照组、大平台对照组和完全睡眠剥夺组.其中正常对照组(NC)5只大鼠,正常笼养;大平台对照组(TC)只有72、96及120 h 3个时间点,每个时间点5只大鼠;完全睡眠剥夺组(TSD)也有72、96及120 h 3个时间点,每个时间点5只大鼠.采用小平台水环境法建立完全睡眠剥夺模型,在不同的时点处死大鼠后运用免疫组化SP法检测各组大鼠海马和皮质caspase-3的分布规律和动态变化.结果:caspase-3在各组的海马和皮质均有表达,随着睡眠剥夺时间的延长,caspase-3的表达逐渐增加,与正常对照组、大平台对照组差异有显著性(P<0.01).结论:完全睡眠剥夺能引起大鼠海马和皮质caspase-3的表达增加.并随睡眠剥夺时间的延长而渐趋明显.可能是完全睡眠剥夺引起大鼠神经元调亡,进而引起大鼠记忆、认知等功能障碍的机制之一.

  11. Effect of protein kinase C alpha, caspase-3, and survivin on apoptosis of oral cancer cells induced by staurosporine

    Institute of Scientific and Technical Information of China (English)

    Yu-xia ZHANG; Shi-bin YU; Jing-ping OU-YANG; Dong XIA; Min WANG; Jin-rong LI

    2005-01-01

    Aim: To elucidate inhibition of protein kinase C α (PKC α) activity by staurosporine on apoptosis of oral cancer cell line tongue squamous cell carcinoma (TSCCa)cells and to clarify the role of survivin and caspase-3 in mediating apoptosis.Methods: TSCCa cell viability was measured by MTT assay after 100 nmol/L staurosporine treatment. Apoptotic cells were identified by using phase contrast microscopy, acridine orange/ethidium bromide staining, and flow cytometry. Level of PKC α and its subcellular location were investigated using Western blot analysis.Expression of survivin and caspase-3 were evaluated using immunocytochemistry.Results: Staurosporine significantly inhibited the cell viability of TSCCa cells in a dose- and time-dependent manner. Marked cell accumulation in G2/M phase was observed after 100 nmol/L staurosporine exposure for 6 h and 12 h. In addition,the percentage of apoptosis increased in a time-dependent manner, from 2.9% in control cultures to approximately 27.4% at 100 nmol/L staurosporine treatment for24 h. Staurosporine displayed difference in inhibitory efficacy between cytosolic and membrance-derived PKC α. The content of PKCα in membrane versus cytosol decreased quickly, from 0.45 in ethanol-treated control cultures to 0.18 after staurosporine exposure for 24 h (P<0.01). After treatment withstaurosporine, a time-dependent reduction of survivin and an activation of caspase-3 were observed in TSCCa cells. Conclusion: Staurosporine inhibited cell viability and promoted apoptosis in TSCCa cells. Inhibition of PKCα activity might be a potential mechanism for staurosporine to induce apoptosis in this cell line. The cleavage of survivin and activation of caspase-3 signaling pathway might contribute to PKC α inhibition-induced apoptosis.

  12. Safrole oxide induces apoptosis by activating caspase-3, -8, and -9 in A549 human lung cancer cells.

    Science.gov (United States)

    Du, Aiying; Zhao, Baoxiang; Yin, Deling; Zhang, Shangli; Miao, Junying

    2006-01-01

    Previously we found that 3,4-(methylenedioxy)-1-(2',3'-epoxypropyl)-benzene (safrole oxide) induced a typical apoptosis in A549 human lung cancer cells. In this study, we further investigated which caspases were activated by safrole oxide during the apoptosis. The data showed that the activity of caspase-3, -8, and -9 was significantly enhanced by the compound, which suggested that safrole oxide might be used as a caspase promoter to initiate lung cancer cell apoptosis.

  13. Scutellaria barbate extract induces apoptosis of hepatoma H22 cells via the mitochondrial pathway involving caspase-3

    Institute of Scientific and Technical Information of China (English)

    Zhi-Jun Dai; Ling-Qin Song; Xi-Jing Wang; Zong-Fang Li; Zong-Zheng Ji; Hong-Tao Ren; Wei Tang; Xiao-Xu Liu; Hua-Feng Kang; Hai-Tao Guan

    2008-01-01

    AIM:To study the growth inhibitory and apoptotic effects of Scutellaria barbata D.Don (S.barbata) and to determine the underlying mechanism of its antitumor activity in mouse liver cancer cell line H22.METHODS:Proliferation of H22 cells was examined by MTr assay.Cellular morphology of PC-2 cells was observed under fluorescence microscope and transmission electron microscope (EM).Mitochondrial transmembrane potential was determined under laser scanning confocal microscope (LSCM) with rhodamine 123 staining.Flow cytometry was performed to analyze the cell cycle of H22 cells with propidium iodide staining.Protein level of cytochrome C and caspase-3 was measured by semi-quantitive RT-PCR and Western blot analysis.Activity of caspase-3 enzyme was measured by spectrofluorometry.RESULTS:M'IF assay showed that extracts from S.barbata (ESB) could inhibit the proliferation of H22 cells in a time-dependent manner.Among the various phases of cell cycle,the percentage of cells in S phase was significantly decreased,while the percentage of cells in G1 phase was increased.Flow cytometry assay also showed that ESB had a positive effect on apoptosis.Typical apoptotic morphologies such as condensation and fragmentation of nuclei and blebbing membrane of apoptotic cells could be observed under transmission electron microscope and fluorescence microscope.To further investige the molecular mechanism behind ESB-induced apoptosis,ESB-treated cells rapidly lost their mitochondrial transmembrane potential,released mitochondrial cytochrome C into cytosol,and induced caspase-3 activity in a dose-dependent manner.CONCLUSION:ESB can effectively inhibit the proliferation and induce apoptosis of H22 cells involving loss of mitochondrial transmembrane potential,release of cytochrome C,and activation of caspase-3.

  14. Diatom-derived oxylipins induce cell death in sea urchin embryos activating caspase-8 and caspase 3/7

    OpenAIRE

    N. Ruocco; Varella, S; Romano, G.; Ianora, A.; Bentley, M. G.; Somma, D.; Leonardi, A.; Mellone, S.; Zuppa, A; Costantini, M

    2016-01-01

    Diatoms are an important class of unicellular algae that produce bioactive secondary metabolites withcytotoxic activity collectively termed oxylipins, including polyunsaturated aldehydes (PUAs), hydroxy-acids (HEPEs), oxo-acids and epoxyalcohols. Previous results showed that at higher concentrations, thePUA decadienal induced apoptosis on copepods and sea urchin embryos via caspase-3 activation; atlower concentrations decadienal affected the expression levels of the caspase-8 gene in embryos ...

  15. Analysis of macrophage apoptosis induced by Brucella melitensis and the effects of caspases 3,8 and 9

    Institute of Scientific and Technical Information of China (English)

    任晓莉

    2013-01-01

    Objective To determine the difference of macrophage RAW264.7 apoptosis induced by Brucella melitensis virulent strain 16M and attenuated strain M5-90 and elucidate the regulatory role of caspases 3,8 and 9.Methods The best multiplicity of infection (MOI) was determined through kinetic analysis of Brucella melitensis strain 16M and M5-90 induced mouse macrophages apop-

  16. Endothelial apoptosis in pulmonary hypertension is controlled by a microRNA/programmed cell death 4/caspase-3 axis.

    Science.gov (United States)

    White, Kevin; Dempsie, Yvonne; Caruso, Paola; Wallace, Emma; McDonald, Robert A; Stevens, Hannah; Hatley, Mark E; Van Rooij, Eva; Morrell, Nicholas W; MacLean, Margaret R; Baker, Andrew H

    2014-07-01

    Pulmonary endothelial cell apoptosis is a transient, yet defining pathogenic event integral to the onset of many pulmonary vascular diseases such as pulmonary hypertension (PH). However, there is a paucity of information concerning the molecular pathway(s) that control pulmonary arterial endothelial cell apoptosis. Here, we introduce a molecular axis that when functionally active seems to induce pulmonary arterial endothelial cell apoptosis in vitro and PH in vivo. In response to apoptotic stimuli, human pulmonary arterial endothelial cells exhibited robust induction of a programmed cell death 4 (PDCD4)/caspase-3/apoptotic pathway that was reversible by direct PDCD4 silencing. Indirectly, this pathway was also repressed by delivery of a microRNA-21 mimic. In vivo, genetic deletion of microRNA-21 in mice (miR-21(-/-) mice) resulted in functional activation of the PDCD4/caspase-3 axis in the pulmonary tissues, leading to the onset of progressive PH. Conversely, microRNA-21-overexpressing mice (CAG-microRNA-21 mice) exhibited reduced PDCD4 expression in pulmonary tissues and were partially resistant to PH in response to chronic hypoxia plus SU 5416 injury. Furthermore, direct PDCD4 knockout in mice (PDCD4(-/-) mice) potently blocked pulmonary caspase-3 activation and the development of chronic hypoxia plus SU 5416 PH, confirming its importance in disease onset. Broadly, these findings support the existence of a microRNA-21-responsive PDCD4/caspase-3 pathway in the pulmonary tissues that when active serves to promote endothelial apoptosis in vitro and PH in vivo.

  17. Preclinical studies identify non-apoptotic low-level caspase-3 as therapeutic target in pemphigus vulgaris.

    Science.gov (United States)

    Luyet, Camille; Schulze, Katja; Sayar, Beyza S; Howald, Denise; Müller, Eliane J; Galichet, Arnaud

    2015-01-01

    The majority of pemphigus vulgaris (PV) patients suffer from a live-threatening loss of intercellular adhesion between keratinocytes (acantholysis). The disease is caused by auto-antibodies that bind to desmosomal cadherins desmoglein (Dsg) 3 or Dsg3 and Dsg1 in mucous membranes and skin. A currently unresolved controversy in PV is whether apoptosis is involved in the pathogenic process. The objective of this study was to perform preclinical studies to investigate apoptotic pathway activation in PV pathogenesis with the goal to assess its potential for clinical therapy. For this purpose, we investigated mouse and human skin keratinocyte cultures treated with PV antibodies (the experimental Dsg3 monospecific antibody AK23 or PV patients IgG), PV mouse models (passive transfer of AK23 or PVIgG into adult and neonatal mice) as well as PV patients' biopsies (n=6). A combination of TUNEL assay, analyses of membrane integrity, early apoptotic markers such as cleaved poly-ADP-ribose polymerase (PARP) and the collapse of actin cytoskeleton failed to provide evidence for apoptosis in PV pathogenesis. However, the in vitro and in vivo PV models, allowing to monitor progression of lesion formation, revealed an early, transient and low-level caspase-3 activation. Pharmacological inhibition confirmed the functional implication of caspase-3 in major events in PV such as shedding of Dsg3, keratin retraction, proliferation including c-Myc induction, p38MAPK activation and acantholysis. Together, these data identify low-level caspase-3 activation downstream of disrupted Dsg3 trans- or cis-adhesion as a major event in PV pathogenesis that is non-synonymous with apoptosis and represents, unlike apoptotic components, a promising target for clinical therapy. At a broader level, these results posit that an impairment of adhesive functions in concert with low-level, non-lethal caspase-3 activation can evoke profound cellular changes which may be of relevance for other diseases including

  18. Preclinical studies identify non-apoptotic low-level caspase-3 as therapeutic target in pemphigus vulgaris.

    Directory of Open Access Journals (Sweden)

    Camille Luyet

    Full Text Available The majority of pemphigus vulgaris (PV patients suffer from a live-threatening loss of intercellular adhesion between keratinocytes (acantholysis. The disease is caused by auto-antibodies that bind to desmosomal cadherins desmoglein (Dsg 3 or Dsg3 and Dsg1 in mucous membranes and skin. A currently unresolved controversy in PV is whether apoptosis is involved in the pathogenic process. The objective of this study was to perform preclinical studies to investigate apoptotic pathway activation in PV pathogenesis with the goal to assess its potential for clinical therapy. For this purpose, we investigated mouse and human skin keratinocyte cultures treated with PV antibodies (the experimental Dsg3 monospecific antibody AK23 or PV patients IgG, PV mouse models (passive transfer of AK23 or PVIgG into adult and neonatal mice as well as PV patients' biopsies (n=6. A combination of TUNEL assay, analyses of membrane integrity, early apoptotic markers such as cleaved poly-ADP-ribose polymerase (PARP and the collapse of actin cytoskeleton failed to provide evidence for apoptosis in PV pathogenesis. However, the in vitro and in vivo PV models, allowing to monitor progression of lesion formation, revealed an early, transient and low-level caspase-3 activation. Pharmacological inhibition confirmed the functional implication of caspase-3 in major events in PV such as shedding of Dsg3, keratin retraction, proliferation including c-Myc induction, p38MAPK activation and acantholysis. Together, these data identify low-level caspase-3 activation downstream of disrupted Dsg3 trans- or cis-adhesion as a major event in PV pathogenesis that is non-synonymous with apoptosis and represents, unlike apoptotic components, a promising target for clinical therapy. At a broader level, these results posit that an impairment of adhesive functions in concert with low-level, non-lethal caspase-3 activation can evoke profound cellular changes which may be of relevance for other

  19. Pathological cyclic strain-induced apoptosis in human periodontal ligament cells through the RhoGDIα/caspase-3/PARP pathway.

    Directory of Open Access Journals (Sweden)

    Li Wang

    Full Text Available AIM: Human periodontal ligament (PDL cells incur changes in morphology and express proteins in response to cyclic strain. However, it is not clear whether cyclic strain, especially excessive cyclic strain, induces PDL cell apoptosis and if so, what mechanism(s are responsible. The aim of the present study was to elucidate the molecular mechanisms by which pathological levels of cyclic strain induce human PDL cell apoptosis. MATERIALS AND METHODS: Human PDL cells were obtained from healthy premolar tissue. After three to five passages in culture, the cells were subjected to 20% cyclic strain at a frequency of 0.1 Hz for 6 or 24 h using an FX-5000T system. Morphological changes of the cells were assessed by inverted phase-contrast microscopy, and apoptosis was detected by fluorescein isothiocyanate (FITC-conjugated annexin V and propidium iodide staining followed by flow cytometry. Protein expression was evaluated by Western blot analysis. RESULTS: The number of apoptotic human PDL cells increased in a time-dependent manner in response to pathological cyclic strain. The stretched cells were oriented parallel to each another with their long axes perpendicular to the strain force vector. Cleaved caspase-3 and poly-ADP-ribose polymerase (PARP protein levels increased in response to pathological cyclic strain over time, while Rho GDP dissociation inhibitor alpha (RhoGDIα decreased. Furthermore, knock-down of RhoGDIα by targeted siRNA transfection increased stretch-induced apoptosis and upregulated cleaved caspase-3 and PARP protein levels. Inhibition of caspase-3 prevented stretch-induced apoptosis, but did not change RhoGDIα protein levels. CONCLUSION: The overall results suggest that pathological-level cyclic strain not only influenced morphology but also induced apoptosis in human PDL cells through the RhoGDIα/caspase-3/PARP pathway. Our findings provide novel insight into the mechanism of apoptosis induced by pathological cyclic strain in

  20. Quercetin induces apoptosis by activating caspase-3 and regulating Bcl-2 and cyclooxygenase-2 pathways in human HL-60 cells.

    Science.gov (United States)

    Niu, Guomin; Yin, Songmei; Xie, Shuangfeng; Li, Yiqing; Nie, Danian; Ma, Liping; Wang, Xiuju; Wu, Yudan

    2011-01-01

    Quercetin is one of the naturally occurring dietary flavonol compounds. It is present abundantly in plants and has chemopreventive and anticancer effects. To investigate its anticancer mechanism, we examined the activity of quercetin against acute leukemia cell line, HL-60. Our results showed that quercetin inhibited cell proliferation and induced apoptosis in a time- and dose-dependent manner. Furthermore, quercetin down-regulated the expression of anti-apoptosis protein Bcl-2 and up-regulated the expression of pro-apoptosis protein Bax. Caspase-3 was also activated by quercetin, which started a caspase-3-depended mitochodrial pathway to induce apoptosis. It was also found that quercetin inhibited the expression of the cycloocygenase-2 (Cox-2) mRNA and Cox-2 protein. Taken together, these findings suggested that quercetin induces apoptosis in a caspase-3-dependent pathway by inhibiting Cox-2 expression and regulates the expression of downstream apoptotic components, including Bcl-2 and Bax. Quercetin can be a potent and promising medicine which might be safely used in leukemia therapy.

  1. Inhibition of Corydalis decumbens Alkaloids on Hydrogen Peroxideinduced Apoptosis of PC12 Cells through Down-regulating Caspase-3 Expression

    Institute of Scientific and Technical Information of China (English)

    YAN Ren-jie; YANG Yi-fang; LUO Yong-ming; WU Chun-zhen

    2011-01-01

    Objective To extract alkaloids from Corydalis decumbens (AsCD) by supercritical CO2 fluid extraction (SFE) and to evaluate protective effects of AsCD against hydrogen peroxide (H2O2)-induced apoptosis in rat PC12 cells.Methods AsCD were extracted by SFE and oxidative damage PC12 cells model was induced by H2O2.The survival rate of the cells was determined by MTT assay; Lactate dehydrogenase release was determined by ultraviolet spectrophotometry; Flow cytometry was used to detect apoptosis; Caspase-3 mRNA and protein were determined by real-time PCR and Western blotting assay,respectively.Results AsCD remarkably reduced the cytotoxicity,prevented membrane damage,and inhibited cell apoptosis.AsCD inhibited Caspase-3 mRNA and protein expression induced by H2O2 in PC12 cells.Conclusion AsCD possess protective effects against H2O2-induced apoptosis in PC12 cells,and the mechanism of AsCD responsible to the inhibition of apoptosis is possibly attributed to thedown-regulating Caspase-3 expression.AsCD might be useful in the treatment of oxidative stress-related neurodegenerative diseases.

  2. Antisense oligonucleotide targeting Livin induces apoptosis of human bladder cancer cell via a mechanism involving caspase 3

    Directory of Open Access Journals (Sweden)

    Weili Zhang

    2010-06-01

    Full Text Available Abstract Background and Aim in recent years, Livin, a new member of IAPs family, is found to be a key molecule in cancers. Researchers consider Livin may become a new target for tumor therapy; however, the role of it in bladder cancer is still unclear. The purpose of this article is to investigate Antisense Oligonucleotide (ASODN of Livin on treating bladder cancer cell and underlying mechanisms. Methods Phosphorathioate modifying was used to synthesize antisense oligonucleotides targeting Livin, followed by transfection into human bladder cancer cell 5637. After transfection, Livin mRNA and protein level, cell proliferation and apoptosis changes, caspase3 level and its effect on human bladder cancer transplantable tumor in nude mice were measured. Result results showed Livin ASODN effectively inhibited Livin expression and tumor cell proliferation, and these effects probably through enhanced caspase3 activity and apoptosis of tumor cells. In nude mice transplantable tumor model, Livin expressions were inhibited meanwhile caspase3 expression was increased. Tumor growth slowed down and apoptosis was enhanced. Conclusion Our data suggest that Livin plays an important role in inhibiting apoptosis of bladder cancer cells. Livin ASODN may promote cell apoptosis, inhibit bladder cancer growth, and become one of the methods of gene therapy for bladder cancer.

  3. Low-dose radiation reverses cisplatin resistance in ovarian cancer cells by changing Survivin and Caspase-3 expression

    Institute of Scientific and Technical Information of China (English)

    Qing Dong; Tao Jiang; Donghai Liang; Xiaoran Liu; Hongsheng Yu

    2016-01-01

    Objective Cisplatin (DDP) is the main chemotherapy drug for ovarian cancer. However, ovarian cancer cel s tend to develop cisplatin resistance in the clinical setting. Tumor cel s are sensitive to low-dose radia-tion (LDR). LDR therapy can improve the ef ects of chemotherapy drugs on ovarian cancer, but the underly-ing mechanisms are not clear. In this study, we explored the impact of low-dose radiation on Survivin and Caspase-3 levels in SKOV3/DDP ovarian cancer cel s that are resistant to cisplatin. Methods Cel viability was examined by cel counting kit-8 (CCK-8) assay, and quantitative PCR was used to detect Caspase-3 and Survivin transcript levels. Flow cytometry was used to detect and quantify apoptotic cel s. Results Cel viability was lower when cel s were treated with LDR and cisplatin than when cel s were treated with conventional radiation and cisplatin, or cisplatin alone (P Conclusion LDR reverses cisplatin resistance in SKOV3/DDP cel s, and may do so by suppressing Survivin expression and increasing Caspase-3 expression.

  4. Diatom-derived oxylipins induce cell death in sea urchin embryos activating caspase-8 and caspase 3/7.

    Science.gov (United States)

    Ruocco, Nadia; Varrella, Stefano; Romano, Giovanna; Ianora, Adrianna; Bentley, Matt G; Somma, Domenico; Leonardi, Antonio; Mellone, Stefano; Zuppa, Antonio; Costantini, Maria

    2016-07-01

    Diatoms are an important class of unicellular algae that produce bioactive secondary metabolites with cytotoxic activity collectively termed oxylipins, including polyunsaturated aldehydes (PUAs), hydroxyacids (HEPEs), oxo-acids and epoxyalcohols. Previous results showed that at higher concentrations, the PUA decadienal induced apoptosis on copepods and sea urchin embryos via caspase-3 activation; at lower concentrations decadienal affected the expression levels of the caspase-8 gene in embryos of the sea urchin Paracentrotus lividus. In the present work, we studied the effects of other common oxylipins produced by diatoms: two PUAs (heptadienal and octadienal) and four hydroxyacids (5-, 9- 11- and 15-HEPE) on P. lividus cell death and caspase activities. Our results showed that (i) at higher concentrations PUAs and HEPEs induced apoptosis in sea urchin embryos, detected by microscopic observation and through the activation of caspase-3/7 and caspase-8 measured by luminescent assays; (ii) at low concentrations, PUAs and HEPEs affected the expression levels of caspase-8 and caspase-3/7 (isolated for the first time here in P. lividus) genes, detected by Real Time qPCR. These findings have interesting implications from the ecological point of view, given the importance of diatom blooms in nutrient-rich aquatic environments.

  5. Bcl-2/caspase 3 mucosal imbalance favors T cell resistance to apoptosis in dogs with inflammatory bowel disease.

    Science.gov (United States)

    Jergens, A; Young, J; Moore, D; Wang, C; Hostetter, J; Augustine, L; Allenspach, K; Schmitz, S; Mosher, C

    2014-04-15

    Canine idiopathic inflammatory bowel disease (IBD) is believed to result from complex interplay between genetic, microbial, and immunologic factors. Abnormal cell death by apoptosis may result in the persistence of activated intestinal T cells that contribute to mucosal inflammation and clinical severity. To test this hypothesis, we investigated the mucosal expression of pro- and anti-apoptotic proteins in different intestinal compartments and their association with inflammatory indices in dogs with IBD. Apoptosis of lamina propria (LP) T cells in duodenal, ileal, and colonic tissues in control and IBD dogs was analyzed by caspase 3/Bcl-2 immunohistochemistry and TUNEL assays. Densities and distributions of LP caspase 3 and Bcl-2 cells were correlated to histopathologic lesions and the clinical activity index (CIBDAI). Compared to control tissues, IBD dogs had significantly (Pdogs, there were significantly greater numbers of Bcl-2 cells at the apical and basilar villus in the duodenum as compared to the colon and to the apical and basilar villus in the ileum (Pdogs compared with controls (Pdogs and the CIBDAI (Pdogs with IBD. Mucosal imbalance of Bcl-2/caspase 3 expression favors T cell resistance to apoptosis which may contribute to T cell accumulation and chronic intestinal inflammation, similar to human IBD.

  6. Quercetin induces apoptosis by activating caspase-3 and regulating Bcl-2 and cyclooxygenase-2 pathways in human HL-60 cells

    Institute of Scientific and Technical Information of China (English)

    Guomin Niu; Songmei Yin; Shuangfeng Xie; Yiqing Li; Danian Nie; Liping Ma; Xiuju Wang; Yudan Wu

    2011-01-01

    Quercetin is one of the naturally occurring dietary flavo-nol compounds. It is present abundantly in plants and has chemopreventive and anticancer effects. To investigate its anticancer mechanism, we examined the activity of quercetin against acute leukemia cell line, HL-60. Our results showed that quercetin inhibited cell proliferation and induced apoptosis in a time- and dose-dependent manner. Furthermore, quercetin down-regulated the expression of anti-apoptosis protein Bcl-2 and up-regulated the expression of pro-apoptosis protein Bax. Caspase-3 was also activated by quercetin, which started a caspase-3-depended mitochodrial pathway to induce apoptosis. It was also found that quercetin inhibited the expression of the cycloocygenase-2 (Cox-2) mRNA and Cox-2 protein. Taken together, these findings suggested that quercetin induces apoptosis in a caspase-3-dependent pathway by inhibiting Cox-2 expression and regulates the expression of downstream apoptotic components, including Bcl-2 and Bax. Quercetin can be a potent and promising medicine which might be safely used in leukemia therapy.

  7. Pfaffosidic Fraction from Hebanthe paniculata Induces Cell Cycle Arrest and Caspase-3-Induced Apoptosis in HepG2 Cells

    Directory of Open Access Journals (Sweden)

    Tereza Cristina da Silva

    2015-01-01

    Full Text Available Hebanthe paniculata roots (formerly Pfaffia paniculata and popularly known as Brazilian ginseng show antineoplastic, chemopreventive, and antiproliferative properties. Functional properties of these roots and their extracts are usually attributed to the pfaffosidic fraction, which is composed mainly by pfaffosides A–F. However, the therapeutic potential of this fraction in cancer cells is not yet entirely understood. This study aimed to analyze the antitumoral effects of the purified pfaffosidic fraction or saponinic fraction on the human hepatocellular carcinoma HepG2 cell line. Cellular viability, proliferation, and apoptosis were evaluated, respectively, by MTT assay, BrdU incorporation, activated caspase-3 immunocytochemistry, and DNA fragmentation assay. Cell cycle was analyzed by flow cytometry and the cell cycle-related proteins were analyzed by quantitative PCR and Western blot. The cells exposed to pfaffosidic fraction had reduced viability and cellular growth, induced G2/M at 48 h or S at 72 h arrest, and increased sub-G1 cell population via cyclin E downregulation, p27KIP1 overexpression, and caspase-3-induced apoptosis, without affecting the DNA integrity. Antitumoral effects of pfaffosidic fraction from H. paniculata in HepG2 cells originated by multimechanisms of action might be associated with cell cycle arrest in the S phase, by CDK2 and cyclin E downregulation and p27KIP1 overexpression, besides induction of apoptosis through caspase-3 activation.

  8. Cell apoptosis in perihematomal brain regions and expression of Caspase-3 protein in patients with hypertensive intracerebral hemorrhage

    Institute of Scientific and Technical Information of China (English)

    Xinqing Zhang; Xiaoliang Yin; Kun Zhang; Zhimin Zhang; Hui Cai; Honglan Xu

    2006-01-01

    BACKGROUND: In patients with intracerebral hemorrhage (ICH), besides the space-occupying effect of hematoma, hematomal component also causes the pathological changes of perihematomal region, including the death of neurons and glial cells, vasogenic brain edema, the destruction of blood brain barrier and so on, which are the important factors to influence the prognosis of patients. Therefore, it is necessary to perform fur ther investigation and study on the pathological characteristics of injury and death of brain nerve cells. OBJECTIVE: To observe the pathological changes of apoptosis and Caspase-3 expression in perihe matomal brain regions in patients with hypertensive ICH (HICH) in different stages of onset, and analyze their relationship. DESIGN: Case-control observation. SETTING: Departments of Neurosurgery and Pathology of Beijing Chuiyangliu Hospital. PARTICIPANTS: Totally 19 patients with HICH, including 12 male, 7 female, aged (58.3±12.8) ranging from 49 to 78 years, whose mean volume of hemorrhage was (48.6±16.4) mL, were involved . All the cases conformed to the diagnostic criteria of intracerebral hemorrhage formulated in the 4th National Cerebrovascular Dis eases Conference and were confirmed by skull CT scanning. Informed consents of operation and specimens were obtained from the patients and relatives.METHODS; ①Patients with HICH who had undergone surgical evacuation of an intracerebral hematoma by traverse temporal lobe approach in the Department of Neurosurgery, Beijing Chuiyangliu Hospital from Jan uary 2004 to July 2005 were involved. Nineteen specimens of brain tissue from perihematomal region of HICH patients in different phases served as patient group. Five specimens were obtained from distant regions of patients in the super-early phase as the control group. According to the time from onset to operation, the 19 cases were divided into 3 groups: 6 cases in super-early phase(onset < 8 hours), 8 cases in early phase (onset about 8 to 24

  9. 多奈哌齐对血管性痴呆小鼠海马caspase-3表达的影响%Effect of donepezil on caspase-3 expression in hippocampus of mice with vascular dementia

    Institute of Scientific and Technical Information of China (English)

    杜江; 张德军; 王勇; 王运杰

    2012-01-01

    Objective To study the effect of donepezil on the expression change of cysteinyl aspirate specific-proteinase 3 (caspase-3) in the hippocampus of mice with vascular dementia (VD). Methods The models of VD mice were established through three times of ischemia-reperfusion in bilateral common carotid arteries. 90 Kunming mice were randomly divided into sham operation(30) ,model (30)and treatment groups (treated with donepezil). The behavioral scores were investigated by step-down and water maze tests at days 29 and 30 respectively. The expression of caspase-3 in the hippocampus of mice was examined by immunohistochemistry and western blot. Results The grades of learning and memory of mice treated with donepezil was obviously increased than those of VD mice(P<0. 05). The expression of caspase-3 in the hippocampus of mice treated with donepezil was lower than in VD mice(P<0. 05). Conclusion Donepezil can down-regulate the expression of caspase-3 in the hippocampus of mice with vascular dementia.%目的 探讨盐酸多奈哌齐对血管性痴呆(VD)小鼠海马半胱氨酸蛋白酶-3(cysteinyl aspirate specific-proteinase 3,caspase-3)表达的调节作用.方法 采用双侧颈总动脉反复缺血-再灌注法制备小鼠VD模型.90只雄性昆明小鼠随机分为3组,假手术组(30只)、模型组(30只)及盐酸多奈哌齐治疗组(30只).在术后第29天、第30天,经跳台试验和水迷宫试验进行行为学测试,用免疫组化和western blot方法检测各组小鼠海马caspase-3的表达变化.结果 盐酸多奈哌齐治疗组小鼠的学习和记忆成绩明显高于VD模型组小鼠(P<0.05);免疫组化和Western blot结果发现,盐酸多奈哌齐治疗组小鼠海马CA1区caspase-3蛋白阳性表达的平均光密度值显著低于VD模型组(P<0.05).结论 盐酸多奈哌齐能够降低VD小鼠海马caspase-3的表达.

  10. GST Caspase-3在非小细胞肺癌(NSCLC)中的表达及意义%The expression and significance of GST Caspase-3 in non-small-cell lung cancer

    Institute of Scientific and Technical Information of China (English)

    姜亦兵; 霍小东

    2009-01-01

    目的 探讨非小细胞肺癌(non-small-cell lung cancer)中谷胱甘肽s-转移酶-Π(glutathione transferase-ΠGST-Π)与半胱氨酸蛋白(Caspase-3)之间的关系以及他们的表达与意义.方法 采用免疫组织化学法检测117例NSCLC中GST-Π的表达水平,流式细胞技术检测Caspase-3的表达;根据肿瘤耐药机制的不同,选择一个耐药元素与一个凋亡促进元素对肿瘤的耐药进行分析评估.结果 GST-Π的阳性率明显高于对照组(P<0.01),GST-Π与肿瘤的分型、分期、转移均无关(P>0.05);在分化表达中随其恶性程度的增高而产生统计学意义.Caspase-3定量分析中,癌组与对照组、鳞癌与腺癌组、高中低分化组、淋巴转移与无淋巴转移组均有不同程度的统计学意义.结论 GST-Π构成的肿瘤耐药与细胞凋亡不直接互为因果,结合Caspas-3定量分析,凋亡指数均有不同程度的衰减.

  11. Evaluation of therapeutic effects of natural killer (NK) cell-based immunotherapy in mice using in vivo apoptosis bioimaging with a caspase-3 sensor.

    Science.gov (United States)

    Lee, Ho Won; Singh, Thoudam Debraj; Lee, Sang-Woo; Ha, Jeoung-Hee; Rehemtulla, Alnawaz; Ahn, Byeong-Cheol; Jeon, Young Hyun; Lee, Jaetae

    2014-07-01

    Natural killer (NK) cell-based immunotherapy is a promising strategy for cancer treatment, and caspase-3 is an important effector molecule in NK cell-mediated apoptosis in cancers. Here, we evaluated the antitumor effects of NK cell-based immunotherapy by serial noninvasive imaging of apoptosis using a caspase-3 sensor in mice with human glioma xenografts. Human glioma cells expressing both a caspase-3 sensor as a surrogate marker for caspase-3 activation and Renilla luciferase (Rluc) as a surrogate marker for cell viability were established and referred to as D54-CR cells. Human NK92 cells were used as effector cells. Treatment with NK92 cells resulted in a time- and effector number-dependent increase in bioluminescence imaging (BLI) activity of the caspase-3 sensor in D54-CR cells in vitro. Caspase-3 activation by NK92 treatment was blocked by Z-VAD treatment in D54-CR cells. Transfusion of NK92 cells induced an increase of the BLI signal by caspase-3 activation in a dose- and time-dependent manner in D54-CR tumor-bearing mice but not in PBS-treated mice. Accordingly, sequential BLI with the Rluc reporter gene revealed marked retardation of tumor growth in the NK92-treatment group but not in the PBS-treatment group. These data suggest that noninvasive imaging of apoptosis with a caspase-3 sensor can be used as an effective tool for evaluation of therapeutic efficacy as well as for optimization of NK cell-based immunotherapy.-Lee, H. W., Singh, T. D., Lee, S.-W., Ha, J.-H., Rehemtulla, A., Ahn, B.-C., Jeon, Y.-H., Lee, J. Evaluation of therapeutic effects of natural killer (NK) cell-based immunotherapy in mice using in vivo apoptosis bioimaging with a caspase-3 sensor.

  12. Effect of infectious hypodermal and haematopoietic necrosis virus (IHHNV) infection on caspase 3c expression and activity in freshwater prawn Macrobrachium rosenbergii.

    Science.gov (United States)

    Arockiaraj, Jesu; Easwvaran, Sarasvathi; Vanaraja, Puganeshwaran; Singh, Arun; Othman, Rofina Yasmin; Bhassu, Subha

    2012-01-01

    Caspase 3c (MrCasp3c) was sequenced from the freshwater giant prawn Macrobrachium rosenbergii using Illumina Solexa Genome Analyzer Technique. MrCasp3c consisted of 2080 bp nucleotide encoded 521 polypeptide with an estimated molecular mass of 59 kDa. MrCasp3c sequence contains caspase family p20 domain profile and caspase family p10 domain profile at 236-367 and 378-468 respectively. The quantitative real time PCR analysis revealed a broad expression of MrCasp3c with the highest expression in haemocyte and the lowest in stomach. The expression of MrCasp3c after challenge with the infectious hypodermal and haematopoietic necrosis virus (IHHNV) was tested in haemocyte. In addition, MrCasp3c was expressed in Escherichia coli by prokaryotic expression plasmid pMAL-c2x. The enzyme activity of MrCasp3c was also found to be up-regulated by IHHNV in haemocyte and hepatopancreas tissues. This study suggested that MrCasp3c may be an effector caspase associated with the induction of apoptosis which is potentially involved in the immune defence of M. rosenbergii.

  13. Avaliação quantitativa da intensidade da transpiração palmar e plantar em pacientes portadores de hiperidrose palmoplantar primária Quantitative assessment of the intensity of palmar and plantar sweating in patients with primary palmoplantar hyperhidrosis

    Directory of Open Access Journals (Sweden)

    Bruno Yoshihiro Parlato Sakiyama

    2012-10-01

    Full Text Available OBJETIVO: Comparar a intensidade de transpiração em palmas das mãos e planta dos pés de indivíduos portadores de hiperidrose com a de um grupo controle. MÉTODOS: Foram selecionados 50 pacientes com diagnóstico clínico de hiperidrose palmoplantar e 25 indivíduos controles. Um método objetivo de quantificação da transpiração foi utilizado com um aparelho eletrônico portátil, não invasivo, com sensores de umidade relativa e de temperatura capazes de quantificar a perda de água transepidérmica. Todos os indivíduos apresentavam índice de massa corpórea de 20-25 kg/cm² e permaneceram em repouso por 20-30 min antes das medições para reduzir a interferência externa. A mensuração foi realizada em sala climatizada com a temperatura de 21-24ºC. Os locais determinados para a aferição foram região hipotenar da face palmar e região medial da face plantar. RESULTADOS: No grupo com hiperidrose palmoplantar, as médias da intensidade de transpiração nas mãos e nos pés foram de, respectivamente, 133,6 ± 51,0 g/m²/h e 71,8 ± 40,3 g/m²/h, enquanto, no grupo controle, essas foram de 37,9 ±18,4 g/m²/h e 27,6 ± 14,3 g /m²/h. As diferenças das médias entre os grupos foram estatisticamente significativas (p OBJECTIVE: To compare individuals with and without hyperhidrosis in terms of the intensity of palmar and plantar sweating. METHODS: We selected 50 patients clinically diagnosed with palmoplantar hyperhidrosis and 25 normal individuals as controls. We quantified sweating using a portable noninvasive electronic device that has relative humidity and temperature sensors to measure transepidermal water loss. All of the individuals had a body mass index of 20-25 kg/cm². Subjects remained at rest for 20-30 min before the measurements in order to reduce external interference. The measurements were carried out in a climate-controlled environment (21-24ºC. Measurements were carried out on the hypothenar region on both hands and

  14. Meta-analysis of the relationship between single nucleotide polymorphism rs72689236 of caspase-3 and Kawasaki disease.

    Science.gov (United States)

    Xing, Yanlin; Wang, Hong; Liu, Xiaomei; Yu, Xianyi; Chen, Rui; Wang, Ce; Yu, Xuexin; Sun, Le

    2014-10-01

    Kawasaki disease is a pediatric systemic vasculitis of unknown etiology, for which a genetic influence is suspected. But whether single nucleotide polymorphism (SNP) of caspase-3 rs72689236 is associated with Kawasaki disease is controversial. The aim of our study is to assess the association between the SNP of caspase-3 and risk for Kawasaki disease. We searched PubMed, MEDLINE, EMBASE, Springer, Elsevier Science Direct, Cochrane Library Google scholar, CNKI (China National Knowledge Infrastructure, in Chinese) and Wanfang database (in Chinese) to identify studies investigating the association between rs72689236 polymorphism and Kawasaki disease occurrence. There were five eligible studies, which included 4,241 (case group 1,560; control group 2,681) participants in this meta-analysis. Pooled odds ratios (ORs) and 95 % confidence intervals (95 % CIs) were calculated in a fixed-effects model (the Mantel-Haenszel method) or a random-effects model (the DerSimonian and Laird method) when appropriate. Significant associations were found under the overall ORs for A-allele comparison (A vs. G, pooled OR 1.33, 95 % CI 1.21-1.46), AA versus GG comparison (pooled OR 1.64, 95 % CI 1.35-2.00), GA versus GG comparison (pooled OR 1.42, 95 % CI 1.24-1.63), recessive model (AA vs. GG + GA, pooled OR 1.37, 95 % CI 1.15-1.64) and dominant model (AA + GA vs. GG, pooled OR 1.47, 95 % CI 1.29-1.67). This meta-analysis suggested that SNP rs72689236 of caspase-3 might be associated with susceptibility of Kawasaki disease and the allele A might increase the risk of Kawasaki disease in Asian samples such as Japanese and Chinese. In addition, individual studies with large sample size are needed to further evaluate the associations in various ethnic populations.

  15. Thimerosal induces DNA breaks, caspase-3 activation, membrane damage, and cell death in cultured human neurons and fibroblasts.

    Science.gov (United States)

    Baskin, David S; Ngo, Hop; Didenko, Vladimir V

    2003-08-01

    Thimerosal is an organic mercurial compound used as a preservative in biomedical preparations. Little is known about the reactions of human neuronal and skin cells to its micro- and nanomolar concentrations, which can occur after using thimerosal-containing products. A useful combination of fluorescent techniques for the assessment of thimerosal toxicity is introduced. Short-term thimerosal toxicity was investigated in cultured human cerebral cortical neurons and in normal human fibroblasts. Cells were incubated with 125-nM to 250-microM concentrations of thimerosal for 45 min to 24 h. A 4', 6-diamidino-2-phenylindole dihydrochloride (DAPI) dye exclusion test was used to identify nonviable cells and terminal transferase-based nick-end labeling (TUNEL) to label DNA damage. Detection of active caspase-3 was performed in live cell cultures using a cell-permeable fluorescent caspase inhibitor. The morphology of fluorescently labeled nuclei was analyzed. After 6 h of incubation, the thimerosal toxicity was observed at 2 microM based on the manual detection of the fluorescent attached cells and at a 1-microM level with the more sensitive GENios Plus Multi-Detection Microplate Reader with Enhanced Fluorescence. The lower limit did not change after 24 h of incubation. Cortical neurons demonstrated higher sensitivity to thimerosal compared to fibroblasts. The first sign of toxicity was an increase in membrane permeability to DAPI after 2 h of incubation with 250 microM thimerosal. A 6-h incubation resulted in failure to exclude DAPI, generation of DNA breaks, caspase-3 activation, and development of morphological signs of apoptosis. We demonstrate that thimerosal in micromolar concentrations rapidly induce membrane and DNA damage and initiate caspase-3-dependent apoptosis in human neurons and fibroblasts. We conclude that a proposed combination of fluorescent techniques can be useful in analyzing the toxicity of thimerosal.

  16. G-CSF prevents caspase 3 activation in Schwann cells after sciatic nerve transection, but does not improve nerve regeneration.

    Science.gov (United States)

    Frost, Hanna K; Kodama, Akira; Ekström, Per; Dahlin, Lars B

    2016-10-15

    Exogenous granulocyte-colony stimulating factor (G-CSF) has emerged as a drug candidate for improving the outcome after peripheral nerve injuries. We raised the question if exogenous G-CSF can improve nerve regeneration following a clinically relevant model - nerve transection and repair - in healthy and diabetic rats. In short-term experiments, distance of axonal regeneration and extent of injury-induced Schwann cell death was quantified by staining for neurofilaments and cleaved caspase 3, respectively, seven days after repair. There was no difference in axonal outgrowth between G-CSF-treated and non-treated rats, regardless if healthy Wistar or diabetic Goto-Kakizaki (GK) rats were examined. However, G-CSF treatment caused a significant 13% decrease of cleaved caspase 3-positive Schwann cells at the lesion site in healthy rats, but only a trend in diabetic rats. In the distal nerve segments of healthy rats a similar trend was observed. In long-term experiments of healthy rats, regeneration outcome was evaluated at 90days after repair by presence of neurofilaments, wet weight of gastrocnemius muscle, and perception of touch (von Frey monofilament testing weekly). The presence of neurofilaments distal to the suture line was similar in G-CSF-treated and non-treated rats. The weight ratio of ipsi-over contralateral gastrocnemius muscles, and perception of touch at any time point, were likewise not affected by G-CSF treatment. In addition, the inflammatory response in short- and long-term experiments was studied by analyzing ED1 stainable macrophages in healthy rats, but in neither case was any attenuation seen at the injury site or distal to it. G-CSF can prevent caspase 3 activation in Schwann cells in the short-term, but does not detectably affect the inflammatory response, nor improve early or late axonal outgrowth or functional recovery.

  17. Phycocyanin for protecting brain ischemia-reperfusion injury and its effect on the expression of Caspase-3 mRNA

    Institute of Scientific and Technical Information of China (English)

    Xuewei Yang; Yunliang Guo; Hongbing Chen

    2006-01-01

    BACKGROUND: Phycocyanin can anti-oxidize and clear free radial. Whether its protective effect on brain is related to Caspase-3, the promoter and operator of apoptosis, is highly concerned.OBJECTIVE: To observe phycocyanin for protecting nerve function and reducing the size of cerebral infarction of rats with brain ischemia-reperfusion and its effect on the expression of Caspase-3 mRNA.DESIGN: A randomized controlled experiment.SETrING: Institute of Cerebrovascular Disease, Affiliated Hospital of Medical College of Qingdao University.MATERIALS: Totally 84 adult healthy female Wistar rats, weighing 210 to 250 g, of clean grade, were provided by the Animal Experimental Center of Shandong University. Phycocyanin (Institute of Oceanography of Chinese Academy of Sciences) was used.METHODS: This experiment was carried out in the Key Laboratory for Prevention and Treatment of Brain Diseases during May to December 2005. ① The rats were randomized into sham-operation group (n=4),control group (n=40) and phycocyanin-treated group (n=40). Middle cerebral artery occlusion/reperfusion (MACO/R) models were created on the rats of control and phycocyanin-treated groups with suture-occluded method by inserting a thread into left side external-internal carotid artery. In the sham-operation group, inserting suture was omitted. After ischemia for 1 hour and reperfusion for 2 hours, suspension of phycocyanin was intragastrically administrated into the rats of the phycocyanin-treated group at 100 mg/kg , and the same volume of normal saline was isochronously administrated into the rats of control group as the same. ② Six rats were chosen respectively from the control group and phycocyanin-treated group, then neurologic impairment degrees of rats were evaluated according to Bederson's grading. ③ Six rats were chosen respectively from the control and phycocyanin-treated groups. The isolated brain tissue was stained with triphenyltetrazolium chloride, and then the size of cerebral

  18. Expression and Significance of Caspase-3 in Lichen planus%扁平苔藓皮损区caspase-3的表达与意义

    Institute of Scientific and Technical Information of China (English)

    郝国华; 白莉

    2008-01-01

    目的:探讨凋亡基因caspase-3在扁平苔藓(Lichen planus ,Lp)病变中的作用.方法:采用免疫组化法对30例扁平苔藓皮损和30例正常皮肤组织凋亡相关蛋白caspase-3的表达情况进行检测.结果:扁平苔藓组caspase-3蛋白表达率高于正常对照组的表达率,差异有统计学意义(P<0.01).结论:caspase-3作为凋亡的最终效应分子,可能在扁平苔藓病变中起到重要作用.

  19. Características quantitativas da carcaça de cordeiros deslanados Morada Nova em função da relação volumoso:concentrado na dieta Quantitative characteristics of the carcass of Morada Nova lambs fed different dietary ratios of forage and concentrate

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    Severino Gonzaga Neto

    2006-08-01

    Full Text Available Com o objetivo de avaliar a influência da relação volumoso:concentrado sobre a composição tecidual e os rendimentos de carcaça e de cortes comerciais de cordeiros Morada Nova em confinamento, utilizaram-se 18 cordeiros com peso vivo (PV inicial de 15 kg, distribuídos em seis grupos de três animais (um em cada dieta, de acordo com a relação volumoso(V:concentrado(C: 40:60, 55:45 e 70:30. Foram avaliados os pesos de corpo vazio (PCV, de carcaça quente (PCQ e de carcaça fria (PCF, a partir dos quais foram calculados os rendimentos biológico (RB, de carcaça quente (RCQ e de carcaça fria (RCF e a perda de peso por resfriamento (PPR. A carcaça foi dividida em cinco cortes cárneos (perna, lombo, costelas, paleta e pescoço, os quais foram pesados para cálculo de seus rendimentos em relação ao peso da meia-carcaça. A perna foi dissecada em músculo, osso e gordura e os pesos desses tecidos foram expressos em peso absoluto e em porcentagem da perna. Calcularam-se as relações músculo:osso e músculo:gordura, o índice de musculosidade da perna (IMP e a área de olho-de-lombo (AOL. O aumento de 30 para 60% de concentrado na dieta elevou os rendimentos de carcaça quente e carcaça fria, assim como o rendimento biológico. Os crescentes teores de concentrado na dieta não afetaram a porcentagem de músculo na perna, mas proporcionaram maior deposição de gordura e maior área de olho-de-lombo.Eighteen lambs averaging initial body weight (BW of 15 kg were allotted to six groups of three animals (one lamb per diet and were fed diets with following forage:concentrate ratios (F:C: 1 40F:60C; 2 55F:45C; or 3 70F:30C. Lambs of each group were slaughtered when the animal fed the diet with the highest concentrate level reached 25 kg of BW. Animals were slaughtered after 16 h of fasting (feed and water. After obtaining the slaughter body weight (SBW, animals were anesthetized by electric discharge and killed by bleeding of the carotid

  20. Expressions of MDM2, Livin and Caspase-3 protein and mRNA in endometrial adenocarcinomas%学位论文摘要

    Institute of Scientific and Technical Information of China (English)

    2011-01-01

    Objective To investigate the relationship of the expression of MDM2,Livin and Caspase-3 protein and mRNA in the development of endometrioid adenocarcinoma (EA). Methods The expression levels of MDM2, Livin and Caspase-3 proteins and mRNA in EA tissues (n = 72), endometrial hyperplasia tissues (n = 60) and normal tissues ( n = 30) were examined by tissue microarray technique, immunohistochemistry( SP method) and in situ hybridization method. Results The positive expression rates of MDM2, Livin and Caspase-3 protein and mRNA in EA were respectively 80. 6% ( 58/72 ), 80. 6% ( 58/72 ), 33.3% ( 24/72 ) and 73.6% ( 53/72 ), 75.0% ( 54/72 ),27.8% (20/72). The positive rates of both MDM2 and Livin protein and mRNA in EA were higher than that in normal endometrium and endometrial hyperplasia( P < 0. 01 ). However, the positive rate of Caspase-3 in EA was lower than that in normal endometrium and endometrial hyperplasia( P < 0. 01 ). The positive expressions of MDM2 protein and mRNA were not related to the histological grade, FIGO stage, depth of invasion and lymph node metastasis. The positive expressions of Livin and Caspase-3 protein and mRNA were related to histological grade (P <0. 01 ,P <0.05 ), but they were not related to FIGO stage, depth of invasion and the lymph node metastasis. The expressions of MDM2, Livin and Caspase-3 protein were positively correlated with their mRNA. The expression of Livin was negatively correlated Caspase-3. Conclusion The expressions of MDM2, Livin and Caspase-3 protein and mRNA correlate with the dedvelopment and progression of EA, which may be valuable biomarkers to detect the early carcinogenesis and prognosis of EA.

  1. Analise quantitativa de diclofenaco em preparações farmaceuticas

    OpenAIRE

    2004-01-01

    Resumo: O diclofenaco é um composto não-esteróide com propriedades anti-reumáticas, antiinflamatórias e antipiréticas. Ele é comercializado na forma de sal de sódio e de potássio. Há na literatura um número significativo de procedimentos analíticos para a quantificação deste princípio ativo, contudo, a maioria destas técnicas é cara e de procedimento relativamente demorado. Existem poucos procedimentos experimentais na região do visível para a sua análise quantitativa. Neste trabalho foram es...

  2. The Growing Complexity of Cancer Cell Response to DNA-Damaging Agents: Caspase 3 Mediates Cell Death or Survival?

    Science.gov (United States)

    Mirzayans, Razmik; Andrais, Bonnie; Kumar, Piyush; Murray, David

    2016-05-11

    It is widely stated that wild-type p53 either mediates the activation of cell cycle checkpoints to facilitate DNA repair and promote cell survival, or orchestrates apoptotic cell death following exposure to cancer therapeutic agents. This reigning paradigm has been challenged by numerous discoveries with different human cell types, including solid tumor-derived cell lines. Thus, activation of the p53 signaling pathway by ionizing radiation and other DNA-damaging agents hinders apoptosis and triggers growth arrest (e.g., through premature senescence) in some genetic backgrounds; such growth arrested cells remain viable, secrete growth-promoting factors, and give rise to progeny with stem cell-like properties. In addition, caspase 3, which is best known for its role in the execution phase of apoptosis, has been recently reported to facilitate (rather than suppress) DNA damage-induced genomic instability and carcinogenesis. This observation is consistent with an earlier report demonstrating that caspase 3 mediates secretion of the pro-survival factor prostaglandin E₂, which in turn promotes enrichment of tumor repopulating cells. In this article, we review these and related discoveries and point out novel cancer therapeutic strategies. One of our objectives is to demonstrate the growing complexity of the DNA damage response beyond the conventional "repair and survive, or die" hypothesis.

  3. Post-Traumatic Caspase-3 Expression in the Adjacent Areas of Growth Plate Injury Site: A Morphological Study

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    Karin Pichler

    2013-07-01

    Full Text Available The epiphyseal plate is a hyaline cartilage plate that sits between the diaphysis and the epiphysis. The objective of this study was to determine the impact of an injury in the growth plate chondrocytes through the study of histological morphology, immunohistochemistry, histomorphometry and Western Blot analyses of the caspase-3 and cleaved PARP-1, and levels of the inflammatory cytokines, Interleukin-6 (IL-6 and Tumor Necrosis Factor alpha (TNF-α, in order to acquire more information about post-injury reactions of physeal cell turnover. In our results, morphological analysis showed that in experimental bones, neo-formed bone trabeculae—resulting from bone formation repair—invaded the growth plate and reached the metaphyseal bone tissue (bone bridge, and this could result in some growth arrest. We demonstrated, by ELISA, increased expression levels of the inflammatory cytokines IL-6 and TNF-α. Immunohistochemistry, histomorphometry and Western Blot analyses of the caspase-3 and cleaved PARP-1 showed that the physeal apoptosis rate of the experimental bones was significantly higher than that of the control ones. In conclusion, we could assume that the inflammation process causes stress to chondrocytes that will die as a biological defense mechanism, and will also increase the survival of new chondrocytes for maintaining cell homeostasis. Nevertheless, the exact stimulus leading to the increased apoptosis rate, observed after injury, needs additional research to understand the possible contribution of chondrocyte apoptosis to growth disturbance.

  4. Post-Traumatic Caspase-3 Expression in the Adjacent Areas of Growth Plate Injury Site: A Morphological Study

    Science.gov (United States)

    Musumeci, Giuseppe; Castrogiovanni, Paola; Loreto, Carla; Castorina, Sergio; Pichler, Karin; Weinberg, Annelie Martina

    2013-01-01

    The epiphyseal plate is a hyaline cartilage plate that sits between the diaphysis and the epiphysis. The objective of this study was to determine the impact of an injury in the growth plate chondrocytes through the study of histological morphology, immunohistochemistry, histomorphometry and Western Blot analyses of the caspase-3 and cleaved PARP-1, and levels of the inflammatory cytokines, Interleukin-6 (IL-6) and Tumor Necrosis Factor alpha (TNF-α), in order to acquire more information about post-injury reactions of physeal cell turnover. In our results, morphological analysis showed that in experimental bones, neo-formed bone trabeculae—resulting from bone formation repair—invaded the growth plate and reached the metaphyseal bone tissue (bone bridge), and this could result in some growth arrest. We demonstrated, by ELISA, increased expression levels of the inflammatory cytokines IL-6 and TNF-α. Immunohistochemistry, histomorphometry and Western Blot analyses of the caspase-3 and cleaved PARP-1 showed that the physeal apoptosis rate of the experimental bones was significantly higher than that of the control ones. In conclusion, we could assume that the inflammation process causes stress to chondrocytes that will die as a biological defense mechanism, and will also increase the survival of new chondrocytes for maintaining cell homeostasis. Nevertheless, the exact stimulus leading to the increased apoptosis rate, observed after injury, needs additional research to understand the possible contribution of chondrocyte apoptosis to growth disturbance. PMID:23899790

  5. In silico identification and crystal structure validation of caspase-3 inhibitors without a P1 aspartic acid moiety

    Science.gov (United States)

    Ganesan, Rajkumar; Jelakovic, Stjepan; Mittl, Peer R. E.; Caflisch, Amedeo; Grütter, Markus G.

    2011-01-01

    Using a fragment-based docking procedure, several small-molecule inhibitors of caspase-3 were identified and tested and the crystal structures of three inhibitor complexes were determined. The crystal structures revealed that one inhibitor (NSC 18508) occupies only the S1 subsite, while two other inhibitors (NSC 89167 and NSC 251810) bind only to the prime part of the substrate-binding site. One of the major conformational changes observed in all three caspase-3–inhibitor complexes is a rotation of the Tyr204 side chain, which blocks the S2 subsite. In addition, the structural variability of the residues shaping the S1–S4 as well as the S1′ subsites supports an induced-fit mechanism for the binding of the inhibitors in the active site. The high-resolution crystal structures reported here provide novel insights into the architecture of the substrate-binding site, which might be useful for the design of more potent caspase inhibitors. PMID:21821879

  6. Hepatitis C virus induced a novel apoptosis-like death of pancreatic beta cells through a caspase 3-dependent pathway.

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    Qian Wang

    Full Text Available Epidemiological and experimental studies have suggested that Hepatitis C virus (HCV infection is associated with the development of type 2 diabetes. Pancreatic beta cell failure is central to the progression of type 2 diabetes. Using virus infection system, we investigate the influence of HCV infection on the fate of the insulinoma cell line, MIN6. Our experiments demonstrate that the HCV virion itself is indispensable and has a dose- and time-dependent cytopathic effect on the cells. HCV infection inhibits cell proliferation and induces death of MIN6 cells with apoptotic characteristics, including cell surface exposure of phosphatidylserine, decreased mitochondrial membrane potential, activation of caspase 3 and poly (ADP-ribose polymerase, and DNA fragmentation in the nucleus. However, the fact that HCV-infected cells exhibit a dilated, low-density nucleus with intact plasma and nuclear membrane indicates that a novel apoptosis-like death occurs. HCV infection also causes endoplasmic reticulum (ER stress. Further, HCV RNA replication was detected in MIN6 cells, although the infection efficiency is very low and no progeny virus particle generates. Taken together, our data suggest that HCV infection induces death of pancreatic beta cells through an ER stress-involved, caspase 3-dependent, special pathway.

  7. Exposure to 1950-MHz TD-SCDMA electromagnetic fields affects the apoptosis of astrocytes via caspase-3-dependent pathway.

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    Yu-xiao Liu

    Full Text Available The usage of mobile phone increases globally. However, there is still a paucity of data about the impact of electromagnetic fields (EMF on human health. This study investigated whether EMF radiation would alter the biology of glial cells and act as a tumor-promoting agent. We exposed rat astrocytes and C6 glioma cells to 1950-MHz TD-SCDMA for 12, 24 and 48 h respectively, and found that EMF exposure had differential effects on rat astroctyes and C6 glioma cells. A 48 h of exposure damaged the mitochondria and induced significant apoptosis of astrocytes. Moreover, caspase-3, a hallmark of apoptosis, was highlighted in astrocytes after 48 h of EMF exposure, accompanied by a significantly increased expression of bax and reduced level of bcl-2. The tumorigenicity assays demonstrated that astrocytes did not form tumors in both control and exposure groups. In contrast, the unexposed and exposed C6 glioma cells show no significant differences in both biological feature and tumor formation ability. Therefore, our results implied that exposure to the EMF of 1950-MHz TD-SCDMA may not promote the tumor formation, but continuous exposure damaged the mitochondria of astrocytes and induce apoptosis through a caspase-3-dependent pathway with the involvement of bax and bcl-2.

  8. Cadmium modulates H-ras expression and caspase-3 apoptotic cell death in breast cancer epithelial MCF-7 cells.

    Science.gov (United States)

    Petanidis, Savvas; Hadzopoulou-Cladaras, Margarita; Salifoglou, Athanasios

    2013-04-01

    Cadmium (Cd) is a well-known metal carcinogen associated with tumor formation and carcinogenesis. It has been shown to induce cancer through various cellular mechanisms involving inhibition of DNA repair, abnormal gene expression, induction of oxidative stress, and triggering apoptosis. It is well-established that the H-ras oncogene is involved in the process of carcinogenesis with direct effects on cellular proliferation and tumorigenesis. Given the biotoxicity of cadmium and its association with carcinogenesis, the effect of that metal ion (Cd(II)) was investigated, in a concentration-dependent fashion, on cell viability, cell proliferation, caspase-3 mediated apoptosis and H-ras gene expression in human breast cancer epithelial MCF-7 cells transfected with the H-ras oncogene (wild type and G12V mutation). The findings show a significant modulation effect of cadmium on H-ras gene expression accompanied by up-regulation of caspase-3-related apoptosis in the concentration range of 100-1000 nΜ cadmium. Concurrently, there is a decrease in MCF-7 proliferation. Collectively, the results a) indicate an interplay of cadmium with H-ras(wt and G12V), with cadmium exhibiting a significant concentration-dependent effect on the modulation of H-ras expression, cell viability and proliferation, and b) project distinctly interwoven roles for both cadmium and H-ras in aberrant physiologies in cancer cells.

  9. Genetically Encoded FRET-Sensor Based on Terbium Chelate and Red Fluorescent Protein for Detection of Caspase-3 Activity

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    Alexander S. Goryashchenko

    2015-07-01

    Full Text Available This article describes the genetically encoded caspase-3 FRET-sensor based on the terbium-binding peptide, cleavable linker with caspase-3 recognition site, and red fluorescent protein TagRFP. The engineered construction performs two induction-resonance energy transfer processes: from tryptophan of the terbium-binding peptide to Tb3+ and from sensitized Tb3+ to acceptor—the chromophore of TagRFP. Long-lived terbium-sensitized emission (microseconds, pulse excitation source, and time-resolved detection were utilized to eliminate directly excited TagRFP fluorescence and background cellular autofluorescence, which lasts a fraction of nanosecond, and thus to improve sensitivity of analyses. Furthermore the technique facilitates selective detection of fluorescence, induced by uncleaved acceptor emission. For the first time it was shown that fluorescence resonance energy transfer between sensitized terbium and TagRFP in the engineered construction can be studied via detection of microsecond TagRFP fluorescence intensities. The lifetime and distance distribution between donor and acceptor were calculated using molecular dynamics simulation. Using this data, quantum yield of terbium ions with binding peptide was estimated.

  10. Clozapine-induced cardiotoxicity in rats: Involvement of tumour necrosis factor alpha, NF-κβ and caspase-3

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    Basel A. Abdel-Wahab

    2014-01-01

    Full Text Available Clozapine, an ideal antipsychotic drug for the treatment of resistant schizophrenia, is considered the most underutilised treatment for schizophrenia. However, safety concerns have been raised about clozapine-induced cardiotoxicity, which may lead to sudden death, particularly in young patients. The exact mechanism of clozapine cardiotoxicity has not yet been thoroughly studied. This study aimed to investigate the possible mechanisms of clozapine-induced cardiotoxicity in a rat model. Young male Wistar rats were treated with clozapine (10, 15 and 25 mg/kg/day, i.p. for 21 days. Haemodynamic and echocardiographic studies were performed for assessment of cardiac functions. Heart sections were studied histopathologically and immunohistochemically. Serum and cardiac markers of cardiotoxicity, oxidative stress, inflammation and apoptosis were evaluated. Heart sections of CLZ-treated animals showed increased cardiac inflammation that correlated with the clozapine dose. Serum levels of CK-MB and LDH levels increased, as did cardiac levels of TNF-α, MDA, NO, myeloperoxidase (MPO, 8-OHdG, caspase-3 and NF-κB p65. In contrast, GSH levels and GSH-Px activity decreased. Furthermore, immunohistochemical examination of the heart sections showed positive immunostaining for both 3-nitrotyrosine and caspase-3 in all clozapine-treated groups. Clozapine, particularly in relatively high doses, has a clear cardiotoxic effect. This cardiotoxicity is accompanied by increased myocardial oxidative stress, inflammatory cytokines, DNA damage and apoptosis with attenuation in antioxidant defences, thus explaining the previously reported myocarditis and pericarditis during clozapine therapy in clinical studies.

  11. Targeting caspase-3 as dual therapeutic benefits by RNAi facilitating brain-targeted nanoparticles in a rat model of Parkinson's disease.

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    Yang Liu

    Full Text Available The activation of caspase-3 is an important hallmark in Parkinson's disease. It could induce neuron death by apoptosis and microglia activation by inflammation. As a result, inhibition the activation of caspase-3 would exert synergistic dual effect in brain in order to prevent the progress of Parkinson's disease. Silencing caspase-3 genes by RNA interference could inhibit the activation of caspase-3. We developed a brain-targeted gene delivery system based on non-viral gene vector, dendrigraft poly-L-lysines. A rabies virus glycoprotein peptide with 29 amino-acid linked to dendrigraft poly-L-lysines could render gene vectors the ability to get across the blood brain barrier by specific receptor mediated transcytosis. The resultant brain-targeted vector was complexed with caspase-3 short hairpin RNA coding plasmid DNA, yielding nanoparticles. In vivo imaging analysis indicated the targeted nanoparticles could accumulate in brain more efficiently than non-targeted ones. A multiple dosing regimen by weekly intravenous administration of the nanoparticles could reduce activated casapse-3 levels, significantly improve locomotor activity and rescue dopaminergic neuronal loss and in Parkinson's disease rats' brain. These results indicated the rabies virus glycoprotein peptide modified brain-targeted nanoparticles were promising gene delivery system for RNA interference to achieve anti-apoptotic and anti-inflammation synergistic therapeutic effects by down-regulation the expression and activation of caspase-3.

  12. Lipopolysaccharide-enhanced early proliferation of insulin secreting NIT-1 cell is associated with nuclear factor-kappaBmediated inhibition of caspase 3 cleavage

    Institute of Scientific and Technical Information of China (English)

    LIU Shan-ying; LIANG Qi-jun; LIN Tian-xin; FAN Xin-lan; LIANG Ying; Uwe Heemann; LI Yan

    2011-01-01

    Background Increased levels of plasma lipopolysaccharide (LPS) have been found in obesity and diabetes patients.This study was to investigate the effect of LPS on pancreatic beta-cell viability and the involvement of caspase 3 in NIT-1 cell line.Methods Mouse insulinoma NIT-1 cells were treated with LPS for the indicated time and dose.Cell viability was measured by cell counting kit-8 reagent.Toll-like receptor 4 (TLR4),caspase 3 and cleaved caspase 3 were detected by Western blotting.Insulin was determined by radioimmunoassay (RIA).Results LPS promoted NIT-1 cell proliferation at 1 μg/ml,peaked at 72 hours of incubation.A reduction in cleavage of caspase 3 was observed upon LPS treatment.Bay11-7082,a specific inhibitor of nuclear factor (NF)-κB,blunted LPS-induced inhibition of caspase 3 cleavage.Reduction in chronic insulin secretion was observed after treatment with LPS at 1 μg/ml for 48 and 72 hours,not for 24 hours.TLR4 protein was upregulated when NIT-1 cells were treated with LPS at 1 pg/ml for 24 hours.Conclusions LPS promotes early NIT-1 cell proliferation in association with NF-κB-mediated inhibition of caspase 3 cleavage.LPS exerts a time-dependent inhibitory effect on chronic insulin secretion from NIT-1 cells.

  13. Flavonoids of Rosa roxburghii Tratt exhibit radioprotection and anti-apoptosis properties via the Bcl-2(Ca(2+))/Caspase-3/PARP-1 pathway.

    Science.gov (United States)

    Xu, Ping; Cai, Xinhua; Zhang, Wenbo; Li, Yana; Qiu, Peiyong; Lu, Dandan; He, Xiaoyang

    2016-10-01

    The objective of our study was to assess the radioprotective effect of flavonoids extracted from Rosa roxburghii Tratt (FRT) and investigate the role of Bcl-2(Ca(2+))/Caspase-3/PARP-1 pathway in radiation-induced apoptosis. Cells and mice were exposed to (60)Co γ-rays at a dose of 6 Gy. The radiation treatment induced significant effects on tissue pathological changes, apoptosis, Ca(2+), ROS, DNA damage, and expression levels of Bcl-2, Caspase-3 (C-Caspase-3), and PARP-1. The results showed that FRT acted as an antioxidant, reduced DNA damage, corrected the pathological changes of the tissue induced by radiation, promoted the formation of spleen nodules, resisted sperm aberration, and protected the thymus. FRT significantly reduced cell apoptosis compared with the irradiation group. The expression of Ca(2+) and C-Caspase-3 was decreased after FRT treatment compared with the radiation-treated group. At the same time, expression of prototype PARP-1 and Bcl-2 increased, leading to a decrease in the percentage of apoptosis cells in FRT treatment groups. We conclude that FRT acts as a radioprotector. Apoptosis signals were activated via the Bcl-2(Ca(2+))/Caspase-3/PARP-1 pathway in irradiated cells and FRT inhibited this pathway of apoptosis by down-regulation of C-Caspase-3 and Ca(2+) and up-regulation of prototype PARP-1 and Bcl-2.

  14. Oridonin induces apoptosis of HeLa cells via altering expres sion of Bcl-2/Bax and activating caspase-3/ICAD pathway

    Institute of Scientific and Technical Information of China (English)

    Chun-ling ZHANG; Li-jun WU; Shin-ichi TASHIRO; Satoshi ONODERA; Takashi IKEJIMA

    2004-01-01

    AIM: To study the mechanisms by which oridonin inhibited HeLa cell growth in vitro. METHODS: Viability of oridonin-induced HeLa cells was measured by MTT assay. Apoptotic cells with condensed nuclei were visualized by phase contrast microscopy. Nucleosomal DNA fragmentation was assayed by agarose gel electrophoresis.Caspase activity was assayed using fiuorometric protease assay. ICAD, Bcl-2, and Bax proteins expression were detected by Western blot analysis. RESULTS: Oridonin induced oligonucleosomal fragmentation of DNA and increased caspase-3 activity, on the other hand, reduced the expression of inhibitor of caspase-3-activated DNase (ICAD), a caspase-3 substrate, at 12 h in HeLa cells. Oridonin-induced DNA fragmentation, caspase-3 activation and down-regulation of ICAD expression were effectively inhibited by a caspase-3 inhibitor, z-DEVD-fmk (z-AspGlu-Val-Asp-fmk). However, pretreatment with an inhibitor of poly (ADP-ribose) polymerase (PARP), 3, 4-dihydro5-[4-(1-piperidinyl)butoxy]-1 (2H)-isoquinolinone (DPQ), did not suppress oridonin-induced HeLa cell death. In addition, oridonin-induced apoptosis was associated with an increase in the expression of the apoptosis inducer Bax, and a significant reduction in expression of the apoptosis suppressor Bcl-2 in mitochondria. CONCLUSION:Oridonin induces HeLa cells apoptosis by altering balance of Bcl-2 and Bax protein expression and activation of caspase-3/ICAD pathway.

  15. EXPRESSION AND CLINICAL SIGNIFICANCE OF SURVIVIN,CASPASE-3, GST AND PGP IN NSCLC%Survivin Caspase-3 GST Pgp在非小细胞肺癌(NSCLC)中的表达及其临床意义

    Institute of Scientific and Technical Information of China (English)

    霍小东; 孙勤暖; 李国华

    2006-01-01

    目的:探讨非小细胞肺癌(non-small-cell lung cancer,NSCLC)中存活素(survivin)、谷胱甘肽S-转移酶-π(glutathions-transferase-π,GST-π)、P-糖蛋白(P-glycoprotein,Pgp)与半胱氨酸蛋白酶(Caspase-3)之间的关系,以及它们的表达与临床各病理指标之间的关系.方法:采用免疫组织化学法检测117例NSCLC中Survivin 、GST-π、Pgp的表达水平,流式细胞术检测Caspase-3的表达FI值;根据肿瘤耐药机制的不同,选择一个凋亡抑制元素、一个凋亡促进元素与两个耐药元素对肿瘤的耐药进行多方面的分析评估.结果:Survivin、GST-π、Pgp的阳性率明显高于对照组(P<0.01),Survivin、Pgp与肿瘤的分型无关(P>0.05),二者在分期分化表达中随其恶性程度增加,表达增强,而具有统计学意义(P<0.05);Pgp在淋巴结转移中高表达,与肿瘤转移有关;Survivin在淋巴结转移中的表达无显著性差异,与肿瘤转移无关;GST-π与肿瘤的分型、分期、转移均无关(P>0.05);在分化表达中与前二者相同.Caspase-3定量分析中,癌组与对照组、鳞癌与腺癌组、高中低分化组、淋巴结转移与无淋巴结转移组之间均有不同程度的统计学意义.结论:Survivin、Pgp高表达时对肿瘤的原发耐药增强,结合Caspase-3定量分析,FI值均有不同程度的衰减.通过生物学行为研究,了解凋亡抑制是肿瘤耐药的主要原因,这有助于临床正确制订初始化疗方案和评估预后.

  16. Efeito da suplementação de selenometionina e vitamina C sobre as características quantitativas e qualitativas do sêmen de coelho = Effect of selenomethionine and vitamin C supplementation on quantitative and qualitative characteristics of rabbit semen

    Directory of Open Access Journals (Sweden)

    Cristiane Arieta Alvarez

    2006-04-01

    Full Text Available Foram avaliadas as características quantitativas e qualitativas do sêmen de coelhos alimentados com rações contendo diferentes níveis de selenometionina e vitamina C (VC. Utilizaram-se 125 coelhos machos, alojados individualmente e distribuídos em um delineamento experimental inteiramente casualizado, com vinte cinco tratamentos e cinco repetições. As colheitas de sêmen foram realizadas uma vez por semana, durante oito semanas. Os parâmetros avaliados foram: volume, cor, pH, motilidade espermática progressiva, vigor espermático e concentração espermática. Foi observado que a cor e a concentração espermática melhoraram à medida que foi adicionado VC na ração. Contudo, volume, pH, motilidade espermática progressiva e vigor espermático não foram significativos (p>0,05 nos tratamentos. O consumo de ração aumentou (pQuantitative and qualitative semen characteristics of rabbits fed with rations containing different selenomethionine and vitamin C (VC levels were studied. 125 male rabbits kept individually in cage were used. They were distributed in a completely randomized design with twenty-five treatments and five replications. Semen was collected once a week. The following parameters were evaluated: volume, color, pH, progressive motility, spermatic vigor, and spermatic concentration. Significant difference (p0.05 among the treatments. The ration intake increased lineally according to the increase of the VC levels, but there was not any effect of selenomethionine levels. Results showed that vitamin C improved the semen color and spermatic concentration and increased the ration intake.

  17. Efeito da suplementação de selenometionina e vitamina C sobre as características quantitativas e qualitativas do sêmen de coelho - DOI: 10.4025/actascianimsci.v28i2.654 Effect of selenomethionine and vitamin C supplementation on quantitative - DOI: 10.4025/actascianimsci.v28i2.654

    Directory of Open Access Journals (Sweden)

    Cláudio Scapinello

    2006-01-01

    Full Text Available Foram avaliadas as características quantitativas e qualitativas do sêmen de coelhos alimentados com rações contendo diferentes níveis de selenometionina e vitamina C (VC. Utilizaram-se 125 coelhos machos, alojados individualmente e distribuídos em um delineamento experimental inteiramente casualizado, com vinte cinco tratamentos e cinco repetições. As colheitas de sêmen foram realizadas uma vez por semana, durante oito semanas. Os parâmetros avaliados foram: volume, cor, pH, motilidade espermática progressiva, vigor espermático e concentração espermática. Foi observado que a cor e a concentração espermática melhoraram à medida que foi adicionado VC na ração. Contudo, volume, pH, motilidade espermática progressiva e vigor espermático não foram significativos (p > 0,05 nos tratamentos. O consumo de ração aumentou (p Quantitative and qualitative semen characteristics of rabbits fed with rations containing different selenomethionine and vitamin C (VC levels were studied. 125 male rabbits kept individually in cage were used. They were distributed in a completely randomized design with twenty-five treatments and five replications. Semen was collected once a week. The following parameters were evaluated: volume, color, pH, progressive motility, spermatic vigor, and spermatic concentration. Significant difference (p 0.05 among the treatments. The ration intake increased lineally according to the increase of the VC levels, but there was not any effect of selenomethionine levels. Results showed that vitamin C improved the semen color and spermatic concentration and increased the ration intake.

  18. Ethanol extracts of Cinnamomum kanehirai Hayata leaves induce apoptosis in human hepatoma cell through caspase-3 cascade

    Directory of Open Access Journals (Sweden)

    Liu YK

    2014-12-01

    Full Text Available Yu-Kuo Liu,1 Kuan-Hsing Chen,2 Yann-Lii Leu,3,4 Tzong-Der Way,5 Ling-Wei Wang,6,7 Yu-Jen Chen,8,9,* Yu-Ming Liu6–8,* 1Department of Chemical and Material Engineering, Chang Gung University, Kwei-Shan, Tao-Yuan, Taiwan; 2Kidney Research Center, Chang Gung Memorial Hospital, School of Medicine, 3Graduate Institute of Natural Products, College of Medicine, 4Chinese Herbal Medicine Research Team, Healthy Aging Research Center, Chang Gung University, Taoyuan, Taiwan; 5Department of Biological Science and Technology, China Medical University, Taichung, Taiwan; 6Division of Radiation Oncology, Department of Oncology, Taipei Veterans General Hospital, Taipei, Taiwan; 7National Yang-Ming University, Taipei, Taiwan; 8School of Medicine, Institute of Traditional Medicine, National Yang Ming University, Taipei, Taiwan; 9Department of Radiation Oncology, Mackay Memorial Hospital, Taipei, Taiwan *These authors contributed equally to this workAbstract: Inducing apoptosis to susceptible cells is the major mechanism of most cytotoxic anticancer drugs in current use. Cinnamomum kanehirai Hayata (Lauraceae, a unique and native tree of Taiwan, is the major host for the medicinal fungus Antrodia cinnamomea which exhibits anti-cancer activity. Because of the scarcity of A. cinnamomea, C. kanehirai Hayata instead, is used as fork medicine in liver cancer. Here we observed the C. kanehirai Hayata ethanol extract could inhibit the cellular viability of both HepG2 and HA22T/VGH human hepatoma cell lines in a dose- and time-dependent manner. We found the mode of cell death was apoptosis according to cell morphological changes by Liu's stain, oligonucleosomal DNA fragmentation by gel electrophoresis, externalization of phosphotidyl serine by detecting Annexin V and hypoploid population by cell cycle analysis. Our results showed that the extracts caused cleavage of caspase-3 and increased enzyme activity of caspase-8 and caspase-9. Caspase 3 inhibitor partially reversed

  19. The forkhead transcription factor FOXO3a controls microglial inflammatory activation and eventual apoptotic injury through caspase 3.

    Science.gov (United States)

    Shang, Yan Chen; Chong, Zhao Zhong; Hou, Jinling; Maiese, Kenneth

    2009-02-01

    Memory loss and cognitive failure are increasingly being identified as potential risks with the recognized increase in life expectancy of the general population. As a result, the development of novel therapeutic strategies for disorders such as Alzheimer's disease have garnered increased attention. The etiologies that can lead to Alzheimer's disease are extremely varied, but a number of therapeutic options are directed against amyloid-beta peptide and inflammatory cell regulation to prevent or halt progressive cognitive loss. In particular, inflammatory microglial cells may have disparate functions that in some scenarios lead to disability through the removal of functional neurovascular cells and in other circumstances foster tissue repair. Given the significance microglial cells hold for neurodegenerative disorders, we therefore examined the function that amyloid (Abeta(1-42)) has upon the microglial cell line EOC 2 and identified a novel role for the forkhead transcription factor FoxO3a and caspase 3. Here we show that Abeta(1-42) leads to progressive injury and apoptotic cell loss in microglial cells that involves both early phosphatidylserine (PS) externalization and late genomic DNA fragmentation over a 24 hour course. Prior to these injury programs, Abeta(1-42) results in the activation and proliferation of microglia as demonstrated by increased proliferating cell nuclear antigen (PCNA) expression and bromodeoxyuridine (BrdU) uptake. Both apoptotic injury as well as the prior activation and proliferation of microglial cells relies upon the presence of FoxO3a, since specific gene silencing of FoxO3a promotes microglial cell protection and prevents the early activation and proliferation of these cells. Furthermore, Abeta(1-42) exposure maintained FoxO3a in an unphosphorylated "active" state and facilitated the cellular trafficking of FoxO3a from the cytoplasm to the cell nucleus to potentially lead to "pro-apoptotic" programs by this transcription factor. One

  20. The caspase 3-dependent apoptotic effect of pycnogenol in human oral squamous cell carcinoma HSC-3 cells.

    Science.gov (United States)

    Yang, In-Hyoung; Shin, Ji-Ae; Kim, Lee-Han; Kwon, Ki Han; Cho, Sung-Dae

    2016-01-01

    In the present study, the apoptotic effect of pycnogenol and its molecular mechanism in human oral squamous cell carcinoma HSC-3 cells were investigated. Pycnogenol significantly inhibited the viability of HSC-3 cells and suppressed neoplastic cell transformation in HSC-3 cells and TPA-treated JB6 cells. It caused caspase-dependent apoptosis evidenced by the increase in cleaved poly (ADP-ribose) polymerase and caspase 3 in a dose-dependent manner. Pycnogenol increased Bak protein by enhancing its protein stability whereas other Bcl-2 family members were not altered. In addition, the treatment with pycnogenol led to the production of reactive oxygen species and N-acetyl-l-cysteine almost blocked pycnogenol-induced reactive oxygen species generation. Taken together, these findings suggest that pycnogenol may be a potential candidate for the chemoprevention or chemotherapy of human oral cancer.

  1. The caspase 3 sensor Phiphilux G2D2 is activated non-specifically in S1 renal proximal tubules

    Science.gov (United States)

    Hato, Takashi; Sandoval, Ruben; Dagher, Pierre C

    2016-01-01

    Tubular cell apoptosis is a major phenotype of cell death in various forms of acute kidney injury. Quantifying apoptosis in fixed tissues is problematic because apoptosis evolves over time and dead cells are rapidly cleared by the phagocytic system. Phiphilux is a fluorescent probe that is activated specifically by caspase 3 and does not inhibit the subsequent activity of this effector caspase. It has been used successfully to quantify apoptosis in cell culture. Here we examined the feasibility of using Phiphilux to measure renal tubular apoptosis progression over time in live animals using intravital 2-photon microscopy. Our results show that Phiphilux can detect apoptosis in S2 tubules but is activated non-specifically in S1 tubules.

  2. Aronia melanocarpa juice induces a redox-sensitive p73-related caspase 3-dependent apoptosis in human leukemia cells.

    Directory of Open Access Journals (Sweden)

    Tanveer Sharif

    Full Text Available Polyphenols are natural compounds widely present in fruits and vegetables, which have antimutagenic and anticancer properties. The aim of the present study was to determine the anticancer effect of a polyphenol-rich Aronia melanocarpa juice (AMJ containing 7.15 g/L of polyphenols in the acute lymphoblastic leukemia Jurkat cell line, and, if so, to clarify the underlying mechanism and to identify the active polyphenols involved. AMJ inhibited cell proliferation, which was associated with cell cycle arrest in G(2/M phase, and caused the induction of apoptosis. These effects were associated with an upregulation of the expression of tumor suppressor p73 and active caspase 3, and a downregulation of the expression of cyclin B1 and the epigenetic integrator UHRF1. AMJ significantly increased the formation of reactive oxygen species (ROS, decreased the mitochondrial membrane potential and caused the release of cytochrome c into the cytoplasm. Treatment with intracellular ROS scavengers prevented the AMJ-induced apoptosis and upregulation of the expression of p73 and active caspase 3. The fractionation of the AMJ and the use of identified isolated compounds indicated that the anticancer activity was associated predominantly with chlorogenic acids, some cyanidin glycosides, and derivatives of quercetin. AMJ treatment also induced apoptosis of different human lymphoblastic leukemia cells (HSB-2, Molt-4 and CCRF-CEM. In addition, AMJ exerted a strong pro-apoptotic effect in human primary lymphoblastic leukemia cells but not in human normal primary T-lymphocytes. Thus, the present findings indicate that AMJ exhibits strong anticancer activity through a redox-sensitive mechanism in the p53-deficient Jurkat cells and that this effect involves several types of polyphenols. They further suggest that AMJ has chemotherapeutic properties against acute lymphoblastic leukemia by selectively targeting lymphoblast-derived tumor cells.

  3. Angelica sinensis polysaccharides promotes apoptosis in human breast cancer cells via CREB-regulated caspase-3 activation

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    Zhou, Wei-Jie; Wang, Sheng [Department of Breast and Thyroid Surgery, Huaihe Hospital, Henan University, Kaifeng 475000 (China); Hu, Zhuang, E-mail: zhuanghu475000@sina.com [Department of Breast and Thyroid Surgery, Huaihe Hospital, Henan University, Kaifeng 475000 (China); Zhengzhou Center for Disease Control and Prevention, Zhengzhou 475000 (China); Zhou, Zhen-Yu; Song, Cai-Juan [Department of Breast and Thyroid Surgery, Huaihe Hospital, Henan University, Kaifeng 475000 (China); Zhengzhou Center for Disease Control and Prevention, Zhengzhou 475000 (China)

    2015-11-20

    Angelica sinensis polysaccharide (ASP) is purified from the fresh roots of Angelica sinensis (AS). This traditional Chinese medicine has been used for thousands of years for treating gynecological diseases and used in functional foods for the prevention and treatment of various diseases, such as inflammation and cancer. The antitumor activity of ASP is related to its biological activities, because it suppresses a variety of pro-proliferative or anti-apoptotic factors that are dramatically expressed in cancer cells of given types. In this study, we show that angelica sinensis polysaccharide induced apoptosis in breast cancer cells of T47D over-expressing the Cyclic AMP response element binding protein (CREB), inducing apoptosis-related signaling pathway activity. The result also found that ASP caused cell death was linked to caspase activity, accompanied by the loss of mitochondrial membrane potential, cytochrome c release, and Bax translocation from the cytosol to the mitochondria. We found that ASP significantly affected the poly-ADP-ribose polymerase (PARP), Bcl-2 Associated X Protein (Bax), Bcl-2, Bcl-xL and apoptotic protease activating facter-1 (Apaf1) protein expression in a dose- and time-dependent manner. DAPI staining and Flow cytometry were used to analyze apoptosis. The nude mice xenograft model was used to evaluate the antitumor effect of ASP in vivo. ASP has profound antitumor effect on T47D cells, probably by inducing apoptosis through CREB signaling pathway. Thus, these results suggest that ASP would be a promising therapeutic agent for breast cancer. - Highlights: • CREB and Caspase-3 signaling pathways are involved in the ASP induced breast cancer cells apoptosis. • ROCK1/Mlc signaling pathway plays a critical role in this ASP-mediated apoptosis. • Angelica sinensis polysaccharide (ASP) affected the PARP, Bax, Bcl-2, Bcl-xL and Apaf1 protein expression. • The activation of CREB and ROCK1 promotes caspase-3 activation and apoptosis induced

  4. LPS inhibits caspase 3-dependent apoptosis in RAW264.7 macrophages induced by the AMPK activator AICAR

    Energy Technology Data Exchange (ETDEWEB)

    Russe, Otto Quintus, E-mail: quintus@russe.eu; Möser, Christine V., E-mail: chmoeser@hotmail.com; Kynast, Katharina L., E-mail: katharina.kynast@googlemail.com; King, Tanya S., E-mail: tanya.sarah.king@googlemail.com; Olbrich, Katrin, E-mail: Katrin.olbrich@gmx.net; Grösch, Sabine, E-mail: groesch@em.uni-frankfurt.de; Geisslinger, Gerd, E-mail: geisslinger@em.uni-frankfurt.de; Niederberger, Ellen, E-mail: e.niederberger@em.uni-frankfurt.de

    2014-05-09

    Highlights: • AMPK-activation induces caspase 3-dependent apoptosis in macrophages. • Apoptosis is associated with decreased mTOR and increased p21 levels. • All effects can be significantly inhibited by the TLR4 agonist lipopolysaccharide. - Abstract: AMP-activated kinase is a cellular energy sensor which is activated in stages of increased ATP consumption. Its activation has been associated with a number of beneficial effects such as decreasing inflammatory processes and the disease progress of diabetes and obesity, respectively. Furthermore, AMPK activation has been linked with induction of cell cycle arrest and apoptosis in cancer and vascular cells, indicating that it might have a therapeutic impact for the treatment of cancer and atherosclerosis. However, the impact of AMPK on the proliferation of macrophages, which also play a key role in the formation of atherosclerotic plaques and in inflammatory processes, has not been focused so far. We have assessed the influence of AICAR- and metformin-induced AMPK activation on cell viability of macrophages with and without inflammatory stimulation, respectively. In cells without inflammatory stimulation, we found a strong induction of caspase 3-dependent apoptosis associated with decreased mTOR levels and increased expression of p21. Interestingly, these effects could be inhibited by co-stimulation with bacterial lipopolysaccharide (LPS) but not by other proinflammatory cytokines suggesting that AICAR induces apoptosis via AMPK in a TLR4-pathway dependent manner. In conclusion, our results revealed that AMPK activation is not only associated with positive effects but might also contribute to risk factors by disturbing important features of macrophages. The fact that LPS is able to restore AMPK-associated apoptosis might indicate an important role of TLR4 agonists in preventing unfavorable cell death of immune cells.

  5. Betaine supplementation protects against renal injury induced by cadmium intoxication in rats: role of oxidative stress and caspase-3.

    Science.gov (United States)

    Hagar, Hanan; Al Malki, Waleed

    2014-03-01

    Cadmium (Cd) is an environmental and industrial pollutant that can induce a broad spectrum of toxicological effects that affect various organs in humans and experimental animals. This study aims to investigate the effect of betaine supplementation on cadmium-induced oxidative impairment in rat kidney. The animals were divided into four groups (n=10 per group): control, cadmium, betaine and betaine+cadmium (1) saline control group; (2) cadmium group in which cadmium chloride (CdCl2) was given orally at a daily dose of 5 mg/kg body weight for four weeks; (3) betaine group, in which betaine was given to rats at a dose of 250 mg/kg/day, orally via gavage for six weeks; (4) cadmium+betaine group in which betaine was given at a dose of 250 mg/kg/day, orally via gavage for two weeks prior to cadmium administration and concurrently during cadmium administration for four weeks. Cadmium nephrotoxicity was indicated by elevated blood urea nitrogen (BUN) and serum creatinine levels. Kidneys from cadmium-treated rats showed an increase in lipid peroxidation measured as thiobarbituric acid-reactive substances (TBARS) concentration and reductions in total antioxidant status (TAS), reduced glutathione (GSH) content, glutathione peroxidase (GSH-Px) activity, superoxide dismutase concentration (SOD) and catalase activity. Caspase-3 activity, a marker of DNA damage was also elevated in renal tissues of cadmium-treated rats. Pre-treatment of rats with betaine substantially attenuated the increase in BUN and serum creatinine levels. Betaine also inhibited the increase in TBARS concentration and reversed the cadmium-induced depletion in total antioxidant status, GSH, GSH-Px, SOD and catalase concentrations in renal tissues. Renal caspase-3 activity was also reduced with betaine supplementation. These data emphasize the importance of oxidative stress and caspase signaling cascade in cadmium nephrotoxicity and suggest that betaine pretreatment reduces severity of cadmium nephrotoxicity

  6. Suppression of granzyme B activity and caspase-3 activation in leukaemia cells constitutively expressing the protease inhibitor 9.

    Science.gov (United States)

    Fritsch, Kristina; Finke, Jürgen; Grüllich, Carsten

    2013-12-01

    Immune surveillance against malignant cells is mediated by cytotoxic T-lymphocytes and NK-cells (CTL/NK) that induce apoptosis through the granzyme-B-dependent pathway. The serine protease inhibitor serpinB9/protease inhibitor-9 (PI-9) is a known inhibitor of granzyme B. Ectopic expression of PI-9 in tumour cells has been reported. However, the impact of PI-9 on granzyme-B-induced apoptosis in tumour cells remains unclear. The aim of this study was to investigate the influence of constitutive PI-9 expression in leukaemia cell lines on the activity of granzyme B and apoptosis induction. PI-9 negative (lymphoblastic Jurkat cells; myeloblastic U937 cells) and PI-9-expressing cell lines (myeloblastic K562 cells, EBV-transformed LCL-1 and LCL-2 B-cells, lymphoblastic Daudi cells, AML-R cells f leukaemia and the U937 subclone U937PI-9(+)). For accurate granzyme B activity determination a quantitative substrate (Ac-IEPD-pNA) cleavage assay was established and caspase-3 activation measured for apoptosis assessment. Cells were treated with a cytotoxic granule isolate that has previously been shown to induce apoptosis through granzyme B signalling. We found a robust correlation between constitutive PI-9 expression levels and the suppression of granzyme B activity. Further, inhibition of granzyme B translated into reduced caspase-3 activation. We conclude, suppression of granzyme B initiated apoptosis in PI-9-expressing cells could contribute to immune evasion and the measurement of granzyme B activity with our assay might be a useful predictive marker in immune-therapeutic approaches against cancer.

  7. THE PARKINSONIAN NEUROTOXIN ROTENONE ACTIVATES CALPAIN AND CASPASE-3 LEADING TO MOTONEURON DEGENERATION IN SPINAL CORD OF LEWIS RATS

    Science.gov (United States)

    SAMANTARAY, S.; KNARYAN, V. H.; GUYTON, M. K.; MATZELLE, D. D.; RAY, S. K.; BANIK, N. L.

    2007-01-01

    Exposure to environmental toxins increases the risk of neurodegenerative diseases including Parkinson’s disease (PD). Rotenone is a neurotoxin that has been used to induce experimental parkinsonism in rats. We used the rotenone model of experimental parkinsonism to explore a novel aspect of extra-nigral degeneration, the neurodegeneration of spinal cord (SC), in PD. Rotenone administration to male Lewis rats caused significant neuronal cell death in cervical and lumbar SC as compared to control animals. Dying neurons were motoneurons as identified by double immunofluorescent labeling for TUNEL+ cells and ChAT-immunoreactivity. Neuronal death was accompanied by abundant astrogliosis and microgliosis as evidenced from GFAP-immunoreactivity and OX-42-immunoreactivity, respectively, implicating an inflammatory component during neurodegeneration in SC. However, the integrity of the white matter in SC was not affected by rotenone administration as evidenced from the non co-localization of any TUNEL+ cells with GFAP-immunoreactivity and MBP-immunoreactivity, the selective markers for astrocytes and oligodendrocytes, respectively. Increased activities of 76 kD active m-calpain and 17/19 kD active caspase-3 further demonstrated involvement of these enzymes in cell death in SC. The finding of ChAT+ cell death also suggested degeneration of SC motoneurons in rotenone-induced experimental parkinsonism. Thus, this is the first report of its kind in which the selective vulnerability of a putative parkinsonian target outside of nigrostriatal system has been tested using an environmental toxin to understand the pathophysiology of PD. Moreover, rotenone-induced degeneration of SC motoneuron in this model of experimental parkinsonism progressed with upregulation of calpain and caspase-3. PMID:17367952

  8. Activity identification of chimeric anti-caspase-3 mRNA hammerhead ribozyme in vitro and in vivo

    Institute of Scientific and Technical Information of China (English)

    XU; Renhuan; (

    2001-01-01

    [1]Andrew, F., Gerard, E., A license to kill, Cell, 1996, 85: 781-784.[2]Thornberry, N. A., Lazebnik, Y., Caspases: Enemies within, Science, 1998, 281: 1312-1316.[3]Kijima, H., Ishida, H., Ohkawa, T. et al., Therapeutic application of ribozymes, Pharmacol. Ther., 1995, 68: 247-264.[4]Phylactou, L. A., Kilpatrick, M. W., Wood, M. J., Ribozymes as therapeutic tools for genetic disease, Hum. Mol. Genet., 1998, 7(10): 1649-1653.[5]Bettrand, E., Pictet, R ., Grange, T., Can heamerhead ribozymes be efficient tools inactivate gene function? Nucleic Acids Res., 1994, 22: 293-300.[6]Lieber, A., Strauss, M., Selection of efficient cleavage sites in target RNAs by using a ribozyme expression library, Mol. Cell Biol., 1995, 15: 540-551.[7]Xu, R. H., Zhou, X. Q., Xie, Q. et al., Preparation and identification of hammerhead ribozyme in vitro against rat caspase-3 mRNA fragment, Chin. J. Hepatol., 2000,8: 361-363.[8]Liu, J., Jin, Y. X., Wang, D. B., A novel vector for abundant expression of antisense RNA, triplex-forming RNA and ribozyme in vivo, High Technology Letters, 2000, 6: 84-88.[9]Sambrook, J., Fritsch, E. F., Maniatis, T., Molecular Cloning: A Laboratory Manual, 2nd ed., New York: Cold Spring Harbor Laboratory Press, 1989.[10]Porter, A. G., J?nicke, R. U., Emerging roles of caspase-3 in apoptosis, Cell Death Differ, 1999, 6: 99-104.[11]Cryns, V., Yuan, J., Proteases to die for, Genes Dev., 1998, 12: 1551-1570.[12]Narendra, K. V., Anikumar, R. K., Fritz, E., Recent developments in the hammerhead ribozyme field, Nucleic Acids Research, 1998, 26: 5237-5242.

  9. Wood dusts induce the production of reactive oxygen species and caspase-3 activity in human bronchial epithelial cells.

    Science.gov (United States)

    Pylkkänen, Lea; Stockmann-Juvala, Helene; Alenius, Harri; Husgafvel-Pursiainen, Kirsti; Savolainen, Kai

    2009-08-21

    Wood dusts are associated with several respiratory symptoms, e.g. impaired lung function and asthma, in exposed workers. However, despite the evidence from epidemiological studies, the underlying mechanisms are not well understood. In the present study, we investigated different wood dusts for their capacity to induce cytotoxicity and production of radical oxygen species (ROS) as well as activation of the apoptotic caspase-3 enzyme in human bronchial epithelial cells (BEAS-2B). Dusts from three different tree species widely used in wood industry were studied; birch and oak represented hardwood species, and pine a common softwood species. All the experiments were carried out in three different concentrations (10, 50, and 500 microg/ml) and the analysis was performed after 0.5, 2, 6, and 24h exposure. All wood dusts studied were cytotoxic to human bronchial epithelial cells in a dose-dependent manner after 2 and 6h treatment. Exposure to pine, birch, or oak dust had a significant stimulating effect on the production of ROS. Also an induction in caspase-3 protease activity, one of the central components of the apoptotic cascade, was seen in BEAS-2B cells after 2 and 6h exposure to each of the wood dusts studied. In summary, we demonstrate that dusts from pine, birch and oak are cytotoxic, able to increase the production of ROS and the apoptotic response in human broncho-epithelial cells in vitro. Thus, our current data suggest oxidative stress by ROS as an important mechanism likely to function in wood dust related pulmonary toxicity although details of the cellular targets and cell-particle interactions remain to be solved. It is though tempting to speculate that redox-regulated transcription factors such as NFkappaB or AP-1 may play a role in this wood dust-evoked process leading to apparently induced apoptosis of target cells.

  10. A PESQUISA EM TURISMO: REFLEXÕES SOBRE AS ABORDAGENS QUALITATIVA E QUANTITATIVA

    Directory of Open Access Journals (Sweden)

    Noémi Marujo

    2013-06-01

    Full Text Available Nas sociedades contemporâneas, o turismo faz parte integrante do consumo do ser humano e, portanto, é difícil imaginar um mundo sem as designadas férias, sem oportunidades para viajar para novos lugares e adquirir novas experiências sobre outras culturas e outras sociedades. Por isso, observar e analisar o fenómeno para compreendê-lo constitui um factor essencial para obter conhecimento sobre o turismo. Por outro lado, e nos estudos turísticos, qualquer metodologia deve ser seleccionada em função dos objectivos de investigação e do tipo de análise que o investigador deseja realizar. A pesquisa qualitativa e quantitativa podem ser apropriadas em diferentes etapas do processo de investigação em turismo. A combinação dos dois métodos pode contribuir para um maior enriquecimento da pesquisa em turismo.

  11. Axonal outgrowth is associated with increased ERK 1/2 activation but decreased caspase 3 linked cell death in Schwann cells after immediate nerve repair in rats

    Directory of Open Access Journals (Sweden)

    Kanje Martin

    2011-01-01

    Full Text Available Abstract Background Extracellular-signal regulated kinase (ERK1/2 is activated by nerve damage and its activation precedes survival and proliferation of Schwann cells. In contrast, activation of caspase 3, a cysteine protease, is considered as a marker for apoptosis in Schwann cells. In the present study, axonal outgrowth, activation of ERK1/2 by phosphorylation (p-ERK 1/2 and immunoreactivity of cleaved caspase 3 were examined after immediate, delayed, or no repair of transected rat sciatic nerves. Results Axonal outgrowth, detected by neurofilament staining, was longer after immediate repair than after either the delayed or no repair conditions. Immediate repair also showed a higher expression of p-ERK 1/2 and a lower number of cleaved caspase 3 stained Schwann cells than after delayed nerve repair. If the transected nerve was not repaired a lower level of p-ERK 1/2 was found than in either the immediate or delayed repair conditions. Axonal outgrowth correlated to p-ERK 1/2, but not clearly with cleaved caspase 3. Contact with regenerating axons affected Schwann cells with respect to p-ERK 1/2 and cleaved caspase 3 after immediate nerve repair only. Conclusion The decreased regenerative capacity that has historically been observed after delayed nerve repair may be related to impaired activation of Schwann cells and increased Schwann cell death. Outgrowing axons influence ERK 1/2 activation and apoptosis of Schwann cells.

  12. The Effects of Magnesium Sulfate on Fetal Rats of FGR and the Expression of Caspase-3 in the Placenta of Maternal Rat

    Institute of Scientific and Technical Information of China (English)

    GAO Hui; ZOU Li

    2005-01-01

    To investigate the effect of magnesium sulfate on the fetal rats of FGR and the expression of caspase-3 in the placenta of maternal rat; To explore the mechanism of using magnesium sulfate to cure the FGR. Establish model of FGR by a way of passive smoking: giving the maternal rats different agent of magnesium sulfate by subcutaneous injection: low agent group (300 mg/kg),high agent group (600 mg/kg). Concentration of magnesium sulfate was monitored. The expres sion of caspase-3 was measured by RT-PCR and immunohistochemistry technology. Both of the concentrations of magnesium sulfate in high and low agents group are higher than the FGR group (P<0. 01); the weight of the placenta and fetal rat in high agent group are higher than the FGR group (P<0.05 and P<0.01); the expression of mRNA and protein of caspase-3 in the two agent group is higher than the FGR group (P<0.05 respectively); concentration of magnesium sulfate in the maternal rat blood correlate to the weight of fetal rat (r=0. 899, P=0. 038) and the expression of caspase-3 in the placenta of maternal rat (r= 0.747, P 0.033; r=-0. 915, P=0.001).The research suggests that the weight of fetal rat could be increased by treatment of magnesium sulfate. Because it would imfrmove the placental function by depressing the expression of caspase-3.

  13. Study on HepG-2 apoptosis induced by saponins isolated from Asparagus and the effects on the activities of caspase-3,8,9

    Institute of Scientific and Technical Information of China (English)

    JI Yu-bin; XU He; JI Chen-feng

    2008-01-01

    Objective To study the effect of saponins of asparagus on apoptosis and the variations of caspaseS, caspase-9 and caspase-3 activity in the process of asparagus induced apoptosis in HepG-2, to investigate the apoptosis mechanism further. Methods Asparagus on apoptosis effects on tumor cells cultured-HepG-2 with different concentrations at different time, IC50 value was measured by MTT assay, the apoptosis rate was determined by FCM with AnnexinV/PI staining, their apoptotic morphology were observed by electron microscopy and Colorimetric method was used to measure caspase-8,9 and caspase-3 activities. Results Experiments of antitumour in vivo showed that saponins of asparagus can inhibit the growth of tumor cell of HepG-2 in evidence, IC50 was 101.15 mg·L-1. Cultured for 72 h, the apoptosis rate had positive increased with concentrations. Apoptotic morphology was observed by electron microscopy. The activities of caspase-8, easpase-9 and caspase-3 had positive increased with concentrations. And have significant difference compared with negative control group(P<0.01). The activities of caspase-8 were high at 24 h, but the activities of caspase-9 and caspase-3 is high at 48 h. Conclusions Aaponins of asparagus can inhibit the growth of tumor cell of HepG2, and the underlying mechanism might be related to up regulation of caspase-8, 9 activity which subsequently transforms caspase-3 into its active form.

  14. Expression of activating transcription factor 3 (ATF 3) and caspase 3 in Schwann cells and axonal outgrowth after sciatic nerve repair in diabetic BB rats.

    Science.gov (United States)

    Stenberg, Lena; Kanje, Martin; Dolezal, Katarina; Dahlin, Lars B

    2012-04-25

    The aim of this study was to evaluate nerve regeneration in relation to the transcription factor, Activating Transcription Factor 3 (ATF 3), and an apoptotic marker, caspase 3, in the Schwann cells of diabetic BB rats (i.e. display type 1 diabetes phenotype). Sciatic nerves in healthy Wistar rats and in diabetic BB rats were transected and immediately repaired. Axonal outgrowth (neurofilament staining) and expression of ATF 3 and caspase 3 were quantified by immunohistochemistry after six days. There was no difference in axonal outgrowth between healthy and diabetic rats. However, the sciatic nerve in the diabetic rats exhibited a larger number of ATF 3 expressing Schwann cells at the site of the lesion and also a higher number of caspase 3 expressing Schwann cells. Similar differences were observed in the distal nerve segment between the healthy and diabetic rats. There were no correlations between the number of Schwann cells expressing ATF 3 and caspase 3. Thus, diabetic BB rats display an increased activation of ATF 3 and also a rise in apoptotic caspase 3 expressing Schwann cells, but with no discrepancy in length of axonal outgrowth after nerve injury and repair at six days. Knowledge about signal transduction mechanisms in diabetes after stress may provide new insights into the development of diabetic neuropathy and neuropathic pain. Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.

  15. Sub-lethal oxidative stress induces lysosome biogenesis via a lysosomal membrane permeabilization-cathepsin-caspase 3-transcription factor EB-dependent pathway.

    Science.gov (United States)

    Leow, San Min; Chua, Shu Xian Serene; Venkatachalam, Gireedhar; Shen, Liang; Luo, Le; Clement, Marie-Veronique

    2016-12-18

    Here we provide evidence to link sub-lethal oxidative stress to lysosomal biogenesis. Exposure of cells to sub-lethal concentrations of exogenously added hydrogen peroxide resulted in cytosol to nuclear translocation of the Transcription Factor EB (TFEB), the master controller of lysosome biogenesis and function. Nuclear translocation of TFEB was dependent upon the activation of a cathepsin-caspase 3 signaling pathway, downstream of a lysosomal membrane permeabilization and accompanied by a significant increase in lysosome numbers as well as induction of TFEB dependent lysosome-associated genes expression such as Ctsl, Lamp2 and its spliced variant Lamp2a, Neu1and Ctsb and Sqstm1 and Atg9b. The effects of sub-lethal oxidative stress on lysosomal gene expression and biogenesis were rescued upon gene silencing of caspase 3 and TFEB. Notably, caspase 3 activation was not associated with phenotypic hallmarks of apoptosis, evidenced by the absence of caspase 3 substrate cleavage, such as PARP, Lamin A/C or gelsolin. Taken together, these data demonstrate for the first time an unexpected and non-canonical role of a cathepsin-caspase 3 axis in the nuclear translocation of TFEB leading to lysosomes biogenesis under conditions of sub-lethal oxidative stress.

  16. A PESQUISA EM TURISMO: REFLEXÕES SOBRE AS ABORDAGENS QUALITATIVA E QUANTITATIVA

    National Research Council Canada - National Science Library

    Noémi Marujo

    2013-01-01

    ... de análise que o investigador deseja realizar. A pesquisa qualitativa e quantitativa podem ser apropriadas em diferentes etapas do processo de investigação em turismo. A combinação dos dois...

  17. Hepatitis B Virus X Protein Sensitizes Primary Mouse Hepatocytes to Ethanol- and TNF-α-Induced Apoptosis by a Caspase-3-Dependent Mechanism

    Institute of Scientific and Technical Information of China (English)

    Won-Ho Kim; Feng Hong; Barbara Jaruga; Zhengsheng Zhang; Saijun Fan; T. Jake Liang; Bin Gao

    2005-01-01

    It is well-documented that alcohol drinking together with hepatitis viral infection accelerates liver injury; however the underlying mechanisms remain unknown. In this paper, we demonstrated that primary hepatocytes from transgenic mice overexpressing hepatitis B virus X protein (HBX) were more susceptible to ethanol- and TNF-α-induced apoptotic killing. Compared to normal control mouse hepatocytes, ethanol and/or TNF-α treatment led to a significant increase in reactive oxygen species, mitochondrial permeability transition, cytochrome C release,caspase-3 activity, and poly (ADP-ribose) polymerase degradation in hepatocytes from HBX transgenic mice.Blocking caspase-3 activity antagonized ethanol- and TNF-α-induced apoptosis in primary hepatocytes from HBX transgenic mice. Taken together, our findings suggest that HBX sensitizes primary mouse hepatocytes to ethanoland TNF-α-induced apoptosis by a caspase-3-dependent mechanism, which may partly explain the synergistic effects of alcohol consumption and hepatitis B virus infection on liver injury.

  18. Active caspase-3 and ultrastructural evidence of apoptosis in spontaneous and induced cell death in bovine in vitro produced pre-implantation embryos

    DEFF Research Database (Denmark)

    Gjørret, Jakob O.; Fabian, Dusan; Avery, Birthe;

    2007-01-01

    In this study we investigated chronological onset and involvement of active caspase-3, apoptotic nuclear morphology, and TUNEL-labeling, as well as ultrastructural evidence of apoptosis, in both spontaneous and induced cell death during pre-implantation development of bovine in vitro produced...... embryos. Pre-implantation embryos (2-cell to Day 8 blastocysts) were cultured with either no supplementation (untreated) or with 10 µM staurosporine for 24 hr (treated). Embryos were subjected to immunohistochemical staining of active caspase-3, TUNEL-reaction for detection of DNA degradation and DAPI......, active caspase-3 and apoptotic nuclear morphology were observed in an untreated 8-cell stage, and TUNEL-labeling was observed from the 16-cell stage. Blastomeres concurrently displaying all apoptotic features were present in a few embryos at 16-cell and morula stages and in all blastocysts. All three...

  19. The role of caspase 3 and BclxL in the action of interleukin 7 (IL-7): a survival factor in activated human T cells

    DEFF Research Database (Denmark)

    Amos, C L; Woetmann, A; Nielsen, M;

    1998-01-01

    by the synthetic glucocorticoid, dexamethasone. Bcl-2 protein expression was uupregulated by IL-7 with or without dexamethasone, but Bc1-2 was expressed at a much lower level than BclxL in these cells. Levels of Bax did not markedly change on either cytokine stimulation or dexamethasone treatment. An unidentified...... cells. Both cytokines abrogated the dexamethasone-induced stimulation of Caspase 3 and prevented the cleavage of poly (ADP-ribose) polymerase (PARP), a substrate for the Caspase 3. IL-7 upregulated the expression of Bc1xL and counteracted the downregulation of this anti-apoptotic protein...... of apoptosis. A clear role for IL-7 as a survival factor for cytokine withdrawal and glucocorticoid induced apoptosis in activated primary hT cells is implicated. In addition, regulation of BclxL and downstream inhibition of Caspase 3 activity may mediate this rescue signal....

  20. Binswanger's disease and quantitative fractional anisotropy Doença de Binswanger e anisotropia fracionada quantitativa

    Directory of Open Access Journals (Sweden)

    Eliasz Engelhardt

    2009-06-01

    Full Text Available OBJECTIVE: To study the integrity of the white matter in Binswanger's disease (BD patients with quantitative fractional anisotropy (DTI-FA. METHOD: Controls (12 and patients with BD (12 were included. Scans performed with MR (GE Signa Horizon/1.5T. Fazekas's score=6 with white matter hyperintensities extension >75% assessed on FLAIR scans. Standard parameters for DTI-FA were used. ROIs placed in symmetrical regions on two axial planes, data pooled in anterior (frontal and posterior (temporo-parieto-occipital regions. Analysis with Functool. Statistics for anterior and posterior regions comparison. RESULTS: DTI-FA showed reduction of anisotropy, reflecting axonal damage and demyelination of fibers, more prominent in anterior in relation to posterior region, in BD patients in comparison to controls. CONCLUSION: Loss of integrity of fiber tracts reflects interruption of neural networks that subserve cognitive, behavioral, and motor integration. The more severely affected frontal region is related to executive dysfunction, a characteristic feature of Binswanger's disease.OBJETIVO: Estudar a integridade da substância branca em pacientes com doença de Binswanger (DB com anisotropia fracionada quantitativa (DTI-FA. MÉTODO: Incluídos controles (12 e pacientes com DB (12. Obtidas imagens de RM (GE Signa Horizon/1,5T. Escore=6 de Fazekas com hiperintensidades da substância branca com extensão >75% avaliados em imagens em FLAIR. Utilizados parâmetros padrão para DTI-FA. ROIs colocados em regiões simétricas de dois planos axiais, dados das regiões anterior (frontal e posterior (têmporo-parieto-occipital reunidos. Análise com Functool. Estatística para comparar regiões anteriores e posteriores. RESULTADOS: DTI-FA mostrou redução da anisotropia, refletindo lesão axonal e desmielinazação de fibras, com predomínio na região anterior em relação à posterior, nos pacientes com DB em comparação aos controles. CONCLUSÃO: Perda da

  1. Investigation of the Relationship between the Intracellular Ca2+ Levels and Caspases-3 and 8 Expression in Rat Mammary Gland Carcinoma Undergoing Apoptosis

    Institute of Scientific and Technical Information of China (English)

    Hui Sun; Jing Zhang; Zhongli Zhan; Baocun Sun; Xishan Hao

    2007-01-01

    OBJECTIVE To investigate the relationship between the level of caspase-3 and 8 expression and intracellular Ca2+ levels in BCML-TA299 breast cancer cells in the process of apoptosis.METHODS Mice were divided into three IFNa-treated groups and one control group as follows: an intratumoral injection, subcutaneous injection, preventive injection, and a control without injection. The cellular DMA content, changes in the cell cycle and the relationship between the cell Ca2+ concentrations and the expression of caspase-3 and 8 were examined. RESULTS After injection of IFN-α-2b by different routes, the morphologic transformation of the breast cancer cells in each group was observed. There was a typical apoptotic response in the intratumoral-injection group. The expression of caspase-3 and 8 was diverse among the experimental groups, and correlated with the cellular Ca2+ concentration. Caspase-3 and 8 expression and the cellular Ca2+ level were higher following intratumoral injection compared to the other treatments (P<0.01). Among the experimental groups, the cell cycle displayed definitive changes.CONCLUSION a) Both caspase-3 and 8 and the intracellular Ca2+ are elevated in the process of cell apoptosis in BCML-TA299 breast cancer tissues. These changes may play important roles in the occurrence and development of breast cancer; b) Variation in the route of IFN-α-2b administration can produce different responses in the expression of caspase-3 and 8 and the concentration of Ca2+ in apoptotic tumor cells.

  2. A utilização da avaliação tipo "teste" on-line como apoio ao ensino presencial: uma abordagem quantitativa sobre a sua contribuição no ensino de ferramentas estatística multivariadas Use of on-line evaluation type "test" as support for presential teaching: a quantitative approach on its contribution to the teaching of multivariate statistics tools

    Directory of Open Access Journals (Sweden)

    Erica Ferreira Marques

    2011-07-01

    Full Text Available Este artigo busca mostrar o quanto a aplicação de uma ferramenta de avaliação tipo "teste" em um ambiente virtual pode contribuir como apoio ao ensino presencial do estudo de ferramentas estatísticas multivariadas para os alunos de graduação em Administração da FEARP/USP, segundo a opinião destes matriculados na disciplina Estatística Aplicada à Administração II. Este trabalho fez parte de um projeto de pesquisa temático desenvolvido na própria instituição, denominado LaViE, que é um ambiente virtual de ensino-aprendizagem de estatística aplicado e desenvolvido para o apoio ao ensino presencial desta área. Baseando-se na importância de um mecanismo de avaliação de conteúdo, o LaViE visou contribuir para o ensino presencial disponibilizando ferramentas de conteúdo, interação e "teste seu conhecimento". Para tanto, esta ferramenta de avaliação foi desenvolvida baseada em questões testes on-line com níveis diferentes de "adaptação" pelo aluno para cada módulo apresentado na disciplina em questão. Neste caso, foram criados três níveis de adaptação: básico (I, intermediário (II e avançado (III. A metodologia utilizada para verificação da eficiência desta ferramenta tipo "teste" no ambiente virtual foi baseada, então, em uma avaliação quantitativa de acordo com a opinião dos alunos (usuários. Nessa pesquisa foram investigados quatro pressupostos. Esses dados foram coletados quando essa disciplina foi ministrada, porém em dois períodos diferentes; no segundo semestre de 2005, como projeto-piloto, e no segundo semestre de 2006. Dessa forma, pôde ser feita uma análise comparativa da opinião dos alunos sobre o sistema em dois momentos diferentes. Para esse levantamento foram utilizados dois questionários aplicados diretamente aos alunos em sala de aula momentos antes da avaliação presencial da disciplina e um outro logo após essa avaliação presencial. Observou-se, por este estudo, a import

  3. Mercury-Induced Externalization of Phosphatidylserine and Caspase 3 Activation in Human Liver Carcinoma (HepG2 Cells

    Directory of Open Access Journals (Sweden)

    Paul B. Tchounwou

    2006-03-01

    Full Text Available Apoptosis arises from the active initiation and propagation of a series of highly orchestrated specific biochemical events leading to the demise of the cell. It is a normal physiological process, which occurs during embryonic development as well as in the maintenance of tissue homeostasis. Diverse groups of molecules are involved in the apoptosis pathway and it functions as a mechanism to eliminate unwanted or irreparably damaged cells. However, inappropriate induction of apoptosis by environmental agents has broad ranging pathologic implications and has been associated with several diseases including cancer. The toxicity of several heavy metals such as mercury has been attributed to their high affinity to sulfhydryl groups of proteins and enzymes, and their ability to disrupt cell cycle progression and/or apoptosis in various tissues. The aim of this study was to assess the potential for mercury to induce early and late-stage apoptosis in human liver carcinoma (HepG2 cells. The Annexin-V and Caspase 3 assays were performed by flow cytometric analysis to determine the extent of phosphatidylserine externalization and Caspase 3 activation in mercury-treated HepG2 cells. Cells were exposed to mercury for 10 and 48 hours respectively at doses of 0, 1, 2, and 3 μg/mL based on previous cytotoxicity results in our laboratory indicating an LD50 of 3.5 ± 0.6 μg/mL for mercury in HepG2 cells. The study data indicated a dose response relationship between mercury exposure and the degree of early and late-stage apoptosis in HepG2 cells. The percentages of cells undergoing early apoptosis were 0.03 ± 0.03%, 5.19 ± 0.04%, 6.36 ± 0.04%, and 8.84 ± 0.02% for 0, 1, 2, and 3 μg/mL of mercury respectively, indicating a gradual increase in apoptotic cells with increasing doses of mercury. The percentages of Caspase 3 positive cells undergoing late apoptosis were 3.58 ± 0.03%, 17.06 ± 0

  4. Developing a powerful In Silico tool for the discovery of novel caspase-3 substrates: a preliminary screening of the human proteome

    Directory of Open Access Journals (Sweden)

    Ayyash Muneef

    2012-01-01

    Full Text Available Abstract Background Caspases are a family of cysteinyl proteases that regulate apoptosis and other biological processes. Caspase-3 is considered the central executioner member of this family with a wide range of substrates. Identification of caspase-3 cellular targets is crucial to gain further insights into the cellular mechanisms that have been implicated in various diseases including: cancer, neurodegenerative, and immunodeficiency diseases. To date, over 200 caspase-3 substrates have been identified experimentally. However, many are still awaiting discovery. Results Here, we describe a powerful bioinformatics tool that can predict the presence of caspase-3 cleavage sites in a given protein sequence using a Position-Specific Scoring Matrix (PSSM approach. The present tool, which we call CAT3, was built using 227 confirmed caspase-3 substrates that were carefully extracted from the literature. Assessing prediction accuracy using 10 fold cross validation, our method shows AUC (area under the ROC curve of 0.94, sensitivity of 88.83%, and specificity of 89.50%. The ability of CAT3 in predicting the precise cleavage site was demonstrated in comparison to existing state-of-the-art tools. In contrast to other tools which were trained on cleavage sites of various caspases as well as other similar proteases, CAT3 showed a significant decrease in the false positive rate. This cost effective and powerful feature makes CAT3 an ideal tool for high-throughput screening to identify novel caspase-3 substrates. The developed tool, CAT3, was used to screen 13,066 human proteins with assigned gene ontology terms. The analyses revealed the presence of many potential caspase-3 substrates that are not yet described. The majority of these proteins are involved in signal transduction, regulation of cell adhesion, cytoskeleton organization, integrity of the nucleus, and development of nerve cells. Conclusions CAT3 is a powerful tool that is a clear improvement over

  5. 子痫前期患者胎盘组织TNF-α及Caspase-3的表达及相关性研究%Study on expression of TNF - α (tumor necrosis factor alpha) and Caspase - 3 and association between them in placenta of pregnant women with preeclampsia

    Institute of Scientific and Technical Information of China (English)

    谭芳女; 乔福元

    2006-01-01

    目的探讨TNF-α及Caspase-3在子痫前期患者胎盘组织中的表达及相关性.方法采用SP法对轻、重度子痫前期组40例和正常妊娠组20例的胎盘组织进行TNF-α和Caspase-3的免疫组化染色,观察各组TNF-α和Caspase-3的定位、分布和表达量的差异.结果重度子痫前期组的TNF-α及Caspase-3的表达量较正常组显著升高(P<0.01).另外,重子痫前期度组TNF-α与Caspase-3的表达量呈正相关(R=0.698,P<0.001).结论子痫前期时胎盘可能是TNF-α的来源之一.子痫前期的发病可能与胎盘局部产生的TNF-α诱导的滋养细胞凋亡(Caspase-3是凋亡通路下游关键蛋白酶)增加有关.

  6. CYP24A1 exacerbated activity during diabetes contributes to kidney tubular apoptosis via caspase-3 increased expression and activation.

    Directory of Open Access Journals (Sweden)

    Alexandre Tourigny

    Full Text Available Decreases in circulating 25,hydroxyl-vitamin D3 (25 OH D3 and 1,25,dihydroxyl-vitamin D3 (1,25 (OH2 D3 have been extensively documented in patients with type 2 diabetes. Nevertheless, the molecular reasons behind this drop, and whether it is a cause or an effect of disease progression is still poorly understood. With the skin and the liver, the kidney is one of the most important sites for vitamin D metabolism. Previous studies have also shown that CYP24A1 (an enzyme implicated in vitamin D metabolism, might play an important role in furthering the progression of kidney lesions during diabetic nephropathy. In this study we show a link between CYP24A1 increase and senescence followed by apoptosis induction in the renal proximal tubules of diabetic kidneys. We show that CYP24A1 expression was increased during diabetic nephropathy progression. This increase derived from protein kinase C activation and increased H(2O(2 cellular production. CYP24A1 increase had a major impact on cellular phenotype, by pushing cells into senescence, and later into apoptosis. Our data suggest that control of CYP24A1 increase during diabetes has a beneficial effect on senescence induction and caspase-3 increased expression. We concluded that diabetes induces an increase in CYP24A1 expression, destabilizing vitamin D metabolism in the renal proximal tubules, leading to cellular instability and apoptosis, and thereby accelerating tubular injury progression during diabetic nephropathy.

  7. Effect of ginsenoside Rh-2 via activation of caspase-3 and Bcl-2-insensitive pathway in ovarian cancer cells.

    Science.gov (United States)

    Kim, Jin Hee; Choi, Jae-Sun

    2016-12-13

    Ginsenoside has been reported to have therapeutic effects for some types of cancer, but its effect on ovarian cancer cells has not been evaluated. In this study, we monitored the effects of ginsenoside-Rh2 (Rh2) on the inhibition of cell proliferation and the apoptotic process in the ovarian cancer cell line SKOV3 using an MTT assay and TUNEL assay. We found that Rh2 inhibited cell proliferation and significantly induced apoptosis. We confirmed the apoptotic effects of Rh2 using western blot analysis of apoptosis-related proteins. Specifically, the levels of cleaved poly ADP ribose polymerase (PARP) and cleaved caspase-3 significantly increased in SKOV3 cells treated with Rh2. Therefore, Rh2 clearly suppressed the growth of SKOV3 cells in vitro, which was associated with induction of the apoptosis pathway. Moreover, the migration assay showed that Rh2 inhibited the invasive ability of SKOV3 cells. Taken together, our results suggest that Rh2 has anticancer effects in SKOV3 cells through inhibition of cell proliferation and induction of apoptosis. Considering the therapeutic potential of Rh2, more studies should be carried out to facilitate the future application of this natural product as a potential anti-cancer agent.

  8. Quercetin derivative induces cell death in glioma cells by modulating NF-κB nuclear translocation and caspase-3 activation.

    Science.gov (United States)

    Kiekow, Cíntia J; Figueiró, Fabrício; Dietrich, Fabrícia; Vechia, Luciana Dalla; Pires, Elisa N S; Jandrey, Elisa H F; Gnoatto, Simone C B; Salbego, Christianne G; Battastini, Ana Maria O; Gosmann, Grace

    2016-03-10

    Treated glioblastoma multiforme (GBM) patients only survive 6 to 14months after diagnosis; therefore, the development of novel therapeutic strategies to treat gliomas remains critically necessary. Considering that phenolic compounds, like quercetin, have the potential to be used in the chemotreatment of gliomas and that some flavonoids exhibit the ability to cross the BBB, in the present study, we investigated the antitumor effect of flavonoids (including chalcones, flavones, flavanones and flavonols). Initially their activities were tested in C6 glioma cells screened using the MTT method, resulting in the selection of chalcone 2 whose feasibility was confirmed by a Trypan Blue exclusion assay in the low μM range on C6 glioma cells. Cell cycle and apoptotic death analyses on C6 glioma cells were also performed, and chalcone 2 increased the apoptosis of the cells but did not alter the cell cycle progression. In addition, treatments with these two compounds were not cytotoxic to hippocampal organotypic cultures, a model of healthy neural cells. Furthermore, the results indicated that 2 induced apoptosis by inhibition of NF-κB and activation of active caspase-3 in glioma cells, suggesting that it is a potential prototype to develop new treatments for GBM in the future. Copyright © 2016 Elsevier B.V. All rights reserved.

  9. Apigenin Protects Endothelial Cells from Lipopolysaccharide (LPS)-Induced Inflammation by Decreasing Caspase-3 Activation and Modulating Mitochondrial Function

    Science.gov (United States)

    Duarte, Silvia; Arango, Daniel; Parihar, Arti; Hamel, Patrice; Yasmeen, Rumana; Doseff, Andrea I.

    2013-01-01

    Acute and chronic inflammation is characterized by increased reactive oxygen species (ROS) production, dysregulation of mitochondrial metabolism and abnormal immune function contributing to cardiovascular diseases and sepsis. Clinical and epidemiological studies suggest potential beneficial effects of dietary interventions in inflammatory diseases but understanding of how nutrients work remains insufficient. In the present study, we evaluated the effects of apigenin, an anti-inflammatory flavonoid abundantly found in our diet, in endothelial cells during inflammation. Here, we show that apigenin reduced lipopolysaccharide (LPS)-induced apoptosis by decreasing ROS production and the activity of caspase-3 in endothelial cells. Apigenin conferred protection against LPS-induced mitochondrial dysfunction and reestablished normal mitochondrial complex I activity, a major site of electron leakage and superoxide production, suggesting its ability to modulate endothelial cell metabolic function during inflammation. Collectively, these findings indicate that the dietary compound apigenin stabilizes mitochondrial function during inflammation preventing endothelial cell damage and thus provide new translational opportunities for the use of dietary components in the prevention and treatment of inflammatory diseases. PMID:23989609

  10. 间变性大细胞淋巴瘤与Caspase-3和Bcl-2

    Institute of Scientific and Technical Information of China (English)

    张丽红; 侯刚强

    2006-01-01

    间变性大细胞淋巴瘤(anaplastic large cell lymphoma,ALCL)是1985年由Stein首先报道的非霍奇金淋巴瘤(NHL)的一种新的独立类型。许多学者认为ALCL的预后与凋亡密切相关。目前与凋亡相关的基因通常分为两大类,促凋亡基因和抗凋亡基因。其中,Caspase(cysteine-asparate protease)家族和Bcl-2(B-cell lymphoma/leukemia-2,Bcl-2)家族起着决定性的作用。本文着重讨论Caspase-3、Bcl-2及其在间变性大细胞淋巴瘤中的作用。

  11. Doxycycline reduces cleaved caspase-3 and microglial activation in an animal model of neonatal hypoxia-ischemia.

    Science.gov (United States)

    Jantzie, Lauren L; Cheung, Po-Yin; Todd, Kathryn G

    2005-03-01

    Neonatal hypoxia-ischemia (HI) is a major contributor to many perinatal neurologic disorders and, thus, the search for therapies and effective treatments for the associated brain damage has become increasingly important. The tetracycline derivative, doxycycline (DOXY), has been reported to be neuroprotective in adult animal models of cerebral ischemia. To investigate the putative neuroprotective effects of DOXY in an animal model of neonatal HI, a time-course study was run such that pups received either DOXY (10 mg/kg) or VEH immediately before hypoxia, 1, 2, or 3 hours after HI (n=6). At 7 days after injury, the pups were euthanized, and the brains were removed and processed for immunohistochemical and Western blot analyses using antibodies against specific markers for neurons, apoptotic markers, microglia, oligodendrocytes, and astrocytes. Results showed that in vulnerable brain regions including the hippocampal formation, thalamus, striatum, cerebral cortex and white matter tracts, DOXY significantly decreased caspase-3 immunoreactivity (a marker of apoptosis), promoted neuronal survival, inhibited microglial activation and reduced reactive astrocytosis compared with VEH-treated HI pups. These effects were found to occur in a time-dependent manner. Taken together, these results strongly suggest that doxycycline has potential as a pharmacological treatment for mild HI in neonates.

  12. Caspase-3-dependent apoptosis of citreamicin ε-induced heLa iells Is associated with reactive oxygen species generation

    KAUST Repository

    Liu, Lingli

    2013-07-15

    Citreamicins, members of the polycyclic xanthone family, are promising antitumor agents that are produced by Streptomyces species. Two diastereomers, citreamicin ε A (1) and B (2), were isolated from a marine-derived Streptomyces species. The relative configurations of these two diastereomers were determined using NMR spectroscopy and successful crystallization of citreamicin ε A (1). Both diastereomers showed potent cytotoxic activity against HeLa (cervical cancer) and HepG2 (hepatic carcinoma) cells with IC 50 values ranging from 30 to 100 nM. The terminal deoxynucleotidyl transferase dUTP nick-end labeling assay confirmed that citreamicin ε A (1) induced cellular apoptosis, and Western blot analysis showed that apoptosis occurred via activation of caspase-3. The 2,7-dichlorofluorescein diacetate assay indicated that citreamicin ε substantially increased the intracellular concentration of reactive oxygen species (ROS). To confirm the hypothesis that citreamicin ε induced apoptosis through an increase in the intracellular ROS concentration, the oxidized products, oxicitreamicin ε A (3) and B (4), were obtained from a one-step reaction catalyzed by Ag 2O. These products, with a reduced capacity to increase the intracellular ROS concentration, exhibited a significantly weakened cytotoxicity in both HeLa and HepG2 cells compared with that of citreamicin ε A (1) and B (2). © 2013 American Chemical Society.

  13. A fusion protein composed of IL-2 and caspase-3 ameliorates the outcome of experimental inflammatory colitis.

    Science.gov (United States)

    Sagiv, Yuval; Kaminitz, Ayelet; Lorberboum-Galski, Haya; Askenasy, Nadir; Yarkoni, Shai

    2009-09-01

    Targeted depletion of immune cells expressing the interleukin-2 (IL-2) receptor can exacerbate inflammatory bowel disease (IBD) through elimination of regulatory T (Treg) cells, or ameliorate its course by depletion of cytotoxic cells. To answer this question we used a fusion protein composed of IL-2 and caspase-3 (IL2-cas) in an experimental model of DSS-induced toxic colitis. In a preventive setting, co-administration of DSS with a daily therapeutic dose of IL2-cas for seven days improved all disease parameters. Although CD4(+)CD25(+) T cells were depleted in the mesenteric lymph nodes, a fractional increase in CD4(+)FoxP3(+) T cells was observed in the spleen. Likewise, IL2-cas therapy improved the outcome of established disease in a chronic model of colitis. These data demonstrate that therapies that use IL-2 as a targeting moiety exert a protective effect over the colon under conditions of inflammation. The efficacy of IL-2-targeted therapy is attributed to reduced activity of reactive T cells, which ameliorates the secondary inflammatory infiltration. IL2-cas evolves as a potential therapeutic tool in IBD.

  14. Análise quantitativa da influência de um novo paradigma ecológico: autocorrelação espacial - DOI: 10.4025/actascibiolsci.v25i1.2113 Quantitative analysis of the influence of a new ecological paradigm: spatial autocorrelation - DOI: 10.4025/actascibiolsci.v25i1.2113

    Directory of Open Access Journals (Sweden)

    Luis Mauricio Bini

    2003-04-01

    Full Text Available O objetivo desse trabalho foi o de avaliar a influência da autocorrelação espacial (ausência de independência estatística de observações obtidas ao longo do espaço geográfico nos estudos ecológicos. Para tanto, uma avaliação dos trabalhos que empregaram os métodos necessários para a quantificação da autocorrelação espacial foi realizada, utilizando os dados fornecidos pelo “Institute for Scientific Information”. Os resultados demonstraram que existe uma tendência crescente da utilização das análises de autocorrelação espacial em estudos ecológicos, e que a presença de autocorrelação espacial significativa foi detectada na maior parte dos estudos. Além disso, esses estudos foram desenvolvidos em vários países, por cientistas de diferentes nacionalidades, com diferentes grupos de organismos e em diferentes tipos de ecossistemas. Dessa forma, pode-se considerar que o reconhecimento explícito da estrutura espacial dos processos naturais, por meio da análise da autocorrelação espacial, é um novo paradigma dos estudos ecológicosThe aim of this paper was to evaluate the influence of the spatial autocorrelation (absence of independence among observations gathered along geographical space in ecological studies. For this task, an evaluation of the studies that used spatial autocorrelation analysis was carried out using the data furnished by the Institute for Scientific Information. There is a positive temporal tendency in the number of studies that used spatial autocorrelation analysis. A significant autocorrelation was detected in most studies. Moreover, scientist of several nationalities carried out these studies in different countries, with different organisms and in different types of ecosystems. In this way, it is possible to consider that the explicit incorporation of the spatial structure of natural processes, through the autocorrelation analysis, is a new ecological paradigm

  15. Huangzhi Oral Liquid Prevents Arrhythmias by Upregulating Caspase-3 and Apoptosis Network Proteins in Myocardial Ischemia-Reperfusion Injury in Rats

    Directory of Open Access Journals (Sweden)

    Xu Ran

    2015-01-01

    Full Text Available To study the effect of Huangzhi oral liquid (HZOL on I/R after 2 h and 4 h and determine its regulatory function on caspase-3 and protein networks. 70 SD male rats were randomly divided into seven groups and established myocardial I/R injury model by ligating the left anterior descending coronary artery. Myocardial infarction model was defined by TTC staining and color of the heart. The levels of CK-MB, CTnI, C-RPL, SOD, and MDA were tested at 2 h and 4 h after reperfusion. HE staining and ultramicrostructural were used to observe the pathological changes. The apoptotic index (AI of cardiomyocyte was marked by TUNEL. The expression levels of caspase-3, p53, fas, Bcl-2, and Bax were tested by immunohistochemistry and western blot. HZOL corrected arrhythmia, improved the pathologic abnormalities, decreased CK-MB, CTnI, C-RPL, MDA, AI, caspase-3, p53, fas, and Bax, and increased SOD ans Bcl-2 with different times of myocardial reperfusion; this result was similar to the ISMOC (P>0.05. HZOL could inhibit arrhythmia at 2 and 4 h after I/R and ameliorate cardiac function, which was more significant at 4 h after reperfusion. This result may be related to decreased expression of caspase-3, p53, and fas and increased Bcl-2/Bax ratio.

  16. Down-modulation of heat shock protein 70 and up-modulation of Caspase-3 during schisandrin B-induced apoptosis in human hepatoma SMMC-7721 cells

    Institute of Scientific and Technical Information of China (English)

    Yi-Feng Wu; Ming-Fu Cao; Yan-Ping Gao; Fei Chen; Tao Wang; Edward P. Zumbika; Kai-Xian Qian

    2004-01-01

    AIM: To investigate the effect of schisandrin B (Sch B) on proliferation and apoptosis of human hepatoma SMMC-7721 cells in vitro and regulation of Hsp70 and Caspases-3, 7, 9 expression by Sch B.METHODS: Human hepatoma cell line SMMC-7721 was cultured and treated with Sch B at various concentrations.Growth suppression was detected with MTT colorimetric assay. Cell apoptosis was confirmed by DNA ladder detection and flow cytometric analysis. The expression of Hsp70,Caspases-3, 7, 9 were analyzed by Western blot analysis.RESULTS: Sch B inhibited the growth of hepatoma SMMC-7721 cells in a dose-dependent manner, leading to a 50% decrease in cell number (LC50) value of 23.50 mg/L. Treatment with Sch B resulted in degradation of chromosomal DNA into small internucleosomal fragments, evidenced by the formation of a 180-200 bp DNA ladder on agarose gels.FCM analysis showed the peak areas of subdiploid at the increased concentration of Sch B. The results of Western bolt analysis showed that Hsp70 was down-regulated and Caspase-3 was up-regulated, while the activity of Caspases-7,-9 had no significant change.CONCLUSION: Sch B is able to inhibit the proliferation of human hepatoma SMMC-7721 cells and induce apoptosis,which goes through Caspase-3-dependent and Caspase-9-independent pathway accompanied with the down-regulation of Hsp70 protein expression at an early event.

  17. Cytochrome c release and caspase-3 activation in retinal ganglion cells following different distance of axotomy of the optic nerve in adult hamsters.

    Science.gov (United States)

    He, M H; Cheung, Z H; Yu, E H; Tay, D K C; So, K F

    2004-11-01

    This study examined the relationship between the distance of axotomy and the death of injured retinal ganglion cells (RGCs) in adult hamsters and the relationship of cytochrome c and caspase-3 on the death pathway of RGCs. The left optic nerve (ON) of adult hamsters was transected either at 1 or 3 mm away from the optic disc, and retrogradely labeled with Flurogold on the ON stump. After a predetermined period of postoperative time, the surviving RGCs were counted by retina flat-mount, and the activation of cytochrome c and caspase-3 were investigated by immunohistochemistry. Cell loss was found to be much faster (P < 0.01), more cells with cytochrome c were observed (P < 0.05) and the activation of caspase-3 was earlier when ON was transected 1 mm away from the optic disc than when was transected 3 mm away from the optic disc. Distance of axotomy affects the axotomized cell death rate where more RGCs died when the ON transection was applied closer to the eye. The timing of activation of caspase-3 in the RGCs may be linked to the distance of axotomy.

  18. Update of the Caspase-3 and Caspase-8 in lichen planus%Caspase-3和 Caspase-8与扁平苔藓的研究进展

    Institute of Scientific and Technical Information of China (English)

    李帅; 栗玉珍

    2016-01-01

    Caspase-3 and Caspase-8 are important proteins in the process of apoptosis. Caspase-8 can start apoptotic reaction which can mediate apoptosis in a variety of ways. Caspase-3 can accelerate cell apop-tosis after activation. The new studies reported high expression of Caspase-3, Caspase-8 in lichen planus le-sions, especially in the basal layer which showed that Caspase-3, Caspase-8 might be involved in the apop-totic response process of lichen planus.%Caspase-3和 Caspase-8作为含半胱氨酸的天冬氨酸蛋白水解酶家族成员,是参与细胞凋亡过程的两个重要蛋白。 Caspase-8是重要的凋亡启动蛋白,可通过多种途径介导凋亡反应的发生, Caspase-3作为凋亡过程中的执行者,活化后可以加速细胞凋亡的发生。最新研究发现 Caspase-3和Caspase-8在扁平苔藓皮损处(特别是基底层处)高表达,表明两者可能参与了扁平苔藓的凋亡过程,并导致了本病的病理变化。

  19. Glutamate-induced apoptosis in primary cortical neurons is inhibited by equine estrogens via down-regulation of caspase-3 and prevention of mitochondrial cytochrome c release

    Directory of Open Access Journals (Sweden)

    Zhang YueMei

    2005-02-01

    Full Text Available Abstract Background Apoptosis plays a key role in cell death observed in neurodegenerative diseases marked by a progressive loss of neurons as seen in Alzheimer's disease. Although the exact cause of apoptosis is not known, a number of factors such as free radicals, insufficient levels of nerve growth factors and excessive levels of glutamate have been implicated. We and others, have previously reported that in a stable HT22 neuronal cell line, glutamate induces apoptosis as indicated by DNA fragmentation and up- and down-regulation of Bax (pro-apoptotic, and Bcl-2 (anti-apoptotic genes respectively. Furthermore, these changes were reversed/inhibited by estrogens. Several lines of evidence also indicate that a family of cysteine proteases (caspases appear to play a critical role in neuronal apoptosis. The purpose of the present study is to determine in primary cultures of cortical cells, if glutamate-induced neuronal apoptosis and its inhibition by estrogens involve changes in caspase-3 protease and whether this process is mediated by Fas receptor and/or mitochondrial signal transduction pathways involving release of cytochrome c. Results In primary cultures of rat cortical cells, glutamate induced apoptosis that was associated with enhanced DNA fragmentation, morphological changes, and up-regulation of pro-caspase-3. Exposure of cortical cells to glutamate resulted in a time-dependent cell death and an increase in caspase-3 protein levels. Although the increase in caspase-3 levels was evident after 3 h, cell death was only significantly increased after 6 h. Treatment of cells for 6 h with 1 to 20 mM glutamate resulted in a 35 to 45% cell death that was associated with a 45 to 65% increase in the expression of caspase-3 protein. Pretreatment with caspase-3-protease inhibitor z-DEVD or pan-caspase inhibitor z-VAD significantly decreased glutamate-induced cell death of cortical cells. Exposure of cells to glutamate for 6 h in the presence or

  20. Dissecting the Opaque-2 regulatory network using transcriptome and proteome approaches along with enzyme activity measurements Análise quantitativa de rna mensageiros, proteínas e atividades enzimáticas no estudo da rede de regulação controlada pelo gene Opaco-2

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    Agnès Lefèvre

    2002-06-01

    Full Text Available The Opaque-2 (O2 gene encodes a transcriptional activator specifically expressed for grain development of maize. o2 mutants have an opaque and chalky kernel, with a decrease in zein storage protein content, and an increase in the proportions of lysine and tryptophan. In this review, we present recent results investigating genetic properties of the O2 network, using transcriptome and proteome approaches, associated with measurements of activities of enzymes of the aspartate pathway and lysine degradation. The structural polymorphism at the O2 locus was investigated by RFLP in a collection of 51 maize inbred lines. Most polymorphic sites were found outside the coding regions. We then searched for relationships between RFLP polymorphism and (i mRNA abundance of O2 and of known or suspected target genes, (ii activity of SDH and (iii amount of zein isoforms. Polymorphic restriction sites in the 5' upstream regions of the O2 gene were found associated with O2 mRNA abundance (three sites and the amount of two 19 kDa alpha-zein isoforms (two sites. One restriction site on the 3' side of the O2 gene was found associated with Lor/Sdh mRNA abundance. Our results indicate relationships between polymorphism at the O2 locus and the expression of some of its target genes. Evidence of these associations has to be confirmed on larger samples, and the analysis of the O2 gene sequence should allow more precise testing of the actual involvement of O2 polymorphism in its own transcriptional expression, and in the expression of its target genes.O gene Opaco-2 (O2, expresso especificamente no grão de milho, transcreve para um fator de transcrição da família "leucine-zipper". Mutantes o2 apresentam grãos opacos, redução na quantidade de zeínas e aumento na proporção de lisina e triptofano. Genes cuja expressão é controlada diretamente pelo O2 são conhecidos (alfa-zeínas de 22 kDa, beta-zeínas de 14 kDa, b-32 e cyPpdk1. Nesta revisão, nós apresentamos

  1. Upregulation and activation of caspase-3 or caspase-8 and elevation of intracellular free calcium mediated apoptosis of indomethacin-induced K562 cells

    Institute of Scientific and Technical Information of China (English)

    张广森; 周光飚; 戴崇文

    2004-01-01

    Background A nonsteroidal anti-inflammatory drug, indomethacin, has been shown to have anti-leukemic activity and induce leukemic cell opoptosis. This study was to elucidate the mechanism of indomethacin-induced K562 cell apoptosis.Methods K562 cells were grown in RPMI 1640 medium and treated with different doses of indomethacin (0 μmol/L, 100 μmol/L, 200 μmol/L, 400 μmol/L, 800 μmol/L) for 72 hours. The cells were harvested, and cell viability or apoptosis was analyzed using MTT assay and AO/EB stain, combining laser scanning confocal microscopy (LSCM) technique separately. For the localization and distribution of intracellular caspase-3 or caspase-8 protein, immunofluorescence assay was carried out. To reveal the activation of caspase-3 or caspase-8 in indomethacin-treated cells, Western blot detection was used. The change in intracellular free calcium was determined by Fluo-3/ Am probe labeling combined with LSCM. Results Indomethacin could lead to K562 cell apoptosis and inhibit cell viability in a concentration-dependent manner. An increased expression of intracellular caspase-3 or caspase-8 was observed at higher doses of indomethacin (400-800 μmol/L). Western blot results showed upregrulation and activation in both caspase-3 and caspase-8 protein. Under indomethacin intervention, the levels of intracellular free calcium showed a significant increase. Blocking the activity of cyclooxygenase did not abolish the effects of indomethacin on K562 cell apoptosis.Conclusions Activation and upregulation of caspase-3 or caspase-8 protein were responsible for Indomethacin-induced K562 cell apoptosis. Variation of intracellular free calcium might switch on the apoptotic pathway and the proapoptotic effect of indomethacin might be cyclooxygenase-independent.

  2. Inhibition of Stat3 activation suppresses caspase-3 and the ubiquitin-proteasome system, leading to preservation of muscle mass in cancer cachexia.

    Science.gov (United States)

    Silva, Kleiton Augusto Santos; Dong, Jiangling; Dong, Yanjun; Dong, Yanlan; Schor, Nestor; Tweardy, David J; Zhang, Liping; Mitch, William E

    2015-04-24

    Cachexia occurs in patients with advanced cancers. Despite the adverse clinical impact of cancer-induced muscle wasting, pathways causing cachexia are controversial, and clinically reliable therapies are not available. A trigger of muscle protein loss is the Jak/Stat pathway, and indeed, we found that conditioned medium from C26 colon carcinoma (C26) or Lewis lung carcinoma cells activates Stat3 (p-Stat3) in C2C12 myotubes. We identified two proteolytic pathways that are activated in muscle by p-Stat3; one is activation of caspase-3, and the other is p-Stat3 to myostatin, MAFbx/Atrogin-1, and MuRF-1 via CAAT/enhancer-binding protein δ (C/EBPδ). Using sequential deletions of the caspase-3 promoter and CHIP assays, we determined that Stat3 activation increases caspase-3 expression in C2C12 cells. Caspase-3 expression and proteolytic activity were stimulated by p-Stat3 in muscles of tumor-bearing mice. In mice with cachexia caused by Lewis lung carcinoma or C26 tumors, knock-out of p-Stat3 in muscle or with a small chemical inhibitor of p-Stat3 suppressed muscle mass losses, improved protein synthesis and degradation in muscle, and increased body weight and grip strength. Activation of p-Stat3 stimulates a pathway from C/EBPδ to myostatin and expression of MAFbx/Atrogin-1 and increases the ubiquitin-proteasome system. Indeed, C/EBPδ KO decreases the expression of MAFbx/Atrogin-1 and myostatin, while increasing muscle mass and grip strength. In conclusion, cancer stimulates p-Stat3 in muscle, activating protein loss by stimulating caspase-3, myostatin, and the ubiquitin-proteasome system. These results could lead to novel strategies for preventing cancer-induced muscle wasting.

  3. Expression of Survivin, CyclinD1, p21WAF1, Caspase-3 in Cervical Cancer and Its Relation with Prognosis

    Institute of Scientific and Technical Information of China (English)

    LU Shi; ZHANG Baohua; WANG Zehua

    2005-01-01

    The implications of Survivin, CyclinD1, p21WAF1, Caspase-3 in the development, progression and prognosis in cervical cancer were investigated. By using immunohistochemical SP method, the expression of Survivin, CyclinD1, p21WAF1 , Caspase-3 was detected in 41 cases of cervical cancer, 17 cases of cervical intraepithelial neoplasia (CIN) and 10 cases of normal tissues, and their relation with pathological grade, clinical stage, metastasis and survival time was analyzed.The results showed that the positive expression rate of Survivin, CyclinD1 in cervical cancer was significantly higher than in CIN group and normal control group (P<0.05). The median survival time in the patients with cervical cancer positive for Survivin and CyclinD1 was significantly shorter than in those with negative expression (P<0.05). The expression of both Survivin and CyclinD1 was not related with tumor grade, clinical stage and metastasis (P>0. 05). The positive expression rate of p21WAF1 , Caspase-3 in cervical ca rcer was significantly lower than in CIN group and normal control group (P<0.05), and had a close relation with tumor grade (P<0.05). The expression of Survivin in cervical cancer in cervical cancer was negatively associated with that of Caspase-3 (P<0.01), but positively with that of CyclinD1 (P<0.01). Cox Multivariate analysis revealed that Survivin was the independent prognostic indicator influencing the survival time of the patients with cervical cancer (P<0.05). It was suggested that the high expression of Survivin or CyclinD1, and low expression of p21WAF1 or Caspase-3 was closely correlated with the development of cervical cancer. Survivin and CyclinD1 could be used as a useful indicator to predict the prognosis of cervical cancer.

  4. Neural networks and statistical analysis for classification of corneal videokeratography maps based on Zernike coefficients: a quantitative comparison Redes neurais e análise estatística para classificação de mapas topográficos da córnea baseados em coeficientes de Zernike: uma comparação quantitativa

    Directory of Open Access Journals (Sweden)

    Luis Alberto Vieira de Carvalho

    2008-06-01

    Full Text Available PURPOSE: The main goal of this study was to develop and compare two different techniques for classification of specific types of corneal shapes when Zernike coefficients are used as inputs. A feed-forward artificial Neural Network (NN and discriminant analysis (DA techniques were used. METHODS: The inputs both for the NN and DA were the first 15 standard Zernike coefficients for 80 previously classified corneal elevation data files from an Eyesys System 2000 Videokeratograph (VK, installed at the Departamento de Oftalmologia of the Escola Paulista de Medicina, São Paulo. The NN had 5 output neurons which were associated with 5 typical corneal shapes: keratoconus, with-the-rule astigmatism, against-the-rule astigmatism, "regular" or "normal" shape and post-PRK. RESULTS: The NN and DA responses were statistically analyzed in terms of precision ([true positive+true negative]/total number of cases. Mean overall results for all cases for the NN and DA techniques were, respectively, 94% and 84.8%. CONCLUSION: Although we used a relatively small database, results obtained in the present study indicate that Zernike polynomials as descriptors of corneal shape may be a reliable parameter as input data for diagnostic automation of VK maps, using either NN or DA.OBJETIVOS: Nosso principal objetivo neste trabalho foi de desenvolver e comparar duas técnicas diferentes para classificação de superfícies corneanas. Uma rede neural artificial alimentada adiante e análise descriminante foram as técnicas de classificação comparadas neste trabalho. MÉTODOS: As entradas para ambos os métodos de classificação foram os primeiros 15 coeficientes de Zernike para 80 córneas mensuradas anteriormente em um topógrafo Eyesys instalado no Departamento de Oftalmologia da Escola Paulista de Medicina - UNIFESP. A rede neural tem 5 saídas que foram associados aos cinco casos típicos contidos na base de dados: ceratocone, astigmatismo a favor da regra, astigmatismo

  5. Análise descritiva quantitativa de edulcorantes em diferentes concentrações

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    CARDELLO Helena Maria André Bolini

    2000-01-01

    Full Text Available Edulcorantes em solução, com a mesma equivalência de doçura, podem apresentar características sensoriais que os tornam diferentes entre si. O presente estudo teve como objetivo realizar Análise Descritiva Quantitativa de soluções de aspartame (APM, extrato de folhas de estévia (SrB e mistura ciclamato/sacarina 2:1 (C/S em diferentes níveis de doçura, ou seja, em equivalência de doçura a uma solução aquosa de sacarose a 3, 10, 20 e 30%. Onze provadores, pré-selecionados através de análise seqüencial, tendo como critério suas habilidades de discriminação, foram treinados após o levantamento da terminologia descritiva. Após o treinamento, os provadores foram selecionados através de seu poder de discriminação, reproducibilidade e concordância com a equipe no uso de escalas. Os termos descritivos dos edulcorantes, para todos os níveis de doçura, gerados através do método rede (Kelly's Repertory Grid Method foram: doçura inicial, doçura residual, amargo inicial, amargo residual, residual de alcaçuz, corpo e acidez. Os resultados obtidos para cada nível de doçura foram analisados através de análise de variância, teste de Tukey e Análise de Componentes Principais. A análise descritiva foi efetiva em caracterizar o perfil sensorial dos edulcorantes em diferentes concentrações, evidenciando as mudanças no perfil com o aumento de suas concentrações.

  6. Mecanismos de busca acadêmica: uma análise quantitativa

    Directory of Open Access Journals (Sweden)

    Diego Buchinger

    2014-04-01

    Full Text Available Os mecanismos de busca consolidaram a pesquisa por informação na internet,sendo os mecanismos de busca acadêmicos (MBAs, hoje, os principais meios para alcançar fontes científicas, como artigos de eventos ou periódicos. Ao invés de gastar horas em uma biblioteca, os MBAs ganham em popularidade pela facilidade de automatizar uma busca e acessar inúmeras bases de dados simultaneamente. Contudo, o caráter incremental das bases de materiais científicos e dos diversos MBAs existentes impõe um desafio aos pesquisadores, que é determinar um MBA que melhor atenda aos seus anseios de pesquisa. Nesse contexto, uma análise quantitativa dos recursos providos pelos MBAs foi realizada com objetivo de determinar quais são os melhores para facilitar as buscas por conteúdo científico, considerando recursos de busca, de refinamento e auxiliares. Dentre um grupo de 40 MBAs iniciais, sete apresentaram-se com σ = 0,5 acima da média nos critérios utilizados, na seguinte ordem: 1o Web of Knowledge; 2o Engineering Village; 3o Scopus SciVerse; 4o IEEE Xplore; 5o ACM Digital Library; 6o Science Direct; e 7o Springer Link. Esses MBAs deveriam ser melhor explorados pelos pesquisadores pois tem inúmeros recursos capazes de facilitar sobremaneira e elevar a qualidade do trabalho de pesquisa.

  7. Avaliação quantitativa das forças laterais da patela: ressonância magnética estática e cinemática Quantitative evaluation of lateral forces on the patella: static and kinematic magnetic resonance imaging

    Directory of Open Access Journals (Sweden)

    Je Hoon Yang

    2007-08-01

    Full Text Available OBJETIVO: Avaliar a validade da ressonância magnética cinemática combinada com a ressonância magnética estática no estudo da articulação femoropatelar. MATERIAIS E MÉTODOS: Foram realizadas ressonância magnética estática e ressonância magnética cinemática em 20 voluntários assintomáticos (40 joelhos e em 23 pacientes (43 joelhos, em aparelho de configuração fechada de 1,5 tesla de campo. Os indivíduos foram posicionados na extremidade da mesa, em 30° de flexão. A translação patelar foi avaliada medindo-se o desvio da bissetriz, o deslocamento lateral da patela e o ângulo de inclinação da patela. Para a comparação entre os estudos estático e cinemático, foi utilizado o teste não-paramétrico de Wilcoxon. Para a comparação entre os voluntários e os pacientes, foi utilizado o teste de Mann-Whitney. RESULTADOS: Houve diferenças significantes entre a ressonância magnética estática e a ressonância magnética cinemática (p OBJECTIVE: To evaluate the usefulness of combining static and kinematic magnetic resonance imaging in the evaluation of the femoropatellar joint. MATERIALS AND METHODS: Twenty healthy volunteers (40 knees and 23 patients (43 knees were submitted to both static and kinematic magnetic resonance imaging on a 1.5 tesla whole-body magnetic resonance scanner. The knees were positioned at 30º flexion with the quadrature knee coil at the inner end of the examination table. The patellar translation was evaluated by measurements of bisect offset, lateral patellar displacement and patellar tilt angle. The nonparametric Wilcoxon test was utilized for statistical analysis of data resulting from the static and kinematic studies in both groups. Nonparametric Mann-Whitney test was utilized in the comparison between healthy volunteers and patients. RESULTS: Statistical analysis demonstrated significant differences (p < 0.05 between static and kinematic magnetic resonance imaging for the three parameters

  8. The effects of ultrasonic scaling duration and replication on caspase-3 expression of Sprague Dawley rat's pulp cells

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    Archadian Nuryanti

    2015-03-01

    Full Text Available Background: Ultrasonic scaling has been used commonly for stain and calculus removal in dental clinic for over 60years. Previous researches even had proved that ultrasonic scaling may give effects on the surface of tooth root. Ultrasonic wave exposure for 20 seconds or more can increase caspase-3 activity as an indicator of increased apoptotic cells associated with tissue damage. Purpose: This research was aimed to investigate the effects of ultrasonic scaling duration and replication on caspace-3 expression in dental pulp cells. Methods: The samples of this research were 54 male Sprague Dawley rats aged 2 months old divided into 2 groups, each of which consisted of 27 mice. The first group was induced with stain, while the second group was not. Each group was divided into 3 subgroups for ultrasonic scaling 1, 3, and 5 times. Each subgroup was divided into 3 sub-subgroups for duration procedure of 15, 30 and 60 seconds respectively. During scaling process, those rats were anesthetized using 0.1 ml of ketamine and 0.1 ml of xylol added to 2 ml of distilled water injected intramuscularly into their right thigh as much as 0.4 ml. Scaling was done on buccal surface of right first maxillary molar from cervical to occlusal. The teeth were decalcified and embedded in paraffin, then their sagittal plane was cut for thickness of 3µm and painted with immunohystochemistry for detecting caspace-3 expression of cell within dental pulp. Results: The results showed that the duration and replication of ultrasonic scaling procedures affected on the expression of caspace-3 cells as analyzed with Univariate Analisis of Variance test (p<0.05. Conclusion: It can be concluded that duration and replication of ultrasonic scaling procedure on teeth with and without stain enhauced the expression of  caspace-3 in dental pulp cells.

  9. Effect of irradiation on the expression of caspase-3 in the submandibular gland of streptozotocin-induced diabetic rats

    Energy Technology Data Exchange (ETDEWEB)

    Lee, Heung Ki; Hwang, Eui Hwan; Lee, Sang Rae [Kyunghee University College of Medicine, Seoul (Korea, Republic of)

    2005-09-15

    To observe the histopathological changes and caspace-3 expression in the submandibular gland in streptozotocin-induced diabetic rats after irradiation. The male Sprague-Dawley rats weighing approximately 250gm were divided into four groups; control, diabetes, irradiation, and diabetes-irradiation groups. Diabetes mellitus was induced in the rats by injecting streptozotocin. Rats in the control and irradiation groups were injected with citrate buffer only. After 5 days, rats in irradiation, and diabetes-irradiation groups were irradiated with a single absorbed dose of 10 Gy to the head and neck region. All the rats were sacrificed at 3, 7, 14, 21, and 28 days after irradiation. The specimen including the submandibular gland were sectioned and observed using histopathological and immunohistochemical methods. In the irradiation group, the condensed nucleus, karyolysis, and degeneration of the acinar cells and atrophy of the duct cells were observed in the early experimental phase. However, the acinar cells were found to be normal at 28 days after irradiation. In the diabetes group, the condensed nucleus, karyolysis, atrophy, and degeneration of the acinar cells were observed in the early experimental phase. However, the acinar cells were found to be normal at 21 days, after diabetic state induction. In the diabetes-irradiation group, the ductal epithelial cells were predominant in their glandular tissues at 28 days after irradiation. In all of the experimental groups, the most prominent change of the acinar cells and ductal cells were observed at 14 days after diabetic state induction and irradiation. The expression of caspase-3 in the acinar cells and ductal cells of the submandibular gland was weak after irradiation, but that in the acinar cells, ductal cells, and fibrous cells of the submandibular gland was prominent after diabetic state induction.

  10. Astilbic Acid Induced COLO 205 cell Apoptosis by Regulating Bcl-2 and Bax Expression and Activating Caspase-3

    Institute of Scientific and Technical Information of China (English)

    ZhengXiao-liang; SunHong-xiang; LiuXue-li; ChenYun-xiang; QianBo-chu

    2005-01-01

    To investigate the effect of astilbic acid (3β,6β-dihydroxyolean-12-en-27-oic acid, AA) on human colorectal carcinoma COLO 205 cell proliferation and apoptosis.Methods Proliferation of COLO 205 cells was measued by MTT assay. Content of DNA in COLO 205 cell was measued by modified diphenylamine assay. AA-induced morphological changes was observed with fluorescence microscope and transmission electron microscope.DNA fragmentation was visualized by agarose gel electrophoresis.Apoptosis rate and cell cycle distribution were deter-mined by flow cytometric analysis.Expressions of Bcl-2 and Bax proteins were visioned by immunohistochemical analysis.The change of relative mitochondral transmembrane potential (MTP) in COLO 205 cell was analyzed with FCM after rhodamine 123 staining. Results The IC50 (96h) of AA for inhibiting COLO 205 cell proliferation was 61.56±0.34 μmol/L.AA induced a marked concentration- and time-dependent inhibition of COLO 205 cell proliferation and reduced the DNA content in COLO 205 cell. Cells treated with AA 64 μmol/L showed typical morphological changes of apoptosis and DNA “ladder” pattern. The cell cycle was arrested in G0/G1 phase, and the apoptosis rate was 28.25% for COLO 205 cells treated with AA 64 μmol/L for 48h. Meanwhile the expression of Bcl-2 protein was decreased while that of Bax was increased and relative MTP was decreased as well. DEVD-CHO 1μmol/L could increase the viability of COLO 205 cells treated with AA for 48h.Conclusion AA showed potent inhibitory activity on COLO 205 cells proliferation,and could induce COLO 205 cells apoptosis through disturbing DNA replication, down-regulating Bcl-2 expression, and up-regulating Bax expression, lowering relative MTP, and activating caspase-3 pathway.

  11. Astilbic acid induced COLO 205 cell apoptosis by regulating Bcl-2 and Bax expression and activating caspase-3

    Institute of Scientific and Technical Information of China (English)

    Xiao-liang ZHENG; Hong-xiang SUN; Xue-li LIU; Yun-xiang CHEN; Bo-chu QIAN

    2004-01-01

    AIM: To investigate the effect of astilbic acid (3β, 6β-dihydroxyolean-12-en-27-oic acid, AA) on human colorectal carcinoma COLO 205 cell proliferation and apoptosis. METHODS: Proliferation of COLO 205 cells was measued by MTT assay. Content of DNA in COLO 205 cell was measued by modified diphenylamine assay. AA-induced morphological changes was observed with fluorescence microscope and transmission electron microscope. DNA fragmentation was visualized by agarose gel electrophoresis. Apoptosis rate and cell cycle distribution were determined by flow cytometric analysis. Expressions of Bcl-2 and Bax proteins were visioned by immunohistochemical analysis. The change of relative mitochondral transmembrane potential (MTP) in COLO 205 cell was analyzed with FCM after rhodamine 123 staining. RESULTS: The ICs0 (96 h) of AA for inhibiting COLO 205 cell proliferation was 61.56±0.34 μmol/L. AA induced a marked concentration- and time-dependent inhibition of COLO 205 cell proliferation and reduced the DNA content in COLO 205 cell. Cells treated with AA 64 μmol/L showed typical morphological changes of apoptosis and DNA "ladder" pattern. The cell cycle was arrested in G0/G1 phase, and the apoptosis rate was 28.25 % for COLO 205 cells treated with AA 64 μmol/L for 48 h. Meanwhile the expression of Bcl-2 protein was decreased while that of Bax was increased and relative MTP was decreased as well. DEVD-CHO 1 μmol/L could increase the viability of COLO 205 cells treated with AA for 48 h. CONCLUSION: AA showed potent inhibitory activity on COLO 205 cells proliferation, and could induce COLO 205 cells apoptosis through disturbing DNA replication, down-regulatin Bcl-2 expression,and up-regulating Bax expression,lowering relative MTP, and activating caspase-3 pathway.

  12. Determinação quantitativa da concentração de silicone em antiespumantes por espectroscopia FT-IR / ATR e calibração multivariada Quantitative determination of silicone in antifoaming products by FT-IR / ATR spectroscopy and multivariate calibration

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    Marcelo H. F. Garcia

    2004-12-01

    Full Text Available Neste trabalho apresentamos uma alternativa para a dosagem do teor de silicone (polidimetilsiloxano em antiespumantes por meio da técnica de espectroscopia no infravermelho com transformada de Fourier (FT-IR, com a utilização do acessório de reflectância total atenuada (ATR. Os espectros foram registrados na faixa espectral de 2500 a 780 cm-1, com resolução de 4 cm-1 e 128 varreduras. A calibração de um modelo linear por meio da utilização do método de mínimos quadrados parciais (PLS aplicado aos espectros foi capaz de determinar satisfatoriamente a concentração de silicone nas amostras. Este método é de extrema importância para indústrias produtoras de antiespumantes siliconados, uma vez que o desempenho de tais produtos geralmente é avaliado em função da viscosidade dos mesmos. Muitas vezes no processo de fabricação de tais produtos ocorre uma homogeneização incompleta do silicone no solvente, o que leva a resultados de viscosidade que não são representativos das amostras analisadas. A determinação da concentração do teor de silicone é uma importante ferramenta para o Controle Estatístico de Processo (CEP, pois evita o desperdício de matérias-primas empregadas na fabricação dos antiespumantes.This work presents an alternative method to determine the concentration of silicone (polydimethylsiloxane in antifoaming products using Fourier Transformed Infrared Spectroscopy (FT-IR with the attenuated total reflectance (ATR accessory. The spectra were recorded in the range from 2500 to 780 cm-1, with a resolution of 4 cm-1 and 128 scans. With calibration of a linear model using PLS regression method applied to spectral data we were able to determine the silicone concentration in the samples. This method may be useful for antifoaming producers since the performance of such products generally is evaluated as a function of their viscosity. Moreover, during manufacturing an incomplete homogenization of silicone in the

  13. Effects of basic fibroblast growth factor on glutamate-induced expression of Caspase-3 and Bcl-2 in the guinea pig retina%碱性成纤维细胞生长因子对谷氨酸诱导的豚鼠视网膜内Caspase-3和Bcl-2表达的影响

    Institute of Scientific and Technical Information of China (English)

    杜长青; 魏丽华; 石运芝; 杜辉; 刘立伟

    2011-01-01

    Objective: To assess the expression of Bcl-2 and Caspase-3 in the retina after superabundant glutamate injury and investigate the neural protective effect of basic fibroblast growth factor (bFGF) on retinal excitotoxicity damage in guinea pigs. Methods: Guinea pigs were randomly divided into a glutamate injury model group, a normal control group and a bFGF treatment group. The expression of Bcl-2 and Caspase-3 in the retina was detected by immunohistochemical method and image analysis. Results: A few of Caspase-3 positive cells were found in the normal control group; the expression of Caspase-3 was significantly increased in the injury group; and Ecl-2 showed weak expression in the retina of the normal control group and there was no significant change after darnage. The expression of Ecl-2 significantly increased and the expression of Caspase-3 reduced in the treatment group received bFGF injection beforehand. There was significant difference between the treatment group and model group. Conclusion: bFGF might selectively reduce the expression of Caspase-3 and up-regulate the expression of Bcl-2 in the retina after superabundant glutamate injury, and one of the possible mechanisms is the inhibition of glutamate-injured RGCs apoptosis by bFGF.%目的:探讨过量谷氨酸毒性损伤后视网膜天冬氨酸蛋白酶-3(Caspase-3)和凋亡相关蛋白Bcl-2的表达及碱性成纤维细胞生长因子(bFGF)对兴奋毒性损伤的保护作用.方法: 豚鼠随机分为正常对照组、谷氨酸损伤组、bFGF治疗组.采用免疫组织化学方法和图像分析技术,对各组豚鼠视网膜内Caspase-3和Bcl-2的表达进行检测.结果: 对照组Caspase-3无明显表达,损伤组在节细胞层、内核层、内界膜和外界膜等处Caspase-3表达阳性面积百分率和密度明显上调,bFGF治疗组Caspase-3表达明显降低.对照组Bcl-2在节细胞层、神经纤维层及内核层内有弱阳性表达,损伤后变化不明显,bFGF治疗组Bcl-2的表

  14. C57BL/6J小鼠初级听皮层神经元凋亡与caspase-3表达的年龄相关性改变%The Age-related Changes of the Expression of Caspase-3 and the apoptosis States of Neurons in Primary Auditory Cortex(AI) of C57BL/6J Mice

    Institute of Scientific and Technical Information of China (English)

    李洪波; 陈继川; 姬长友

    2009-01-01

    目的 探讨不同月龄C57BL/6J小鼠初级听皮层中caspase-3的表达及初级听皮层神经元凋亡情况,探讨两者之间的关系以及caspase-3、凋亡在老年性聋发生、发展中的作用.方法 分别选取2月龄(1 5~20克)和10月龄(45~60克)C57BL/6J小鼠各15只,免疫组织化学法染色检测两组C57BL/6J小鼠初级听皮层caspase-3的表达情况,末端转移酶介导的原位缺口末端标记染色(TUNEL)技术检测两组小鼠仞级听皮层神经元凋亡状况.结果 与2月龄组C57BL/6J小鼠相比,10月龄组C57BL/6J小鼠初级听皮层中caspase-3的表达显著增多,初级听皮层神经元凋亡数目明显增多(P值均<0.01),且caspase-3的表达与神经元的凋亡呈正相关(r=0.5202).结论 caspase-3的表达可能在老年性聋的发生、发展过程中起重要作用,它参与了小鼠初级听皮层神经元的凋亡调控过程,可能是老年性聋的发病机制中一个重要因素.%Objective This study is to study the age related changes of the expression of caspase-3 and the apoptosis states of neurons in primary auditory cortex of 15 young C57BL/6J mice(2 months, 15~20 g) and 15 old C57BL/6J mice(10 months, 50~60 g) and to determine probable physical effects underlying these changes. This paper also discusses the relationship of caspase-3 and apotosis states in primary auditory cortex, the possible role of caspase-3 in primary auditory cortex and the pathogenesis of presbycusis. Methods The immunohistochemical methods were applied to explore the differences of the expression of caspase-3 and the apoptosis states determined by TUNEI. method in the primary auditory cortex between young and old C57BL/6J mice. Results The expression of caspase-3 and apoptosis in Al of old C57BL/6J mice was significantly higher than that in the counterpart of young C57BL/6J mice. Conclusion The results presented a direct morphological evidence for the strengthening of caspase-3 in the primary auditory cortex in

  15. Avaliação quantitativa da sustentabilidade estrutural de um latossolo vermelho-amarelo cultivado com eucalipto na região de Peçanha-MG Quantitative evaluation of the structural sustainability of a red-yellow latosol cultivated with eucalyptus in Peçanha-MG

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    Moacir de Souza Dias Junior

    2003-06-01

    Full Text Available Uma das limitações para que o desenvolvimento florestal sustentável seja atingido está relacionado com a questão do tráfego durante a realização das operações de colheita e transporte de madeira, pois elas podem causar degradação da estrutura do solo. Visando obter uma solução para este problema, os objetivos deste estudo foram: a propor um modelo de sustentabilidade da estrutura dos solos cultivados com Eucalyptus spp., na região de Peçanha-MG, em função da pressão de preconsolidação (sigmap e da umidade (U; e b determinar, através do uso deste modelo, a influência das operações realizadas com o Feller-Büncher e Skidder (Pneu 30.5L.32, Feller-Büncher e Skidder (Pneu 66.43.00.26 e Harvester e Forwarder na estrutura do solo. Este estudo foi conduzido em um Latossolo Vermelho-Amarelo (LV, sob Eucalyptus, utilizando amostras indeformadas, coletadas antes e após as operações da colheita, na profundidade de 0,10 a 0,125 m. As amostras indeformadas foram utilizadas nos ensaios de compressão uniaxial. Foram determinadas também a textura e a densidade de partículas do solo. O modelo de sustentabilidade da estrutura do Latossolo Vermelho-Amarelo obtido para os projetos deste estudo é expresso pela seguinte equação: sigmap = 10(2,72-1,29 U. O uso dos modelos de sustentabilidade da estrutura e da pressão de preconsolidação, determinada após o tráfego, permitiu quantificar os efeitos causados pelo Feller-Büncher e Skidder (Pneu 30.5L.32, Feller-Büncher e Skidder (Pneu 66.43.00.26 e Harvester e Forwarder na estrutura do Latossolo Vermelho-Amarelo. Nos projetos Buriti e São Leonardo, as operações realizadas com o Harvester e Forwarder foram as que mais degradaram a estrutura do solo, enquanto no projeto Dourado foram as realizadas com o Feller-Büncher e Skidder (Pneu 30.5L.32 e Feller-Büncher e Skidder (Pneu 66.43.00.26.One of the limitations for sustainable forest development is related to wheel traffic during

  16. 龙葵碱对HepG2细胞内caspase-3及Bcl-2蛋白含量的影响%Effect of Solanine on the Contents of Caspase-3 and Bcl-2 in HepG2

    Institute of Scientific and Technical Information of China (English)

    高世勇; 王秋娟; 季宇彬

    2006-01-01

    目的:观察龙葵碱对HepG2细胞内caspase-3及Bcl-2蛋白含量的影响,阐明龙葵碱诱导肿瘤细胞凋亡的作用机制.方法:采用激光共聚焦扫描显微术和Western blot法检测caspase-3和Bcl-2蛋白含量,并对二者在细胞内的位置进行定位.结果:龙葵碱能够显著升高HepG2细胞内caspase-3蛋白含量,降低Bcl-2的含量,并且均具有剂量依赖性.Bcl-2蛋白和caspase-3蛋白均在细胞浆内呈不均匀分布,在细胞核中无分布,龙葵碱对二者的分布没有影响.结论:龙葵碱通过抑制Bcl-2的活性,激活caspase-3蛋白,诱导细胞凋亡的发生.

  17. Double staining techniques in the study of caspase-3 and the apoptosis after traumatic brain injury in rats%双标染色技术在大鼠脑创伤后caspase-3与神经细胞凋亡研究中的应用

    Institute of Scientific and Technical Information of China (English)

    宋朝彦; 董晓辉; 谢东

    2014-01-01

    目的 探讨末端标记(TUNEL)法与caspase-3免疫组织化学法双标染色技术在研究大鼠神经细胞凋亡机制中应用的优越性.方法 雄性Wistar大鼠12只随机分成两组,用Marmarou方法造成大鼠重型弥漫性颅脑创伤.切片先按照试剂盒提供的方法行TUNEL染色显示凋亡,再按caspase-3免疫组化试剂盒检测方法继续染色.结果 双标染色显色满意,大鼠海马区大部分TUNEL阳性细胞同时也表现caspase-3阳性.结论 caspase-3与TUNEL双标染色能更好的反映出大鼠脑创伤后caspase-3与凋亡之间的关系,体现了双标染色技术的优越性.

  18. Análise quali-quantitativa da ictiofauna acompanhante na pesca do camarão sete-barbas, na Armação do Itapocoroy, Penha, Santa Catarina Quali-quantitative analysis of sea-bob-shrimp's ichthyofauna bycatch, at Armação do Itapocoroy, Penha, Santa Catarina

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    Joaquim O. Branco

    2006-06-01

    Full Text Available A ictiofauna acompanhante na pesca de camarão sete-barbas foi estudada mensalmente entre agosto 1996 a julho 2003 nas áreas de atuação da pesca artesanal na Armação do Itapocoroy, Penha. Foram utilizadas duas redes-de-arrasto com portas, malha de 3,0 cm na manga e corpo e de 2,0 cm no ensacador, tracionada por baleeira com velocidade média de 2,0 nós. A temperatura e salinidade média da água de fundo mantiveram um padrão uniforme de flutuação sazonal, sem diferença significativa entre os anos de coleta. Das 60 espécies capturadas, 22 foram comuns ao longo do período de coleta. As famílias Sciaenidae, Carangidae e Clupeidae apresentaram o maior número de espécies contribuindo com 92,0% das capturas, onde Stellifer spp., Paralonchurus brasiliensis e Isopisthus parvipinnis foram mais abundantes nas coletas, apresentando ocorrência regular ao longo dos anos. Ocorreram flutuações sazonais na abundância e diversidade da ictiofauna com diferença significativa entre os anos de coleta e biomassa, bem como em número de exemplares. O monitoramento da pesca de arrasto direcionada ao camarão sete-barbas poderá contribuir na manutenção dos estoques e assegurar a subsistência das comunidades tradicionais no litoral catarinense.The ichthyofauna bycatch in sea-bob-shrimp's fishery was monthly studied from August 1996 to July 2003, in the artisanal fishing grounds at Armação do Itapocoroy, Penha. It had been used two net-of-drag with doors, mesh of 3.0 cm in the sleeve and body and of 2.0 cm in the sacker, pushed for whaleboat with average speed of 2.0 km/h. Mean values of bottom water temperature and salinity remained uniform in a seasonal fluctuation, showing no significant differences among the studied years. A total of 60 species were collected, in which 22 were considered common species along the sampled period. Sciaenidae, Carangidae and Clupeidae families presented the highest number of specimens, accounting to 92% of

  19. Experimental model in the qualitative and quantitative assessment of non-Helicobacter gastric microflora under proton pump inhibitors action Modelo experimental na avaliação qualitativa e quantitativa da microflora gástrica não-Helicobacter sob ação de inibidores de bomba de próton

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    Augusto Diogo Filho

    2006-10-01

    Full Text Available PURPOSE: To evaluate models of gastric material collection from Wistar rats with and without using proton pump inhibitors(PPIs. METHODS: Twenty-four rats underwent intraperitoneal omeprazol treatment, and other 12 received similar treatment with 0.9% saline. All animals underwent collection of gastric material samples, after stomach removal, by either biopsies, or aspirates, or swabs. Samples were bacteriologically processed in order to identify species and strains. Values are described as natural logarithm of colony former units per mL [Ln(CFU/mL]. Kruskal-Wallis and Mann-Whitney non-parametric tests were used, and pOBJETIVO: Avaliar modelos de coleta de material gástrico de ratos da linhagem Wistar, com e sem o uso de inibidores de bomba de próton (IBPs. MÉTODOS: 24 ratos foram submetidos a tratamento com omeprazol intraperitoneal e 12 outros ratos receberam tratamento semelhante com solução salina a 0,9%. Os animais foram submetidos a coleta de amostras de material gástrico, após retirada do estômago, utilizando-se de biópsias, aspirados ou swabs. Os materiais obtidos foram processados bacteriologicamente para identificação de espécimes quanto ao gênero. Os valores são descritos em logaritmo natural das unidades formadoras de colônias por mL [Ln(UFC/mL]. Utilizou-se os testes não-paramétricos de Kruskal-Wallis e Mann-Whitney, considerando-se p<0,05 como estatisticamente significativo. RESULTADOS: Não se observou diferença significativa da quantidade de Ln(UFC/mL entre os três métodos de coleta, independente do uso de omeprazol. Também não se observou diferença significativa de Ln(UFC/mL ao comparar-se os métodos individualmente entre si nas condições de uso de omeprazol ou placebo. Houve aumento significativo da variedade de gêneros de bactérias com o uso de IBP, nos 3 métodos de coleta, sendo isto mais perceptível na biópsia e swab. CONCLUSÃO: Não houve diferença entre os três métodos de coleta de

  20. Comparação quantitativa da comunidade de aves de um fragmento de floresta semidecidual do interior do Estado de São Paulo em intervalo de 30 anos A 30-year quantitative comparison of the bird community of a semideciduous forest remnant in the state of São Paulo

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    Vagner Cavarzere

    2012-12-01

    Full Text Available Poucos estudos avaliaram em longo prazo a variação no tamanho populacional das espécies de aves em fragmentos florestais. Para avaliar a riqueza e a abundância específica da comunidade de aves de um remanescente de mata semidecidual do interior do Estado de São Paulo, sudeste do Brasil, foi conduzido o censo da avifauna florestal utilizando-se a metodologia de contagem em transecção. Estes resultados foram comparados com levantamento realizado na mesma localidade 30 anos antes, e as aves foram classificadas de acordo com suas categorias alimentares com a finalidade de associá-las à tendência ao aumento/diminuição de suas abundâncias após este intervalo de tempo. Embora tenha havido predominância de espécies com diminuição populacional, todas as categorias tróficas analisadas apresentaram também espécies com aumento em suas abundâncias. A maioria das espécies com propensão a deslocarem-se entre fragmentos apresentou diminuição em suas abundâncias. Sugerimos que, em relação a suas abundâncias específicas, as categorias tróficas são igualmente afetadas pelos processos da fragmentação, e que a regeneração florestal sofrida pelo remanescente pode ter resultado na perda de espécies de bordas. Espécies cujas abundâncias tenham reduzido neste intervalo de tempo podem sofrer extinção local futuramente.Few studies have evaluated long-term changes in avian abundance in forest remnants. To compare both species richness and abundance of the bird community in a forest fragment located in the municipality of Gália, state of São Paulo, southeastern Brazil, we surveyed forest birds using transect counts. We compared our results with a survey conducted 30 years earlier at the same locality and further classified bird species according to their food habits to eventually predict fluctuations of specific abundance. Although species with population declines predominated in the community, all trophic categories had species

  1. Análise genético-quantitativa de pesos aos 8, 12, 18 e 24 meses de idade em um rebanho da raça Guzerá Quantitative and genetic analysis of weights at 8, 12, 18 and 24 months of age in a Guzerat breed herd

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    Talita Buttarello Mucari

    2003-12-01

    Full Text Available O objetivo deste trabalho foi obter estimativas de herdabilidades, correlações genéticas, tendências genéticas e correlações de "rank" dos touros, para os pesos aos 8, 12, 18 e 24 meses de idade, no rebanho Guzerá do Campus da UNESP, Ilha Solteira, SP. As herdabilidades e os valores genéticos dos animais foram estimados por modelo animal, usando o programa computacional MTDFREML. As correlações genéticas (Pearson e de "rank" dos touros (Spearman foram obtidas pelo procedimento CORR do SAS, utilizando os valores genéticos dos animais, enquanto as tendências genéticas foram calculadas pelo procedimento REG do SAS, utilizando a média anual dos referidos valores genéticos. O modelo estatístico para a análise genética incluiu os efeitos fixos de grupo contemporâneo e a covariável idade da vaca ao parto (efeitos linear e quadrático, e os efeitos aleatórios genético aditivo direto, genético aditivo materno, de ambiente permanente da vaca e residual. As estimativas obtidas para a herdabilidade direta foram 0,14; 0,08; 0,08 e 0,13 e para materna, 0,01; 0,02; 0,02 e 0,05, respectivamente, para os pesos aos 8, 12, 18 e 24 meses de idade. As estimativas dos coeficientes de correlação genética foram positivas e de alta magnitude entre todos os pesos estudados. As tendências genéticas anuais foram baixas e significativas apenas para os pesos aos 8 e 18 meses de idade. As correlações de "rank" dos touros foram moderadas e significativas, implicando em razoável manutenção de posição de classificação dos touros, quando se comparam, dois a dois, os pesos estudados.The objective of this study was to estimate heritabilities, genetic correlations, genetic trends and rank correlations of the bulls, for weights at 8, 12, 18 and 24 months of age, in the Guzerat herd from UNESP Campus, Ilha Solteira, SP. The heritabilities and the breeding values were estimated by animal model, using the MTDFREML computational program. The genetic

  2. 大鼠脑出血后细胞凋亡与Caspase-3、Ref-1表达的相关性%Correlation between expression of Caspase-3、 Ref-1 and apoptosis after intracerebral hemorrhage in rats

    Institute of Scientific and Technical Information of China (English)

    洪丽蓉; 沈国理; 喻森明; 胡晓华

    2007-01-01

    [目的]研究脑出血(intracerebral hemorrhage,ICH)血肿周围缺血半暗带区中半胱氨酸蛋白酶(Caspase-3)与氧化还原因子-1(redox factor-1,Ref-1)的表达与细胞凋亡的关系.[方法]采取立体定向技术,将SD大鼠自体不凝血50 μl注入大脑尾状核区制备脑出血模型,将大鼠随机分为正常组和出血组,分别在不同时间点断头取脑,连续切片作TUNEL、Ref-1和Caspase-3免疫组化染色.[结果]脑出血后血肿周围缺血半暗带区中细胞凋亡与Caspase-3表达呈正相关(r=0.466,P<0.01),与Ref-1表达呈负相关(r=-0.195,P<0.05);且Caspase-3表达在开始及高峰时间上先于细胞凋亡的发生,Ref-1表达明显下降和下降谷底时间均早于细胞凋亡出现的时间.[结论]缺血半暗带区脑组织中细胞凋亡与Ref-1及Caspase-3表达相关,且caspase-3表达的高峰、Ref-1表达的下降均先于细胞凋亡的发生.

  3. Time lapse imaging analysis of the effect of ER stress modulators on apoptotic cell assessed by caspase3/7 activation in NG108-15 cells

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    Ayako Saito

    2016-03-01

    Full Text Available This paper reports the data from the long term time lapse imaging of neuronal cell line NG108-15 that were treated with apoptosis inducer or various ER stress inducers. Use of the fluorescent reporter for activated caspase3/7 in combination with the conventional light microscope allowed us to investigate the time course of apoptosis induction at the single cell level. Quantitative as well as qualitative data are presented here to show the effect of two different ER stress modulating chemical compounds on caspase3/7-dependent apoptosis in neuronal cell line NG108-15 cells. Additional results and interpretation of our data concerning ER stress and apoptosis in NG108-15 cells can be found in Suga et al. (2015 [1] and in Suga et al. (2015 [2].

  4. Expressions of Caspase-3 and Bcl-2 in Related Encephalic Region of Rats with Poisoning of Methamphetamine%甲基苯丙胺中毒大鼠相关脑区Caspase-3和Bcl-2表达的研究

    Institute of Scientific and Technical Information of China (English)

    段晓飞; 邓冲; 曾晓锋; 赵永和; 王尚文; 李桢

    2011-01-01

    目的:研究凋亡相关因子Caspase-3和Bcl-2在甲基苯丙胺神经毒性机制中所发挥的作用.方法:40只健康雄性SD大鼠随机分为对照组(n=10)和实验组(又分为3个亚组,分别为注射1天后、4天后、7天后,n=10).实验组给予20mg/kg的甲基苯丙胺腹腔注射,对照组给予相同剂量的生理盐水注射.用免疫组化检测中毒大鼠相关脑区Caspase-3和Bcl-2的表达,用图像分析技术对检测结果进行分析.结果:Caspase-3在中毒大鼠不同脑区表达逐渐增加并可见明显的阳性信号;Bcl-2在中毒大鼠不同脑区表达逐渐减弱.结论:凋亡相关因子Caspase-3和Bcl-2参与了甲基苯丙胺神经毒性机制.%Objective: To study the function of Caspase-3 and Bcl-2 protein in the methamphetamine mechanism. Methods: Forty male Sprague-Dawley rats were randomly divided into control group (n=10) and experimental group(dividing it into the first, fourth and seventh group after the injection, n=10). Rats in the experimental group were intraperitoneal injected with methamphetamine hydrochloride (20mg/kg), and those in the control group were injected with saline with the same volume. Then examining the protein expression of Caspase-3 and Bcl-2 of poisoning rats in related encephalic region by immunohistochemistry technique and analyzing the test results through image. Results: Expression of Caspase-3 increased significantly in the related encephalicregion in rat poisoning of methamphetamine where exist more clearly positive signal. Express of Bcl-2 the reduced gruadually in the related encephalicregion in rat poisoning of methamphetamine. Conclusion: The abnormal expression of Caspase-3 and Bcl-2 shows that they take part in the Methamphetamine-lnduced Neurotoxicity.

  5. Expression of PDCD5 and caspase 3 in the cochlea of different age of C57BL/6J mice%PDCD5和caspase3在不同年龄段C57BL/6J小鼠耳蜗中的表达

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    王燕; 褚汉启; 周良强; 陈金; 李志勇; 刘云; 张平; 黄孝文; 崔永华

    2011-01-01

    Objective To investigate the age related changes of the expression of programmed cell death 5 (PDCDS) and caspase 3 in the cochlea of the different age of C57 BL/6J mice. The relationship of PDCD5 and caspase 3 and the possible roles in the pathogenesis of presbycusis were also discussed.Methods C57 mice of 3, 6, 9 and 12 months old were selected and divided into 4 groups, with 15 mice in each group. Auditory function of C57BL/6J mice was measured by auditory brainstem response (ABR) respectively. The changes of PDCD5 and Caspase 3 protein in the cochlea were detected by immunohistochemistry and Western blotting, the changes of PDCD5 mRNA and caspase 3 mRNA were detected using RT-PCR. Results With the increase of age, the mean value for ABR thresholds in response to click, 4 kHz and 8 kHz sound stimulus of C57 mice gradually increased,the expression of PDCD5 and caspase 3 were increased also. At 3 months and 6 months of age in the cochlea of C57, all sorts of expression of PDCD5 and caspase 3 and the expression were enhanced with age. There was an evident expression at 9 months age, but the highest expression was detected at 12 months age. The PDCD5 and Caspase 3 expression were statistically different in each group ( P < 0. 05 ). The changes of PDCD5 and caspase 3 mRNA expression were in accordance with that of PDCD5 and Caspase 3 protein expression by the realtime PCR. Conclusions The expression levels of PDCD5 and caspase 3 in the cochlea of C57 mice increase with age, the results suggested that the expression of PDCD5 and caspase 3 might play an important role in the pathogenic mechanism of presbycusis.%目的 检测程序化细胞死亡分子5(programmed cell death 5,PDCD5)和半胱氨酸天冬氨酸蛋白酶3(caspase 3)在不同年龄段C57BL/6J小鼠耳蜗中的表达,初步探讨其在年龄相关性听力下降发生、发展中的作用。方法 选择3、6、9及12月龄段C57BL/6J小鼠各15只,即按月龄分为四组。

  6. Constitutive nitric oxide synthase-mediated caspase-3 S-nitrosylation in ghrelin protection against Porphyromonas gingivalis-induced salivary gland acinar cell apoptosis.

    Science.gov (United States)

    Slomiany, B L; Slomiany, A

    2010-06-01

    Recent advances in identifying the salivary constituents capable of influencing the oral mucosal inflammatory responses have brought to focus the importance of a peptide hormone, ghrelin. Here, we report on the involvement of ghrelin in controlling the apoptotic processes induced in sublingual salivary gland acinar cells by the lipopolysaccharide (LPS) of a periodontopathic bacterium, Porphyromonas gingivalis. We show that the countering effect of ghrelin on the LPS-induced acinar cell apoptosis was associated with the increase in constitutive nitric oxide synthase (cNOS) activity, and the reduction in caspase-3 and inducible nitric oxide synthase (iNOS). The loss in countering effect of ghrelin on the LPS-induced changes in apoptosis and caspase-3 activity was attained with Src kinase inhibitor, PP2, as well as Akt inhibitor, SH-5, and cNOS inhibitor, L-NAME, but not the iNOS inhibitor, 1400W. The effect of ghrelin on the LPS-induced changes in cNOS activity, moreover, was reflected in the increased cNOS phosphorylation that was sensitive to PP2 as well as SH-5. Furthermore, the ghrelin-induced up-regulation in cNOS activity was associated with the increase in caspase-3 S-nitrosylation that was susceptible to the blockage by SH-5 and L-NAME. The findings point to the involvement of ghrelin in Src/Akt kinase-mediated cNOS activation and the apoptogenic signal inhibition through the NO-induced caspase-3 S-nitrosylation.

  7. Glutamine supplementation prevents exercise-induced neutrophil apoptosis and reduces p38 MAPK and JNK phosphorylation and p53 and caspase 3 expression.

    Science.gov (United States)

    Lagranha, Claudia J; Hirabara, Sandro M; Curi, Rui; Pithon-Curi, Tania C

    2007-01-01

    We have previously shown that a single session of exercise induces DNA fragmentation, mitochondrial membrane depolarization, increases expression of pro-apoptotic genes (bax and bcl-xS) and decreases expression of anti-apoptotic genes (bcl-xL) in rat neutrophils. Glutamine supplementation had a protective effect in the apoptosis induced by a single session of exercise. The mechanism involved in the effect of single session of exercise to induce apoptosis was investigated by measuring expression of p53 and caspase 3 and phosphorylation of p38 mitogen-activated protein kinases (MAPK) and cJun NH(2)-terminal kinase (JNK) in neutrophils from rats supplemented or not with glutamine. Exercise was carried out on a treadmill for 1 h and the rats were killed by decapitation. Neutrophils were obtained by intraperitoneal (i.p.) lavage with PBS, 4 h after injection of oyster glycogen solution. Glutamine supplementation (1g per Kg b.w.) was given by gavage 1 h before the exercise session. Gene expression and protein phosphorylation were then analyzed by reverse transcriptase chain reaction (RT-PCR) and Western blotting, respectively. A single session of exercise increased p38 MAPK and JNK phosphorylation and p53 and caspase 3 expression. Glutamine supplementation partially prevented the increase in p38 MAPK and JNK phosphorylation and p53 expression, and fully abolished the increase in caspase 3 expression. Thus, neutrophil apoptosis induced by a single session of exercise is accompanied by increased p53 and caspase 3 expression and p38 MAPK and JNK phosphorylation. Glutamine supplementation prevents these effects of exercise and reduces apoptosis.

  8. The Effect of Silver Nanoparticles on the Biochemical Parameters of Liver Function in Serum, and the Expression of Caspase-3 in the Liver Tissues of Male Rats

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    Pourhamzeh

    2016-06-01

    Full Text Available Background Silver nanoparticles have antibacterial properties and their use is growing in different industries. Since the toxicity of nanosilver is not well known, it is essential to examine its safety. Objectives This experiment was undertaken to study the effects of nanosilver on rat liver function with an evaluation of blood biochemistry parameters and caspase-3 expression in the liver. Materials and Methods In this experimental study, 40 adult male Sprague-Dawley rats were divided into five groups. In the four experimental groups, nanosilver particles were given orally for 28 consecutive days at doses of 30, 125, 300, or 700 mg/kg. Rats in the control group received equal volumes of deionized water. To evaluate the expression of caspase-3 in liver tissue, the real-time PCR method was used. Albumin, total protein, total bilirubin, alanine aminotransferase, aspartate aminotransferase, and alkaline phosphatase were measured with an RA-1000 autoanalyzer. Results The results indicated that caspase-3 was upregulated in the treated groups compared to the control group (P 0.05. Conclusions Based on the present study, it can be concluded that oral administration of silver nanoparticles for 28 days had no effect on rat liver function, but probably led to early apoptotic stages.

  9. Aflatoxin B1 affects apoptosis and expression of Bax, Bcl-2, and Caspase-3 in thymus and bursa of fabricius in broiler chickens.

    Science.gov (United States)

    Peng, Xi; Chen, Kejie; Chen, Jin; Fang, Jing; Cui, Hengmin; Zuo, Zhicai; Deng, Junliang; Chen, Zhengli; Geng, Yi; Lai, Weimin

    2016-09-01

    Aflatoxin B1 is known as a mycotoxin that develops various health problems of animals, the effects of AFB1 on thymus and bursa of Fabricius in chickens are not clear. The objective of this study was to investigate the apoptosis of thymus and bursa of Fabricius in broilers fed with AFB1 . Two hundred Avian broilers were randomly divided into four groups of 50 each, namely control group and three AFB1 groups fed with 0.15 mg, 0.3 mg, and 0.6 mg AFB1 /kg diet, respectively. In this study, flow cytometer and immunohistochemical approaches were used to determine the percentage of apoptotic cells and the expression of Bax, Bcl-2, and Caspase-3. The results showed that consumption of AFB1 diets results in increased percentage of apoptotic cells and increased expression of Caspase-3 in both thymus and bursa of Fabricius. The expression of Bax was increased and the expression of Bcl-2 was decreased in the thymus, but no significant changes in Bax and Bcl-2 expression were observed in the bursa of Fabricius when broilers fed with AFB1 . These findings suggest that adverse effects of AFB1 on thymus and bursa of Fabricius in broilers were confirmed by increased apoptotic cells and abnormal expression of Caspase-3. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1113-1120, 2016.

  10. Caspase-3 dependent nitrergic neuronal apoptosis following cavernous nerve injury is mediated via RhoA and ROCK activation in major pelvic ganglion.

    Science.gov (United States)

    Hannan, Johanna L; Matsui, Hotaka; Sopko, Nikolai A; Liu, Xiaopu; Weyne, Emmanuel; Albersen, Maarten; Watson, Joseph W; Hoke, Ahmet; Burnett, Arthur L; Bivalacqua, Trinity J

    2016-07-08

    Axonal injury due to prostatectomy leads to Wallerian degeneration of the cavernous nerve (CN) and erectile dysfunction (ED). Return of potency is dependent on axonal regeneration and reinnervation of the penis. Following CN injury (CNI), RhoA and Rho-associated protein kinase (ROCK) increase in penile endothelial and smooth muscle cells. Previous studies indicate that nerve regeneration is hampered by activation of RhoA/ROCK pathway. We evaluated the role of RhoA/ROCK pathway in CN regulation following CNI using a validated rat model. CNI upregulated gene and protein expression of RhoA/ROCK and caspase-3 mediated apoptosis in the major pelvic ganglion (MPG). ROCK inhibitor (ROCK-I) prevented upregulation of RhoA/ROCK pathway as well as activation of caspase-3 in the MPG. Following CNI, there was decrease in the dimer to monomer ratio of neuronal nitric oxide synthase (nNOS) protein and lowered NOS activity in the MPG, which were prevented by ROCK-I. CNI lowered intracavernous pressure and impaired non-adrenergic non-cholinergic-mediated relaxation in the penis, consistent with ED. ROCK-I maintained the intracavernous pressure and non-adrenergic non-cholinergic-mediated relaxation in the penis following CNI. These results suggest that activation of RhoA/ROCK pathway mediates caspase-3 dependent apoptosis of nitrergic neurons in the MPG following CNI and that ROCK-I can prevent post-prostatectomy ED.

  11. Possible involvement of caspase-6 and -7 but not caspase-3 in the regulation of hypoxia-induced apoptosis in tube-forming endothelial cells.

    Science.gov (United States)

    Eguchi, Ryoji; Toné, Shigenobu; Suzuki, Akio; Fujimori, Yoshihiro; Nakano, Takashi; Kaji, Kazuhiko; Ohta, Toshiro

    2009-01-15

    We recently reported that a broad-spectrum caspase inhibitor zVAD-fmk failed, while p38 inhibitor SB203580 succeeded, to prevent chromatin condensation and nuclear fragmentation induced by hypoxia in tube-forming HUVECs. In this study, we investigated the reasons for zVAD-fmk's inability to inhibit these morphological changes at the molecular level. The inhibitor effectively inhibited DNA ladder formation and activation of caspase-3 and -6, but it surprisingly failed to inhibit caspase-7 activation. On the other hand, SB203580 successfully inhibited all of these molecular events. When zLEHD-fmk, which specifically inhibits initiator caspase-9 upstream of caspase-3, was used, it inhibited caspase-3 activation but failed to inhibit caspase-6 and -7 activation. It also failed to inhibit hypoxia-induced chromatin condensation, nuclear fragmentation and DNA ladder formation. Taken together, our results indicate that, during hypoxia, caspase-7 is responsible for chromatin condensation and nuclear fragmentation while caspase-6 is responsible for DNA ladder formation.

  12. 核桃青皮提取物对Hela细胞增殖及caspase-3蛋白表达的影响

    Institute of Scientific and Technical Information of China (English)

    陈默然; 赵行宇; 沈楠; 张巍

    2011-01-01

    目的观察不同浓度核桃青皮提取物对Hela细胞增殖的抑制作用,探讨提取物对Hela细胞生长的影响。方法光镜下观察Hela细胞形态的改变,MTT法测定细胞生长抑制率,免疫组化观察caspase-3蛋白的表达。结果随提取物浓度的增加,细胞形态的改变明显,细胞变圆,折光率增加,细胞逐渐减少,对细胞生长的抑制作用增强,caspase-3蛋白的表达增加。结论提取物可抑制Hela细胞的生长,上调Hela细胞caspase-3蛋白的表达。

  13. [Salidroside protects cultured rat subventricular zone neural stem cells against hypoxia injury by inhibiting Bax, Bcl-2 and caspase-3 expressions].

    Science.gov (United States)

    Qi, Cunfang; Zhang, Junfeng; Chen, Xinlin; Zhang, JiansHui; Yang, Pengbo; Jiao, Qian; Zhang, Pengbo; Lu, Hai-Xia; Liu, Yong

    2013-07-01

    To explore the effects of salidroside (sal) on the expressions of Bcl-2, Bax and caspases-3 proteins in cultured rat subventricular zone (SVZ) neural stem cells (NSCs) exposed to hypoxia injury. Primarily cultured SVZ NSCs from adult SD rats were incubated with salidroside (120 and 240 µmol/L) for 24 h prior to exposure to hypoxia. The cell viability was assessed with MTT assay, and the cell apoptosis was analyzed using TUNEL staining and flow cytometry. Western blotting was performed to detect the expressions of Bcl-2, Bax and caspase-3 in the cells. Salidroside pretreatment of the cells for 24 h resulted in an obvious resistance to hypoxia-induced cell apoptosis and decrement of cell viability (PSalidroside also antagonized the effect of hypoxia exposure in lowering Bcl-2/Bax ratio apoptosis of rat neural stem cells and decreased the expression of caspases-3 protein (PSalidroside can significantly resist hypoxia-induced. The neuroprotective effect of salidroside may be related to the modulation of expressions of apoptosis-related proteins.

  14. Hepatitis B Virus X Protein Sensitizes Primary Mouse Hepatocytes to Ethanol-and TNF-α-Induced Apoptosis by a Caspase-3-Dependent Mechanism

    Institute of Scientific and Technical Information of China (English)

    Won-HoKim; FengHong; BarbaraJaruga; ZhengshengZhang; SaijunFan; T.JakeLiang; BinGao

    2005-01-01

    It is well-documented that alcohol drinking together with hepatitis viral infection accelerates liver injury; however the underlying mechanisms remain unknown. In this paper, we demonstrated that primary hepatocytes from transgenic mice overexpressing hepatitis B virus X protein (HBX) were more susceptible to ethanol- and TNF-α-induced apoptotic killing. Compared to normal control mouse hepatocytes, ethanol and/or TNF-α treatment led to a significant increase in reactive oxygen species, mitochondrial permeability transition, cytochrome C release, caspase-3 activity, and poly (ADP-ribose) polymerase degradation in hepatocytes from HBX transgenic mice. Blocking caspase-3 activity antagonized ethanol- and TNF-α-induced apoptosis in primary hepatocytes from HBX transgenic mice. Taken together, our findings suggest that HBX sensitizes primary mouse hepatocytes to ethanoland TNF-α-induced apoptosis by a caspase-3-dependent mechanism, which may partly explain the synergistic effects of alcohol consumption and hepatitis B virus infection on liver injury. Cellular & Molecular Immunology. 2005;2(1):40-48.

  15. Changes in Ca2+ concentration and caspase-3 expression and their relationship in Raji cells exposed to electromagnetic radiation%电磁辐射致Raji细胞内Ca2+浓度和Caspase-3蛋白表达的改变及其相互关系

    Institute of Scientific and Technical Information of China (English)

    王炜; 刘换新; 王德文; 左红艳; 彭瑞云

    2013-01-01

    目的 研究电磁脉冲(electromagnetic pulse,EMP)、S波段高功率微波(S-band high power microwave,S-HPM)、X波段高功率微波(X-band high power microwave,X-HPM)3种不同波段电磁辐射对Raji细胞内Ca2+浓度、Caspase-3蛋白表达的影响及两者之间的相互关系,探讨电磁辐射损伤的相关调控机制.方法 常规培养Raji细胞,用S-HPM、X-HPM和EMP 3种不同波段的电磁辐射照射Raji细胞,设假照射为对照组.于对数生长期照射,照射后6h收集细胞,采用激光扫描共聚焦显微镜检测细胞内Ca2+浓度的改变,采用免疫印迹法(Western blot)检测细胞内Caspase-3蛋白的表达.结果 3种不同波段的电磁辐射照射后6h,Raji细胞内Ca2+浓度增加(EMP组、S-HPM组、X-HPM组Ca2+荧光强度分别为69.56±1.71、50.06±1.89、70.68±1.59),Caspase-3蛋白表达水平上调(EMP组、S-HPM组、X-HPM组Caspase-3蛋白表达水平分别为0.964±0.12、0.586±0.16、0.970±0.07),各辐射组Raji细胞Ca2+荧光强度、Caspase-3蛋白表达水平均明显高于对照组(分别为43.08±2.08、0.444 ±0.13),差异均有统计学意义(P<0.01);EMP组和X-HPM组Ca2+荧光强度、Caspase-3蛋白表达水平均明显高于S-HPM组,差异亦有统计学意义(P<0.01):EMP与X-HPM组间的差异无统计学意义(P>0.05),但均较S-HPM组上调明显,差异均有统计学意义(P<0.01).辐射组Ca2+浓度增加的规律和Caspase-3蛋白表达上调的规律具有一致性,线性回归分析结果显示,Caspase-3蛋白的表达随着Ca2+浓度的增高而上调,二者具有一定的相关性(P<0.01).结论 3种辐射损伤效应可能是通过增加细胞内Ca2+浓度进一步诱导Caspase-3过表达所致.%Objective To study the effects of electromagnetic pulse (EMP),S-band high power microwave (S-HPM),and X-band high power microwave (X-HPM) on the Ca2+ concentration and caspase-3 expression in Raji cells and the rclationship between Ca2+ concentration and caspase-3 expression

  16. Estudo Transcultural sobre Liderança em Relações Públicas e Gestão da Comunicação: análise quantitativa dos temas de maior importância para os brasileiros / Transcultural Study on Leadership in Public Relations and Communication Management: quantitative analysis of the issues of greatest importance to the Brazilians

    Directory of Open Access Journals (Sweden)

    Andréia Athaydes

    2013-12-01

    Full Text Available Este artigo apresenta o relato parcial de uma investigação realizada em âmbito internacional, sob coordenação da Universidade do Alabama e do The Plank Center Institute (EUA, a fim de compreender como as constantes e rápidas mudanças políticas, econômicas e sociais impactam nas rotinas e no desempenho dos gestores em RP e Comunicação. As impressões foram obtidas através de questionário online e entrevistas em profundidade com gestores de RP e Comunicação de diferentes organizações. Aqui, são detalhados os dados quantitativos referentes à primeira seção do questionário online, que buscou conhecer, entre 10 temas pré-definidos, quais os de maior importância para os participantes. Estar preparado para lidar eficazmente com as crises, lidar com a velocidade e volume do fluxo de informação e melhorar os indicadores de comunicação para demonstrar o valor do trabalho realizado são as principais preocupações dos gestores. / This article presents part of an investigation into an international, coordinated by the University of Alabama and The Plank Center Institute (USA in order to understand how the constant and rapidly changing political, economic and social impact in the routines and performance of managers in PR and Communications. The prints were obtained through online questionnaire and in-depth interviews with managers of PR and Communication of different organizations. Here are detailed quantitative data relating to the first section of the online questionnaire, which sought to know from 10 predefined themes, which are the most important to the participants. Be prepared to deal effectively with crises, handle the speed and volume of information flow and improve communication indicators to demonstrate the value of the work performed are the main concerns of managers.

  17. Análise anatômica qualitativa e quantitativa e densidade básica da madeira de Alchornea sidifolia Müll. Arg. (Euphorbiaceae. Qualitative and quantitative anatomical analysis of the wood of Alchornea sidifolia Müll. Arg. (Euphorbiaceae

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    Luiz SANTINI JUNIOR

    2010-12-01

    Full Text Available Objetivou-se caracterizar a anatomia e densidade básica da madeira de Alchornea sidifolia (tapiá-guaçu, e investigar a variação radial destas características. O material de estudo procede do lote cinco do Rodoanel trecho sul no Estado de São Paulo, onde cinco árvores foram identificadas no campo, retirando-se discos do DAP com média de 30 cm de diâmetro. As análises das amostras seguiram os métodos usualmente empregados em estudo de anatomia de madeiras. A espécie apresentou camadas de crescimento distintas; vasos solitários e múltiplos; placas de perfuração simples; parênquima axial apotraqueal em linhas; raios unisseriados, alternando células quadradas e eretas com inclusões cristalíferas e amido; fibras libriformes, não septadas e gelatinosas; canais radiais; máculas medulares; densidade básica baixa (0,38 g cm-3. Ocorreu aumento no sentido medula-casca apenas para o comprimento das fibras, diâmetro dos vasos e diâmetro das pontoações intervasculares, características que denotam madeira homogênea e de baixa resistência. Fibras com maiores lume estão associadas a menores valores de densidade básica.In this paper, we characterize the anatomy and basic density of wood in Alchornea sidifolia (tapiá-guaçu, and investigate the radial variation of these characteristics. Five trees were cut in the “Rodoanel” section southern in São Paulo state, were removed disks with 30 cm at breast height. Six samples were taken from the pith to bark, which were analyzed according to the usual methods in anatomy and properties of wood. The results revealed the presence of layer growth, solitary and multiples vessels; simple perforation plate; axial parenchyma in lines up three cells wide; rays exclusively uniseriate, all ray cells upright and/or square with inclusions crystals and starch; ground tissue fibers, non-septate and gelatinous; radial canals; pith fleck present; low basic density (0.38 g cm-3. There was an increase

  18. OBSTRUCTIVE NEPHROPATHY OF KIDNEY STONE: The Role of Caspase-3, Transforming Growth Factor-β and Tumor Necrosis Factor-α in Kidney Fibrosis

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    A. A. G. Oka

    2014-08-01

    Full Text Available Objective: Kidney stones are one of the causes of obstructive nephropathy and chronic kidney disease (CKD, characterized by the presence of hydronephrosis and kidney dysfunction. The histhopathologycharacteristics of CKD is kidney fibrosis, which consists of glomerulosclerosis (GS, tubulointerstitial fibrosis (TIF and tubular atrophia (TA.This research aims to find out that the role of caspase-3, TGF-β and TNF-α as risk factors for kidney fibrosis in obstructive nephropathy of kidney stone. Methode: A total of 82 samples of kidney biopsy patientsduring kidney stone surgery in Sanglah General Hospital Denpasar-Bali from February antil December 2013 wereapplied match paired based on age and sex divided in to 41 samples as case and 41 samples as control. Determination of caspase-3, TNF-α and TGF-β1 were carried out using Kit Methode and kidney fibrosis stained with Masson's Trichrome. Results: Bivariate analysis test indicates that OR caspase-3 activity and kidney fibrosis were 3.50; 95%CI (2.95-4.19; p=0.001, OR levels of TGF-β and kidney fibrosis were 2.00; 95% CI (1.72-2.34; p=0.04 andOR levels of TNF-α and kidney fibrosis were 0.60; 95% CI (0.53-0.68; p=0.14. On the other hand, multivariate analysis test OR for caspase-3 activity , levels of TGF-β, levels of TNF-α and kidney fibrosis after adjustment of hydronephrosis, eLFG, and Hb were 2.43; 95% CI (0.86 – 6.90; p=0.09,  2.14; 95% CI (0.74-6.18; p=0.16 and 1.12; 95% CI (0.40-3.16; p=0.82. Conclusion: In obstructive nephropathy of kidney stone patients with high activity of caspase-3 have 3.5 fold risk to gain kidney fibrosis and with high levels of TGF-β have a 2 fold risk to gain kidney fibrosis. However, high levels of TNF-α was not as a risk factor to gain kidney fibrosis. These results can be used as a based data to run further research for obtaining new strategy for managing kidney fibrosis.  

  19. Precursor of advanced glycation end products mediates ER-stress-induced caspase-3 activation of human dermal fibroblasts through NAD(PH oxidase 4.

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    Danielle T Loughlin

    Full Text Available BACKGROUND: The precursor for advanced glycation end products, 3-deoxyglucosone (3DG is highly upregulated in skin explants of diabetic cutaneous wounds, and has been shown to negatively impact dermal fibroblasts, which are crucial in wound remodeling. 3DG induces apoptosis however; the mechanisms involved in the apoptotic action of 3DG in the pathogenesis of diabetic chronic wounds are poorly understood. Therefore, we sought to delineate novel mechanisms involved with the 3DG-collagen induced apoptosis. METHODOLOGY/PRINCIPAL FINDINGS: Using human dermal fibroblasts, we demonstrated that 3DG-modified collagen induces oxidative stress and caspase-3 activation. Oxidative stress was found to be dependent on the upregulation of NAD(PH oxidase 4 (Nox4, a reactive oxygen species (ROS Nox homologue, triggering endoplasmic reticulum (ER stress, as assessed by the ER stress-induced apoptosis marker Growth Arrest and DNA Damage-inducible gene 153 (GADD153. We demonstrated that 3DG-collagen activated GADD153 via phosphorylation of p38 mitogen activated protein kinase (MAPK, and this was dependent on upstream ROS. Inhibition of ROS and/or p38 MAPK abrogated 3DG-collagen induced caspase-3 activation. Our investigations also demonstrated that 3DG-collagen-induced caspase-3 activation did not signal through the canonical receptor for advanced glycation end products (RAGE but through integrin alpha1beta1. To further verify the role of integrins, neutralization of integrins alpha1beta1 prevented 3DG-collagen-induced upregulation of ROS, GADD153, and caspase-3 activation; suggesting that 3DG-collagen signaling to the fibroblast is dependent on integrins alpha1beta1. CONCLUSIONS/SIGNIFICANCE: Taken together, these findings demonstrate for the first time that a RAGE independent mechanism is involved in 3DG-collagen-induced apoptosis. Moreover, the ER stress pathway through activation of Nox4 by integrins alpha1beta1 plays a key role in 3DG-collagen-induced caspase

  20. THE ROLE OF PROTEIN OXIDATIVE MODIFICATION IN REDOX-REGULATION OF CASPASE-3 ACTIVITY IN BLOOD LYMPHOCYTES DURING OXIDATIVE STRESS IN VITRO

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    O. L. Nosareva

    2015-01-01

    Full Text Available The formation of oxidative stress lies at the heart of many frequent and socially-important diseases. Blood lymphocytes are the cells which provide immunological control of our organism. As a result of their function implementation blood lymphocytes contact with different endogenic and exogenic factors, which can lead to active oxygen species production activation, macromolecules oxidative modification and to cell survival alteration. At the present time it is essential to expand and deepen the fundamental knowledge of blood lymphocytes apoptosis regulation peculiarities. The research objective was to establish the interaction among alterations of glutathione system condition, carbonylation level, protein glutathionylation and caspase-3 activity in blood lymphocytes during oxidative stress in vitro.Material and Methods. The material for research was blood lymphocytes cultivated with addition of hydrogen peroxide in final concentration of 0,5 mmol and/or protein SH-group inhibitor N-ethylmaleimide – 5 mmol, protector – 5 mmol – 1,4-dithioerythritol. Reduced, oxidized and protein-bound glutathione concentration was measured by method of spectropho-tometry, additionally, the ratio size of reduced to oxidized thiol fraction was estimated. With help of enzymoimmunoassay the level of protein carbonyl derivatives was evaluated; caspase-3 activity was registered by spectrofluorometric method.Results. Protein SH-group blocking in blood lymphocytes during oxidative stress in vitro was accompanied by protein-bound glutathione concentration rapid decrease in connection with increase of protein carbonyl derivatives content and caspase-3 activity. Protein SH-group protection in blood lymphocytes during oxidative stress in vitro was accompanied by concentration increase of protein-bound glutathione and protein carbonyl derivatives under comparable values of enzyme activity under study.Conclusion. The carried out research shows that caspase-3 and protein

  1. Nerve growth factor downregulates c-jun mRNA and Caspase-3 in striate cortex of rats after transient global cerebral ischemia/reperfusion

    Institute of Scientific and Technical Information of China (English)

    Dacheng Jin; Tiemin Wang; Xiubin Fang

    2006-01-01

    BACKGROUND: Immediate early gene (LEG) c-jun is a sensitive marker for functional status of nerve cells.Caspase-3 is a cysteine protease,which is a critical regulator of apoptosis. The effect of exogenous nerve growth factor (NGF) on the expression of c-jun Mrna and Caspase-3 protein in striate cortex of rats with transient global cerebral ischemia/reperfusion (IR) is unclear.OBJECTIVE: To study the protective effect of exogenous NGF on the brain of rats with transient global cerebral IR and its effecting pathway by observing the expression of c-jun Mrna and Caspase-3 protein.DESIGN: Randomized controlled animal trial.SETTING: Department of Neural Anatomy, Institute of Brain,China Medical University.MATERTALS:Eighteen healthy male SD rats of clean grade, aged 1 to 3 months, with body mass of 250 to 300 g, were involved in this study. NGF was provided by Dalian Svate Pharmaceutical Co.,Ltd, c-jun in situ hybridization detection kit, Caspase-3 antibody and SABC kit were purchased from Boster Biotechnology Co. ,Ltd.METHODS: This trial was carried out in the Department of Neural Anatomy, Institute of Brain, China Medical University during September 2003 to April 2005. ①Experimental animals were randomized into three groups with 6 in each: sham-operation group,IR group and NGF group. ②After the rats were anesthetized,the bilateral common carotid arteries and right external carotid arteries of rats were bluntly dissected and bilateral common carotid arteries were clamped for 30 minutes with bulldog clamps. Reperfusion began after buldog clamps were removed. Normal saline of 1mL and NGF (1×106 U/L) of 1 Ml was injected into the common carotid artery of rats via right external carotid arteries in the IR group and NGF group respectively.The injection was conducted within 30 minutes, and then the right external carotid arteries were ligated. In the sham-operation group, occlusion of bilateral common carotid arteries and administration of drugs were phosphate buffer

  2. Mapeamento de locos de características quantitativas associados à composição de carcaça, no cromossomo seis de suíno

    OpenAIRE

    A.V. Pires; P.S. Lopes; GUIMARÃES, S. E. F.; C.T. Guimarães; J.O. Peixoto

    2008-01-01

    Uma população de suínos, composta de 550 animais F2, foi produzida a partir do intercruzamento da geração F1, obtida pelo cruzamento divergente de dois machos da raça nativa brasileira Piau com 18 fêmeas comerciais. O objetivo do trabalho foi mapear locos de características quantitativas (QTL) associados a cortes de carcaça. Os animais foram genotipados para 13 marcadores microssatélites, distribuídos no cromossomo 6 de suínos. As características avaliadas foram: peso total do pernil, peso do...

  3. Ezh2、Runx3和caspase-3在子宫内膜样腺癌中的表达及相关性%Expression and relationship of Ezh2, Runx3 and caspase-3 in endometrial adenocarcinoma

    Institute of Scientific and Technical Information of China (English)

    郭真理; 陈柯; 王晓秋; 胡闻

    2011-01-01

    目的 探讨Ezh2、Runx3和caspase-3在正常子宫内膜、子宫内膜增殖症和子宫内膜样腺癌组织中的表达及相关性.方法 采用组织芯片技术和免疫组织化学SP法检测30例正常子宫内膜组织、30例单纯性增生内膜、30例复杂伴不典型增生内膜、72例子宫内膜样腺癌组织中Ezh2、Runx3和caspase-3蛋白的表达.结果 Ezh2、Runx3和caspase-3蛋白在子宫内膜样腺癌组织中的阳性表达率分别为83.3%(60/72)、26.4%(19/72)、33.3%(24/72);Ezh2在腺癌组织中的阳性率明显高于正常和各型增生内膜(16.7%、33.3%、63.3%;P0.05);caspase-3的阳性表达与组织分级有关(P0.05).Ezh2与Runx3的蛋白表达呈负相关(rs=-0.262,P0.05). Positive expression of caspase-3 protein was related to the histological grade(P0.05). The expression of Ezh2 protein was negatively correlated to that of Runx3 (rs=-0.262, P<0.05). Conclusions Abnormal expression of Ezh2, Runx3 and caspase-3 proteins is associated with the development and progression of endometrioid adenocarcinoma. Combined analysis of Ezh2, Runx3 and caspase-3 may offer prognostic information for patients with endometrial cancer.

  4. 活血益智片对血管性痴呆小鼠凋亡蛋白p53及caspase-3表达的影响%Effect of Huoxueyizhi slice on p53 and caspase-3 protein expression in hippocampus of vascular dementia mice

    Institute of Scientific and Technical Information of China (English)

    王丽红; 文慧; 齐越; 贾冬; 周绮; 鄂蕴娟

    2011-01-01

    Objective To observe the effect of Huoxueyizhi slice on memory impairment and expression of p53 and caspase -3 in hippocampus of vascular dementia ( VaD ) mice. Methods Vascular dementia (VaD) mice model was made by two times of transient ischemia and reperfusion of bilateral common carotid arteries plus tail bleeding. The memory test, p53 and caspase -3 protein expression in hippocampus of VaD mice treated with and without Huoxueyizhi slice(2. 24,4. 48,9. 96 g · kg-1) and hydergine (7. 8 mg· kg-1), continuous administration of 28 days, were measured. Results Compared with model group, Huoxueyizhi slice significantly improved memory impairment. Meanwhile, based on the determination in hippocampus through method of the western blot, Huoxueyizhi slice decreased the expression of the proapoptotic protein p53 and caspase -3. Conclusion Huoxueyizhi slice can exert antiapoptotic effect through regulating p53 and caspase -3 protein expression.%目的 研究活血益智片(抗血管性痴呆中成药)对血管性痴呆小鼠(Van)认知功能及海马神经元凋亡蛋白的影响.方法 用反复缺血再灌注小鼠模型,模拟VaD行为及病理过程,观察3个剂量(2.24,4.48,9.96 g·kg-1)活血益智片组及氢麦角碱组7.8mg·kg-1连续给药28 d后,对海马区学习记忆成绩、p53及caspase - 3蛋白表达的影响.结果 活血益智片组与模型组比较,可提高小鼠的学习记忆能力;降低p53及caspase - 3蛋白表达.结论 活血益智片可通过调节p53及caspase-3蛋白表达而发挥抗凋亡作用.

  5. Características quantitativas e qualitativas da carcaça de novilhas alimentadas com diferentes fontes energéticas em dietas à base de cascas de algodão e de soja Quantitative and qualitative carcass characteristics of heifers fed different energy sources on a cottonseed hulls and soybean hulls based diet

    Directory of Open Access Journals (Sweden)

    icardo Kazama

    2008-02-01

    Full Text Available Objetivou-se avaliar as características de carcaça e a composição físico-química e de ácidos graxos do músculo Longissimus de novilhas em confinamento alimentadas com cascas de algodão (CAL e de soja (CSO como volumosos e com diferentes fontes energéticas, como milho moído, farelo de gérmen de milho (FGM ou farelo de arroz integral (FAR. Foram utilizadas 24 novilhas mestiças com peso corporal de abate de 350 ± 35 kg, distribuídas em delineamento inteiramente casualizado, com três tratamentos (dietas e oito repetições (animais. As novilhas foram abatidas após jejum de sólidos de 24 horas. As carcaças foram identificadas, pesadas e armazenadas em câmara fria a 2ºC até a manhã seguinte para as avaliações de carcaça. Os animais alimentados com as dietas experimentais não diferiram quanto às características de carcaça; os valores médios de rendimento de carcaça quente foram de 51,4%, os de área de olho-de-lombo, 50,8 cm², e os de espessura de gordura de cobertura, 3,3 mm. As dietas também não influenciaram a composição físico-química do músculo Longissimus, que apresentou em média 73,4% de umidade, 23,0% de PB, 48,2 mg/100g de colesterol e 4,84 kgf/cm³ de força de cisalhamento. Animais alimentados com a dieta FGM apresentaram maiores concentrações de CLA (0,48%, trans-vacênico (7,69% e pior razão ômega 6/ômega 3 no músculo Longissimus, em virtude da maior quantidade de ácido linoléico desta dieta. Animais alimentados com a dieta FAR apresentaram maior concentração de ácido esteárico no músculo Longissimus, provavelmente em razão do maior conteúdo de ácido oléico desta dieta em relação às demais.This work was carried out to evaluate carcass characteristics, physicochemical and fatty acids compositions of Longissimus muscle from feedlot heifers fed cottonseed hulls (CSH and soybean hulls (SBH as forage, and different energy sources: ground corn (COR, corn germ meal (CGM or whole rice

  6. Effect of Quercetin on Caspase-3 of A549 Cell Induced by Influenza Virus H1N1%槲皮素对甲型H1N1流感病毒诱导的A549细胞凋亡效应酶Caspase-3的影响

    Institute of Scientific and Technical Information of China (English)

    万巧凤; 吴莉; 杨美玲; 马锐; 梁军; 顾立刚

    2011-01-01

    Objective To investigate the influence of quercetin on Caspase-3 of lung epithelial tumour A549 infected with H1N1. Methods MTT method was adopted to determine H1N1 virulence, quercetin cytotoxicity, inhibitory effect of quercetin on A549 cytopathic with H1N1 cause. H1N1 of 100 TCID50 was used to infect A549 for 2 h and then change quercetin fluid containing 10 mg/L to cultivate. After 4, 12, 24, 48 h, cells were collected to extract total protein. By adopting the Western-blot method and applying Image-Pro Plus, the grayscale value of Caspase-3 and fl-actin were determined. The gray ratio was relative expression quantity of Caspase-3. Results The TCIDso of H1N1 to A549 was IO-4175, the highest non-toxic concentration of quercetin to A549 (Tco) was 40 mg/L, the least effective concentration (MEC) of quercetin inhibition to A549 cytopathic with H1N1 cause was 10 mg/L. After the infection of H1N1, quercetin significantly inhibited Caspase-3 expression within 4-48 h, showing that quercetin plays an obvious antiapoptotic role. Conclusion Quercetin can play antiviral infectious functions by inhibiting the content or activity of Caspase-3.%目的 观察银杏叶主要活性成分槲皮素对甲型H1N1流感病毒感染的人肺上皮瘤细胞A549凋亡效应酶Caspase-3的影响.方法 采用MTT法测定H1N1毒力、槲皮素细胞毒性作用、槲皮素对H1N1致A549细胞病变的抑制作用.然后用100 TCID50甲型H1N1感染A549 2 h后换用含10 mg/L槲皮素维持液继续培养,于感染后4、12、24、48 h收集细胞,提取细胞总蛋白,采用Western blot方法,应用Image-Pro Plus测定Caspase-3与β-actin灰度值,两者灰度比值为Caspase-3的相对表达量.结果 甲型H1N1 A549的TCID50为10-4.75,槲皮素A549的最大无毒浓度为40 mg/L,槲皮素抑制甲型H1N1致A549细胞病变的最小有效浓度为10 mg/L.在甲型H1N1感染A549细胞后4~ 48 h内,槲皮素可显著抑制Caspase-3蛋白表达,具有明显的抗凋亡作用.结论 槲皮素可通过抑制Caspase

  7. IL-6 Induces Pancreatic β Cell Apoptosis via Down-regulation of Sirt1 and Activation of p53/caspase-3 Pathway%IL-6通过Sirt1/p53/caspase-3通路介导胰岛β细胞凋亡

    Institute of Scientific and Technical Information of China (English)

    刘波; 林雅军; 黎健

    2011-01-01

    目的 探讨炎性因子IL-6是否通过Sirt1/p53/caspase-3通路介导胰岛β细胞凋亡.方法 Western 印迹检测Sirt1在小鼠各组织器官和胰岛β细胞系NIT-1细胞中的表达,免疫荧光法检测Sirt1在细胞中的定位.IL-6(10 ng/ml)处理NIT-1细胞48 h,Hoechst3334染色及流式细胞仪检测细胞凋亡,Western印迹检测细胞内Sirt1、P53、乙酰化P53(acety-P53)、caspase-3和cleaved caspase-3的水平变化.结果 Sirt1在小鼠各组织器官和胰岛β细胞中均有表达,主要定位于细胞核.IL-6处理NIT-1细胞后,伴随Sirt1表达的显著减少,acety-P53明显上调,p53/caspase-3通路活化,NIT-1细胞凋亡增加.结论 IL-6通过下调Sirt1进而激活p53/caspase-3信号通路引起胰岛β细胞凋亡.%Objective To investigate whether IL-6 induces pancreatic β cell apoptosis through down-regulation of Sirt1 and activation of p53/caspase-3 pathway. Methods Sirt1 expression in diverse mouse organs and pancreatic β cell line NIT-1 was detected by Western blot. The location of Sirtl in NIT-1 cells was observed by immunofluorescence. After treated with 1Ong/ml IL-6 for 48 h,apoptosis of NIT-1 cells was detected by Hoechst3334 staining and flow cytometry. Western blot was used to analyze the levels of Sirt1, p53, acety-p53, caspase-3 and cleaved caspase-3, respectively. Results Sirt1 was expressed in diverse mouse organs and pancreatic β cell line NIT-1. and mainly located in the nucleus. Treatment of NIT-1 cells with 1Ong/ml IL-6 for 48 h induced apoptosis, accompanied with decreased Sirt1 level. enhanced acety-p53 and activation of p53/caspase3. Conclusion IL-6 induces NIT-1 cell apoptosis via down-regulation of Sirt1 and activation of p53/caspase -3 pathway.

  8. Expressions of X-linked inhibitor of apoptosis protein, caspase-3 and Smac/DIABLO in nasopharyngeal carcinoma tissues%鼻咽癌组织中X连锁凋亡抑制蛋白及caspase-3和Smac/DIABLO的表达及意义

    Institute of Scientific and Technical Information of China (English)

    李栋才; 杨贵; 姚春苑

    2010-01-01

    目的 采用组织芯片技术研究鼻咽癌组织中X连锁凋亡抑制蛋白(XIAP)、caspase-3与其拮抗蛋白Smae/DIABLO的表达及其意义.方法 以50例鼻咽癌患者鼻咽部活体组织石蜡标本和20例鼻咽部慢性炎症组织石蜡标本为研究对象,利用组织芯片技术同时采用SP免疫组织化学方法检测鼻咽癌组织芯片中XIAP、caspase-3和Smac/DIAB-LO的表达.结果 鼻咽癌组织中XIAP的阳性表达高于鼻咽部慢性炎症组织,caspase-3和Smac/DIABLO的阳性表达明显低于鼻咽部慢性炎症组织,差别均有统计学意义(P<0.05).XIAP阳性的鼻咽癌石蜡标本Smac/DIABLO阳性表达率低于XIAP阴性的鼻咽癌石蜡标本Smac/DIABLO阳性表达率,鼻咽癌组织中XIAP与Smac/DIABLO表达呈负相关(P<0.05).结论 XIAP在鼻咽癌中高表达,caspase-3、Smac/DIABLO在鼻咽癌中低表达,鼻咽癌中XIAP和Smac/DI-ABLO表达呈负相关.XIAP、caspase-3和Smac/DIABLO可能是鼻咽癌细胞凋亡信号传导网络中的重要一环,与鼻咽癌的发生、发展密切相关.

  9. 右美沙芬对糖尿病视网膜神经病变中Caspase-3表达的影响%Effects of dextromethorphan on expression of Caspase-3 in retinal nerve cell of rats with diabetic retinopathy

    Institute of Scientific and Technical Information of China (English)

    马岚; 张奕霞; 练海东; 郝小英

    2015-01-01

    目的 观察右美沙芬对糖尿病大鼠视网膜神经病变中Caspase-3表达的影响.方法 健康成年Sprague Dawley大鼠48只,12只作为正常对照组(D组),36只大鼠链脲佐菌素腹腔注射建立糖尿病大鼠模型,并随机分为糖尿病模型组(A组),10 mg·kg-1右美沙芬治疗组(B组;造模成功后一次性腹腔注射10 mg·kg-1右美沙芬),30 mg·kg-1右美沙芬治疗组(C组;造模成功后一次性腹腔注射30 mg·kg-1右美沙芬),每组各12只.各组分别于4周末、8周末各处死大鼠6只,行免疫组织化学法检测视网膜组织Caspase-3蛋白的表达.结果 视网膜Caspase-3光密度值:A组4周、8周时分别为0.256±0.031和0.374±0.234,B组分别为0.244±0.122和0.352±0.152,C组分别为0.174±0.021和0.256±0.138,D组分别为0.074±0.018和0.072±0.233.A组4周Caspase-3蛋白表达主要见于神经节细胞层,8周时阳性表达进一步增加,扩展到内核层.C组Caspase-3表达变化规律与A组基本一致,但4周、8周时阳性表达量均低于A组(均为P<0.05),A组与B组间4周和8周时Caspase-3阳性表达量差异均无统计学意义(均为P>0.05).D组4周、8周时大鼠视网膜上几乎无Caspase-3阳性表达.结论 早期糖尿病大鼠视网膜Caspase-3参与了糖尿病视网膜病变的发病机制,右美沙芬可以抑制Caspase-3蛋白的表达,对糖尿病视网膜病变有一定的治疗作用.

  10. GDNF基因修饰的神经干细胞抑制脑卒中后大鼠的Caspase-3表达%The grafting neural stem cells modified by GDNF gene inhibits the expression of Caspase-3 in rats subjected to cerebral ischemia reperfusion

    Institute of Scientific and Technical Information of China (English)

    陈贵军; 高小青; 杨朝鲜; 谭树凯; 袁琼兰

    2012-01-01

    目的 研究胶质源性神经营养因子(glial cell line-derived neural factor,GDNF)基因修饰的神经干细胞(neural stem cells,NSCs)移植对脑卒中后大鼠缺血侧脑组织内半胱氨酰天冬氨酸特异性蛋白酶-3(cysteinyl aspartate specific proteinase-3,caspase-3)表达的影响,探讨GDNF基因修饰的神经干细胞(GDNF/NSCs)移植对大鼠脑卒中的神经保护作用机制.方法 取新生大鼠脑组织分离培养NSCs,收集第6代前的NSCs备用.用重组腺病毒GDNF转染神经干细胞,制备GDNF/NSCs.暂时性阻塞大鼠大脑中动脉制备脑卒中模型,3d后,用脑立体定位仪向卒中侧侧脑室分别给予NSCs、GDNF/NSCs和生理盐水.再灌注时间1周、2周、3周、5周、7周后处死大鼠(n=3).裂解卒中侧脑组织,离心后得到脑组织蛋白样品,通过蛋白免疫印迹( Western Blotting)检测Caspase-3表达.结果 GDNF/NSCs、NSCs、NS各组caspase-3的表达在1周、2周、3周、5周、7周各时间点均逐渐降低.NSCs组、GDNF/NSCs组显著低于NS组(P<0.01;P<0.001);GDNF/NSCs组明显低于NSCs组(P<0.01).结论 GDNF/NSCs移植治疗脑卒中的机制可能与抑制caspase-3表达有关.%Objective To study the effects of grafting neural stem cells( NSCs) modified by glial cell line-derived neural factor(GDNF) gene (GDNF/NSCs) on the expression of Caspase-3 in rats subjected to cerebral ischemia reperfusion. Methods NSCs were cultured from newborn rats and NSCs before 6 generation were used for grafting use. NSCs were infected by recombinant GDNF adenovirus to prepare NSCs - overexpressing GDNF( GDNF/NSCs). Rat stroke was performed by occluding the middle cerebral artery occlusion for 2 h and reperfusion. At 3 days after reperfusion, NSCs, GDNF/NSCs and saline was infused into ipsilateral ventricle respectively. According to different reperfusion time, each group was subdivide into 5 groups: 1,2,3,5,7 weeks ( n = 3 ). At each time point, rats were sacrificed and brains were

  11. 海藻糖对液氮深低温保存的同种瓣caspase-3表达的影响%Study the expression of caspase-3 on trehalose as cryoprotectant for preserving aortic valve homograft in liquid nitrogen

    Institute of Scientific and Technical Information of China (English)

    程晨晨; 常青; 徐平; 高洪波

    2011-01-01

    目的 探讨海藻糖作为深低温冷冻保护剂对液氮保存的同种带瓣大动脉组织细胞caspase-3表达的影响.方法 在液氮中冻存时间点分为12、15、18个月,依据冻存液不同分为5组,分别是:0.1 mol/L DMSO(对照组),0.1 mol/L海藻糖(实验组1),0.1 mol/L海藻糖+0.1 mol/L DMSO (实验组2),0.2mol/L海藻糖+0.1 mol/LDMSO(实验组3),0.3 moL/L海藻糖+0.1 mol/L DMSO(实验组4).采用RT-PCR和Western Blot方法分别测定caspase-3的相对表达量.新鲜组作为阴性对照.结果 在每个时间点下(P<0.05),细胞凋亡最轻的是新鲜组,较好的是实验组2和实验组3,其次依次为实验组4、实验组1,凋亡最严重的是对照组.结论 海藻糖和DMSO的联合运用能够很好的抑制caspase-3的表达.0.1 mol/L海藻糖+0.1 mol/LDMSO和0.2mol/L海藻糖+0.1 mol/L DMSO 能最大限度的抑制caspase-3的表达.%Objective To observe the expression of caspase-3 on the trehalose as cryoprotectant for preserving aortic valve homograft in liquid nitrogen.Methods The aortic valve homograft was divided into 5groups,namely:0.1 mol/L DMSO(control group),0.1 mol/L trehalose(experimental group 1),0.1 mol/L trehalose+0.1 mol/L DMSO(experimental group 2),0.2 mol/L trehalose+0.1 mol/L DMSO(experimental group 3),0.3 mol/L trehalose+0.1 mol/L DMSO(experimental group4).At the time of 12 months,15 months and 18 months when preserved in liquid nitrogen,relative expression of caspase-3 of the aortic valve homograft was measured by RT-PCR and Western Blot.Fresh group was a negative control group.Results At the same time(P<0.05),the expression of caspase-3 of fresh aortic tissue was slightest.The experimental group 2 was in accord with the experiment group 3,which was of a sort compare with the fresh group.The experimental group 4,which was worse than the experimental group 2 and 3,ranked above the experimental group 1.The worst was the control group.Conclusions The joint use of trehalose and DMSO could well inhibit the

  12. 吗啡缺氧预处理对兔缺氧/复氧肾脏损伤的保护作用和caspase-3的影响%Protection Against Acute Hypoxic/Reoxygenation Injury to Kidney for Rabbit with Morphine Hypoxic Preconditioning by Observing the Expression of Caspase-3 Protein

    Institute of Scientific and Technical Information of China (English)

    江晓琴; 倪娟; 杨沛; 黄蔚; 罗金凤; 李华凤; 王亚平

    2011-01-01

    The maintenance of the balance between oxygen supply and oxygen consumption is a key measure in preventing acute kidney hypoxic/reoxygenation injury. Morphine can inhibit metabolism and reduce the oxygen consumption. We tried to investigate the protective effects of morphine hypoxic preconditioning on acute kidney hypoxic/reoxygenation injury in rabbit and its influence on expression of caspase-3 protein. Kidney hypoxic and reoxy-genation were induced by making the tested rabbits inhale 8% oxygen for three hours firstly, and then putting them in the air to breathe in normal oxygen for another three hours. Morphine hypoxic preconditioning was induced by administering morphine 3mg/kg, and then hypoxic of 8% oxygen was induced. Caspase-3 protein expression in renal tissue was assessed by immunohistochemical method. In the present study, the expressions of caspase-3 protein were significantly higher in saline-control hypoxic group than in morphine hypoxic preconditioning group((29. 3±5. T)% vs. (12. 16 + 1. 23)%, P<0. 05). These observations suggested that morphine hypoxic preconditioning can protect rabbit against acute kidney hypoxic/reoxygenation injury by decreasing expression of caspase-3 protein.%维持机体氧供与氧耗相对平衡是防治缺氧/复氧损伤的重要措施.吗啡可降低代谢,降低氧耗.细胞凋亡是缺血再灌注损伤发病机制中的重要环节,半胱氨酸蛋白酶3(caspase-3)是凋亡的关键酶和执行者.本文拟通过观察缺氧/复氧损伤肾脏组织中caspase-3蛋白的变化,探讨吗啡缺氧预处理发挥肾脏保护作用的可能机制.经自制面罩吸入8% O2 3 h后吸入空气复氧3 h,建立全身缺氧/复氧肾损伤兔动物模型,缺氧/复氧结束后分别取肾脏组织,采用Envision两步法行caspase-3免疫组化染色.实验结果表明:正常对照组肾脏组织中极少的caspase-3阳性细胞表达.单纯缺氧/复氧组和吗啡缺氧预处理组阳性蛋白表达指数分别为(29.3±5

  13. Imatinib induces H2AX phosphorylation and apoptosis in chronic myelogenous leukemia cells in vitro via caspase-3/Mst1 pathway

    Institute of Scientific and Technical Information of China (English)

    Yan-jun ZHANG; Lian-ning DUAN; Cheng-rong LU; Yan CAO; Yuan LUO; Rong-feng BAO; Shu YAN; Mei XUE; Feng ZHU; Zhe WANG

    2012-01-01

    Aim:Histone H2AX is a novel tumor suppressor and its phosphorylation at the C terminus (Ser139 and Tyr142)is required for tumor cell apoptosis.The aim of the present study was to elucidate the mechanisms underlying imatinib-induced C-terminal phosphorylation of H2AX in chronic myelogenous leukemia cells in vitro.Methods:BCR-ABL-positive K562 cells were used.Microscopy,Western blotting and flow cytometry were used to study the signaling pathways that regulate imatinib-induced H2AX phosphorylation and the apoptotic mechanisms.Results:Treatment of K562 cells with imatinib (1-8 μmol/L)induced phosphorylation of H2AX at Ser139 and Tyr142 in time-and dose-dependent manners.In contrast,imatinib at the same concentrations did not affect H2AX acetylation at Lys 5,and the acetylated H2AX maintained a higher level in the cells.Meanwhile,imatinib (1-8 μmol/L)activated caspase-3 and its downstream mammalian STE20-like kinase 1 (Mst1),and induced apoptosis of K562 cells.The caspase-3 inhibitor Z-VAD (40 μmol/L)reduced imatinibinduced H2AX phosphorylation at Ser139 and Tyr142 and blocked imatinib-induced apoptosis of K562 cells.Imatinib (4 μmol/L)induced expression of Williams-Beuren syndrome transcription factor (WSTF),but not wild-type p53-induced phosphatase 1 (Wip1)in K562 cells.Conclusion:The caspase-3/Mst1 pathway is required for H2AX C-terminal phosphorylation at Ser139 and Tyr142 and subsequent apoptosis in Bcr-Abl-positive K562 cells induced by imatinib.

  14. Antrodia camphorata Potentiates Neuroprotection against Cerebral Ischemia in Rats via Downregulation of iNOS/HO-1/Bax and Activated Caspase-3 and Inhibition of Hydroxyl Radical Formation

    Directory of Open Access Journals (Sweden)

    Po-Sheng Yang

    2015-01-01

    Full Text Available Antrodia camphorata (A. camphorata is a fungus generally used in Chinese folk medicine for treatment of viral hepatitis and cancer. Our previous study found A. camphorata has neuroprotective properties and could reduce stroke injury in cerebral ischemia animal models. In this study, we sought to investigate the molecular mechanisms of neuroprotective effects of A. camphorata in middle cerebral artery occlusion (MCAO rats. A selective occlusion of the middle cerebral artery (MCA with whole blood clots was used to induce ischemic stroke in rats and they were orally treated with A. camphorata (0.25 and 0.75 g/kg/day alone or combined with aspirin (5 mg/kg/day. To provide insight into the functions of A. camphorata mediated neuroprotection, the expression of Bax, inducible nitric oxide synthase (iNOS, haem oxygenase-1 (HO-1, and activated caspase-3 was determined by Western blot assay. Treatment of aspirin alone significantly reduced the expressions of HO-1 (P<0.001, iNOS (P<0.001, and Bax (P<0.01 in ischemic regions. The reduction of these expressions was more potentiated when rats treated by aspirin combined with A. camphorata (0.75 g/kg/day. Combination treatment also reduced apoptosis as measured by a significant reduction in active caspase-3 expression in the ischemic brain compared to MCAO group (P<0.01. Moreover, treatment of A. camphorata significantly (P<0.05 reduced fenton reaction-induced hydroxyl radical (OH• formation at a dose of 40 mg/mL. Taken together, A. camphorata has shown neuroprotective effects in embolic rats, and the molecular mechanisms may correlate with the downregulation of Bax, iNOS, HO-1, and activated caspase-3 and the inhibition of OH• signals.

  15. Garlic (Allium sativum Fresh Juice Induces Apoptosis in Human Oral Squamous Cell Carcinoma: The Involvement of Caspase-3, Bax and Bcl-2

    Directory of Open Access Journals (Sweden)

    Farrokh Farhadi

    2015-12-01

    Full Text Available Background and aims. There is no report on the apoptotic impact of Allium sativum L.(Garlic on the oral squamous cell carcinoma (KB; hence, this study was designed to survey the apoptotic effects of garlic fresh juice (GFJ on the KB cells. Materials and methods. MTTassay (MicrocultureTetrazolium Assay was carried out to evaluate the cytotoxicity of GFJ on KB cells. Furthermore, TUNEL(Terminal deoxynucleotidyltransferase-mediated dUTP nick end labelingand DNA fragmentation tests were performed to determine if GFJ is able to induce apoptosis in KB cells. Also a standard kit was used to assess caspase-3 activity in KB cells. Also western blotting was employed to evaluate the effect of GFJ on Bax:Bcl-2 ratio. Results. Significant cytotoxic effects were observed for the minimum used concentration (1g/mL as calculated to be 77.97±2.3% for 24 h and 818±3.1% for 36h of incubation (P < 0.001. Furthermore, TUNEL and DNA fragmentation tests corroborated the apoptosis inducing activity of GFJ. Consistently, after treating KB cells with GFJ(1μg/mL, caspase-3 activity and Bax:Bcl-2 ratio were raised by 7.3±0.6 and (P <0.001 folds, respectively. Conclusion. The results of this study advanced that GFJ induces apoptosis in the KB cells through increasing caspase-3 activity and Bax:Bcl2 ratio which could be attributed to its organo-sulfurcomponents.

  16. Tetramethylpyrazine suppresses HIF-1α, TNF-α, and activated caspase-3 expression in middle cerebral artery occlusion-induced brain ischemia in rats

    Institute of Scientific and Technical Information of China (English)

    Yi CHANG; George HSIAO; Seu-hwa CHEN; Yi-cheng CHEN; Jiing-han LIN; Kuang-hung LIN; Duen-suey CHOU; Joen-rong SHEU

    2007-01-01

    Aim: To examine the detailed mechanisms underlying the inhibitory effect of tetramethylpyrazine (TMPZ) in inflammatory and apoptotic responses induced by middle cerebral artery occlusion (MCAO) in rats. Methods: MCAO-induced focal cerebral ischemia in rats was used in this study. The hypoxia-inducible factor-1α (HIF-1α), activation of caspase-3, and TNF- mRNA transcription in ischemic regions were detected by immunoblotting and RT-PCR, respectively.Anti-oxidative activity was investigated using a thiobarbituric acid-reactive sub-stance (TBARS) test in rat brain homogenate preparations. Results: We showed the statistical results of the infarct areas of solvent- and TMPZ (20 mg/kg)-treated groups at various distances from the frontal pole in MCAO-induced focal cerebral ischemia in rats. Treatment with TMPZ (20 mg/kg) markedly reduced the infarct area in all regions, especially in the third to fifth sections. MCAO-induced focal cerebral ischemia was associated with increases in HIF-1α and the activation of caspase-3, as well as TNF-α transcription in ischemic regions. These expressions were markedly inhibited by treatment with TMPZ (20 mg/kg). However, TMPZ (0.5-5 mmol/L) did not significantly inhibit TBARS reaction in rat brain homogenates.Conclusion: The neuroprotective effect of TMPZ may be mediated at least by a portion of the inhibition of HIF-let and TNF-α activations, followed by the inhibi-tion of apoptosis formation (active caspase-3), resulting in a reduction in the infarct volume in ischemia-reperfusion brain injury. Thus, TMPZ treatment may represent an ideal approach to lowering the risk of or improving function in is-chemia-reperfusion brain injury-related disorders.

  17. Disturbance of Bcl-2, Bax, Caspase-3, Ki-67 and C-myc expression in acute and subchronic exposure to benzo(a)pyrene in cervix.

    Science.gov (United States)

    Gao, Meili; Li, Yongfei; Ji, Xiaoying; Xue, Xiaochang; Chen, Lan; Feng, Guodong; Zhang, Huqin; Wang, Huichun; Shah, Walayat; Hou, Zhanwu; Kong, Yu

    2016-03-01

    Epidemiological studies have demonstrated that cigarette smoking is an important cofactor or an independent risk factor for the development of cervical cancer. Benzo(a)pyrene (BaP) is one of the most potent tobacco smoke carcinogens in tobacco smoke. BaP induced DNA damage and over expression in p53 cervical tissue of mice as demonstrated in our previous study. Here we present the findings of exposure to BaP on the expression of Bcl-2, C-myc, Ki-67, Caspase-3 and Bax genes in mouse cervix. Acute intraperitoneal administration of BaP (12.5, 25, 50, 100mg/kg body weight) to ICR female mice induced a significant increase in Bcl-2, C-myc, Ki-67 mRNA and protein level till 72h except in Bcl-2 at 24h with 12.5, 25, 50mg/kg as well as at 48h with 12.5mg/kg body weight post treatment. A significant increase was also seen in Caspase-3 and Bax mRNA and protein level with peak level at 24h and gradual decrease till 72h, however, the expression of caspase-3 increased while that of Bax decreased with increasing dose of Bap after 24h. In sub chronic intraperitoneal and oral gavage administration of BaP (2.5, 5, 10mg/kg body weight), similar significant increase was observed for all the examined genes as compared to the control and vehicle groups, however the expression of Bax decreased in a dose dependent manner. The findings of this study will help in further understanding the molecular mechanism of BaP induced carcinogenesis of cervical cancer.

  18. Growth inhibitory effect of KYKZL-1 on Hep G{sub 2} cells via inhibition of AA metabolites and caspase-3 pathway and cell cycle arrest

    Energy Technology Data Exchange (ETDEWEB)

    Cheng, Jing; Du, Yi-Fang; Xiao, Zhi-Yi; Pan, Li-Li; Li, Wei; Huan, Lin; Gong, Zhu-Nan [Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing (China); Wei, Shao-Hua [College of Chemistry and Materials Science, Nanjing Normal University, Nanjing (China); Huang, Shi-Qian; Xun, Wei; Zhang, Yi; Chang, Lei-Lei; Xie, Meng-Yu [Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing (China); Ao, Gui-Zhen [Department of Medicinal Chemistry, School of Pharmacy, Soochow University, Jiangsu (China); Cai, Jie; Qiu, Ting; Wu, Hao; Sun, Ting [Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing (China); Xu, Guang-Lin, E-mail: xudunlop@126.com [Jiangsu Key Laboratory for Molecular and Medical Biotechnology, College of Life Science, Nanjing Normal University, Nanjing (China); Department of Pharmacology, University of Michigan, Ann Arbor (United States)

    2014-01-01

    KYKZL-1, a newly synthesized compound with COX/5-LOX dual inhibition, was subjected to the inhibitory activity test on Hep G{sub 2} growth. We found that KYKZL-1 inhibited the growth of Hep G{sub 2} cells via inducing apoptosis. Further studies showed that KYKZL-1 activated caspase-3 through cytochrome c release from mitochondria and down regulation of Bcl-2/Bax ratio and reduced the high level of COX-2 and 5-LOX. As shown in its anti-inflammatory effect, KYKZL-1 also exhibited inhibitory effect on the PGE{sub 2} and LTB{sub 4} production in Hep G{sub 2} cells. Accordingly, exogenous addition of PGE{sub 2} or LTB{sub 4} reversed the decreases in cell viability. In addition, KYKZL-1 caused cell cycle arrest at the S–G{sub 2} checkpoint via the activation of p21{sup CIP1} protein and down-regulation of cyclin A expression. These data indicate that the growth inhibitory effect of KYKZL-1 is associated with inhibition of AA metabolites and caspase-3 pathway and cell cycle arrest. Combined with our previous findings, KYKZL-1 exhibiting COX/5-LOX inhibition may be a promising potential agent not only for inflammation control but also for cancer prevention/therapy with an enhanced gastric safety profile. - Highlights: • KYKZL-1 is designed to exhibit COX/5-LOX dual inhibition. • KYKZL-1 resulted in apoptosis of Hep G{sub 2} cells. • KYKZL-1 activated caspase-3 through cytochrome c and bcl-2/bax ratio. • KYKZL-1 caused cell cycle arrest via modulation of p21{sup CIP1} and cyclin A level.

  19. Long-term exposure of human gingival fibroblasts to cigarette smoke condensate reduces cell growth by modulating Bax, caspase-3 and p53 expression.

    Science.gov (United States)

    Alamri, A; Semlali, A; Jacques, É; Alanazi, M; Zakrzewski, A; Chmielewski, W; Rouabhia, M

    2015-08-01

    Smoking cigarettes increases the risk of oral tissue damage leading to periodontal disease. Gingival fibroblasts, the predominant cell type inhabiting gingival connective tissue, play a critical role in remodeling and maintaining gingival structure. The objective of this study was to investigate the effect of long-term exposure to cigarette smoke on human gingival fibroblast survival/apoptosis and the molecular pathways involved in these cell responses. Human gingival fibroblasts were extracted from healthy non-smokers and cultured in the presence of cigarette smoke condensate (CSC). At the end of each time point, cell growth was evaluated by means of MTT assay. Apoptotic and necrotic gene's expression was investigated by polymerase chain reaction array and by annexin V/propidium iodide staining and cell cycle assays. Western blot was used to investigate Bax and p53 proteins. These tests were supported by caspase 3 activity analyses. High levels of CSC decreased cell growth and deregulated cell cycle progression by increasing the G(0)/G(1) and reducing the S and G(2)/M phases of the gingival fibroblasts. Polymerase chain reaction arrays revealed the activation of several apoptotic genes by CSC, including TNF receptors, caspases, Bax and p53. This was supported by increases in the Bax and p53 protein levels as well as by an elevated activity of caspase-3 in the CSC-exposed cells. Immunofluorescence staining demonstrated that both Bax and caspase-3 displayed a cytosolic and mitochondrial distribution in the CSC-exposed gingival fibroblasts, compared to controls. The damaging effect of CSC on gingival fibroblast growth was also supported by the decrease in interleukin 6 and 8 secretion by the gingival fibroblasts. These results suggest that CSC may contribute to deregulating fibroblast functions. This can compromise fibroblast-epithelial cell interactions, which ultimately increases the risk of gingival tissue damage and the onset of periodontitis. © 2014 John Wiley

  20. P53-mediated cell cycle arrest and apoptosis through a caspase-3-independent, but caspase-9-dependent pathway in oridonin-treated MCF-7 human breast cancer cells

    Institute of Scientific and Technical Information of China (English)

    Qiao CUI; Jing-hua YU; Jin-nan WU; Shin-ichi TASHIRO; Satoshi ONODERA; Mutsuhiko MINAMI; Takashi IKEJIMA

    2007-01-01

    Aim: To study the caspase-3-independent mechanisms in oridonin-induced MCF-7 human breast cancer cell apoptosis in vitro. Methods: The viability of oridonin-treated MCF-7 cells was measured by MTT (thiazole blue) assay. Apoptotic cells with condensed nuclei were visualized by phase contrast microscopy. Nucleoso-mal DNA fragmentation was assayed by agarose gel electrophoresis. The apoptotic ratio was determined by lactate dehydrogenase assay. Cell cycle alternation and mitochondrial membrane potential were measured by flow cytometric analysis. Bax, Bcl-2, caspase-3, caspase-9, heat shock protein (Hsp)90, p53, p-p53, p21, Poly (ADP-ribose) polymerase (PARP), and the inhibitor of caspase-activated Dnase (ICAD) protein expressions were detected by Western blot analysis. Results: Oridonin inhibited cell growth in a time- and dose-dependent manner. Cell cycle was altered through the upregulation of p53 and p21 protein expressions. Pan-caspase inhibitor Z-VAD-fmk and calpain inhibitor Ⅱ both decreased cell death ratio. Nucleosomal DNA fragmentation and the downregulation of △ψmit were detected in oridonin-induced MCF-7 cell apoptosis, which was involved in a postmitochondrial caspase-9-dependent pathway. Decreased Bcl-2 and Hsp90 expression levels and increased Bax and p21 expression levels were positively correlated with elevated levels of phosphorylated p53 phosphorylation. Moreover, PARP was partially cleaved by calpain rather than by capase-3. Conclusion: DNA damage provoked alternations in the mitochondrial and caspase-9 pathways as well as p53-mediated cell cycle arrest, but was not related to caspase-3 activity in oridonin-induced MCF-7 cells.

  1. Erythropoietin inhibits gamma-irradiation-induced apoptosis by upregulation of Bcl-2 and decreasing the activation of caspase 3 in human UT-7/erythropoietin cell line.

    Science.gov (United States)

    Liu, Yuan-Yuan; She, Zhen-Jue; Yao, Ming-Hui

    2010-05-01

    1. Erythropoietin (EPO) can reverse radiotherapy-induced anaemia by stimulating bone marrow cells to produce erythrocytes. However, there are limited studies that address the mechanisms by which EPO exerts its beneficial effects in radiotherapy-induced anaemia. In the present study, we used a human bone marrow-derived EPO-dependent leukaemia cell line UT-7/EPO that progressed further in erythroid development to evaluate the anti-apoptotic effects of EPO on irradiated human erythroid progenitor. 2. The UT-7/EPO cells exposed to gamma-irradiation were cultured in the presence or absence of EPO at a concentration of 7 U/mL. The cell viability, cell apoptosis and the expression of apoptosis-related proteins Bcl-2, Bax and caspase 3 were examined. 3. The results showed that EPO protected the viability of human UT-7/EPO cells exposed to gamma-irradiation. EPO significantly inhibited gamma-irradiation-induced apoptosis in human UT-7/EPO cells: a significant decrease in the percentage of apoptotic cells was observed (62, 69 and 62% at 24, 48 and 72 h, respectively). Furthermore, EPO significantly increased the expression of Bcl-2 protein and the relative Bcl-2/Bax ratio, and decreased the activation of caspase 3 and formation of the p17 and p12 cleavage in similar conditions. 4. In conclusion, EPO exerts anti-apoptotic effects on irradiated human UT-7/EPO cells through upregulation of Bcl-2 protein and the relative Bcl-2/Bax ratio, and by decreasing the activation of caspase 3. These findings may contribute to our understanding of the beneficial function of EPO in radiotherapy-induced anaemia.

  2. Effects of acupoint versus non-acupoint electroacupuncture on cerebral cortical neuronal Bcl-2,Bax and caspase-3 expression in a rat model of focal cerebral ischemia

    Institute of Scientific and Technical Information of China (English)

    Jun Wang; Junming Fan; Yongshu Dong; Xia Huang; Hongxia Zhang

    2008-01-01

    BACKGROUND: Several studies have demonstrated that electroacupuncture by acupoint selection can inhibit cerebral cortical neuronal apoptosis following cerebral ischemia/reperfusion.OBJECTIVE: To validate the effects of electroacupuncture by acupoint selection on the expression level of cortical neuronal anti-apoptotic Bcl-2 protein and the apoptotic executive protein, caspase-3, in rat models of focal cerebral ischemia/reperfusion.DESIGN, TIME AND SETTING: This randomized grouping, neural cell and molecular biology animal experiment was performed at the Laboratory of Pharmacology of Traditional Chinese Medicine and the Laboratory Animal Center of Henan Institute of Traditional Chinese Medicine between November 2006 and May 2007.MATERIALS: Atotal of 40 healthy male adult Sprague-Dawley rats were randomly and evenly divided into four groups: sham-operated, model, electroacupuncture and non-acupoint control. G6895 electro-acupuncture instruments were purchased from Shanghai Huayi Instrument Factory, China. Caspase-3, Bcl-2 and Bax kits were provided by Wuhan Boster Bioengineering Co., Ltd., China.METHODS: Middle cerebral artery occlusion was induced in the model, electroacupuncture and non-acupoint groups. In the electroacupuncture group, the acupoints Jianyu (LI15), Waiguan (SJ5), Biguan (ST31), and Zusanli (ST36) were given electroacupuncture. In the non-acupoint control group, at each time point (immediately after ischemia and after reperfusion, or 2 hours after reperfusion), electroacupuncture was performed at the midpoints of Tianquan (PC2)-Quze (PC 3) line, Quze (PC 3)-Ximen (PC4) line, Zuwuli (LRlO)-Yinbao (LRg) line, and Xiguan (LR7)-Zhongdu (LR6) line. Electroacupuncture parameters were set with a continuous wave with a frequency of 10 Hz, wave width 0.6 ms, voltage 1.5-3.0 V, and a duration of 10 minutes. The sham-operated and model groups received only animal fixation without electroacupuncture procedure.MAIN OUTCOME MEASURES: Five rats were selected from

  3. PCSK9 siRNA inhibits HUVEC apoptosis induced by ox-LDL via Bcl/Bax-caspase9-caspase3 pathway.

    Science.gov (United States)

    Wu, Chun-Yan; Tang, Zhi-Han; Jiang, Lu; Li, Xue-Fei; Jiang, Zhi-Sheng; Liu, Lu-Shan

    2012-01-01

    This paper investigated the effects of ox-LDL on PCSK9, and the molecular mechanisms of PCSK9 siRNA-inhibited apoptosis induced by ox-LDL in human umbilical vein endothelial cells (HUVECs), to clarify the role of PCSK9 in atherosclerogenesis. HUVECs were incubated with ox-LDL for 24 h. The apoptosis was observed by Hoechst 33258 staining. The expression of PCSK9, LOX-1 mRNAs and proteins was detected by RT-PCR, western blot, respectively. The PCSK9 siRNAs labeled with fluorescence were transfected into HUVECs by Lipofectamine 2000. After transfection for 24 h, cells were treated with ox-LDL for 24 h, HUVECs apoptosis transfected siRNA was detected by Hoechst 33258 staining and flow cytometer. The expression of Bcl-2, Bax, caspase3, 8, 9 was detected by western blot. The activity of caspase3, 9 was detected by kits. Our results showed that apoptosis of HUVECs and the expressions of PCSK9 and LOX-1 were upregulated secondary to induction by ox-LDL in a concentration-dependent manner. However, ox-LDL-induced HUVEC apoptosis and PCSK9 expression, but not LOX-1 expression, were significantly reduced by PCSK9 siRNA. These results demonstrate a linkage between HUVEC apoptosis and PCSK9 expression. Furthermore, we detected the possible pathway involved in apoptotic regulation by PCSK9 siRNA; our results showed that the expression of Bcl-2 decreased, whereas that of Bax increased. In addition, ox-LDL enhanced the activity of caspase9 and then caspase3. Pretreatment of HUVECs with PCSK9 siRNA blocked these effects of ox-LDL. These findings suggest that ox-LDL-induced HUVECs apoptosis could be inhibited by PCSK9 siRNA, in which Bcl/Bax-caspase9-caspase3 pathway maybe was involved through reducing the Bcl-2/Bax ratio and inhibited the activation of both caspase9 and 3.

  4. Methotrexate induces poly(ADP-ribose) polymerase-dependent, caspase 3-independent apoptosis in subsets of proliferating CD4+ T cells

    DEFF Research Database (Denmark)

    Nielsen, C H; Albertsen, L; Bendtzen, K;

    2007-01-01

    . Exposure of CA-stimulated PBMC to MTX significantly increased their level of cleaved poly(ADP-ribose) polymerase (PARP), and a similar tendency was observed in TT-stimulated cells. Unlike CA and TT, the mitogen phytohaemagglutinin (PHA) induced proliferation of both CD4- and CD4+ T cells, and induced...... apoptosis in both undivided and divided Th cells. PHA-induced apoptosis involved activation of caspase-3 and the anti-apoptotic protein Bcl-2 in addition to PARP cleavage, suggesting that PHA induces apoptosis via different pathways than CA and TT. We suggest that the latter are more representative...

  5. Polyphenol Compounds of Mahkota Dewa (Phaleria macrocarpa[Scheff.] Boerl Up-regulated Caspase-3 and Apoptosis Index in Balb/c Strain Mice

    Directory of Open Access Journals (Sweden)

    Indranila KS

    2016-04-01

    Full Text Available Background: Polyphenol compounds of Mahkota Dewa (Phaleria macrocarpa[Scheff.] Boerl (PMD can potentially be used as ant cancer treatment by scavanging radical molecules. The effect in vivois still limited to Indonesia. Purpose: This research was aimed to validate the activity of PMD in increasingcaspase-3 expression and apoptosis in Balb/c mice, induced by Benzo(apyrene (BaP. Methods: A posttest control group was implemented and used by 40 Balb/c mice at the age of 1-2 weeks, with the body weight of 20-30 g. The tumor induction was administered to the mice using BaP. The animals were randomized into two groups called the control group and the PMD treatment group, the latter of which was given a dosage of 50mg. Lung tumor growth was assessed through surgery at week 8, 17, and 26. The results of caspase-3expression and apoptotic index from IHC-TUNEL staining were analyzed using Kruskal-Wallis, Mann-Whitney, One-way ANOVA, and Post hoc test LSD with significant levels of p<α (0,05.This research was approved by Ethical Clearance. Results: Oral administration of 50mg PMD significantly increased caspase-3 expression and apoptotic index in the treatment group animals at weeks 8, 17, and 26. Carcinogenesis incidence in the control group were respectively found at2,32±0,26 and 3,93±0,46 at weeks 8 and 26, while those of the treatment group were 1,88±0,38 and 0,88±0,22 (p=0,001. The apoptotic index in the control group was0,00±0,00 at 8 weeksand0,92+0,22at 26 weeks, whereas the indexes of the treatment group were 1,12±0,71 and 2,02±1,05 (p=0,001. In the control group, the caspase-3 expression at weeks 8 and 26 were 0,28±0,17 and 0,56±0,16, while those in the treatment group were 0,60±0,14 at week 8 and 2,52±0,33 at week 26 (p=0,001. Conclusion: The treatment of PMD effectively induced cell apoptosis in the Balb/c mice via up- regulation of the caspase-3 expression, thereby increasing the apoptotic index. This shows that PMD has anticancer

  6. Effect of tenuigenin on the Caspase-3 and Par-4 expression of neural stem cells induced by beta-amyloid protein%远志总皂苷干预β-淀粉样蛋白致伤神经干细胞Caspase-3及Par-4的表达

    Institute of Scientific and Technical Information of China (English)

    张晓梅; 孙光涛; 黄作义; 吴成吉; 戚询中; 朱晓峰

    2011-01-01

    BACKGROUND: The effect of tenuigenin which has a good nerve protection to neural stem cells has not been reported. OBJECTIVE: To investigate the protective effect and mechanism of tenuigenin for neural stem cells in the hippocampus impaired by β-amyloid protein.METHODS: The third passage neural stem cells generated from the hippocampi of Kunming mice which injured by β-amyloid protein in vitro were incubated with different concentrations of tenuigenin.RESULTS AND CONCLUSION: Immunocytochemical technique was used to detect Caspase3-positive neural stem cells. The study revealed that the expression of Par-4 and caspase-3 positive neural stem cells in the tenuigenin group were significantly lower than that in the control group with statistical significance (P < 0.05). Tenuigenin can reduce the expression of Par-4 and Caspase-3 in neural stem cells impaired by β-amyloid protein.%背景:远志总皂苷具有良好的神经保护作用.目的:分析远志总皂苷对β-淀粉样蛋白致伤海马神经干细胞的保护作用及机制.方法:自昆明小鼠海马分离培养神经干细胞,取第3代神经干细胞,用含不同质量浓度远志总皂苷与β-淀粉样蛋白致伤体外培养的神经干细胞共孵育.结果:应用免疫组织化学法检测Caspase-3阳性神经干细胞,与对照组相比,远志总皂苷组Caspase-3阳性细胞率及Par-4的表达明显降低,差异具有显著性意义(P<0.05).提示远志总皂苷能够降低β-淀粉样蛋白致伤神经干细胞中Caspase-3及Par-4的表达.

  7. Effect of 1, 25 (OH)2D3 on Apoptosis of Human Mesangial Cells and Caspase-3 Expression%活性维生素D3对人系膜细胞凋亡及Caspase-3表达的影响

    Institute of Scientific and Technical Information of China (English)

    张昊; 尹璇; 陈建平; 马莉; 张春江; 刘春

    2015-01-01

    目的 探讨活性维生素D3[1,25 (OH)2D3]对人系膜细胞凋亡的影响,并初步探究其作用机制.方法 体外培养人系膜细胞,随机分为4组:正常对照组,表皮生长因子(EGF)组,1,25 (OH)2D3组,EGF联合1,25 (OH)2D3组,采用流式细胞术检测各组系膜细胞的凋亡率,Western blotting检测各组半胱氨酸蛋白酶-3(Caspase-3)蛋白表达的情况.结果 与正常对照组比较,EGF组系膜细胞凋亡率降低,Caspase-3表达量降低(P<0.05),1,25 (OH)2D3组系膜细胞的凋亡率升高,Caspase-3表达量升高(P<0.05);与EGF组比较,EGF联合1,25 (OH)2D3组系膜细胞的凋亡率升高,Caspase-3表达量升高(P<0.05);EGF联合1,25 (OH)2D3组与正常对照组系膜细胞的凋亡率和Caspase-3表达量间差异均无统计学意义(P>0.05).结论 1,25 (OH)2D3可能是通过上调Caspase-3的表达而诱导系膜细胞的凋亡.EGF可抑制人系膜细胞的凋亡,1,25 (OH)2D3可逆转EGF对系膜细胞凋亡的抑制作用.

  8. Effects of erythropoietin on neuron apoptosis and expression of caspase-3 after excitotoxic brain injury in mice during different developmental stages%促红细胞生成素对发育期小鼠脑损伤后神经细胞凋亡和caspase-3表达的影响

    Institute of Scientific and Technical Information of China (English)

    金宝; 张育才

    2013-01-01

    目的 研究促红细胞生成素(erythropoietin,EPO)对鹅膏蕈氨酸(ibotenic acid,Ibo)致发育期小鼠兴奋性脑损伤后神经细胞凋亡和caspase-3表达的影响.方法 以144只健康雌性昆明小白鼠作为实验动物,其中7日龄(P7)、21日龄(p21)和42日龄(P42)小鼠各48只;再将同日龄小鼠随机(随机数字法)分为假手术组(n=16)、Ibo模型组(n=16)和EPO治疗组(n=16).采用颅内微量注射法向小鼠左侧海马注射Ibo 1μl(5μg)建立脑损伤模型.EPO治疗组小鼠海马注射Ibo后,腹腔注射5000 U/ (kg·d) EPO,连续3 d;Ibo模型组和假手术组腹腔注射等体积生理盐水.各组小鼠在建模后3d处死,Nissl染色观察小鼠海马神经细胞凋亡病理改变;双抗体夹心ABC-ELISA法检测caspase-3含量变化;分光光度法检测caspase-3活性改变.结果 光学显微镜观察结果显示,与假手术组相比,Ibo模型组小鼠海马区神经细胞发生明显变性和死亡,神经细胞数量明显减少;EPO治疗组小鼠海马区神经细胞凋亡程度较轻.Ibo模型组与假手术组相比,caspase-3含量和活性明显升高(P <0.05);EPO治疗组caspase-3含量和活性的升高幅度明显低于Ibo模型组(P<0.05).结论 Ibo致脑损伤时,脑组织caspase-3的表达增强,导致神经细胞凋亡.EPO可减轻脑损伤后神经细胞凋亡,起到脑保护的作用,其机制可能与下调caspase-3的表达有关.%Objective To investigate the effects of erythropoietin (EPO) on neuron apoptosis and expression of caspase-3 after excitotoxic brain injury induced by ibotenic acid (Ibo) in mice during different developmental stages.Methods A total of 144 healthy KM mice aged 7 d (n =48),21 d (n =48) and 42 d (n =48) were selected,and those of the same age were randomly (random number) divided into 3 groups:sham surgery group (n =16),Ibo group (n =16) and EPO treated group (n =16).Brain injury model was established by Ibo 1μl (5 μg) injected into left hippocampus.In EPO

  9. Expression of Caspase-3 mRNA in frontal cortex and hippocampus of chronic stress-induced depression rats treated by electro-acupuncture%电针对慢性应激抑郁模型大鼠额叶皮层及海马Caspase-3基因表达的影响

    Institute of Scientific and Technical Information of China (English)

    梁佳; 卢峻; 崔善福; 王俊仁; 图娅

    2012-01-01

    目的 观察电针对慢性应激抑郁模型大鼠额叶皮层及海马半胱氨酸蛋白酶-3(Cysteinyl aspartate specific protease-3,Caspase-3)表达的影响,探讨电针治疗抑郁症的作用机理.方法 将Sprague-Dawley (SD)大鼠48只随机数字表法分为4组:空白组、模型组、模型+电针组(电针组)、模型+阳性药组(帕罗西汀组),12只/组;采用旷场实验进行行为学评价;采用实时荧光定量PCR(RT-PCR)方法检测大鼠额叶皮层及海马Caspase-3 mRNA含量.结果 ①旷场实验:与空白组大鼠[分别为水平(66±13)格、竖立(10±2)次]相比,模型组大鼠水平穿越格数[(29±7)格]、竖立次数[(6±2)次]均有减少(P<0.05、P>0.05).与模型组相比,电针组大鼠水平穿越格数[(61±9)格]、竖立次数[(13±1)次]均明显提高(P<0.01);帕罗西汀组大鼠水平穿越格数[(39±10)格]、竖立次数[(8±1)次]有所增加,但差异无统计学意义(P>0.05).②RT-PCR:与空白组相比,模型组大鼠额叶皮层、海马Caspase-3 mRNA含量明显升高(P<0.05).与模型组相比,电针组与帕罗西汀组大鼠额叶皮层Caspase-3 mRNA含量明显下降,差异有统计学意义(P<0.05);电针组与帕罗西汀组大鼠海马Caspase-3 mRNA含量有下降趋势,但差异无统计学意义(P>0.05).结论 慢性应激抑郁模型大鼠额叶皮层及海马Caspase-3 mRNA含量增加,电针可以从基因水平下调其表达,这可能是电针发挥抗抑郁治疗作用的途径之一.%Objective To observe the expression of Caspase-3 mRNA in prefrontal cortex and hippocampus of chronic stress-induced depression rats,and to detect the machnisms of antidepression by electro-acupuncture.Methods Sprague-Dawley rats were randomly divided into four groups:control group,model group,model +electro-acupuncture group and model + paroxetine group,12 rats in each group.Open-field test was used to observe the changes of movements,and real-time fluorescence quantitative PCR (RT

  10. Quercetin-induced apoptosis acts through mitochondrial- and caspase-3-dependent pathways in human breast cancer MDA-MB-231 cells.

    Science.gov (United States)

    Chien, Su-Yu; Wu, Yao-Chung; Chung, Jing-Gung; Yang, Jai-Sing; Lu, Hsu-Feng; Tsou, Mei-Fen; Wood, W G; Kuo, Shou-Jen; Chen, Dar-Ren

    2009-08-01

    There has been considerable evidence recently demonstrating the anti-tumour effects of flavonols. Quercetin, an ubiquitous bioactive flavonol, inhibits cells proliferation, induces cell cycle arrest and apoptosis in different cancer cell types. The precise molecular mechanism of quercetin-induced apoptosis in human breast cancer cells is unclear. The purpose of this study was to investigate effects of quercetin on cell viability and to determine its underlying mechanism in human breast cancer MDA-MB-231 cells. Quercetin decreased the percentage of viable cells in a dose- and time-dependent manner, which was associated with cell cycle arrest and apoptosis. Quercetin did not increase reactive oxygen species generation but increased cytosolic Ca(2+) levels and reduced the mitochondrial membrane potential (DeltaPsi(m)). Quercetin treatment promoted activation of caspase-3, -8 and -9 in MDA-MB-231 cells. Caspase inhibitors prevented the quercetin-induced loss of cell viability. Quercetin increased abundance of the pro-apoptotic protein Bax and decreased the levels of anti-apoptotic protein Bcl-2. Confocal laser microscope examination indicated that quercetin promoted apoptosis-inducing factor (AIF) release from mitochondria and stimulated translocation to the nucleus. Taken together, these findings suggest that quercetin results in human breast cancer MDA-MB-231 cell death through mitochondrial- and caspase-3-dependent pathways.

  11. Apoptosis induction-related cytosolic calcium responses revealed by the dual FRET imaging of calcium signals and caspase-3 activation in a single cell.

    Science.gov (United States)

    Miyamoto, Akitoshi; Miyauchi, Hiroshi; Kogure, Takako; Miyawaki, Atsushi; Michikawa, Takayuki; Mikoshiba, Katsuhiko

    2015-04-24

    Stimulus-induced changes in the intracellular Ca(2+) concentration control cell fate decision, including apoptosis. However, the precise patterns of the cytosolic Ca(2+) signals that are associated with apoptotic induction remain unknown. We have developed a novel genetically encoded sensor of activated caspase-3 that can be applied in combination with a genetically encoded sensor of the Ca(2+) concentration and have established a dual imaging system that enables the imaging of both cytosolic Ca(2+) signals and caspase-3 activation, which is an indicator of apoptosis, in the same cell. Using this system, we identified differences in the cytosolic Ca(2+) signals of apoptotic and surviving DT40 B lymphocytes after B cell receptor (BCR) stimulation. In surviving cells, BCR stimulation evoked larger initial Ca(2+) spikes followed by a larger sustained elevation of the Ca(2+) concentration than those in apoptotic cells; BCR stimulation also resulted in repetitive transient Ca(2+) spikes, which were mediated by the influx of Ca(2+) from the extracellular space. Our results indicate that the observation of both Ca(2+) signals and cells fate in same cell is crucial to gain an accurate understanding of the function of intracellular Ca(2+) signals in apoptotic induction.

  12. Fucose-containing sulfated polysaccharides from brown seaweeds inhibit proliferation of melanoma cells and induce apoptosis by activation of caspase-3 in vitro.

    Science.gov (United States)

    Ale, Marcel Tutor; Maruyama, Hiroko; Tamauchi, Hidekazu; Mikkelsen, Jørn D; Meyer, Anne S

    2011-12-01

    Fucose-containing sulfated polysaccharides (FCSPs) extracted from seaweeds, especially brown macro-algae, are known to possess essential bioactive properties, notably growth inhibitory effects on tumor cells. In this work, we conducted a series of in vitro studies to examine the influence of FCSPs products from Sargassumhenslowianum C. Agardh (FSAR) and Fucus vesiculosus (FVES), respectively, on proliferation of melanoma B16 cells and to investigate the underlying apoptosis promoting mechanisms. Cell viability analysis showed that both FCSPs products, i.e., FSAR and FVES, decreased the proliferation of the melanoma cells in a dose-response fashion, with FSAR being more potent at lower dosages, and FVES being relatively more anti-proliferative than FSAR at higher dosages. Flow cytometric analysis by Annexin V staining of the melanoma cells exposed to the FCSPs products confirmed that both FSAR and FVES induced apoptosis. The FCSPs-induced apoptosis was evidenced by loss of plasma membrane asymmetry and translocation of the cell membrane phospholipids and was accompanied by the activation of caspase-3. The FCSPs bioactivity is proposed to be attributable to distinct structural features of the FCSPs, particularly the presence of sulfated galactofucans (notably in S.henslowianum) and sulfated fucans (notably in F. vesiculosus). This study thus indicates that unfractionated FCSPs may exert bioactive effects on skin cancer cells via induction of apoptosis through cascades of reactions that involve activation of caspase-3.

  13. [Modified alkaloids from Chelidonium majus L. induce G2/M arrest, caspase-3 activation, and apoptosis in human acute lymphoblastic leukemia MT-4 cells].

    Science.gov (United States)

    Fil'chenkov, O O; Zavelevych, M P; Khranovs'ka, N M; Zaïka, L A; Potopal's'kyĭ, A I

    2006-01-01

    The novel anticancer drug amitosine representing the mixture of thiophosphamide-modified alkaloids from Chelidonium majus L. has been reported to inhibit growth of various solid tumors in vivo. However, its antileukemic activity as well as the mechanisms of anticancer action have not been yet extensively examined. In this study, amitosine treatment at a dose of 100-250 microg/mL for 24 h resulted in dose-dependent inhibition of MT-4 cell proliferation in vitro with apoptosis induction in the setting of the significant G2/M phase arrest (up to 70% of cells). While amitosine induced caspase-3 activation in MT-4 cells, the increase in the number of cells containing the active caspase-3 did not correlate with the increase of apoptotic cell percentage. Western blotting data revealed the accumulation of cytochrome c in cytosolic fraction of MT-4 cells within 6 h after treatment with 100 microg/mL amitosine. To sum up, amitosine has been shown to possess strong antiproliferative and apoptosis-inducing activities in MT-4 cells in vitro, which seem to be mediated partially through caspase-dependent mitochondrial death pathways.

  14. Fucose-Containing Sulfated Polysaccharides from Brown Seaweeds Inhibit Proliferation of Melanoma Cells and Induce Apoptosis by Activation of Caspase-3 in Vitro

    Directory of Open Access Journals (Sweden)

    Anne S. Meyer

    2011-12-01

    Full Text Available Fucose-containing sulfated polysaccharides (FCSPs extracted from seaweeds, especially brown macro-algae, are known to possess essential bioactive properties, notably growth inhibitory effects on tumor cells. In this work, we conducted a series of in vitro studies to examine the influence of FCSPs products from Sargassum henslowianum C. Agardh (FSAR and Fucus vesiculosus (FVES, respectively, on proliferation of melanoma B16 cells and to investigate the underlying apoptosis promoting mechanisms. Cell viability analysis showed that both FCSPs products, i.e., FSAR and FVES, decreased the proliferation of the melanoma cells in a dose-response fashion, with FSAR being more potent at lower dosages, and FVES being relatively more anti-proliferative than FSAR at higher dosages. Flow cytometric analysis by Annexin V staining of the melanoma cells exposed to the FCSPs products confirmed that both FSAR and FVES induced apoptosis. The FCSPs-induced apoptosis was evidenced by loss of plasma membrane asymmetry and translocation of the cell membrane phospholipids and was accompanied by the activation of caspase-3. The FCSPs bioactivity is proposed to be attributable to distinct structural features of the FCSPs, particularly the presence of sulfated galactofucans (notably in S. henslowianum and sulfated fucans (notably in F. vesiculosus. This study thus indicates that unfractionated FCSPs may exert bioactive effects on skin cancer cells via induction of apoptosis through cascades of reactions that involve activation of caspase-3.

  15. Molecular and immunohistochemical expression of apoptotic proteins Bax, Bcl-2 and Caspase 3 in infantile hemangioma tissues as an effect of propranolol treatment.

    Science.gov (United States)

    Wnęk, Aneta; Andrzejewska, Ewa; Kobos, Józef; Taran, Katarzyna; Przewratil, Przemysław

    2017-05-01

    Infantile hemangiomas (IHs) are the most common benign tumors of childhood. They are characterized by a unique clinical course with two phases, proliferation and involution, which are followed by regression. The therapy of infantile hemangiomas was revolutionized in 2008 by the introduction of propranolol, however, the mechanism of its influence on hemangiomas remains unclear. The study included 71 patients with IHs, 27 of whom were treated with propranolol while the remaining 44 were used as a comparative group. The expression of Bcl-2, Bax and Caspase3 was determined with immunohistochemistry and mRNA of Bax, Bcl-2 and Caspase3 were assessed with the use of RT-PCR. Both methods revealed a statistically significant decrease in Bcl-2 expression and an increase in Bax in IHs tissues after propranolol treatment. The results obtained for Bax and Bcl-2 proteins may indicate a link between the effect of propranolol and apoptosis. Higher Bax and lower Bcl-2 expression in the propranolol treated group indicates a strong pro- apoptotic action countering any anti-apoptotic activity; apoptosis was indicted in IH tissue as a potential result of propranolol treatment, with potential clinical impact in other tumors. Copyright © 2017 European Federation of Immunological Societies. Published by Elsevier B.V. All rights reserved.

  16. A recombinant matriptase causes an increase in caspase-3 activity in a small-intestinal epithelial IEC-6 line cultured on fibronectin-coated plates.

    Science.gov (United States)

    Mochida, Seiya; Tsuzuki, Satoshi; Inouye, Kuniyo; Fushiki, Tohru

    2014-05-01

    Matriptase is an epithelial-derived type-II transmembrane serine protease. This protease is expressed prominently in the villus tip of small-intestinal epithelia at which senescent cells undergo shedding and/or apoptosis. The basement membrane of epithelial cells, including small-intestinal epithelial cells, contains extracellular matrix (ECM) proteins such as fibronectin and laminin. We found previously that high concentrations of a recombinant matriptase catalytic domain (r-MatCD) (e.g. 1 μM) caused an increased detachment of and increases in the activity of apoptotic effector caspase-3 in a rat small-intestinal epithelial IEC-6 line cultured on laminin-coated plates and proposed that at sites with its high level of expression, matriptase contributes to promoting shedding and/or detachment-induced death of epithelial cells through a mechanism mediating loss of cell-ECM adhesion. In this study, we found that even without increasing cell detachment, a high concentration of r-MatCD causes an increase in caspase-3 activity in IEC-6 cells cultured on fibronectin-coated plates, suggesting that the recombinant matriptase can cause apoptosis by a mechanism unrelated to cell detachment. Also, r-MatCD-treated IEC-6 cells on fibronectin were found to display spindle-like morphological changes. We suggest that r-MatCD causes apoptosis of IEC-6 on fibronectin by a mechanism involving the disruption of cell integrity.

  17. Matrine inhibits diethylnitrosamine-induced HCC proliferation in rats through inducing apoptosis via p53, Bax-dependent caspase-3 activation pathway and down-regulating MLCK overexpression

    Science.gov (United States)

    Zhang, Xiaolin; Yu, Hao

    2016-01-01

    The proliferation of hepatocellular carcinoma (HCC) cells is one of the leading causes of liver cancer mortality in humans. The inhibiting effects of matrine on HCC cell proliferation have been studied, but the mechanism of that inhibition has not been fully elucidated. Since, apoptosis plays an important role in HCC cell proliferation. We examined the apoptosis-inducing effect of matrine on tumor cells. Western blot analysis of p53, Bax, cleaved caspase-3 and myosin light chain kinase (MLCK) revealed that matrine induced tumor cell apoptosis by controlling anoikis. It activated p53, Bax-dependent caspase-3 and blocked the ECM-integrin mediated cell survival pathway through down-regulating MLCK over-expression in the liver of rats with diethyl nitrosamine (DENA)-induced HCC. Our results suggest that matrine can inhibit the proliferation of HCC cells through inducing tumor cell apoptosis via activation of the p53 pathway and inhibition of MLCK overexpression. Matrine may thus be used as a potentially promising reagent to inhibit HCC cell proliferation and MLCK may be a novel target for the treatment of HCC. PMID:27642320

  18. TAF15 and the leukemia-associated fusion protein TAF15-CIZ/NMP4 are cleaved by caspases-3 and -7

    Energy Technology Data Exchange (ETDEWEB)

    Alves, Juliano, E-mail: jalves@gnf.org [Department of Molecular Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (United States); Wurdak, Heiko [Department of Chemistry, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (United States); Garay-Malpartida, Humberto M. [Department of Pharmacology, Institute of Biomedical Sciences, University of Sao Paulo, Av. Lineu Prestes 1524, Sao Paulo, SP, CEP 05508-900 (Brazil); Harris, Jennifer L. [Department of Molecular Biology, The Scripps Research Institute, 10550 N. Torrey Pines Road, La Jolla, CA 92037 (United States); Protease Biochemistry, Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 92121 (United States); Occhiucci, Joao M.; Belizario, Jose E. [Department of Pharmacology, Institute of Biomedical Sciences, University of Sao Paulo, Av. Lineu Prestes 1524, Sao Paulo, SP, CEP 05508-900 (Brazil); Li, Jun, E-mail: jli2@gnf.org [Protease Biochemistry, Genomics Institute of the Novartis Research Foundation, 10675 John Jay Hopkins Drive, San Diego, CA 92121 (United States)

    2009-07-10

    Caspases are central players in proteolytic pathways that regulate cellular processes such as apoptosis and differentiation. To accelerate the discovery of novel caspase substrates we developed a method combining in silico screening and in vitro validation. With this approach, we identified TAF15 as a novel caspase substrate in a trial study. We find that TAF15 was specifically cleaved by caspases-3 and -7. Site-directed mutagenesis revealed the consensus sequence {sup 106}DQPD/Y{sup 110} as the only site recognized by these caspases. Surprisingly, TAF15 was cleaved at more than one site in staurosporine-treated Jurkat cells. In addition, we generated two oncogenic TAF15-CIZ/NMP4-fused proteins which have been found in acute myeloid leukemia and demonstrate that caspases-3 and -7 cleave the fusion proteins at one single site. Broad application of this combination approach should expedite identification of novel caspase-interacting proteins and provide new insights into the regulation of caspase pathways leading to cell death in normal and cancer cells.

  19. 7-ketocholesterol induces apoptosis of MC3T3-E1 cells associated with reactive oxygen species generation, endoplasmic reticulum stress and caspase-3/7 dependent pathway

    Directory of Open Access Journals (Sweden)

    Yuta Sato

    2017-03-01

    Full Text Available Type 2 diabetes mellitus (T2DM is associated with an increased risk of bone fractures without reduction of bone mineral density. The cholesterol oxide 7-ketocholesterol (7KCHO has been implicated in numerous diseases such as atherosclerosis, Alzheimer's disease, Parkinson's disease, cancer, age-related macular degeneration and T2DM. In the present study, 7KCHO decreased the viability of MC3T3-E1 cells, increased reactive oxygen species (ROS production and apoptotic rate, and upregulated the caspase-3/7 pathway. Furthermore, these effects of 7KCHO were abolished by pre-incubation of the cells with N-acetylcysteine (NAC, an ROS inhibitor. Also, 7KCHO enhanced the mRNA expression of two endoplasmic reticulum (ER stress markers; CHOP and GRP78, in MC3T3-E1 cells. Pre-incubation of the cells with NAC suppressed the 7KCHO-induced upregulation of CHOP, but not GRP78. In conclusion, we demonstrated that 7KCHO induced apoptosis of MC3T3-E1 cells associated with ROS generation, ER stress, and caspase-3/7 activity, and the effects of 7KCHO were abolished by the ROS inhibitor NAC. These findings may provide new insight into the relationship between oxysterol and pathophysiology of osteoporosis seen in T2DM.

  20. Decreased rate of protein synthesis, caspase-3 activity, and ubiquitin-proteasome proteolysis in soleus muscles from growing rats fed a low-protein, high-carbohydrate diet.

    Science.gov (United States)

    Batistela, Emanuele; Pereira, Mayara Peron; Siqueira, Juliany Torres; Paula-Gomes, Silvia; Zanon, Neusa Maria; Oliveira, Eduardo Brandt; Navegantes, Luiz Carlos Carvalho; Kettelhut, Isis C; Andrade, Claudia Marlise Balbinotti; Kawashita, Nair Honda; Baviera, Amanda Martins

    2014-06-01

    The aim of this study was to investigate the changes in the rates of both protein synthesis and breakdown, and the activation of intracellular effectors that control these processes in soleus muscles from growing rats fed a low-protein, high-carbohydrate (LPHC) diet for 15 days. The mass and the protein content, as well as the rate of protein synthesis, were decreased in the soleus from LPHC-fed rats. The availability of amino acids was diminished, since the levels of various essential amino acids were decreased in the plasma of LPHC-fed rats. Overall rate of proteolysis was also decreased, explained by reductions in the mRNA levels of atrogin-1 and MuRF-1, ubiquitin conjugates, proteasome activity, and in the activity of caspase-3. Soleus muscles from LPHC-fed rats showed increased insulin sensitivity, with increased levels of insulin receptor and phosphorylation levels of AKT, which probably explains the inhibition of both the caspase-3 activity and the ubiquitin-proteasome system. The fall of muscle proteolysis seems to represent an adaptive response that contributes to spare proteins in a condition of diminished availability of dietary amino acids. Furthermore, the decreased rate of protein synthesis may be the driving factor to the lower muscle mass gain in growing rats fed the LPHC diet.

  1. Addition of exogenous NAD+ prevents mefloquine-induced neuroaxonal and hair cell degeneration through reduction of caspase-3-mediated apoptosis in cochlear organotypic cultures.

    Directory of Open Access Journals (Sweden)

    Dalian Ding

    Full Text Available BACKGROUND: Mefloquine is widely used for the treatment of malaria. However, this drug is known to induce neurological side effects including depression, anxiety, balance disorder, and sensorineural hearing loss. Yet, there is currently no treatment for these side effects. PRINCIPAL FINDINGS: In this study, we show that the coenzyme NAD(+, known to play a critical role in maintaining the appropriate cellular redox environment, protects cochlear axons and sensory hair cells from mefloquine-induced degeneration in cultured rat cochleae. Mefloquine alone destroyed hair cells and nerve fiber axons in rat cochlear organotypics cultures in a dose-dependent manner, while treatment with NAD(+ protected axons and hair cells from mefloquine-induced degeneration. Furthermore, cochlear organs treated with mefloquine showed increased oxidative stress marker levels, including superoxide and protein carbonyl, and increased apoptosis marker levels, including TUNEL-positive nuclei and caspases-3. Treatment with NAD(+ reduced the levels of these oxidative stress and apoptosis markers. CONCLUSIONS/SIGNIFICANCE: Taken together, our findings suggest that that mefloquine disrupts the cellular redox environment and induces oxidative stress in cochlear hair cells and nerve fibers leading to caspases-3-mediated apoptosis of these structures. Exogenous NAD(+ suppresses mefloquine-induced oxidative stress and prevents the degeneration of cochlear axons and sensory hair cells caused by mefloquine treatment.

  2. Liquiritin (LT) exhibits suppressive effects against the growth of human cervical cancer cells through activating Caspase-3 in vitro and xenograft mice in vivo.

    Science.gov (United States)

    He, She-Hong; Liu, Hong-Gai; Zhou, Yu-Fei; Yue, Qing-Fen

    2017-08-01

    Cervical cancer is one of the most common female malignancies worldwide. Liquiritin (LT), a major constituent of Glycyrrhiza Radix, possesses a variety of pharmacological activities, including anti-cancer, anti-oxidative, anti-inflammatory and neuro-protective effects. However, its role in human cervical cancer remains to be elusive. In our study, we found that LT suppressed cervical cancer cell migration, invasion and cloning ability with little cytotoxicity to human normal cells. In addition, apoptosis was induced by LT in cervical cancer cells through activation of Caspase-3 and poly ADP-ribose polymerase (PARP) cleavage. LT-triggered apoptosis was dependent on extrinsic and intrinsic pathways, which were relied on Fas-associated protein with death domain (FADD)- and Bcl-2/Bax-regulated pathways, leading to Caspase-8 and Caspase-9 cleavage, respectively. LT was found to increase FADD expression, while reduce Bcl-2 expression, contributing to Caspase-3 cleavage. And tumor suppressors, p21 and p53, were enhanced after LT treatment, inhibiting the growth of cervical cancer cells in vitro. Significantly, in vivo study suggested that tumor growth was impeded by LT in a dose-dependent manner through enhancing apoptosis. Together, the data here revealed that LT was an effective and promising candidate for preventing human cervical cancer progression via apoptosis enhancement. Copyright © 2017. Published by Elsevier Masson SAS.

  3. Matrine inhibits diethylnitrosamine-induced HCC proliferation in rats through inducing apoptosis via p53, Bax-dependent caspase-3 activation pathway and down-regulating MLCK overexpression.

    Science.gov (United States)

    Zhang, Xiaolin; Yu, Hao

    2016-01-01

    The proliferation of hepatocellular carcinoma (HCC) cells is one of the leading causes of liver cancer mortality in humans. The inhibiting effects of matrine on HCC cell proliferation have been studied, but the mechanism of that inhibition has not been fully elucidated. Since, apoptosis plays an important role in HCC cell proliferation. We examined the apoptosis-inducing effect of matrine on tumor cells. Western blot analysis of p53, Bax, cleaved caspase-3 and myosin light chain kinase (MLCK) revealed that matrine induced tumor cell apoptosis by controlling anoikis. It activated p53, Bax-dependent caspase-3 and blocked the ECM-integrin mediated cell survival pathway through down-regulating MLCK over-expression in the liver of rats with diethyl nitrosamine (DENA)-induced HCC. Our results suggest that matrine can inhibit the proliferation of HCC cells through inducing tumor cell apoptosis via activation of the p53 pathway and inhibition of MLCK overexpression. Matrine may thus be used as a potentially promising reagent to inhibit HCC cell proliferation and MLCK may be a novel target for the treatment of HCC.

  4. Linoleic acid derivative DCP-LA protects neurons from oxidative stress-induced apoptosis by inhibiting caspase-3/-9 activation.

    Science.gov (United States)

    Yaguchi, Takahiro; Fujikawa, Hirokazu; Nishizaki, Tomoyuki

    2010-05-01

    The present study aimed at understanding the effect of the linoleic acid derivative 8-[2-(2-pentyl-cyclopropylmethyl)-cyclopropyl]-octanoic acid (DCP-LA) on oxidative stress-induced neuronal death. Sodium nitroprusside (SNP; 1 mM) reduced viability of cultured rat cerebral cortical neurons to 50% of basal levels, but DCP-LA significantly prevented the SNP effect in a concentration (1-100 nM)-dependent manner. In addition, DCP-LA (100 nM) rescued neurons from SNP-induced degradation. SNP (1 mM) activated caspase-3 and -9 in cultured rat cerebral cortical neurons, but DCP-LA (100 nM) abolished the caspase activation. For a mouse model of middle cerebral artery occlusion, oral administration with DCP-LA (1 mg/kg) significantly diminished degraded area due to cerebral infarction. The results of the present study, thus, demonstrate that DCP-LA protects neurons at least in part from oxidative stress-induced apoptosis by inhibiting activation of caspase-3/-9.

  5. Tracing the accumulation and effects of mercury uptake in the previtellogenic ovary of crucian carp, Carassius auratus gibelio by autometallography and caspase-3 immunohistochemistry.

    Science.gov (United States)

    Zarnescu, Otilia

    2009-02-01

    The aims of the present study were to apply the AMG technique for localization of mercury at the light and electron microscopic level in the ovary of crucian carp after exposure to mercuric chloride and to find out if this heavy metal induces expression of caspase-3. Depending on the stage of ovarian follicle development, two patterns of mercury accumulation have been found in previtellogenic ovary of crucian carp. The first mercury accumulation pattern has been found in the early previtellogenic oocyte without zona radiata. In these oocytes, mercury accumulates into an ooplasmic region that seems to correspond to the Balbiani body (32-65 microm oocyte diameter), throughout the cytoplasm (84-116 microm oocyte diameter) and in the cortical cytoplasm (approximately 180 microm oocyte diameter). The second mercury accumulation pattern has been found in the late previtellogenic oocyte with cortical alveoli (229-330 microm oocyte diameter). Ultrastructural observations have shown grains of silver-enhanced mercury inside coated vesicles, the cortical lysosome-like bodies or multivesicular bodies and cortical alveoli. Immunohistochemistry reaction for caspase-3 was positive in nuclei of the early previtellogenic oocyte and Balbiani body.

  6. nNOS抑制剂下调局灶性脑缺血半暗带和核心区Calpain和Caspase-3的活化

    Institute of Scientific and Technical Information of China (English)

    孙明; 赵育梅; 徐超

    2008-01-01

    目的:一氧化氮合酶(NOS)分为三种亚型,即神经元型(nNOS)、内皮细胞型(eNOS)和诱导型(iNOS)。7-硝基吲唑(7-NI)对纯化的三种亚型的NOS都有抑制作用,但在体内,它对nNOS具有较高的选择性。体外研究表明一氧化氮(NO)可影响半胱氨酸蛋白酶Calpain和Caspase-3的活性,本研究观察了7-NI对局灶性脑缺血后Calpain及Caspase-3活性的影响。方法:采用动脉腔内插线制备大鼠局灶性脑缺血模型,

  7. Peperphentonamine hydrochloride protects against gentamicin-induced cochlea damage by lowering cochlear caspase-3 expression in guinea pigs%盐酸椒苯酮胺通过降低caspase-3表达减轻庆大霉素豚鼠耳蜗损伤

    Institute of Scientific and Technical Information of China (English)

    陈浩; 谢民强; 吴剑; 李威; 李永贺

    2014-01-01

    Objective To study the protective effect of peperphentonamine hydrochloride (PPTA) against gentamicin-induced cochlear damage and its mechanism to inhibit cell apoptosis. Methods Guinea pigs with normal hearing were randomized into control, gentamicin, and PPTA treatment groups, and the guinea pigs models of gentamicin-induced cochlear damage received intraperitoneal injection of PPTA. The changes of hearing of the guinea pigs were evaluated with auditory brainstem response (ABR) test, and the protein expression of caspase-3 in the cochlear tissue was detected using Western blotting. TUNEL staining, scanning and transmission electron microscopy were performed to observe the morphological changes of the cochlea. Results The threshold in ABR in PPTA treatment group was significantly higher than that in the control group (P<0.05) but significantly lower than that in gentamicin group. Western blotting showed a significantly increased caspase-3 expression in gentamicin group (P<0.001); caspase-3 expression in PPTA group was obviously higher than that in the control group but much lower than that in gentamicin group (P<0.001). TUNEL assay and electron microscopy revealed serious damages of the hair cells in gentamicin group with numerous apoptotic cells in the organ of Corti, stria vascularis and spiral ganglion, and such cochlear damages were obviously alleviated in PPTA group. Conclusion PPTA can protect against gentamicin-induced cochlear damage in guinea pigs by decreasing the protein expression of caspase-3 to inhibit cell apoptosis.%目的:研究盐酸椒苯酮胺(PPTA)对庆大霉素耳蜗损伤的保护作用及抗凋亡机制。方法听力正常豚鼠分3组:正常组、GM组和PPTA组。采用庆大霉素致豚鼠耳蜗损伤模型,PPTA腹腔注射,ABR分析听力变化,Western blot检测耳蜗组织中caspase-3蛋白表达,TUNEL染色、扫描电镜和透射电镜观察形态学改变。结果 ABR反应阈:GM组、PPTA组显

  8. 补肾益冲抗衰汤对卵巢储备功能下降大鼠卵巢颗粒细胞凋亡因子Bcl-2、Bax、Caspase-3的影响%The Effect of Bushenyichongkangshuai Decoction on Ovarian Granule Cell Apoptosis Factors Bcl-2,Bax,Caspase-3 of DOR Rat Model

    Institute of Scientific and Technical Information of China (English)

    张妙; 邱晓晓; 朱长玲; 朱程芬; 程泾

    2014-01-01

    [目的]观察补肾益冲抗衰汤对卵巢储备功能下降模型大鼠卵巢颗粒细胞凋亡因子Bcl-2、Bax、Caspase-3蛋白表达的影响。[方法]以70只8周龄雌性SD大鼠为研究对象,采用雷公藤多苷片制作卵巢储备功能下降大鼠模型。观察大鼠一般情况、卵巢指数、卵巢组织病理形态、血清E2、FSH、FSH/LH水平及卵巢颗粒细胞凋亡因子Bcl-2、Bax、Caspase-3蛋白表达。[结果]补肾益冲抗衰汤能使卵巢储备功能下降大鼠动情周期缩短、子宫、卵巢重量增加,大鼠血清FSH水平、FSH/LH降低,血清E2水平增高(P<0.05或P<0.01),使卵巢颗粒细胞抑制细胞凋亡因子Bcl-2蛋白表达增加,促进细胞凋亡因子Bax、Caspase-3蛋白表达减少(P<0.05或P<0.01)。[结论]雷公藤多甙片致DOR大鼠模型与人类卵巢储备功能下降的临床表现基本一致。细胞凋亡调节因子Bcl-2、Bax、Caspase-3蛋白与卵巢储备功能密切相关,补肾益冲抗衰汤有抑制卵巢颗粒细胞凋亡、减少卵泡闭锁从而改善卵巢储备功能的作用。%[Objective]To investigate the effect of Bushenyichongkangshuai decoction on ovarian granule cellapoptosis Bcl-2, Bax, Caspase-3 of DOR rat modeled by Tripterygium. [Methods]8-week old 70 healthy female rats as the research object ,rat models were made of Tripterygium,the rats estrus cycle, fol icle stimulating hormone(FSH), luteinizing hormone(LH), estrogen(E2), the morphological changes of the ovary and uterus, ovarian granule cellapopto-sis factors Bcl-2, Bax and Caspase-3 were detected. [Results] Bushenyichongkangshuai can increase emotional cycles, uterine, ovarian weight, change serum FSH, FSH/LH, E2(P<0.05 or P<0.01),and impact ovarian granule cells apoptosis factors Bcl-2, and Bax in caspase-3 has significant differences( P<0.05 or P<0.01). [Conclusion] The diminished ovarian reserve model rats set up with Tripterygium wilfordii is corresponding to DOR typical performance of

  9. 丹参注射液对缺氧神经干细胞凋亡和Caspase-3活性的影响%Effects of salvia miltiorrhizae injection on hypoxia-induced apoptosis or cultured rat neuronal stem cells and activity of Caspase-3

    Institute of Scientific and Technical Information of China (English)

    黄涛; 韩富; 张志强; 谭齐家; 谢才军; 谢绍盈; 朱灿辉

    2008-01-01

    Objective To explore the effects of salvia miltiorrhizae (SM) injection on the apoptosis of cultured rat neuronal stem cells induced by hypoxia and the activity of Caspase-3, in order to provide the further evidence for the molecular mechanism of neuroprotection of SM injection. Methods The neuronal stem cells from neonatal rat hippocampus were cultured and divided randomly into normal control group, hypoxia group and SM treatment group. After Hoechst staining, the apoptotic morphological change and apoptosis percentage were observed under fluorescence microscope. The activities of Caspase-3 in the 3 groups were evaluated by the colorimetric assay. Results Compared with normal control group [(2.75±0.28)%, 1.16±0.07], the percentage of apoptosis and the activity of Caspase-3 were increased significantly in neuronal stem cells cultured in hypoxia [(30.12%±2.09)%,3.85±0.41, P<0.05). Application of SM injection reduced markedly the percentage of apoptosis and the activity of Caspase-3 of the neuronal stem cells cultured in hypoxia [(9.16±1.34)%, 1.50±0.09, P<0.05].Conclusion SM injection can depress the apoptosis of the rat neuronal stem cells induced by hypoxia,so as to exert the neuroprotection.%目的 探讨丹参注射液对缺氧培养大鼠神经干细胞的凋亡及Caspase-3活性的影响,以进一步明确丹参注射液神经保护作用的分子机制.方法 体外培养新生大鼠海马神经干细胞,将其分为正常对照组,缺氧培养组及丹参注射液处理组.Hoechst染色后荧光显微镜下观察并计算细胞凋亡率:比色法检测各组细胞Caspase-3的相对活性.结果 缺氧培养大鼠神经干细胞的细胞凋亡率(30.12%±2.09%)及Caspase-3活性(3.85±0.41)均较正常对照组(2.75%±0.28%,1.16±0.07)明显升高,差异有统计学意义(P<0.05);施加丹参注射液后,大鼠神经干细胞的细胞凋亡率(9.16%±1.34%)和Caspase-3活性(1.50±0.09)均较缺氧培养组明显下降,差异有统计学意义(P<0

  10. 舒肝解郁胶囊对抑郁模型大鼠海马神经元凋亡及脑组织caspase-3蛋白表达的影响%Effect of Shuganjieyu capsules on neuronal apoptosis in hippocampal CA3 area and the expression of caspase-3 in the brain of rat depression model

    Institute of Scientific and Technical Information of China (English)

    傅锦华; 刘勇; 王清勇; 赵靖平

    2012-01-01

    Objective: To evaluate the effect of "Shuganjieyu" (SGJY) capsules on neuronal apoptosis in hippocampal CA3 area and the expression of caspase-3 in the brain of rat depression model, and to investigate its pharmacological mechanisms in depression treatment. Methods: Adult male SD rats were randomly divided into 4 groups: a control, a model, a SGJY and a fluoxetine group. The rat depression model was established under chronic unpredictable mild stress (CUMS) and separate feeding. The behaviors were measured by open-field test, sucrose consumption and forced swimming test. We observed the neuronal morphology structure and neuronal apoptosis in the hippocampal CA3 area. We detected the rat caspase-3 expression level of medial prefrontal cortex ( mPFC) and hippocampal CA3 area by Western blot. Results: After 21-day stress, compared with the model group, spontaneous activity and sucrose consumption and preference percentage of the rats in the SGJY group significantly increased, while the immobility time in forced swimming test, the number of apoptotic cells and the protein levels of caspase-3 significantly reduced (P0.05). Conclusion: SGJY capsules can reduce the depression symptoms of CUMS and help to increase hippocampal neuron generation, survival and neogenesis, reduce the protein levels of caspase-3, and reverse neurocyte apoptosis in the rat depression model with the same efficacy as fluoxetine.%目的:研究舒肝解郁胶囊对抑郁模型大鼠海马神经元凋亡及脑组织caspase-3蛋白表达的影响,探讨其治疗抑郁症的作用机制.方法:将雄性SD大鼠随机分为正常对照组、模型组、舒肝解郁组和氟西汀组四组;采用慢性轻度不可预见性应激(CUMS)结合孤养建立抑郁大鼠模型,并用旷场、糖水消耗和强迫游泳试验评价大鼠的行为学改变,观察海马CA3区神经元的形态结构及凋亡,应用蛋白印记分析检测脑组织caspase-3蛋白的表达.结果:与模型组比较,舒肝解郁

  11. 丹参酮Ⅱa对实验性自身免疫性脑脊髓炎大鼠大脑GFAP和Caspase-3表达的影响%Effects of Tanshinone ⅡA on the expression of GFAP and caspase-3 in the brain of experimental autoim- mune encephalomyelitis rats

    Institute of Scientific and Technical Information of China (English)

    闫俊; 杨雪; 冯娟

    2016-01-01

    目的 观察丹参酮ⅡA对实验性自身免疫性脑脊髓炎(EAE)大鼠的治疗作用以及对大脑中胶质纤维酸性蛋白(GFAP)和半胱氨酸天冬氨酸蛋白酶3(Caspase-3)表达的影响.方法 把40只大鼠随机分成4组,空白对照组(Naive组)、EAE高剂量用药组(50 mg/kg,TSⅡA-H组)、EAE低剂量用药组(25 mg/kg,TSⅡA-L组)和EAE溶剂对照组(Vehicle组).EAE模型诱导后每天对大鼠进行神经功能评分和体质量测量.发病高峰期(第18天)处死大鼠并取材,分别进行H E染色和劳克坚牢蓝(LFB)染色检测大鼠脑组织炎性浸润改变和脊髓脱髓鞘情况.采用免疫组化法和免疫印迹法(West-ern Blot)检测各组大鼠脑组织中GFAP和Caspase-3的表达水平.结果 用药后TSⅡA-H组和TSⅡA-L组大鼠临床症状好转,大脑炎性细胞浸润减少,脊髓脱髓鞘的改变减轻,大脑中GFAP和Caspase-3的表达水平下降.结论 丹参酮ⅡA对EAE大鼠有治疗作用,其作用机制可能是通过下调GFAP和Caspase-3的表达来参与炎性反应和凋亡过程的调节.%Objective To observe the treatment effect of Tanshinone ⅡA on experimental autoim-mune encephalomyelitis rats and the expression of GFAP and caspase -3 in the brain.Methods Forty rats were randomly divided into four groups ,blank control group (Naive) ,high dose treatment group (TSⅡA -H) ,low dose treatment group (TSⅡA -L) and solvent control group (Vehicle).After in-duction of EAE model ,nerve function scores and weight were measured everyday.All the rats were kill-ed and drawn at the peak of the disease (the 18th day).HE statining and Lauck Fast Blue(LFB) staining were separately performed to observe the inflammatory infiltration and spinal cord demyelination situa-tion.The level of GFAP and caspase-3 in brain was detected via immunohistochemistry and immunobo-ltting (western blot).Results Compared to the Vehicle group rats ,better clinical symptoms ,less infil-tration of inflammatory cells and lower

  12. Effects of Compound Radix Sophorae Flavescentis injection on proliferation, apoptosis and Caspase-3 expression in adenoid cystic carcinoma ACC-2 cells%复方苦参注射液对泪腺腺样囊性癌ACC-2细胞增殖、凋亡及Caspase-3蛋白表达的影响

    Institute of Scientific and Technical Information of China (English)

    石博; 徐慧

    2012-01-01

    目的 探讨复方苦参注射液对泪腺腺样囊性癌ACC-2细胞增殖、凋亡及Caspases-3蛋白表达的影响.方法 体外培养人泪腺腺样囊性癌ACC-2细胞,应用MTT法检测细胞增殖;Annexin V/PI双染色流式细胞仪检测细胞凋亡和细胞周期;ELISA法检测Caspases-3蛋白表达.结果 复方苦参注射液对ACC-2细胞的体外增殖具有抑制作用,量效关系显著,与对照组比较有统计学差异(P<0.01),半数抑制浓度(IC50)为0.84 g/ml.经流式细胞仪检测表明,复方苦参注射液能使ACC-2细胞G0-G1期逐渐增加,G2-M期和S期逐渐减少,并且随着剂量的增加,ACC-2细胞凋亡率明显增加(P<0.05或P<0.01).复方苦参注射液能增强ACC-2细胞Caspases-3蛋白的表达(P<0.05或P<0.01),并呈剂量依赖性.结论 复方苦参注射液能有效抑制人泪腺腺样囊性癌ACC-2细胞Caspases-3蛋白表达,诱导ACC-2细胞凋亡,抑制肿瘤细胞增殖.%Objective To investigate the effects of compound Radix Sophorae Flavescentis injection on proliferation, apoptosis and Caspase-3 expression in human adenoid cystic carcinoma ACC-2 cells. Methods ACC-2 cells were cultured in vitro. MTT assay was used to measure the cell proliferative effect. The Annexin V/PI double staining analysis by flow cytometry was used to evaluate apoptotic rate and the cell cycle. The expression of Caspases-3 protein was detected by enzyme-linked immunosorbent assay ( ELISA ). Results Compound Radix Sophorae Flavescentis injection could inhibit the proliferation of ACC-2 cells in vitro,and dose-effect relationship was significant P<0. 01 ). IC50 of ACC-2 was 0. 84 g/ml. The flow cytometry test indicated, compound Radix Sophorae Flavescentis injection could make ACC-2 cells Go-G, phase gradually increasing, G2-M period and S phase reduce gradually, and with the increase of the dose, ACC-2 cell apoptosis rate increased significantly ( P<0. 05 or P<0.01 ). Compound Radix Sophorae Flavescentis injection could enhance ACC-2

  13. Clinicopathological significance and relations of Caspase-3 expression, cell proliferation and apoptosis in gastric cancer and the precancerous lesions%乳腺癌钼靶X线表现特征(118例分析)

    Institute of Scientific and Technical Information of China (English)

    XiaoBin Hu; Jing Zhao; Lin Yang; Yan Xin

    2009-01-01

    Objective: we investigated the relationship between the expression of Caspase-3. cell proiferation and apoptosis in gastric cancer and their precancerous lesions, to explore the tumorigenesis of the stomach mucosa. Methods: Caspase-3 expression in 13 normal gastric mucosa, 6 chronic atrophic gastritis (CAG), 31 intestinal metaplasia (IM), 114 dysplasia (DYS) and 20 gastric carcinomas were investigated immunohistochemically. Cell proliferation was evaluated with anti-Ki-67immunostaining and apoptosis was evaluated using DNA fragmentation in situ by TdT-mediated dUTP biotin nick end label-ing (TUNEL) method. Results: Caspase-3 mild-moderately positive expression was observed in most of normal superficial epithelia, its positively polar distribution in normal mucosa, CAG, IM, DYS and gastric carcinomas changed as seen in TU-NEL, and so did the positive rate. Caspase-3 protein expression showed significantly positive correlation with the number of apoptotic cells labeled with TUNEL (correlation coefficient r=0.94; P 0.05). Conclusion: Caspase-3 protein expression was up-regulated from CAG to IM and mild-moderate atypical dysplasia, but down-regulated in severe dysplasia and gastric carcinoma, indicating that inactivity or reduced expression of Caspase-3 is closely correlated with carcinogenesis of the stomach mucosa.

  14. 过氧化氢体外诱导细粒棘球蚴原头节细胞Caspase-3表达及超微结构改变的影响%Effect of hydrogen peroxide on the expression of caspase-3 and changes of ultra- structures in protoscolex cells of Echinococcus granulosus in vitro

    Institute of Scientific and Technical Information of China (English)

    胡汉华; 康金凤; 陈蓉; 白山别克; 艾赛提; 汤建安; 谢风莲

    2009-01-01

    目的 探讨过氧化氢(H2O2)体外诱导细粒棘球蚴原头节细胞凋亡、Caspase-3表达和细胞超微结构的影响.方法 RPMI1640添加谷氨酰胺组即体外培养细粒棘球蚴原头节,用5mmol/L H2O2诱导8h,使其发生细胞凋亡.用原位末端脱氧核糖核苷酸转移酶标记技术(TUNEL 法)检测原头节细胞凋亡情况,用过氧化物酶标记链霉卵白素(SP)染色半胱天冬氨酸蛋白酶-3(caspase-3).在透射电镜下观察原头节细胞超微结构的变化.结果 过氧化氢诱导原头节细胞凋亡细胞增加,caspase-3表达增加,电镜观察原头节细胞异染色质增加,部分细胞染色质异常浓缩呈现凋亡细胞征象.结论 H2O2可诱导细粒棘球蚴原头节细胞凋亡,且caspase-3参与原头节细胞的凋亡.

  15. Effect of antisense oligodeoxyribonucleotides targeting at HER-2 mRNA on Caspase-3 protein expression in SK-BR-3 breast cancer cells%靶向HER-2 mRNA反义寡核苷酸对SK-BR-3乳腺癌细胞Caspase-3蛋白表达的影响

    Institute of Scientific and Technical Information of China (English)

    杨栓平; 宋海峰; 宋三泰; 郑晓玲; 张凤霞

    2003-01-01

    目的检测HER-2特异性的反义寡核苷酸HA824对SK-BR-3乳腺癌细胞Caspase-3蛋白表达的影响.方法选用HER-2 高表达的SK-BR-3乳腺癌细胞株作为试验的细胞株.HA824合成如前所述,HA4为已报道的阳性序列,Scramble设定为随机对照序列.上述药物分别以浓度为200 nmol*L-1孵育SK-BR-3细胞8 h和36 h,之后采用逆转录PCR法与免疫细胞化学技术检测SK-BR-3细胞HER-2 mRNA 及Caspase-3蛋白表达水平.结果与Scramble对照序列相比,HA824、HA4能抑制HER-2 mRNA的表达,HA824作用更强;与Scramble 相比,HA824、HA4对Caspase-3蛋白表达水平则无影响.结论 HA824、HA4这两种靶向HER-2 mRNA反义药物对SK-BR-3乳腺癌细胞株增殖的抑制作用可能与激活Caspase-3蛋白无关.

  16. Effects of Modified Gandou Decoction on Protein and mRNA Expression of X-linked Inhibitor of Apoptosis, Caspase-9, and Caspase-3 in Brain Tissue of TX Mice%肝豆汤改良方对TX小鼠脑组织XIAP、Caspase-9和Caspase-3蛋白及其mRNA表达的影响

    Institute of Scientific and Technical Information of China (English)

    赵雯; 马艳红; 韩咏竹; 程楠; 饶娆; 王训

    2015-01-01

    目的 探究肝豆汤改良方抑制肝豆状核变性(Wilson's disease,WD)脑神经细胞凋亡的分子机制.方法 选取1月龄TX小鼠120只,随机分为TX模型组(40只)、肝豆汤改良方组(40只)、丁苯酞组(20只),另选DL小鼠40只作为正常组;每组再分成2组,各20只,分别予以生理盐水、肝豆汤改良方、丁苯酞灌胃2个月和4个月.分别采用免疫组织化学法和Western blot法检测小鼠脑组织中X-连锁凋亡抑制蛋白(X-linked inhibitor of apoptosis,XIAP)、胱冬肽酶-9(Caspase-9)及胱冬肽酶-3(Caspase-3)蛋白表达水平,采用RT-PCR技术检测小鼠脑组织中XIAP、Caspase-9及Caspase-3 mRNA表达水平.结果 免疫组织化学检测显示,肝豆汤改良方组XIAP表达水平较同月龄模型组明显增多,Caspase-9和Caspase-3表达水平较同月龄模型组下降,5月龄和3月龄肝豆汤改良方组XIAP和Caspase-3表达水平有所差异.Western blot检测结果显示,月龄因素对脑组织XIAP、Caspase-9、Caspase-3蛋白表达水平的主效应均无统计学意义(P>0.05);月龄因素和分组因素对3种蛋白表达水平的交互作用均无统计学意义(P>0.05);对于同月龄小鼠,模型组脑组织XIAP表达水平显著低于正常组(P<0.05),Caspase-9和Caspase-3蛋白表达水平显著高于正常组(P<0.05);肝豆汤改良方组XIAP表达水平显著高于模型组(P<0.05),Caspase-9和Caspase-3蛋白表达水平显著低于模型组(P<0.05).RT-PCR检测结果显示,月龄因素对小鼠脑组织XIAP mRNA表达水平的主效应无统计学意义(P>0.05),但对Caspase-9、Caspase-3 mRNA表达水平的主效应具有统计学意义(P<0.05);月龄因素和分组因素对XIAP、Caspase-9、Caspase-3 mRNA表达水平的交互作用均无统计学意义(P>0.05).对于相同月龄小鼠,模型组脑组织XIAP mRNA表达水平显著低于正常组(P<0.05),而Caspase-9、Caspase-3 mRNA表达水平显著高于正常组(P<0.05);与模型组比较,丁苯酞

  17. 止咳化痰苗药对慢性阻塞性肺疾病大鼠肺组织Caspase-3蛋白及mRNA表达的影响%Effects of the traditional Chinese MIAO Nationality's herbs of expectorant cough on Caspase-3 expression in rats with COPD

    Institute of Scientific and Technical Information of China (English)

    孔令雯; 葛正行; 李常

    2012-01-01

    目的:研究Caspase-3基因在大鼠慢性阻塞性肺疾病(COPD)模型肺组织中的表达同时给予苗药干预治疗,探讨Caspase-3基因在COPD发病中的作用与意义及止咳化痰苗药作用机制.方法:雄性SD大鼠60只,随机分为慢性阻塞性肺疾病组(COPD组)、苗药治疗组(苗药组)和正常对照组(对照组),每组20只.COPD组、苗药组采用每日熏香烟和每月2次气管内滴入脂多糖法制作COPD大鼠模型,苗药组在造模30d起给苗药灌胃治疗.通过半定量病理分析的方法观察大鼠肺组织损伤的变化;用免疫组化和RT-PCR检测肺中Caspase-3蛋白及其mRNA表达.结果:COPD组及苗药组平均肺内衬间隔(MLI)较对照组明显增加,平均肺泡数(MAN)较对照组明显下降,COPD组与苗药组比较有明显差异(P<0.05);COPD模型组大鼠肺Caspase-3蛋白阳性表达率明显高于对照组及苗药组;Caspase-3 mRNA半定量检测显示COPD组高于对照组及苗药组,苗药组与COPD组相比较有明显差异(P<0.05).各组Caspase-3 mRNA的表达与MLI成正相关,与MAN表达成负相关.结论:Caspase-3与慢性阻塞性肺疾病的病理过程的形成可能有关,止咳化痰苗药有可能通过抑制细胞凋亡而缓解COPD病理进程.%Objective: To investigate the expression and significance of caspase-3 in rats with chronic obstructive pulmonary(COPD), meanwhile, to study the expression changes of caspase-3 after treaded with Miao Nationality's herbs and fing out the mechanism of action of it.Methods: 60 male SD rats were randomly divided into the COPD group, Miao herb group and control group. The COPD rat model was established by exposure to cigarette smoking and inhalation of lipopolysaccharide twice a month, Miao herb group was treated with gavage of Miao herbs 30 days. By semi-quantitative pathological analysis, we tnesure the pathological changes of lung tissues.Caspase-3 protein and mRNA expression were detected by immunohistochemistry and RT

  18. The Influence of Saffron in Expression of Apoptosis Related Protein Caspase-3, Bcl-2 in Rats with Liver Fibrosis%藏红花对肝纤维化大鼠肝脏组织中Caspase-3,Bcl-2表达的影响

    Institute of Scientific and Technical Information of China (English)

    梅夏齐; 汪云; 王风秀; 杨培青

    2016-01-01

    目的:研究藏红花(Saffron)对肝纤维化大鼠肝组织中半胱氨酸天冬氨酸蛋白酶-3(Caspase-3),B细胞淋巴瘤/白血病-2(Bcl-2)表达的影响.方法:45只雄性SD大鼠随机均分为3组:正常组、模型组、藏红花组,除正常组外,另外两组给予40%四氯化碳橄榄油溶液腹腔注射制备肝纤维化大鼠模型,在造模的同时,藏红花组给予藏红花溶液5 mL·kg-1·d-1灌胃.8周后处死大鼠,留取肝脏组织,通过Masson染色观察大鼠肝纤维化的形成,免疫组化方法检测肝组织中Caspase-3及Bcl-2蛋白表达.结果:模型组大鼠肝纤维化程度最重,藏红花组较模型组纤维化程度轻,正常组大鼠肝脏无纤维化.与模型组比,藏红花组Caspase-3在肝实质内呈弱阳性表达,而纤维间隔内表达增加;Bcl-2的表达明显减少(P<0.05),正常组Caspase-3及Bcl-2几乎无表达.结论:藏红花具有抗肝纤维化作用,其机制可能与调节Caspase-3,Bcl-2的表达有关.

  19. Correlation of Thyroid Peroxidase Antibody and the Expression of TRAIL,Caspase-3 in Hashimoto Thyroiditis%桥本甲状腺炎中TRAIL、caspase-3的表达与甲状腺过氧化物酶抗体的相关性研究

    Institute of Scientific and Technical Information of China (English)

    黄凌宁; 杨立勇; 张声; 郑俊敏; 林华; 陈于鹏

    2011-01-01

    目的 分析肿瘤坏死因子相关凋亡诱导配体(tumor necrosis factor related apoptosis inducing ligand,TRAIL)、半胱天冬酶-3(caspase-3)在桥本甲状腺炎(Hashimoto thyroiditis,HT)中的表达情况,以及与甲状腺过氧化物酶抗体(Thyroid peroxidase antibody TPOAb)的相关性研究.方法采用链霉抗生物素蛋白-过氧化物酶法检测51例桥本甲状腺炎(Hashimoto thyroiditis,HT)患者的甲状腺标本的TRAIL、caspase-3的表达情况,同时检测血清TPOAb,并以32份正常甲状腺标本作为对照,比较2组的差异.结果①HT的甲状腺滤泡上皮细胞中TRAIL、caspase-3的表达阳性率分别为88.2%、96.1%,高于正常对照的21.8%、12.5%.(P<0.05)②HT患者血清的TPOAb的阳性检出率分别是90.2%,明显高于正常对照组12.5%.(P<0.05)③将HT的甲状腺滤泡上皮细胞中TRAIL、caspase-3的表达程度分级,与TPOAb滴度高低进行相关性分析,发现二者无相关关系.结论 HT患者中,TRAIL及caspase-3可能共同参与甲状腺滤泡细胞的破坏,参与HT的发病及病理发展过程.HT患者中血清TPOAb能否检出比其滴度高低更有意义.

  20. 宫颈癌术前介入化疗前后组织中Survivin、Caspase-3和Caspase-7的表达变化及意义%The significance of different expression of survivin,caspase-3 and caspase-7 in cervical carcinoma cells before and after the ameba femoralis interventional chemotherapy

    Institute of Scientific and Technical Information of China (English)

    杨斌; 陈赛英; 史佃云; 张佃乾

    2009-01-01

    目的:研究宫颈癌术前行动脉介入化疗前后组织中细胞凋亡相关蛋白Survivin、caspase-3、Caspase-7的表达变化及其临床意义.方法:对54例宫颈癌患者术前行介入化疗前宫颈活检组织和介入化疗后手术切除的标本,免疫组化检测Survivin、Caspase-3、Caspase-7的表达,同时检测38例正常宫颈组织以及28例宫颈上皮内瘤变组织作为对照.结果:Survivin在介入化疗前宫颈癌组织中呈高表达,介入化疗后Survivin呈现不同程度的下降,而凋亡相关蛋白Caspase-3、Caspase-7的表达则呈不同程度的上升.相关性分析表明,介入化疗后Survivin的表达变化与Caspase-3及Capase-7的表达变化呈负相关,介入前后3种蛋白表达变化越大,临床有效率越高.结论:Survivin高表达及Caspase-3、Caspase-7低表达在官颈癌的发生发展中有一定作用,宫颈癌患者术前动脉介入化疗可以通过促进肿瘤细胞的凋亡来达到抑制肿瘤生长、缩小肿瘤体积进而抑制其远处转移.

  1. Effects of electro-acupuncture on Noxa and caspase-3 expression in hippocampal CA1 region of a vascular dementia rat model

    Institute of Scientific and Technical Information of China (English)

    Yanzhen Zhu; Qiang Wu; Ling Lin

    2008-01-01

    BACKGROUND: Noxa, a pro-apoptotic member of the Bcl-2 protein family, has been shown to induce the mitochondrial pathway of apoptosis and to mediate hypoxic cell death in a rat model of cerebral ischcmia.This suggests that Noxa could participate in apoptosis during vascular dementia (VD).OBJECTIVE,: To detect Noxa and caspase-3 expression after electro-acupuncture in VD rats to further validate the mechanism of electro-acupuncture-induced effects in the treatment of VD.DESIGN, TIME AND SETTING: A randomized, controlled study was performed at the Center for the Neurobiology of Fujian Medical University between January 2006 and March 2007.MATERIALS: A total of forty adult, male, Sprague Dawley rats were included in this study. The following equipment was used: confocal laser scanning microscope (SpS, Leica, Germany), water matte (Bejing Suntendy Science and Technology Co., Ltd., China), and SDZ-Ⅱ electronic acupuncture treatment instruments (Suzhou Medical Appliance Factory, China).METHODS: Thirty-eight rats with sufficient learning and memory abilities were selected by Morris water maze criteria. Twelve rats received sham-surgery; the remaining 26 rats were used to establish a VD model by bilateral occlusion of the common carotid arteries. The rats that survived the occlusion procedure were randomly assigned into an electro-acupuncture group (n = 11) and a VD model group (n = 12).MAIN OUTCOME MEASURES: Neuropathological changes were observed with hematoxylin-eosin staining of the hippocampus and expression of Noxa and caspasc-3 in the hippocampal CAI region was analyzed by confocal laser scanning microscope following immunofluorescence staining.RESULTS: Expressions of Noxa and caspase-3 in the electro-acupuncture group and sham-operated group were less than in the VD model group (P < 0.01). Electro-acupuncture reduced the amount of apoptotic neurons in hippocampal CA1 area of rats with VD. The average latency in the Morris water maze test was significantly shorter

  2. FoxO3a governs early microglial proliferation and employs mitochondrial depolarization with caspase 3, 8, and 9 cleavage during oxidant induced apoptosis.

    Science.gov (United States)

    Shang, Yan Chen; Chong, Zhao Zhong; Hou, Jinling; Maiese, Kenneth

    2009-11-01

    Microglia of the central nervous system have a dual role in the ability to influence the survival of neighboring cells. During inflammatory cell activation, microglia can lead to the disposal of toxic cellular products and permit tissue regeneration, but microglia also may lead to cellular destruction with phagocytic removal. For these reasons, it is essential to elucidate not only the underlying pathways that control microglial activation and proliferation, but also the factors that determine microglial survival. In this regard, we investigated in the EOC 2 microglial cell line with an oxygen-glucose deprivation (OGD) injury model of oxidative stress the role of the "O" class forkhead transcription factor FoxO3a that in some scenarios is closely linked to immune system function. We demonstrate that FoxO3a is a necessary element in the control of early and late apoptotic injury programs that involve membrane phosphatidylserine externalization and nuclear DNA degradation, since transient knockdown of FoxO3a in microglia preserves cellular survival 24 hours following OGD exposure. However, prior to the onset of apoptotic injury, FoxO3a facilitates the activation and proliferation of microglia as early as 3 hours following OGD exposure that occurs in conjunction with the trafficking of the unphosphorylated and active post-translational form of FoxO3a from the cytoplasm to the cell nucleus. FoxO3a also can modulate apoptotic mitochondrial signal transduction pathways in microglia, since transient knockdown of FoxO3a prevents mitochondrial membrane depolarization as well as the release of cytochrome c during OGD. Control of this apoptotic cascade also extends to progressive caspase activation as early as 1 hour following OGD exposure. The presence of FoxO3a is necessary for the expression of cleaved (active) caspase 3, 8, and 9, since loss of FoxO3a abrogates the induction of caspase activity. Interestingly, elimination of FoxO3a reduced caspase 9 activity to a lesser

  3. Influence of cadmium on Bcl-2, Bax and Caspase-3 mRNA transcription in rat's primary cultured cerebral cortical neurons%镉对大鼠大脑皮质神经细胞Bcl-2,Bax和Caspase-3mRNA转录的影响

    Institute of Scientific and Technical Information of China (English)

    袁燕; 孙娅; 江辰阳; 刘学忠; 顾建红; 卞建春; 刘宗平

    2012-01-01

    To investigate the effects of cadmium on transcription levels of apoptosis-related genes Bcl-2,Bax and Caspase-3 mRNA in primary cerebral cortical neurons of SD rats,the neurons were exposured to cadmium at different concentrations (0,5,10,20μmol/L ) for 12 h. MTT assay was applied to test the cell viability,and real-time fluorescent quantitative PCR was used to detect the transcription levels of Bcl-2,Bax and Caspase-3 mRNA. In comparison with the control group, the results showed that the cell viability decreased significantly with the higher concentration cadmium (P〈0.05 or P〈0.01). Additionally, transcription levels of Bax and Caspase-3 mRNA increased significantly (P〈0.01),but transcription level of Bcl-2 mRNA and Bcl-2/Bax decreased signifi- cantly (P〈0. 05,P〈0. 01). It was concluded that cadmium can inhibit the growth of neurons and regulate the transcription of apoptosis-related genes,including Bcl-2,Bax and Caspase-3.%为研究镉对大鼠大脑皮质神经细胞凋亡相关基因Bcl-2、Bax和Caspase-3mRNA转录水平的影响,选用不同浓度(0、5、10、20μmol/L)醋酸镉染毒大鼠大脑皮质神经细胞12h,用MTT法检测细胞存活率,实时荧光定量PCR检测Bcl-2、Bax和Caspase-3mRNA的转录水平,并计算Bcl-2/Bax的比值。结果表明,与对照组相比,各染毒组细胞存活率随染毒剂量增加而显著下降(P〈0.05或P〈0.01);Bax和Caspase-3mRNA转录水平极显著升高(P〈0.01),而Bcl-2mRNA转录水平显著降低(P〈0.05),Bcl-2/Bax极显著降低(P〈0.01)。说明镉能抑制大鼠大脑皮质神经细胞的生长,并可调节凋亡相关基因Bcl2、Bax和Caspase-3mRNA的转录。

  4. NO induces a cGMP-independent release of cytochrome c from mitochondria which precedes caspase 3 activation in insulin producing RINm5F cells.

    Science.gov (United States)

    Tejedo, J; Bernabé, J C; Ramírez, R; Sobrino, F; Bedoya, F J

    1999-10-08

    Exposure of RINm5F cells to interleukin-1beta and to several chemical NO donors such as sodium nitroprusside (SNP), SIN-1 and SNAP induce apoptotic events such as the release of cytochrome c from mitochondria, caspase 3 activation, Bcl-2 downregulation and DNA fragmentation. SNP exposure led to transient activation of soluble guanylate cyclase (sGC) and prolonged protein kinase G (PKG) activation but apoptotic events were not attenuated by inhibition of the sGC/PKG pathway. Prolonged activation of the cGMP pathway by exposing cells to the dibutyryl analogue of cGMP for 12 h induced both apoptosis and necrosis, a response that was abolished by the PKG inhibitor KT5823. These results suggest that NO-induced apoptosis in the pancreatic beta-cell line is independent of acute activation of the cGMP pathway.

  5. Ziyuglycoside II-induced apoptosis in human gastric carcinoma BGC-823 cells by regulating Bax/Bcl-2 expression and activating caspase-3 pathway

    Energy Technology Data Exchange (ETDEWEB)

    Zhu, A.K. [Department of General Surgery, Nanjing Medical University, Affiliated Hangzhou Hospital, Hangzhou (China); Zhou, H.; Xia, J.Z. [Department of General Surgery, Nanjing Medical University, Affiliated Wuxi Second Hospital, Wuxi (China); Jin, H.C. [Department of General Surgery, Nanjing Medical University, Affiliated Hangzhou Hospital, Hangzhou (China); Wang, K. [Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu Province (China); Yan, J.; Zuo, J.B. [Department of General Surgery, Nanjing Medical University, Affiliated Wuxi Second Hospital, Wuxi (China); Zhu, X. [Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu Province (China); Shan, T. [Department of General Surgery, Nanjing Medical University, Affiliated Wuxi Second Hospital, Wuxi (China)

    2013-08-13

    Ziyuglycoside II is an active compound of Sanguisorba officinalis L. that has anti-inflammation, antioxidation, antibiosis, and homeostasis properties. We report here on the anticancer effect of ziyuglycoside II on human gastric carcinoma BGC-823 cells. We investigated the effects of ziyuglycoside II on cell growth, cell cycle, and cell apoptosis of this cell line. Our results revealed that ziyuglycoside II could inhibit the proliferation of BGC-823 cells by inducing apoptosis but not cell cycle arrest, which was associated with regulation of Bax/Bcl-2 expression, and activation of the caspase-3 pathway. Our study is the first to report the antitumor potential of ziyuglycoside II in BGC-823 gastric cancer cells. Ziyuglycoside II may become a potential therapeutic agent against gastric cancer in the future.

  6. Hepatoprotective Role of Hydrangea macrophylla against Sodium Arsenite-Induced Mitochondrial-Dependent Oxidative Stress via the Inhibition of MAPK/Caspase-3 Pathways

    Directory of Open Access Journals (Sweden)

    Md Rashedunnabi Akanda

    2017-07-01

    Full Text Available Sodium arsenite (NaAsO2 has been recognized as a worldwide health concern. Hydrangea macrophylla (HM is used as traditional Chinese medicine possessing antioxidant activities. The study was performed to investigate the therapeutic role and underlying molecular mechanism of HM on NaAsO2-induced toxicity in human liver cancer (HepG2 cells and liver in mice. The hepatoprotective role of HM in HepG2 cells was assessed by using 3-(4,5-dimethylthiazol-2-Yl-2,5-diphenyltetrazolium bromide (MTT, reactive oxygen species (ROS, and lactate dehydrogenase (LDH assays. Histopathology, lipid peroxidation, serum biochemistry, quantitative real-time polymerase chain reaction (qPCR and Western blot analyses were performed to determine the protective role of HM against NaAsO2 intoxication in liver tissue. In this study, we found that co-treatment with HM significantly attenuated the NaAsO2-induced cell viability loss, intracellular ROS, and LDH release in HepG2 cells in a dose-dependent manner. Hepatic histopathology, lipid peroxidation, and the serum biochemical parameters alanine aminotransferase (ALT and aspartate aminotransferase (AST were notably improved by HM. HM effectively downregulated the both gene and protein expression level of the mitogen-activated protein kinase (MAPK cascade. Moreover, HM well-regulated the Bcl-2-associated X protein (Bax/B-cell lymphoma-2 (Bcl-2 ratio, remarkably suppressed the release of cytochrome c, and blocked the expression of the post-apoptotic transcription factor caspase-3. Therefore, our study provides new insights into the hepatoprotective role of HM through its reduction in apoptosis, which likely involves in the modulation of MAPK/caspase-3 signaling pathways.

  7. Grb7 and Hax1 may colocalize partially to mitochondria in EGF treated SKBR3 cells and their interaction can affect Caspase3 cleavage of Hax1

    Science.gov (United States)

    Qian, Lei; Bradford, Andrew M.; Cooke, Peter H.; Lyons, Barbara A.

    2017-01-01

    Growth factor receptor bound protein 7 (Grb7) is a signal transducing adaptor protein that mediates specific protein-protein interactions in multiple signaling pathways. Grb7, with Grb10 and Grb14, are members of the Grb7 protein family. The topology of the Grb7 family members contains several protein-binding domains that facilitate the formation of protein complexes and high signal transduction efficiency. Grb7 has been found overexpressed in several types of cancers and cancer cell lines, and is presumed involved in cancer progression through promotion of cell proliferation and migration via interactions with the ErbB2 (HER2) receptor, FAK (focal adhesion kinase), Ras-GTPases, and other signaling partners. We previously reported Grb7 binds to Hax1 (HS1 associated protein X1) isoform 1, an anti-apoptotic protein also involved in cell proliferation and calcium homeostasis. In this study, we confirm the in vitro Grb7/Hax1 interaction is exclusive to these two proteins and their interaction does not depend on Grb7 dimerization state. In addition, we report Grb7 and Hax1 isoform 1 may colocalize partially to mitochondria in EGF treated SKBR3 cells and growth conditions can affect this colocalization. Moreover, Grb7 can affect Caspase3 cleavage of the Hax1 isoform 1 in vitro, and Grb7 expression may slow the Caspase3 cleavage of Hax1 isoform 1 in apoptotic HeLa cells. Finally, Grb7 is shown to increase cell viability in apoptotic HeLa cells in a time dependent manner. Taken together, these discoveries provide clues for the role of a Grb7/Hax1 protein interaction in apoptosis pathways involving Hax1. PMID:26869103

  8. Caspase-3 feedback loop enhances Bid-induced AIF/endoG and Bak activation in Bax and p53-independent manner.

    Science.gov (United States)

    Guo, W; Zhang, Y; Ling, Z; Liu, X; Zhao, X; Yuan, Z; Nie, C; Wei, Y

    2015-10-15

    Chemoresistance in cancer has previously been attributed to gene mutations or deficiencies. Bax or p53 deficiency can lead to resistance to cancer drugs. We aimed to find an agent to overcome chemoresistance induced by Bax or p53 deficiency. Here, we used immunoblot, flow-cytometry analysis, gene interference, etc. to show that genistein, a major component of isoflavone that is known to have anti-tumor activities in a variety of models, induces Bax/p53-independent cell death in HCT116 Bax knockout (KO), HCT116 p53 KO, DU145 Bax KO, or DU145 p53 KO cells that express wild-type (WT) Bak. Bak knockdown (KD) only partially attenuated genistein-induced apoptosis. Further results indicated that the release of AIF and endoG also contributes to genistein-induced cell death, which is independent of Bak activation. Conversely, AIF and endoG knockdown had little effect on Bak activation. Knockdown of either AIF or endoG alone could not efficiently inhibit apoptosis in cells treated with genistein, whereas an AIF, endoG, and Bak triple knockdown almost completely attenuated apoptosis. Next, we found that the Akt-Bid pathway mediates Bak-induced caspase-dependent and AIF- and endoG-induced caspase-independent cell death. Moreover, downstream caspase-3 could enhance the release of AIF and endoG as well as Bak activation via a positive feedback loop. Taken together, our data elaborate the detailed mechanisms of genistein in Bax/p53-independent apoptosis and indicate that caspase-3-enhanced Bid activation initiates the cell death pathway. Our results also suggest that genistein may be an effective agent for overcoming chemoresistance in cancers with dysfunctional Bax and p53.

  9. Apoptosis induced by copper oxide quantum dots in cultured C2C12 cells via caspase 3 and caspase 7: a study on cytotoxicity assessment.

    Science.gov (United States)

    Amna, Touseef; Van Ba, Hoa; Vaseem, M; Hassan, M Shamshi; Khil, Myung-Seob; Hahn, Y B; Lee, Hak-Kyo; Hwang, I H

    2013-06-01

    We report herein the synthesis and characterization of copper oxide quantum dots and their cytotoxic impact on mouse C2C12 cells. The utilized CuO quantum dots were prepared by the one-pot wet chemical method using copper acetate and hexamethylenetetramine as precursors. The physicochemical characterization of the synthesized CuO quantum dots was carried out using X-ray diffraction, energy-dispersive X-ray analysis, and transmission electron microscopy. To examine the in vitro cytotoxicity, C2C12 cell lines were treated with different concentrations of as-prepared quantum dots and the viability of cells was analyzed using Cell Counting Kit-8 assay at regular time intervals. The morphology of the treated C2C12 cells was observed under a phase-contrast microscope, whereas the quantification of cell viability was carried out via confocal laser scanning microscopy. To gain insight into the mechanism of cell death, we examined the effect of CuO quantum dots on the candidate genes such as caspases 3 and 7, which are key mediators of apoptotic events. In vitro investigations of the biological effect of CuO quantum dots have shown that it binds genomic DNA, decreases significantly the viability of cells in culture in a concentration (10-20 μg/mL) dependent manner, and inhibits mitochondrial caspases 3 and 7. To sum up, the elucidation of the pathways is to help in understanding CuO quantum dot-induced effects and evaluating CuO quantum dot-related hazards to human health.

  10. Extracellular TDP-43 aggregates target MAPK/MAK/MRK overlapping kinase (MOK) and trigger caspase-3/IL-18 signaling in microglia.

    Science.gov (United States)

    Leal-Lasarte, María M; Franco, Jaime M; Labrador-Garrido, Adahir; Pozo, David; Roodveldt, Cintia

    2017-07-01

    Dysregulated microglial responses are central in neurodegenerative proteinopathies, including amyotrophic lateral sclerosis (ALS) and frontotemporal lobar disease (FTLD). Pathologic TDP-43, which is typically found in intracellular inclusions, is a misfolding protein with emerging roles in ALS and FTLD. Recently, TDP-43 species have been found in extracellular fluids of patients; however, the overall implications of TDP-43-mediated signaling linked to neuroinflammation are poorly understood. Our work-the first, to our knowledge, to focus on innate immunity responses to TDP-43 aggregates-shows that such species are internalized by microglia and cause abnormal mobilization of endogenous TDP-43. Exposure to TDP-43 aggregates elicited not only IL-1β, but also NLRP3-dependent and noncanonical IL-18 processing. Moreover, we report a link between TDP-43 and neuronal loss via the apoptosis-independent emerging roles of caspase-3 in neurotoxic inflammation. Our results further support the view of noncell autonomous neurodegenerative mechanisms in ALS. Remarkably, we demonstrate that TDP-43 aggregates bind to and colocalize with MAPK/MAK/MRK overlapping kinase (MOK) and show that its phosphorylation status is disrupted. Finally, we show that this TDP-43-caused activation state can be altered by exogenous Hsp27 and Hsp70 chaperones. Our study provides new insight into the immune phenotype, mechanisms, and signaling pathways that operate in microglial neurotoxic activation in ALS.-Leal-Lasarte, M. M., Franco, J. M., Labrador-Garrido, A., Pozo, D., Roodveldt, C. Extracellular TDP-43 aggregates target MAPK/MAK/MRK overlapping kinase (MOK) and trigger caspase-3/IL-18 signaling in microglia. © FASEB.

  11. Doxorubicin in vivo rapidly alters expression and translation of myocardial electron transport chain genes, leads to ATP loss and caspase 3 activation.

    Directory of Open Access Journals (Sweden)

    Amy V Pointon

    Full Text Available BACKGROUND: Doxorubicin is one of the most effective anti-cancer drugs but its use is limited by cumulative cardiotoxicity that restricts lifetime dose. Redox damage is one of the most accepted mechanisms of toxicity, but not fully substantiated. Moreover doxorubicin is not an efficient redox cycling compound due to its low redox potential. Here we used genomic and chemical systems approaches in vivo to investigate the mechanisms of doxorubicin cardiotoxicity, and specifically test the hypothesis of redox cycling mediated cardiotoxicity. METHODOLOGY/PRINCIPAL FINDINGS: Mice were treated with an acute dose of either doxorubicin (DOX (15 mg/kg or 2,3-dimethoxy-1,4-naphthoquinone (DMNQ (25 mg/kg. DMNQ is a more efficient redox cycling agent than DOX but unlike DOX has limited ability to inhibit gene transcription and DNA replication. This allowed specific testing of the redox hypothesis for cardiotoxicity. An acute dose was used to avoid pathophysiological effects in the genomic analysis. However similar data were obtained with a chronic model, but are not specifically presented. All data are deposited in the Gene Expression Omnibus (GEO. Pathway and biochemical analysis of cardiac global gene transcription and mRNA translation data derived at time points from 5 min after an acute exposure in vivo showed a pronounced effect on electron transport chain activity. This led to loss of ATP, increased AMPK expression, mitochondrial genome amplification and activation of caspase 3. No data gathered with either compound indicated general redox damage, though site specific redox damage in mitochondria cannot be entirely discounted. CONCLUSIONS/SIGNIFICANCE: These data indicate the major mechanism of doxorubicin cardiotoxicity is via damage or inhibition of the electron transport chain and not general redox stress. There is a rapid response at transcriptional and translational level of many of the genes coding for proteins of the electron transport chain

  12. Quercetin nanoparticles induced autophagy and apoptosis through AKT/ERK/Caspase-3 signaling pathway in human neuroglioma cells: In vitro and in vivo.

    Science.gov (United States)

    Lou, Miao; Zhang, Li-Na; Ji, Pei-Gang; Feng, Fu-Qiang; Liu, Jing-Hui; Yang, Chen; Li, Bao-Fu; Wang, Liang

    2016-12-01

    Neuroglioma is a complex neuroglial tumor involving dysregulation of many biological pathways at multiple levels. Quercetin is a potent cancer therapeutic agent presented in fruit and vegetables, preventing tumor proliferation, and is a well known cancer therapeutic agent and autophagy mediator. Recent studies showed that drug delivery by nanoparticles have enhanced efficacy with reduced side effects. In this regard, gold-quercetin into poly (dl-lactide-co-glycolide) nanoparticles was examined. In the present study, quercetin nanoparticle induced cell autophagy and apoptosis in human neuroglioma cell was investigated. Quercetin nanoparticle administrated to animals displayed suppressed role in tumor growth. The cell viability was deterined through CCK8 assay. Transmission electron microscopy was utilized to observe the formation of autophagosome. The cell apoptosis was assessed by annexin V-PI staining. The protein expression of cell autophagy regulators and tumor suppressors were analyzed via western blot and RT-PCR. Treatment of human neuroglioma cell with quercetin nanoparticle induced cell death in a dose-and time-dependent manner. The flow cytometry results showed that the proportion of the apoptosis cells had gained after quercetin nanoparticle treatment compared to untreatment group. Moreover, the expression of activated PI3K/AKT and Bcl-2 were down-regulated upon quercetin nanoparticle treatment in human neuroglioma cells. The expression level of LC3 and ERK as well as cytoplasm p53, cleaved Caspase-3 and PARP was positively correlated with the concentration of quercetin nanoparticle. In addition, p-mTOR and GAIP were obviously down-regulated by quercetin nanoparticle treatment in a dose-dependent manner. These results indicated that quercetin nanoparticle could induce autophagy and apoptosis in human neuroglioma cells, the underlying molecular mechanisms, at least partly, through activation LC3/ERK/Caspase-3 and suppression AKT/mTOR signaling.

  13. New avenue in the treatment of temporal lobe epilepsy by classical anti-epileptics: A hypothetical establishment of executioner Caspase 3 inactivation by molecular modeling

    Directory of Open Access Journals (Sweden)

    M Vijey Aanandhi

    2015-01-01

    Full Text Available Patients with temporal lobe epilepsy (TLE are prescribed first-line antiepileptic drugs and surgery to the management of this disorder. Unfortunately, the surgical treatment has been shown to be beneficial for the selected patients but fails to provide a seizure-free outcome in 20-30% of TLE patients. In our present study, we investigate the possibilities of marketed antiepileptic drugs in a different manner to improve the present situation in TLE. Molecular docking simulation study and various open source computational tools were used to perform the study. AutoDock 4.2 MGL tools, Pymol visualize tools, Patch dock server, and Swarm Dock servers (protein-protein docking were used to perform the molecular modeling. FTsite and computed atlas of surface topography of protein open source server were used to understand the pocket and ligand binding information respectively. Toxtree application was used to determine the toxicity profile of the drug by Cramers rule. The obtained molecular docking models (Caspase 3, Procaspase 8, and Fas-associated death domain [FADD] with selected compounds (Clonazepam, Clobazepam, and Retigabine showed promising trio blocking event of FADD, Caspase 3, and Procaspase 8 (−6.66 kcal, −8.1 kcal, 6.46 kcal by Clonazepam respectively. Protein-protein interaction study (Swarm Dock, Patch Dock server indicated promising results that helped to establish our hypothesis. Toxtree showed a quantitative structure toxicity relationship report that helps to clarify the toxicity of the selected compounds. Clonazepam showed a trio inhibition property that may lead to develop a new era of the new generation benzodiazepine prototype drugs in the future. Filtered compounds will further process for higher in vitro, in vivo models for better understanding of the mechanism.

  14. Effect of myocardial reperfusion on cardiocyte apoptosis and expression of bcl-2, bax and caspase-3 in rats with depression%心肌再灌注对抑郁大鼠心肌细胞凋亡以及bcl-2、bax和caspase-3表达的影响

    Institute of Scientific and Technical Information of China (English)

    刘淑珍; 尤鑫; 熊小栓; 刘兴德

    2012-01-01

    目的:探讨心肌缺血再灌注(I/R)对抑郁大鼠心肌细胞凋亡及凋亡基因bcl-2、bax和caspase-3的影响.方法:Wistar大鼠32只,随机分为4组,每组各8只.A组:非抑郁大鼠假手术组;B组:抑郁大鼠假手术组;C组:非抑郁大鼠心肌I/R组;D组:抑郁大鼠心肌I/R组.采用慢性轻度不可预知性应激结合孤养制备抑郁模型,用敞箱实验和液体消耗实验观察大鼠行为改变;运用结扎左冠状动脉前降支的方法复制心肌I/R模型.运用TUNEL法检测心肌凋亡细胞;运用免疫组化法和逆转录-聚合酶链反应(RT- PCR)方法检测bcl-2、bax和caspase-3的表达.结果:(1)与A、B组比较,C、D组心肌细胞凋亡数量显著增加(P<0.01),A、B两组间比较无显著差异;与C组比较,D组心肌细胞凋亡数量显著增加(P<0.05).(2)与A、B组比较,C、D组Bcl-2、Bax和caspase-3蛋白和mRNA表达显著增加(P<0.01),A、B两组间比较无显著差异;与C组比较,D组Bcl-2蛋白和mRNA表达显著减少(P<0.05),而Bax和caspase-3蛋白和mRNA表达显著增加(P<0.05).结论:心肌缺血再灌注可加重抑郁大鼠缺血心肌细胞凋亡,其机制可能与上调bax和caspase-3基因表达、下调bcl-2基因表达有关.%AIM:To explore ihe effecl of ischemia/reperfusion (I/R) on cardiac myocyle apoplosis and ihe expression of bcl -2, bax and caspase - 3 in ihe rals wilh depression. METHODS; Thirty - Lwo Wislar rals were randomly divided inlo 4 groups,including group A; sham operation group; group B; ihe rals wilh depression undergoing sham opera-lion; group C; ihe rals wilh myocardial I/R operalion; group D; ihe rals wilh depression undergoing myocardial I/R opera-lion. The ral model of depression was produced by chronic mild unpredictable slress and separalion. The behaviors of ihe rals were delecled by open field lesl and fluid consumption lesl. The myocardial I/R model was made by ligaling ihe lefl an-lerior descending branch of coronary artery in ihe rals. The

  15. 阿托伐他汀对大鼠脑缺血再灌注 PERK/elfR2a通路及 Caspase-3表达的影响%The Study of PERK/eIF2a Pathway and the Expression of Caspase-3 in Ischemia-reperfusion Rats and Atorvastatin Intervention

    Institute of Scientific and Technical Information of China (English)

    彭文娟; 杨剑文; 刘湘玉; 杨期明

    2016-01-01

    目的:研究蛋白激酶 R 样内质网激酶(PERK)/ eIFR2a 通路及 Caspase-3在大鼠脑缺血再灌注损伤中的作用机制及阿托伐他汀对其的影响。方法采用大脑中动脉线栓塞法制作大鼠脑缺血再灌注模型;随机分为缺血再灌注组、假手术组、阿托伐他汀组、阿托伐他汀+ Salubrinal 抑制剂组,大体标本采用 TTC 染色,釆用 West-ern-blot 法检测 PERK、Caspase-3蛋白表达及 eIF2a 蛋白磷酸化。结果与假手术组相比,大鼠缺血再灌注后PERK 蛋白表达及 eIF2a 的磷酸化增加, Caspase-3表达的活性增强(P <0.01);阿托伐他汀干预可以减轻 PERK 蛋白表达及 eIF2a 蛋白磷酸化(P <0.05)。给予特异性 eIF2a 磷酸化抑制剂 Salubrinal 后可抑制 eIF2a 的磷酸化及Caspase-3表达的活性(P <0.05),对 PERK 蛋白表达无影响。形态学上从 TTC 染色提示:在缺血再灌注组 TTC 染色可见大片脑梗死组织。 Salubrinal 抑制剂及阿托伐他汀干预后脑梗死体积明显缩小(P <0.05)。结论内质网应激通过 PERK/ eIF2a/ Caspase-3途径促进细胞凋亡,阿托伐他汀干预可以减轻脑缺血再灌注损伤。%Objective To study the PERK/ eIF2a pathways and Caspase 3 in the mechanism of action of ischemia reperfusion injury in rats and the effect of atorvastatin. Methods Produce the ischemia reperfusion model rats by Middle cerebral artery embolism method which were divided into the ischemia reperfusion group,control group,atorvastatin interven-tion group and eIF2a suppression group. To observe the changes of ischemic brain,specimens were treated with TTC stai-ning,the PERK,Caspase-3 protein expression and protein phosphorylation eIF2a were detected by western-blot. Results Compared with the control group,after ischemia reperfusion,PERK protein expression and protein phosphorylation eIF2a in-creased,the expression of Caspase 3 was enhanced,and the expression of PERK protein and phosphorylation eIF2a was re

  16. ANÁLISE QUANTITATIVA DOS IMPACTOS DO PROCESSO DE ACULTURAÇÃO NA FUSÃO DAS EMPRESAS Y E Z

    Directory of Open Access Journals (Sweden)

    Guilherme Carneiro Felippe dos Santos

    2015-02-01

    Full Text Available O presente estudo teve o objetivo de analisar o processo de integração cultural de uma fusão envolvendo empresas brasileiras (denominadas Y e Z por meio de uma análise quantitativa. Com intuito de mapear diferenças entre as percepções dos funcionários originários das empresas Y e Z, utilizou-se questionário eletrônico baseado no modelo conceitual de pesquisa desenvolvido por Liao e Yen (2003, contendo questões referentes à cultura organizacional e às atitudes de trabalho dos empregados. O índice de resposta da pesquisa foi de 67%. A análise dos dados foi efetuada por intermédio do teste estatístico não paramétrico de Mann-Whitney (teste-U. Os resultados apontam para a existência de percepções culturais distintas entre os membros originários de cada empresa, sugerindo que o processo de integração da fusão está incompleto e que a organização se encontra num período de estresse da aculturação, no qual não há concordância entre os funcionários sobre o melhor modo de integração.

  17. A política de identidades na sociolingüística quantitativa e na geolingüística monodimensional: o informante The politcs of identities in the quantitative sociolinguistics and in the monodimensional geolinguistics: the informer

    OpenAIRE

    Maria Isabel Borges

    2009-01-01

    Neste trabalho, procuramos apontar alguns aspectos das políticas de identidades e de representação em torno da figura do informante, em estudos feitos a partir das bases de dados do VARSUL (Variação Lingüística Urbana na Região Sul do Brasil) e do ALERS (Atlas Lingüístico-etnográfico da Região Sul do Brasil). Na primeira base de dados, os pressupostos teóricos e metodológicos são da sociolingüística quantitativa enquanto, na segunda base de dados, os pressupostos teóricos e metodológicos são ...

  18. 电针对骶髓损伤后神经源性膀胱大鼠尿流动力学及膀胱组织中Caspase-3表达的影响%Effect of Electrical Acupuncture on Urodynamics and Expression of Caspase-3 in Bladder Tissue in Rats with Neurogenic Bladder ;after Sacral Spinal Sord Injury

    Institute of Scientific and Technical Information of China (English)

    艾坤; 张泓; 张雨辰; 鲍秋影; 李景兴; 匡静之

    2013-01-01

    Objective To explore the effects of electrical acupuncture at Ciliao (BL32), Zhongji (RN03), Sanyinjiao (SP06) acupoints on the expression of caspase-3 in bladder tissue and the urodynamics of rats with neurogenic bladder (detrusor areflexia type) after sacral spinal cord injury. Methods 10 rats were randomly selected as blank control group from 40 female Sprague-Dawley rats. The other 30 rats were randomly divided into model group, acupoint group and control group after modeling, with 10 rats in each group. The urodynamics was mea-sured on the 14th and 22nd day after modeling. The control group and acupoint group received electrical acupuncture 7 times at the acu-points and other points nearby, respectively, from the 15th day. The expression of caspase-3 in bladder tissue was detected with en-zyme-linked immuno sorbent assay (ELISA) method. Results Compared with the model group, the maximum bladder capacity, bladder com-pliance and expression of caspase-3 in bladder tissue significantly decreased after treatment (P<0.01), the filling bladder pressure significant-ly increased (P<0.05) in the acupoint group. And the acupuncture group was better than the control group (P<0.05). Conclusion Electrical acupuncture at Ciliao, Zhongji and Sanyinjiao can reduce the maximum bladder capacity in neurogenic bladder rats and inhabit the expres-sion of caspase-3 in bladder tissue.%目的:观察电针次髎、中极、三阴交穴对骶髓损伤后神经源性膀胱(逼尿肌无反射型)大鼠尿流动力学以及膀胱组织中caspase-3表达的影响。方法雌性Sprague-Dawley大鼠40只,随机抽取10只为空白组,其余造模后随机分为模型组、穴位组和对照点组,每组10只。术后第15天起按组分别施以7次治疗。测量术后第14天及干预治疗7次后1 d的大鼠尿流动力学参数;治疗结束后取膀胱组织采用ELISA法检测在膀胱组织中caspase-3的表达。结果电针治疗后大鼠最大膀胱容量显著降低(P<0

  19. Caracterização de linhagens endogâmicas recombinantes e mapeamento de locos de características quantitativas associados a ciclo e produtividade do feijoeiro-comum

    Directory of Open Access Journals (Sweden)

    Faleiro Fábio Gelape

    2003-01-01

    Full Text Available O objetivo deste trabalho foi caracterizar 154 linhagens endogâmicas recombinantes por meio da avaliação de características quantitativas, morfológicas, moleculares e de resistência a doenças e mapear locos de características quantitativas associados a ciclo e produtividade do feijoeiro-comum. Adotando o valor do limite de detecção (LOD de 4,0 e uma freqüência máxima de recombinação de 0,40, foram mapeados 43 marcadores em nove grupos de ligação cobrindo uma distância de recombinação total de 247,8 cM. A distância entre marcadores adjacentes variou entre 0 e 28 cM, com média de 7,3 cM. Os grupos de ligação variaram em tamanho de 2,3 a 61,2 cM. Os genes de resistência à ferrugem e à antracnose ficaram localizados no mesmo grupo de ligação. Foram mapeados locos associados às oito características quantitativas estudadas, e a explicação da variância fenotípica pelos marcadores variou de 14,03% a 40,14%. Os resultados encontrados lançam bases para o desenvolvimento de mapas específicos saturados e de utilidade em programas de melhoramento do feijoeiro-comum.

  20. 龙葵碱调控Bcl-2与Bax蛋白表达及caspase-3活性诱导HepG2细胞凋亡的研究%Induction of solanine on HepG2 cell apoptosis by regulation of Bcl-2/Bax expression and caspase-3 activity

    Institute of Scientific and Technical Information of China (English)

    高世勇; 徐丽丽; 季宇彬

    2009-01-01

    目的 探讨龙葵碱诱导HepG2细胞凋亡的作用机制.方法 透射电镜观察凋亡细胞形态变化,原位缺口末端榆测法(TUNEL法)检测DNA断裂情况,流式细胞术检测细胞凋亡率,间接免疫荧光法激光共聚焦扫描显微术检测Bcl-2与Bax蛋白表达,比色法检测caspase-3活性的变化.结果 在透射电镜下观察,龙葵碱组细胞出现细胞固缩,染色质致密,核凝聚固缩,染色体断裂形成核碎块,凋亡小体形成等细胞凋亡特征形态.TUNEL法发现龙葵碱高、中、低剂量组HepG2细胞均有绿色荧光,阴性对照组无荧光.流式细胞术分析表明0.4、2、10μmol/L龙葵碱作用HepG2细胞24 h凋亡率分别为4.0%、8.5%、20.1%.同时,龙葵碱升高caspase-3活性,下调Bcl-2蛋白表达,上调Bax蛋白表达.结论 龙葵碱通过降低Bcl-2/Bax的值,激活caspase-3酶活性诱导HepG2细胞凋亡.

  1. Effect of Blockade of Co- stimulatory Signal CD86 on the Expression of Caspase-3 and Laminin B Proteins and Pregnancy Outcome%阻断协同刺激分子对大鼠母胎界面中Caspase-3、Laminin B 的表达及妊娠结局的影响

    Institute of Scientific and Technical Information of China (English)

    赵富玺; 李拴明; 张源源; 刘润花; 刘丽华; 崔克

    2007-01-01

    目的:探讨阻断协同刺激分子CD86对自然流产模型孕鼠母胎界面Caspase-3和Laminin B的表达及妊娠结局的影响.方法:实验组于妊娠第4.5天腹腔注射大鼠抗小鼠CD86单克隆抗体,对照组注射大鼠同型IgG2b,而正常妊娠组不作任何处理.于妊娠第13.5天计算胚胎吸收率,用免疫组化法测定Caspase-3和Laminin B 的表达.结果:实验组的胚胎吸收率显著低于对照组(χ2=7.441,P<0.05);实验组中Caspase-3蛋白阳性表达率明显低于对照组(P<0.05),实验组中Laminin B 蛋白阳性表达率明显高于对照组(P<0.05).结论:妊娠早期阻断协同刺激分子CD86可使母胎界面中的Caspase-3和Laminin B 分别通过各自不同的途径来发挥免疫耐受作用,并使自然流产模型孕鼠的胚胎吸收率降低至正常妊娠水平.

  2. 流产鼠Caspase-3与laminin B和PAI-1的表达及对妊娠结局的影响%Study of the expression of caspase - 3, laminin B and PAI- 1 proteins and outcome of pregnancy in murine abortion- prone model

    Institute of Scientific and Technical Information of China (English)

    李拴明; 赵富玺; 刘润花; 杨秀兰; 闫平

    2007-01-01

    目的:研究阻断CD86协同刺激分子对自然流产模型孕鼠母胎界面Caspase-3、laminin B和PAI-1的表达及对妊娠结局的影响.方法:实验组于妊娠第4.5天腹腔注射大鼠抗小鼠CD86单抗,实验对照组注射大鼠同型IgG2b,正常妊娠组不作任何处理.于妊娠第13.5天计算胚胎吸收率,用免疫组化测定Caspase-3、laminin B和PAI-1的表达,并进行图像分析、检测免疫组化染色灰度值(A).结果:①实验组的胚胎吸收率显著低于实验对照组(P<0.05);②实验组中Caspase-3蛋白灰度值明显高于实验对照组(P<0.05),实验组中laminin B和PAI-1蛋白灰度值均明显低于实验对照组(P<0.05).结论:妊娠早期阻断CD86协同刺激分子可使母胎界面中的Caspase-3、laminin B和PAI-1分别通过各自不同的途径发挥免疫耐受作用并且使自然流产模型孕鼠的胚胎吸收率降低至正常妊娠水平.

  3. 甲泼尼龙预干预脊髓缺血再灌注损伤对Caspase-3表达的影响%Effect of methylprednisolone pretreatment of ischemia-reperfusion injury of spinal cord on expression of caspase-3 in rats

    Institute of Scientific and Technical Information of China (English)

    夏磊; 孟志强; 王凯; 王肖虎; 陈宗霖; 丁勇

    2011-01-01

    Objective To investigate the effect of methylprednisolone (MP) pretreatment of ischemia-reperfusion injury of spinal cord (SCII) on expression of caspase-3 in rats. Methods One hundred and twenty healthy, purebred, adult male SD rats were randomly divided into 3 equal groups (n = 40).Croup A was a normal one in which only the abdominal aorta was exposed without establishing an injury model. Croup B was an SCII model in which the abdominal aorta was first obstructed for 30 minutes and then relaxed and next reperfused. After reperfusion for 3 hours, the rats were killed to harvest samples of lumbar spinal cord. Group C was an MP pretreatment one in which tail intravenous injection of MP (30mg/kg) was conducted 30 minutes before establishment of an SCII model. Pathological changes of the injured spinal cord were observed by HE staining, and expressions of caspase-3 in the spinal cord tissue were observed by immunohistochemical staining. Results In group A, the outlines of neuronal cells were clear with polarity and a round nucleus. Ramified Nissl bodies were evenly arranged around the nucleus. Cytoplasm was stained uniformly dark. Organizational structure was integrated and clear without neutrophile granulocyte infiltration,hemorrhage or edema. In group B, neuronal cells were swelling and decreased in quantity, with blunt polarity and mild atrophy of nucleolus. Cranuloses of Nissl bodies were observed with pale staining. There was neutrophile granulocyte infiltration. In group C, mild edema was observed in part of neuronal cells, nucleoli were still clear and there was no neutrophile granulocyte infiltration. Caspase-3 was highly expressed in the spinal cord tissue in group B, with significant differences compared with group A ( P < 0. 05). Expression of caspase-3 in the spinal cord was slightly increased in group C, but the difference was significant compared with group B (P < 0. 05) . Conclusion Since MP pretreatment can reduce expression of caspase-3 in SCII rats

  4. 中药复方对戊四氮致痫大鼠海马神经元caspase-3表达的影响%Effect of Chinese Medicine Compound on the Expression of Caspase-3 in Hippocampus of Epileptic Rats Induced by Pentetrazole

    Institute of Scientific and Technical Information of China (English)

    王玢; 迟华基; 袁方曜

    2010-01-01

    目的:观察戊四氮(PTZ)致痫大鼠海马神经元caspase-3表达以及中药复方AAP的脑保护作用.方法: 144只健康成年雄性Wistar大鼠随机分为对照组(CK组)、模型组(PTZ组)、中药大剂量组(AAPl组)、中药中剂量组(AAPm组)、中药小剂量组(AAPs组)和丙戊酸钠组(VPA组);每组各6只.CK组和PTZ组分别给予生理盐水(4mL/kg·d)灌胃;中药各组分别给予中药复方大、中、小剂量(10.26g/kg、5.13g/kg、2.56g/kg)灌胃,每天1次;VPA组腹腔注射VPA(20mg/kg·d).造模第一天,除CK组外,其余各组大鼠均腹腔注射戊四氮(PTZ)75mg/kg,观察记录大鼠行为学变化;于致痫后12h、2d、5d、7d相应时间点取材,制备脑标本;免疫组化检测caspase-3表达.结果:致痫后,除CK组外,其余各组海马区caspase-3阳性表达增强;7天,与PTZ组相比,AAPl组、AAPm组和AAPs组海马CA3区caspase-3阳性表达减弱 (P<0.05).结论:caspase-3参与致痫大鼠海马神经元凋亡过程;AAP能降低caspase-3表达,减少神经元凋亡,有神经保护作用.

  5. Mitochondrial and nuclear damages and caspase-3 expression in the hippicampal CA3 region of rats with kainic acid induced status epilepticus

    Institute of Scientific and Technical Information of China (English)

    Shuhai Tang; Jianying Sun; Xiaojun Pan; Li Zhang

    2006-01-01

    BACKGROUND: Some scholars believed that the neuronal injury after status epilepticus is apoptosis,the main evidence is the changes of expressions of various apoptosis releted genes,such as immediate-early gene,p53 gene and genes of bcl-2 family,etc.But there is still no ultrastructural evidence for apoptosis.OBJECTIVE: To observe the ultrastructural damages of mitochondrion and nucleus and the changes of caspase expression in neurons of hippocampal CA3 region in rats with status epilepticus induced by kainic acid.DESIGN: A randomized controlled study.SETTING: Department of Anesthesiology and Department of Neurology,Qilu Hospital of Shandong University.MATERIALS: Seventy-five adult male Wistar rats of 250-300 g.clean degree,were provided by the experimental animal center of Shandong University.Kainic acid was purchased from Sigma Company (USA);rabbit anti-rat polyclonal antibody caspase-3 from Santa Cruz Company(USA).METHODS:The experiments were carried out in the Department of Anesthesiology,Qilu Hospital of Shandong University from October 2005 to February 2006.①The 75 rats were randomly divided into experimental group (n=45)and control group(n=30).②Model establishment,convulsion grading and the judging standards for status epilepticus:Rats in the experimental group were given intraperitoneal injection of kainic acid(10 mg/kg),and those in the control group were injected with saline of the same volume.The time of seizure was recorded and their behavioral manifestations were observed,and the seizure was terminated by intraperitoneal injection of diazepam(10 mg/kg).③Observation under electron microscope:At 3, 12 and 24 hours after status epilepticus respectively,bilateral hippocampal tissues were taken out,semithin sections of about 75 nm were prepared after fixation,dehydration and embedding,and then observed under H-800 transmission electron microscope.④Immunohistochemical detection:Bilateral hippocampi were removed at 3,12 and 24 hours after status

  6. Hydrogen-rich saline attenuates isoflurane-induced caspase-3 activation and cognitive impairment via inhibition of isoflurane-induced oxidative stress, mitochondrial dysfunction, and reduction in ATP levels

    Science.gov (United States)

    Li, Cheng; Hou, Lengchen; Chen, Dan; Lin, Fuqing; Chang, Tao; Li, Mengzhu; Zhang, Lingling; Niu, Xiaoyin; Wang, Huiying; Fu, Shukun; Zheng, Junhua

    2017-01-01

    Objectives: The inhaled general anesthetic isoflurane has been shown to induce caspase-3 activation in vitro and in vivo. The underlying mechanisms and functional consequences of this activity remain unclear. Isoflurane can induce caspase-3 activation by causing accumulation of reactive oxygen species (ROS), mitochondrial dysfunction, and reduction in adenosine triphosphate (ATP) levels. This study aimed to investigate the protective effect of hydrogen, a novel antioxidant, against isoflurane-induced caspase-3 activation and cognitive impairment. Methods: H4 human neuroglioma cells overexpressing human amyloid precursor protein were treated with saline or hydrogen-rich saline (HS, 300 μM), with or without 2% isoflurane, for 6 h or 3 h. Western blot analysis, fluorescence assays, and a mitochondrial swelling assay were used to evaluate caspase-3 activation, levels of ROS and ATP, and mitochondrial function. The effect of the interaction of isoflurane (1.4% for 2 h) and HS (5 mL/kg) on cognitive function in mice was also evaluated using a fear conditioning test. Results: We found that HS attenuated isoflurane-induced caspase-3 activation. Moreover, HS treatment mitigated isoflurane-induced ROS accumulation, opening of mitochondrial permeability transition pores, reduction in mitochondrial membrane potential, and reduction in cellular ATP levels. Finally, HS significantly alleviated isoflurane-induced cognitive impairment in mice. Conclusions: Our results suggest that HS attenuates isoflurane-induced caspase-3 activation and cognitive impairment via inhibition of isoflurane-induced oxidative stress, mitochondrial dysfunction, and reduction in ATP levels. These findings warrant further research into the underlying mechanisms of this activity, and indicate that HS has the potential to attenuate anesthesia neurotoxicity.

  7. M867, a novel selective inhibitor of caspase-3 enhances cell death and extends tumor growth delay in irradiated lung cancer models.

    Directory of Open Access Journals (Sweden)

    Kwang Woon Kim

    Full Text Available BACKGROUND: Lung cancer remains the leading cause of cancer death worldwide. Radioresistance of lung cancer cells results in unacceptable rate of loco-regional failure. Although radiation is known to induce apoptosis, our recent study showed that knockdown of pro-apoptotic proteins Bak and Bax resulted in an increase in autophagic cell death and lung cancer radiosensitivity in vitro. To further explore the potential of apoptosis inhibition as a way to sensitize lung cancer for therapy, we tested M867, a novel chemical and reversible caspase-3 inhibitor, in combination with ionizing radiation in vivo and in vitro. METHODS AND FINDINGS: M867 reduced clonogenic survival in H460 lung cancer cells (DER = 1.27, p = 0.007 compared to the vehicle-treated treated cells. We found that administration of M867 with ionizing radiation in an in vivo mouse hind limb lung cancer model was well tolerated, and produced a significant tumor growth delay compared to radiation alone. A dramatic decrease in tumor vasculature was observed with M867 and radiation using von Willebrand factor staining. In addition, Ki67 index showed >5-fold reduction of tumor proliferation in the combination therapy group, despite the reduced levels of apoptosis observed with terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling staining. Radiosensitizing effect of M867 through inhibiting caspases was validated using caspase-3/-7 double-knockout (DKO mouse embryonic fibroblasts (MEF cell model. Consistent with our previous study, autophagy contributed to the mechanism of increased cell death, following inhibition of apoptosis. In addition, matrigel assay showed a decrease in in vitro endothelial tubule formation during the M867/radiation combination treatment. CONCLUSIONS: M867 enhances the cytotoxic effects of radiation on lung cancer and its vasculature both in vitro and in vivo. M867 has the potential to prolong tumor growth delay by inhibiting tumor proliferation

  8. Neuroprotective effects of puerarin derivative on focal cerebral ischemia and reperfusion and its effect on expression of caspase-3 in rats%葛根素衍生物对局灶性脑缺血再灌注的保护作用及对caspase-3表达的影响

    Institute of Scientific and Technical Information of China (English)

    张宏伟; 张帆; 董志; 朱毅

    2016-01-01

    Objective: To observe the protection effect of puerarin derivative (G20) on focal cerebral ischemia and reperfusion injury in rats, and explore whether its mechanism is associated with the expression of caspase-3.Methods: Model of focal cerebral ischemia and reperfusion was induced using an intraluminal monofilament blockade by Longa. The rats were randomly divided into six groups, which were the sham-operation group, the model group, G group (puerarin 25.0 mg·kg-1), G20 groups