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Sample records for inha enoil-acp redutase

  1. Efeito das mutações I16T, I21V, I47T e S94A na afinidade da enzima 2-trans-Enoil-ACP (CoA) redutase de Mycobacterium tuberculosis pelo cofator NADH : estudos por simulação pela dinâmica molecular e docking molecular

    OpenAIRE

    Evelyn Koeche Schroeder

    2004-01-01

    aumento do número de casos de tuberculose e o surgimento de cepas de Mycobacterium tuberculosis (MTB) resistentes a múltiplas drogas, entre elas a isoniazida (INH) representa um sério problema de saúde pública. A enzima InhA, ou 2-trans-Enoil-ACP (CoA) redutase de MTB, catalisa a redução NADHdependente de ácidos graxos α,β-insaturados, precursores dos ácidos micólicos (importantes componentes do envelope celular do MTB). Mutações no gene estrutural da InhA estão associadas à resistência in vi...

  2. Inhibin alpha-subunit (INHA expression in adrenocortical cancer is linked to genetic and epigenetic INHA promoter variation.

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    Johannes Hofland

    Full Text Available Adrenocortical carcinoma (ACC is a rare, but highly malignant tumor of unknown origin. Inhibin α-subunit (Inha knockout mice develop ACCs following gonadectomy. In man, INHA expression varies widely within ACC tissues and its circulating peptide inhibin pro-αC has been described as a novel tumor marker for ACC. We investigated whether genetic and epigenetic changes of the INHA gene in human ACC cause loss or variation of INHA expression. To this end, analyses of INHA sequence, promoter methylation and mRNA expression were performed in human adrenocortical tissues. Serum inhibin pro-αC levels were also measured in ACC patients. INHA genetic analysis in 37 unique ACCs revealed 10 novel, heterozygous rare variants. Of the 3 coding bases affected, one variant was synonymous and two were missense variants: S72F and S184F. The minor allele of rs11893842 at -124 bp was observed at a low frequency (24% in ACC samples and was associated with decreased INHA mRNA levels: 4.7±1.9 arbitrary units for AA, compared to 26±11 for AG/GG genotypes (P = 0.034. The methylation of four proximal INHA promoter CpGs was aberrantly increased in five ACCs (47.7±3.9%, compared to normal adrenals (18.4±0.6%, P = 0.0052, whereas the other 14 ACCs studied showed diminished promoter methylation (9.8±1.1%, P = 0.020. CpG methylation was inversely correlated to INHA mRNA levels in ACCs (r = -0.701, p = 0.0036, but not associated with serum inhibin pro-αC levels. In conclusion, aberrant methylation and common genetic variation in the INHA promoter occur in human ACCs and are associated with decreased INHA expression.

  3. Inhibin Alpha-Subunit (INHA) Expression in Adrenocortical Cancer Is Linked to Genetic and Epigenetic INHA Promoter Variation

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    Hofland, Johannes; Steenbergen, Jacobie; Voorsluijs, Jacoba M.; Verbiest, Michael M. P. J.; de Krijger, Ronald R.; Hofland, Leo J.; de Herder, Wouter W.; Uitterlinden, Andre G.; Feelders, Richard A.; de Jong, Frank H.

    2014-01-01

    Adrenocortical carcinoma (ACC) is a rare, but highly malignant tumor of unknown origin. Inhibin α-subunit (Inha) knockout mice develop ACCs following gonadectomy. In man, INHA expression varies widely within ACC tissues and its circulating peptide inhibin pro-αC has been described as a novel tumor marker for ACC. We investigated whether genetic and epigenetic changes of the INHA gene in human ACC cause loss or variation of INHA expression. To this end, analyses of INHA sequence, promoter methylation and mRNA expression were performed in human adrenocortical tissues. Serum inhibin pro-αC levels were also measured in ACC patients. INHA genetic analysis in 37 unique ACCs revealed 10 novel, heterozygous rare variants. Of the 3 coding bases affected, one variant was synonymous and two were missense variants: S72F and S184F. The minor allele of rs11893842 at −124 bp was observed at a low frequency (24%) in ACC samples and was associated with decreased INHA mRNA levels: 4.7±1.9 arbitrary units for AA, compared to 26±11 for AG/GG genotypes (P = 0.034). The methylation of four proximal INHA promoter CpGs was aberrantly increased in five ACCs (47.7±3.9%), compared to normal adrenals (18.4±0.6%, P = 0.0052), whereas the other 14 ACCs studied showed diminished promoter methylation (9.8±1.1%, P = 0.020). CpG methylation was inversely correlated to INHA mRNA levels in ACCs (r = −0.701, p = 0.0036), but not associated with serum inhibin pro-αC levels. In conclusion, aberrant methylation and common genetic variation in the INHA promoter occur in human ACCs and are associated with decreased INHA expression. PMID:25111790

  4. INHA promoter polymorphisms are associated with premature ovarian failure.

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    Harris, Sarah E; Chand, Ashwini L; Winship, Ingrid M; Gersak, Ksenija; Nishi, Yoshihiro; Yanase, Toshihiko; Nawata, Hajime; Shelling, Andrew N

    2005-11-01

    Inhibin is an important glycoprotein that is involved in folliculogenesis. INHA, the gene encoding the inhibin alpha subunit, was recently proposed as a candidate for premature ovarian failure (POF), a syndrome that leads to the cessation of ovarian function under the age of 40 years. 70 POF patients and 70 controls were screened for the previously identified INHA -16C>T transition mutation. The T allele was found in 31/70 (44.3%) of controls, but only 18/70 (25.7%) of POF patients. This result indicates that the T allele is significantly underrepresented in the POF patient population (Fisher's exact test, two-tail: P = 0.033). Sequence analysis of the INHA promoter in 50 POF patients and 50 controls identified a highly polymorphic imperfect TG repeat at approximately -300 bp, that consisted of four common haplotypes (A, B, C and D). The -16T allele is linked to the shortest repeat haplotype (haplotype C). Despite the association between haplotype C and POF, no significant difference was found between the promoter activity of a luciferase reporter construct containing haplotype C, and most of the other haplotypes tested. Interestingly, haplotype B failed to show any promoter activity. We conclude that the inheritance of specific INHA promoter haplotypes predispose to the development of premature ovarian failure.

  5. High Affinity Inha Inhibitors with Activity Against Drug-Resistant Strains of Mycobacterium Tuberculosis

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    Sullivan,T.; Truglio, J.; Boyne, M.; Novichenok, P.; Zhang, X.; Stratton, C.; Li, H.; Kaur, T.; Amin, A.; et al.

    2006-01-01

    Novel chemotherapeutics for treating multidrug-resistant (MDR) strains of Mycobacterium tuberculosis (MTB) are required to combat the spread of tuberculosis, a disease that kills more than 2 million people annually. Using structure-based drug design, we have developed a series of alkyl diphenyl ethers that are uncompetitive inhibitors of InhA, the enoyl reductase enzyme in the MTB fatty acid biosynthesis pathway. The most potent compound has a Ki{prime} value of 1 nM for InhA and MIC{sub 99} values of 2-3 {micro}g mL{sup -1} (6-10 {micro}M) for both drug-sensitive and drug-resistant strains of MTB. Overexpression of InhA in MTB results in a 9-12-fold increase in MIC{sub 99}, consistent with the belief that these compounds target InhA within the cell. In addition, transcriptional response studies reveal that the alkyl diphenyl ethers fail to upregulate a putative efflux pump and aromatic dioxygenase, detoxification mechanisms that are triggered by the lead compound triclosan. These diphenyl ether-based InhA inhibitors do not require activation by the mycobacterial KatG enzyme, thereby circumventing the normal mechanism of resistance to the front line drug isoniazid (INH) and thus accounting for their activity against INH-resistant strains of MTB.

  6. Rastreio virtual na descoberta de possíveis inibidores da 5 α-redutase

    OpenAIRE

    Sousa, Joana Filipa Monteiro de

    2013-01-01

    Dissertação de mestrado em Química Farmacêutica Industrial, apresentada à Faculdade de Farmácia da Universidade de Coimbra A 5α-redutase é uma proteína microssomal, que converte a testosterona em dihidrotestosterona (DHT) um androgénio mais potente que tem como função induzir a diferenciação durante o desenvolvimento do feto que conduz ao desenvolvimento dos genitais externos masculinos. Quando ocorrem distúrbios no desempenho desta enzima, podem surgir distúrbios como o pseudo...

  7. Antitubercular drugs for an old target: GSK693 as a promising InhA direct inhibitor

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    María Martínez-Hoyos

    2016-06-01

    Full Text Available Despite being one of the first antitubercular agents identified, isoniazid (INH is still the most prescribed drug for prophylaxis and tuberculosis (TB treatment and, together with rifampicin, the pillars of current chemotherapy. A high percentage of isoniazid resistance is linked to mutations in the pro-drug activating enzyme KatG, so the discovery of direct inhibitors (DI of the enoyl-ACP reductase (InhA has been pursued by many groups leading to the identification of different enzyme inhibitors, active against Mycobacterium tuberculosis (Mtb, but with poor physicochemical properties to be considered as preclinical candidates. Here, we present a series of InhA DI active against multidrug (MDR and extensively (XDR drug-resistant clinical isolates as well as in TB murine models when orally dosed that can be a promising foundation for a future treatment.

  8. Phosphorylation of InhA inhibits mycolic acid biosynthesis and growth of Mycobacterium tuberculosis

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    Molle, Virginie; Gulten, Gulcin; Vilchèze, Catherine; Veyron-Churlet, Romain; Zanella-Cléon, Isabelle; Sacchettini, James C.; Jacobs, Jr, William R.; Kremer, Laurent (CNRS-UMR); (Einstein); (TAM)

    2011-08-24

    The remarkable survival ability of Mycobacterium tuberculosis in infected hosts is related to the presence of cell wall-associated mycolic acids. Despite their importance, the mechanisms that modulate expression of these lipids in response to environmental changes are unknown. Here we demonstrate that the enoyl-ACP reductase activity of InhA, an essential enzyme of the mycolic acid biosynthetic pathway and the primary target of the anti-tubercular drug isoniazid, is controlled via phosphorylation. Thr-266 is the unique kinase phosphoacceptor, both in vitro and in vivo. The physiological relevance of Thr-266 phosphorylation was demonstrated using inhA phosphoablative (T266A) or phosphomimetic (T266D/E) mutants. Enoyl reductase activity was severely impaired in the mimetic mutants in vitro, as a consequence of a reduced binding affinity to NADH. Importantly, introduction of inhA{_}T266D/E failed to complement growth and mycolic acid defects of an inhA-thermosensitive Mycobacterium smegmatis strain, in a similar manner to what is observed following isoniazid treatment. This study suggests that phosphorylation of InhA may represent an unusual mechanism that allows M. tuberculosis to regulate its mycolic acid content, thus offering a new approach to future anti-tuberculosis drug development.

  9. Mutações no gene da metilenotetrahidrofolato redutase e síndrome de Down

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    Grillo Laura Brunelli das Neves

    2002-01-01

    Full Text Available Sindrome de Down (SD é uma alteração genética e metabólica complexa atribuída à presença de três cópias do cromossomo 21. O cromossomo extra em 93% dos casos é de origem materna e é resultante de uma segregação anormal durante a meiose (não-disjunção. Com exceção da idade materna avançada, fatores de risco para a não-disjunção meiótica não estão bem estabelecidos. Um estudo preliminar sugeriu que o metabolismo anormal do folato e a mutação 677 (C->T no gene da metilenotetrahidrofolato redutase (MTHFR podem ser fatores de risco maternos para a SD. A freqüência das mutações MTHFR 677 (C->T e 1.298 (A->C foram avaliadas em 36 mães de crianças com SD e em 200 indivíduos-controle. Os resultados demonstraram que as mutações 677 (C->T e 1.298 (A->C são mais prevalentes entre mães de crianças com SD do que nos controles. A heterozigose das duas mutações foi a combinação mais freqüente. O resultado desse estudo inicial sugere que mutações no gene da MTHFR seriam um fator de risco para a SD.

  10. Mutações no gene da metilenotetrahidrofolato redutase e síndrome de Down

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    Laura Brunelli das Neves Grillo

    2002-12-01

    Full Text Available Sindrome de Down (SD é uma alteração genética e metabólica complexa atribuída à presença de três cópias do cromossomo 21. O cromossomo extra em 93% dos casos é de origem materna e é resultante de uma segregação anormal durante a meiose (não-disjunção. Com exceção da idade materna avançada, fatores de risco para a não-disjunção meiótica não estão bem estabelecidos. Um estudo preliminar sugeriu que o metabolismo anormal do folato e a mutação 677 (C->T no gene da metilenotetrahidrofolato redutase (MTHFR podem ser fatores de risco maternos para a SD. A freqüência das mutações MTHFR 677 (C->T e 1.298 (A->C foram avaliadas em 36 mães de crianças com SD e em 200 indivíduos-controle. Os resultados demonstraram que as mutações 677 (C->T e 1.298 (A->C são mais prevalentes entre mães de crianças com SD do que nos controles. A heterozigose das duas mutações foi a combinação mais freqüente. O resultado desse estudo inicial sugere que mutações no gene da MTHFR seriam um fator de risco para a SD.

  11. Controle da atividade da nitrato redutase em plantas de abacaxizeiro submetidas a baixas temperaturas em diferentes fases do ciclo diurno

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    Aline Tiemi Matsumura

    2013-01-01

    O nitrato é uma das principais fontes de nitrogênio disponível para as plantas, sendo a nitrato redutase (NR) a enzima responsável pela sua redução a nitrito. O nitrito é considerado tóxico em altas concentrações e, por esse motivo, a atividade da NR possui uma regulação complexa, principalmente em nível transcricional e pós-traducional. Trabalhos anteriores do nosso grupo, utilizando plantas de abacaxizeiro cultivadas in vitro, demonstraram que, em condições de termoperíodo de 28ºC dia/15ºC ...

  12. AMPLIFIKASI DAN IDENTIFIKASI MUTASI REGIO PROMOTER inhA PADA ISOLAT Mycobacterium tuberculosis MULTIDRUG RESISTANCE DENGAN TEKNIK POLYMERASE CHAIN REACTION

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    Devita Kusdianingrum

    2014-10-01

    Full Text Available ABSTRAK: Sekitar 8-20% isolate M. tuberculosis yang resisten terhadap isoniazid diketahui telah mengalami mutasi pada posisi regio promoter inhA [1]. Untuk memperoleh titik mutasi pada regio promoter, maka amplifikasi fragmen target perlu untuk dilakukan. Tujuan dilakukannya penelitian ini adalah untuk mengamplifikasi regio promoter inhA, mengetahui ada tidaknya mutasi dan jenis mutasi pada isolat 134 MDR-TB. Tahap isolasi DNA dilakukan menggunakan metode Boom yang telah dimodifikasi. Fragmen target diamplifikasi dengan teknik PCR menggunakan sepasang primer (forward primer 5’ ACATACCTGCTGCGCAAT 3’ dan reverse primer 5’ CTCCGGTAACCAGGACT GAA 3’. Amplikon disekuensing secara satu arah menggunakan forward primer. Analisis homologi dilakukan menggunakan program online BLASTn, sementara identifikasi mutasi dilakukan menggunakan software MEGA4. Hasil penelitian menunjukkan bahwa analisis homologi isolate 134 terhadap M. tuberculosis H37Rv adalah sebesar 99%. Tahap analisis mutasi menemukan terjadinya perubahan sitosin menjadi timin (CàT pada posisi -15 isolat 134 MDR-TB ABSTRACT: Approximately 8-20% M. tuberculosis isolates that are resistant to isoniazid habe been known to have a mutation in inhA promoter region [1]. To find the mutation in inhA promoter region, it is necessary to carry out the amplification of the target fragment. The purpose of this research were to amplify the inhA promoter region and to find out if there is a mutation and type of mutation at MDR-TB isolate. DNA isolation was done by a modified Boom method. Target fragment was amplified by a pair primer (forward primer 5’ ACATACCTGCTGCGCAAT 3’ and reverse primer 5’ CTCCGGTAACCAGGACT GAA 3’ using Polymerase Chain Reaction (PCR technique. Amplicon was sequenced in one forward direction. Homology analysis was conducted by online BLASTn program, while the mutation was identified by MEGA4. The result of this research showed that homology analysis of 134 was homolog

  13. Atividade da redutase do nitrato e fluorescência da clorofila a em mamoeiro Nitrate reductase activity and chlorophyll a fluorescence in papaya

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    Renata Venturim Fontes

    2008-03-01

    Full Text Available O objetivo do presente trabalho foi correlacionar a atividade da redutase do nitrato e a eficiência fotoquímica máxima do fotossistema II (FSII, expressa pela razão F V/F M (F V = fluorescência variável e F M = fluorescência máxima, em mamoeiro (Carica papaya L. cv. Tainung 01 e Sunrise Solo 72/12 em condições de campo. O potencial fotoquímico do FSII foi medido in situ em folhas adaptadas ao escuro. Depois, nas mesmas folhas, foi medida a atividade da enzima. Não houve diferença significativa entre a eficiência fotoquímica máxima do FSII entre as cultivares Tainung 01 e Sunrise Solo 72/12, porém a atividade da redutase do nitrato foi notoriamente maior na primeira. A atividade da redutase do nitrato foi altamente correlacionada à eficiência fotoquímica máxima do FS II tanto na cv. Tainung 01 (coeficiente de correlação r= 0,740 e coeficiente de determinação r²= 0,706 quanto na Sunrise Solo 72/12 (coeficiente de correlação r= 0,960 e coeficiente de determinação r²= 0,945. Esses resultados sugerem que há uma correlação entre a fluorescência da clorofila a e a atividade da redutase do nitrato nessas plantas.The objective of the present work was to connect the nitrate reductase activity and the photochemical efficiency of the photosystem II (PSII, measured as F V/F M, in papaya (Carica papaya L. cv. Tainung 01 and Sunrise Solo 72/12 in field conditions. The photochemical potential of PSII was measured in situ on attached leaves that had been dark adapted, prior to the fluorescence measurement. After, in same leaves, the enzyme activity was measured. There was not significant difference among the photochemical efficiency of PSII between cv. Tainung 01 and Sunrise Solo 72/12, even so, the activity of the enzyme was markedly higher in cv. Tainung 01 than in cv. Sunrise Solo 72/12. The nitrate redutase activity was highly correlated with the photochemical efficiency of the PSII in both cv. Tainung 01 (coefficient of

  14. High Prevalence of inhA Promoter Mutations among Patients with Drug-Resistant Tuberculosis in KwaZulu-Natal, South Africa.

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    Abraham J Niehaus

    Full Text Available Drug-resistant tuberculosis (TB remains extremely difficult to treat because there are often few remaining active medications and limited diagnostic options to detect resistance. Resistance to isoniazid is typically caused by mutations in either katG or the inhA promoter. inhA mutations confer low-level resistance to isoniazid and cross-resistance to ethionamide while katG mutations confer high-level isoniazid resistance and no cross-resistance. Line Probe Assays (LPAs that detect mutations in katG and inhA are currently performed on all positive TB cultures in KwaZulu-Natal province, South Africa, but the frequency of inhA mutations in drug-resistant TB patients has not been examined.We sought to determine the proportion of patients who could potentially benefit from high-dose isoniazid and who may be resistant to ethionamide. We reviewed 994 LPA (Hain MTBDRplus results at the TB reference laboratory in KwaZulu-Natal to determine the frequency of mutations in either katG or the inhA promoter. We stratified these results by drug-resistance category (i.e., MDR-TB, pre-XDR-TB, and XDR-TB as determined by phenotypic drug-susceptibility testing.Among MDR- and XDR-TB isolates, the prevalence of inhA mutations without a concurrent katG mutation was 14.8% and 10.3% respectively. The prevalence of inhA mutations with OR without a katG mutation was 30.3% and 82.8%, respectively.More than 10% of patients with MDR- and XDR-TB may benefit from high-dose isoniazid. Although ethionamide is empirically included in all MDR- and XDR-TB regimens, nearly a third of MDR-TB patients and a majority of XDR-TB patients likely have resistance to ethionamide. Laboratories performing line probe assays should report specific band patterns so that clinicians may adjust treatment regimens accordingly.

  15. High Prevalence of inhA Promoter Mutations among Patients with Drug-Resistant Tuberculosis in KwaZulu-Natal, South Africa

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    Niehaus, Abraham J.; Mlisana, Koleka; Gandhi, Neel R.; Mathema, Barun; Brust, James C. M.

    2015-01-01

    Background Drug-resistant tuberculosis (TB) remains extremely difficult to treat because there are often few remaining active medications and limited diagnostic options to detect resistance. Resistance to isoniazid is typically caused by mutations in either katG or the inhA promoter. inhA mutations confer low-level resistance to isoniazid and cross-resistance to ethionamide while katG mutations confer high-level isoniazid resistance and no cross-resistance. Line Probe Assays (LPAs) that detect mutations in katG and inhA are currently performed on all positive TB cultures in KwaZulu-Natal province, South Africa, but the frequency of inhA mutations in drug-resistant TB patients has not been examined. Methods We sought to determine the proportion of patients who could potentially benefit from high-dose isoniazid and who may be resistant to ethionamide. We reviewed 994 LPA (Hain MTBDRplus) results at the TB reference laboratory in KwaZulu-Natal to determine the frequency of mutations in either katG or the inhA promoter. We stratified these results by drug-resistance category (i.e., MDR-TB, pre-XDR-TB, and XDR-TB) as determined by phenotypic drug-susceptibility testing. Results Among MDR- and XDR-TB isolates, the prevalence of inhA mutations without a concurrent katG mutation was 14.8% and 10.3% respectively. The prevalence of inhA mutations with OR without a katG mutation was 30.3% and 82.8%, respectively. Conclusion More than 10% of patients with MDR- and XDR-TB may benefit from high-dose isoniazid. Although ethionamide is empirically included in all MDR- and XDR-TB regimens, nearly a third of MDR-TB patients and a majority of XDR-TB patients likely have resistance to ethionamide. Laboratories performing line probe assays should report specific band patterns so that clinicians may adjust treatment regimens accordingly. PMID:26332235

  16. Cross-docking study on InhA inhibitors: a combination of Autodock Vina and PM6-DH2 simulations to retrieve bio-active conformations.

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    Stigliani, Jean-Luc; Bernardes-Génisson, Vania; Bernadou, Jean; Pratviel, Geneviève

    2012-08-21

    InhA, the NADH-dependent enoyl-acyl carrier protein reductase from Mycobacterium tuberculosis (Mtb) is the proposed main target of the first-line antituberculosis drug isoniazid (INH). INH activity is dependent on activation by the catalase peroxidase KatG, a Mtb enzyme whose mutations are linked to clinical resistance to INH. Other inhibitors of InhA that do not require any preliminary activation are known. The design of such direct potent inhibitors represents a promising approach to circumvent this resistance mechanism. An ensemble-docking process with four known InhA X-ray crystal structures and employing the Autodock Vina software was performed. Five InhA inhibitors whose bioactive conformations are known were sequentially docked in the substrate cavity of each protein. The efficiency of the docking was assessed and validated by comparing the calculated conformations to the crystallographic structures. For a same inhibitor, the docking results differed from one InhA conformation to another; however, docking poses that matched correctly or were very close to the expected bioactive conformations could be identified. The expected conformations were not systematically well ranked by the Autodock Vina scoring function. A post-docking optimization was carried out on all the docked conformations with the AMMP force field implemented on the VEGAZZ software, followed by a single point calculation of the interaction energy, using the MOPAC PM6-DH2 semi-empirical quantum chemistry method. The conformations were subsequently submitted to a PM6-DH2 optimization in partially flexible cavities. The resulting interaction energies combined with the multiple receptor conformations approach allowed us to retrieve the bioactive conformation of each ligand.

  17. Obtaining partial purified xylose reductase from Candida guilliermondii Obtenção de xilose redutase de Candida guilliermondii parcialmente purificada

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    Ester Junko Tomotani

    2009-09-01

    Full Text Available The enzymatic bioconversion of xylose into xylitol by xylose reductase (XR is an alternative for chemical and microbiological processes. The partial purified XR was obtained by using the following three procedures: an agarose column, a membrane reactor or an Amicon Ultra-15 50K Centrifugal Filter device at yields of 40%, 7% and 67%, respectively.A bioconversão enzimática da xilose em xilitol pela xilose redutase (XR é uma alternativa para as vias química e microbiológica. Avaliouse a purificação parcial da XR, utilizando os três seguintes procedimentos: uma coluna de agarose, um reator com membrana ou tubos de ultracentrifugação Amicon Ultra-15 50K, com rendimento de 40%, 7% ou 67%, respectivamente.

  18. Miopatia por deficiência de succinato-citocromo-C-redutase: possível defeito no complexo II da cadeia respiratória

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    Lineu Cesar Werneck

    1989-12-01

    Full Text Available Relato do caso de mulher de 24 anos de idade que apresentava astenia desde a puberdade, com agravamento nos últimos anos, cuja biópsia muscular revelou grande acúmulo de mitocôndrias. As dosagens dos enzimas mitocondriais mostrou importante redução da succinato-citocromo-C-redutase, sugerindo defeito na cadeia respiratória a nível do complexo II. Medicada com altas doses de vitamina C e K, melhorou da força muscular. São feitas considerações a respeito das principais síndromes com miopatias mitocondriais, bem como a respeito dos métodos de investigação em defeitos da cadeia respiratória.

  19. Detection of the mutations in katG315 and inhA-15 of Mycobacterium tuberculosis strains isolated from Chinese patients

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    @@ The situation of tuberculosis and M. tuberculosis (MTB) drug resistance is severe in China. Among the MTB isolates, 27.8% were resistant to one primary antituberculosis drug at least. Isoniazid (INH) is one of the first-line anti-TB drugs. The rates of primary resistance and acquired resistance to INH were 11.0% and 31.0%, respectively.1 MTB strains resistance to INH is mainly caused by the alterations in several genes encoding the molecular targets as follows: catalase peroxidase (katG), the enoyl-acyl carrier protein reductase (inhA), the alkyl hydroperoxide reductase (ahpC), beta-ketoacyl-acyl carrier protein synthase (kasA), and NADH dehydrogenase (ndh).2-4 However, the mutations of katG and inhA promoter region, especially in codon 315 of katG, were found in the majority of INH-resistant clinical isolates.

  20. Biological evaluation of potent triclosan-derived inhibitors of the enoyl-acyl carrier protein reductase InhA in drug-sensitive and drug-resistant strains of Mycobacterium tuberculosis.

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    Stec, Jozef; Vilchèze, Catherine; Lun, Shichun; Perryman, Alexander L; Wang, Xin; Freundlich, Joel S; Bishai, William; Jacobs, William R; Kozikowski, Alan P

    2014-11-01

    New triclosan (TRC) analogues were evaluated for their activity against the enoyl-acyl carrier protein reductase InhA in Mycobacterium tuberculosis (Mtb). TRC is a well-known inhibitor of InhA, and specific modifications to its positions 5 and 4' afforded 27 derivatives; of these compounds, seven derivatives showed improved potency over that of TRC. These analogues were active against both drug-susceptible and drug-resistant Mtb strains. The most active compound in this series, 4-(n-butyl)-1,2,3-triazolyl TRC derivative 3, had an MIC value of 0.6 μg mL(-1) (1.5 μM) against wild-type Mtb. At a concentration equal to its MIC, this compound inhibited purified InhA by 98 %, and showed an IC50 value of 90 nM. Compound 3 and the 5-methylisoxazole-modified TRC 14 were able to inhibit the biosynthesis of mycolic acids. Furthermore, mc(2) 4914, an Mtb strain overexpressing inhA, was found to be less susceptible to compounds 3 and 14, supporting the notion that InhA is the likely molecular target of the TRC derivatives presented herein. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  1. Detection of katG and inhA mutations to guide isoniazid and ethionamide use for drug-resistant tuberculosis

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    Bollela, V. R.; Namburete, E. I.; Feliciano, C. S.; Macheque, D.; Harrison, L. H.; Caminero, J. A.

    2017-01-01

    SUMMARY BACKGROUND Depending on the presence of mutations that determine isoniazid (INH) susceptibility (katG and inhA), Mycobacterium tuberculosis may be susceptible to high doses of INH or ethionamide (ETH). OBJECTIVE To describe the INH resistance profile and association of katG mutation with previous INH treatment and level of drug resistance based on rapid molecular drug susceptibility testing (DST) in southern Brazil and central Mozambique. DESIGN Descriptive study of 311 isolates from Ribeirão Preto, São Paulo, Brazil (2011–2014) and 155 isolates from Beira, Mozambique (2014–2015). Drug resistance patterns and specific gene mutations were determined using GenoType® MTBDRplus. RESULTS katG gene mutations were detected in 12/22 (54.5%) Brazilian and 32/38 (84.2%) Mozambican isolates. inhA mutations were observed in 9/22 (40.9%) isolates in Brazil and in 4/38 (10.5%) in Mozambique. Both katG and inhA mutations were detected in respectively 1/22 (5%) and 2/38 (5.2%). The difference in the frequency of katG mutations in Brazil and Mozambique was statistically significant (P = 0.04). katG mutations were present in 68.8% (33/48) of patients previously treated with INH and 31.2% (15/48) of patients without previous INH. This difference was not statistically significant (P = 0.223). CONCLUSION INH mutations varied geographically; molecular DST can be used to guide and accelerate decision making in the use of ETH or high doses of INH. PMID:27393546

  2. Radiolabelling and positron emission tomography of PT70, a time-dependent inhibitor of InhA, the Mycobacterium tuberculosis enoyl-ACP reductase.

    Science.gov (United States)

    Wang, Hui; Liu, Li; Lu, Yang; Pan, Pan; Hooker, Jacob M; Fowler, Joanna S; Tonge, Peter J

    2015-11-01

    PT70 is a diaryl ether inhibitor of InhA, the enoyl-ACP reductase in the Mycobacterium tuberculosis fatty acid biosynthesis pathway. It has a residence time of 24 min on the target, and also shows antibacterial activity in a mouse model of tuberculosis infection. Due to the interest in studying target tissue pharmacokinetics of PT70, we developed a method to radiolabel PT70 with carbon-11 and have studied its pharmacokinetics in mice and baboons using positron emission tomography. Copyright © 2015 Elsevier Ltd. All rights reserved.

  3. Function of heterologous Mycobacterium tuberculosis InhA, a type 2 fatty acid synthase enzyme involved in extending C20 fatty acids to C60-to-C90 mycolic acids, during de novo lipoic acid synthesis in Saccharomyces cerevisiae.

    Science.gov (United States)

    Gurvitz, Aner; Hiltunen, J Kalervo; Kastaniotis, Alexander J

    2008-08-01

    We describe the physiological function of heterologously expressed Mycobacterium tuberculosis InhA during de novo lipoic acid synthesis in yeast (Saccharomyces cerevisiae) mitochondria. InhA, representing 2-trans-enoyl-acyl carrier protein reductase and the target for the front-line antituberculous drug isoniazid, is involved in the activity of dissociative type 2 fatty acid synthase (FASII) that extends associative type 1 fatty acid synthase (FASI)-derived C(20) fatty acids to form C(60)-to-C(90) mycolic acids. Mycolic acids are major constituents of the protective layer around the pathogen that contribute to virulence and resistance to certain antimicrobials. Unlike FASI, FASII is thought to be incapable of de novo biosynthesis of fatty acids. Here, the genes for InhA (Rv1484) and four similar proteins (Rv0927c, Rv3485c, Rv3530c, and Rv3559c) were expressed in S. cerevisiae etr1Delta cells lacking mitochondrial 2-trans-enoyl-thioester reductase activity. The phenotype of the yeast mutants includes the inability to produce sufficient levels of lipoic acid, form mitochondrial cytochromes, respire, or grow on nonfermentable carbon sources. Yeast etr1Delta cells expressing mitochondrial InhA were able to respire, grow on glycerol, and produce lipoic acid. Commensurate with a role in mitochondrial de novo fatty acid biosynthesis, InhA could accept in vivo much shorter acyl-thioesters (C(4) to C(8)) than was previously thought (>C(12)). Moreover, InhA functioned in the absence of AcpM or protein-protein interactions with its native FASII partners KasA, KasB, FabD, and FabH. None of the four proteins similar to InhA complemented the yeast mutant phenotype. We discuss the implications of our findings with reference to lipoic acid synthesis in M. tuberculosis and the potential use of yeast FASII mutants for investigating the physiological function of drug-targeted pathogen enzymes involved in fatty acid biosynthesis.

  4. Atividade da enzima redutase do nitrato em algodoeiro submetido ao déficit hídrico e posterior recuperação da turgescência

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    Marur Celso Jamil

    2000-01-01

    Full Text Available A atividade da redutase do nitrato foi estudada em plantas de algodão (Gossypium hirsutum, L., cvs IAC 20 e IAC 13-1, submetidas a déficits crescentes de água, e posterior recuperação da turgescência. A atividade fisiológica das folhas, representada pela atividade da enzima, sofreu redução quando o y a foi menor que -1,5 MPa. Não ocorreram diferenças significativas entre os dois cultivares em relação ao estabelecimento do estresse, nem tampouco quanto ao padrão de recuperação da turgescência em plantas estressadas. O ensaio in vivo da atividade da redutase do nitrato mostrou ser um método sensível para a avaliação do efeito do estresse hídrico sobre o crescimento das plantas, constituindo-se em um parâmetro indicativo do nível geral do metabolismo na planta.

  5. Atividades de nitrogenase e redutase de nitrato e produtividade do feijoeiro “Ouro Negro” em resposta à adubação foliar com molibdênio

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    A. C. S. Pessoa

    2001-03-01

    Full Text Available A efetividade do suprimento de nitrogênio ao feijoeiro, por meio de fertilizante e, ou, de fixação biológica, e a dependência desses processos à disponibilidade de molibdênio constituem, ainda, um problema mal resolvido. Objetivando avaliar os efeitos da aplicação foliar de Mo na atividade das enzimas nitrogenase e redutase do nitrato e na produtividade do feijoeiro, realizou-se um experimento, em condições de campo, em um Podzólico Vermelho-Amarelo do município de Coimbra (MG. Os tratamentos constituíram-se de doses crescentes de Mo (0, 40, 80 e 120 g ha-1 de Mo, aplicadas em adubação foliar aos 25 dias da emergência. Na adubação de base, usaram-se 20 kg ha-1 de N e 60 kg ha-1 de K2O. Não foi aplicado nitrogênio em cobertura. A aplicação foliar de Mo aumentou as atividades da nitrogenase e da redutase do nitrato, mantendo-as em patamares mais altos durante o ciclo da cultura, proporcionando maiores teores de N nas folhas e maior produtividade. A eficiência máxima foi alcançada com 80 g ha-1 de Mo, com produtividade de 1.893 kg ha-1 de grãos, 3,23 vezes maior que a da testemunha (sem Mo. A aplicação foliar de Mo aumentou, também, em 1,54 vez o número de vagens por planta, em 0,23 o número de grãos por vagem e em 0,15 o peso de 100 grãos, em comparação ao tratamento que não recebeu Mo.

  6. Normal bone density in male pseudohermaphroditism due to 5alpha- reductase 2 deficiency Pseudohermafroditas masculinos por deficiência de 5alfa-redutase 2 apresentam densidade óssea normal

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    Elaine Maria Frade Costa

    2001-10-01

    Full Text Available Bone is an androgen-dependent tissue, but it is not clear whether the androgen action in bone depends on testosterone or on dihydrotestosterone. Patients with 5alpha-reductase 2 deficiency present normal levels of testosterone and low levels of dihydrotestosterone, providing an in vivo human model for the analysis of the effect of testosterone on bone. OBJECTIVE: To analyze bone mineral density in 4 adult patients with male pseudohermaphroditism due to 5alpha-reductase 2 deficiency. RESULTS: Three patients presented normal bone mineral density of the lumbar column (L1-L4 and femur neck, and the other patient presented a slight osteopenia in the lumbar column. CONCLUSION: Patients with dihydrotestosterone deficiency present normal bone mineral density, suggesting that dihydrotestosterone is not the main androgen acting in bone.O tecido ósseo é um tecido andrógeno-dependente porém não está claro se a ação androgênica depende da testosterona ou da diidrotestosterona. Os pacientes portadores de deficiência de 5alfa-redutase tipo 2, constituem um modelo natural para avaliar o efeito isolado da testosterona sobre a massa óssea. OBJETIVO: Avaliar a densidade mineral óssea em quatro pacientes adultos portadores de pseudohermafroditismo masculino por deficiência da 5alfa-redutase tipo 2. RESULTADOS: Três pacientes apresentaram densidade mineral óssea normal na coluna lombar e fêmur e o quarto paciente apresentou osteopenia leve em coluna lombar. CONCLUSÃO: Pacientes com deficiência de diidrotestosterona apresentam densidade mineral óssea normal sugerindo que a diidrotestosterona não é o andrógeno que age sobre o osso.

  7. Efeito da aplicação de piraclostrobina na taxa fotossintética, respiração, atividade da enzima nitrato redutase e produtividade de grãos de soja

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    Evandro Binotto Fagan

    2010-12-01

    Full Text Available Com objetivo de avaliar o efeito da aplicação da piraclostrobina em variáveis fisiológicas e fenométricas da cultura de soja, realizou-se um experimento de campo entre novembro de 2005 e maio de 2006. Foram realizadas avaliações de taxa fotossintética e respiração. Também foi avaliada a atividade da enzima nitrato redutase, massa de mil grãos e produtividade. O delineamento experimental foi de blocos ao acaso com três tratamentos (T1 - sem aplicação de fungicida, T2 - duas aplicações da piraclostrobina e T3 - duas aplicações de tebuconazol - triazol com quatro repetições. A aplicação da piraclostrobina ocasionou incremento na taxa fotossintética. A atividade da enzima nitrato redutase foliar somente foi incrementada pela aplicação de estrobilurina nos 15 dias após a primeira aplicação. As plantas tratadas com estrobilurina tiveram o acréscimo de 7% e 8% na massa de mil grãos e de 1080 e 468 kg ha-1na produtividade quando comparado à testemunha sem aplicação e ao tratamento com triazol (tebuconazol respectivamente. Os resultados indicam que a piraclostrobina (estrobilurina afeta a taxa de assimilação de carbono e de nitrogênio na cultura de soja, o que é refletido na produtividade de grãos.

  8. Mutações no gene da metilenotetrahidrofolato redutase e síndrome de Down Mutations in the methylene-tetrahydrofolate reductase gene and Down syndrome

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    Laura Brunelli das Neves Grillo

    2002-12-01

    Full Text Available Sindrome de Down (SD é uma alteração genética e metabólica complexa atribuída à presença de três cópias do cromossomo 21. O cromossomo extra em 93% dos casos é de origem materna e é resultante de uma segregação anormal durante a meiose (não-disjunção. Com exceção da idade materna avançada, fatores de risco para a não-disjunção meiótica não estão bem estabelecidos. Um estudo preliminar sugeriu que o metabolismo anormal do folato e a mutação 677 (C->T no gene da metilenotetrahidrofolato redutase (MTHFR podem ser fatores de risco maternos para a SD. A freqüência das mutações MTHFR 677 (C->T e 1.298 (A->C foram avaliadas em 36 mães de crianças com SD e em 200 indivíduos-controle. Os resultados demonstraram que as mutações 677 (C->T e 1.298 (A->C são mais prevalentes entre mães de crianças com SD do que nos controles. A heterozigose das duas mutações foi a combinação mais freqüente. O resultado desse estudo inicial sugere que mutações no gene da MTHFR seriam um fator de risco para a SD.Down syndrome (DS is a complex genetic and metabolic disorder attributed to the presence of three copies of chromosome 21. The extra chromosome derives from the mother in 93% of cases and is due to abnormal chromosome segregation during meiosis (nondisjunction. Except for advanced age at conception, maternal risk factors for meiotic nondisjunction are not well established. A recent preliminary study suggested that abnormal folate metabolism and the 677 (C->T mutation in the methylene-tetrahydrofolate reductase (MTHFR gene may be maternal risk factors for DS. Frequency of the MTHFR 677 (C->T and 1298 (A->C mutations was evaluated in 36 mothers of children with DS and in 200 controls. The results are consistent with the observation that the MTHFR 677 (C->T and 1298 (A->C mutations are more prevalent among mothers of children with DS than controls. In addition, the most prevalent genotype was the combination of both mutations

  9. Correlations of mutations in katG, oxyR-ahpC and inhA genes and in vitro susceptibility in Mycobacterium tuberculosis clinical strains segregated by spoligotype families from tuberculosis prevalent countries in South America

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    Suffys Philip N

    2009-02-01

    Full Text Available Abstract Background Mutations associated with resistance to rifampin or streptomycin have been reported for W/Beijing and Latin American Mediterranean (LAM strain families of Mycobacterium tuberculosis. A few studies with limited sample sizes have separately evaluated mutations in katG, ahpC and inhA genes that are associated with isoniazid (INH resistance. Increasing prevalence of INH resistance, especially in high tuberculosis (TB prevalent countries is worsening the burden of TB control programs, since similar transmission rates are noted for INH susceptible and resistant M. tuberculosis strains. Results We, therefore, conducted a comprehensive evaluation of INH resistant M. tuberculosis strains (n = 224 from three South American countries with high burden of drug resistant TB to characterize mutations in katG, ahpC and inhA gene loci and correlate with minimal inhibitory concentrations (MIC levels and spoligotype strain family. Mutations in katG were observed in 181 (80.8% of the isolates of which 178 (98.3% was contributed by the katG S315T mutation. Additional mutations seen included oxyR-ahpC; inhA regulatory region and inhA structural gene. The S315T katG mutation was significantly more likely to be associated with MIC for INH ≥2 μg/mL. The S315T katG mutation was also more frequent in Haarlem family strains than LAM (n = 81 and T strain families. Conclusion Our data suggests that genetic screening for the S315T katG mutation may provide rapid information for anti-TB regimen selection, epidemiological monitoring of INH resistance and, possibly, to track transmission of INH resistant strains.

  10. ACCUMULATION OF NITROGEN COMPOUNDS AND NITRATE REDUCTASE ACTIVITY IN LETTUCE CULTIVATED IN DIFFERENT CROP SYSTEMS ACÚMULO DE COMPOSTOS NITROGENADOS E ATIVIDADE DA REDUTASE DO NITRATO EM ALFACE PRODUZIDA EM DIFERENTES SISTEMAS DE CULTIVO

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    Fernanda Nunes Ibrahim

    2008-09-01

    limits recommended by literature.

    KEY-WORDS: Vegetables; quality; nitrite; metabolism of nitrogen.

    O teor de nitrato é um importante índice da qualidade dos alimentos, pois quando ingerido nitrato é reduzido a nitrito, o qual pode gerar compostos prejudiciais ao organismo humano. Diante disso, o presente trabalho teve como objetivo estudar o transporte e acúmulo de compostos nitrogenados e a atividade da enzima redutase do nitrato em alface cultivar 'Vera' produzida em Registro (SP em diferentes sistemas de cultivo. Foram coletadas amostras da seiva do xilema, da parte aérea e da raiz das plantas para quantificação de compostos nitrogenados e atividade da redutase do nitrato in vivo. A concentração de nitrato na seiva xilema, os teores de nitrato e aminoácidos, bem como a atividade da redutase do nitrato, demonstraram transporte, acumulação e assimilação mais intensos em plantas cultivadas em hidroponia, seguido do sistema convencional e por último, do orgâ

  11. N-partitioning, nitrate reductase and glutamine synthetase activities in two contrasting varieties of maize Partição de nitrogênio e atividade das enzimas nitrato redutase e glutamina sintetase em duas cultivares contrastantes de milho

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    Altair Toledo Machado

    2001-02-01

    Full Text Available In order to identify useful parameters for maize genetic breeding programs aiming at a more efficient use of N, two maize varieties of contrasting N efficiency, Sol da Manhã NF (efficient and Catetão (inefficient were compared. Experiments were carried out under field and greenhouse conditions, at low and high N levels. The parameters analysed included total and relative plant and grain N content, biomass and the activities of nitrate reductase and glutamine synthetase in different parts of the plant. It was found that the translocation efficiency of N and photoassimilates to the developing seeds and the source-sink relations were significantly different for the two varieties. N content of the whole plant and grain, cob weight and the relative ear dry weight were useful parameters for characterizing the variety Sol da Manhã NF as to its efficient use of N. Enzymes activity of glutamine synthetase (transferase reaction and nitrate reductase did not differ among the varieties.Com o objetivo de identificar parâmetros que possam ser utilizados em programas de melhoramento genético em milho para uso eficiente de N, duas cultivares de milho contrastantes quanto ao uso deste nutriente, Sol da Manhã NF (eficiente e Catetão (não-eficiente, foram avaliadas em dois experimentos conduzidos no campo e em casa de vegetação, respectivamente, sob nível baixo e alto de N. Os caracteres avaliados foram: teor e conteúdo de N em diferentes partes da planta; massa seca; peso dos grãos e de diferentes partes da planta; biomassa, e atividade das enzimas nitrato redutase e glutamina sintetase. O mecanismo de translocação de N e de fotoassimilados para os grãos e a relação fonte/dreno foram importantes para diferenciar a cultivar eficiente da não-eficiente. Conteúdo de N nos grãos e total das plantas, peso do sabugo e relação peso de espiga/matéria seca foram importantes para caracterizar a cultivar Sol da Manhã NF eficiente no uso do N. A

  12. Methylenetetrahydrofolate reductase gene polymorphism is not related to the risk of ischemic cerebrovascular disease in a Brazilian population Polimorfismo no gene de metilenetetrahidrofolato redutase não está relacionado com o risco de doença cerebrovascular isquêmica em uma população brasileira

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    Samuel Katsuyuki Shinjo

    2007-01-01

    Full Text Available PURPOSE: Data are conflicting concerning the risk for ischemic stroke associated with a common polymorphism in the gene encoding 5,10-methylenetetrahydrofolate reductase C677T, which predisposes carriers to hyperhomocysteinemia. A meta-analysis study suggested that the 5,10-methylenetetrahydrofolate reductase 677TT genotype might have a small influence in determining susceptibility to ischemic stroke. METHODS: We analyzed the 5,10-methylenetetrahydrofolate reductase 677TT genotype polymorphism in Brazilian subjects with ischemic stroke, using a case-control design. RESULTS: We compared 5,10-methylenetetrahydrofolate reductase genotypes in groups of subjects presenting ischemic stroke (n = 127 and normal control (n = 126 and found an odds ratio of 1.97 (95% CI, 0.84-4.64 in a multivariate analysis in which results were adjusted to baseline clinical characteristics of study participants. CONCLUSION: We found that the homozygous 5,10-methylenetetrahydrofolate reductase C677T genotype was not a risk factor for ischemic stroke in these Brazilian subjects.OBJETIVO: Os dados são conflitantes em relação a risco de acidente cerebrovascular associado a polimorfismo do gene 5,10-metilenetetrahidrofolato redutase C677T, o qual predispõe a hiperhomocisteinemia. Um estudo de meta-análise sugere que o genotipo 5,10-metilenetetrahidrofolato redutase 677TT poderia ter uma pequena influência em determinar susceptibilidade a acidente cerebrovascular. MÉTODOS: Analisamos este polimorfismo em indivíduos brasileiros com acidente cerebrovascular isquêmico, baseando-se em um estudo de caso-controle. RESULTADOS: Comparamos os genótipos 5,10-metilenetetrahidrofolato redutase em grupos de indivíduos com acidente cerebrovascular isquêmico (n=127 e controle normal (n=126, e encontramos Odds Ratio de 1,97 (IC 95% 0,84 - 4,64 em uma análise multivariada, na qual os resultados foram ajustados a características clínicas basais dos indivíduos estudados. DISCUSS

  13. Terapia combinada de colestiramina e inibidores da HMG-CoA redutase na prevenção secundária da doença coronária

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    Novazzi José Paulo

    1998-01-01

    Full Text Available OBJETIVO: Avaliar as alterações obtidas no perfil lipídico de coronariopatas dislipidêmicos, após a adição de colestiramina, em pacientes tratados com inibidores da HMG-CoA redutase, e que não atingiram os valores ideais de LDL-colesterol. MÉTODOS: Vinte coronariopatas (12 submetidos à revascularização do miocárdio, 3 à angioplastia coronária e 5 mantidos sob tratamento clínico, com média de idade de 60,78 anos, que já realizavam dieta hipolipemiante e eram medicados com lovastatina 20mg/dia ou sinvastatina 10mg/dia, receberam também colestiramina na dose de 8 a 16g/dia durante 8 semanas, com o objetivo de reduzir LDL-colesterol para valores inferiores a 100mg/dl. RESULTADOS: Houve significante redução do colesterol total (valor médio inicial 239,52mg/dL e ao final 199,00mg/dL, obtendo-se um decréscimo percentual médio de 16,92%. O valor médio de LDL-colesterol também se reduziu, significantemente, de 172,73mg/dL para 118,26mg/dL, com decréscimo percentual médio de 31,53%. A trigliceridemia média aumentou, ainda dentro da faixa de referência normal, de 145,05mg/dL para 162,00mg/dL, (diferença percentual média de 11,69%. Houve significante aumento da fração HDL-colesterol de um valor médio inicial de 38,00mg/dL para um valor médio final de 48,21mg/dL (diferença média percentual 26,87%. Não houve efeitos adversos que impedissem a continuidade do tratamento. CONCLUSÃO: A associação de colestiramina a doses baixas de vastatinas em pacientes com hipercolesterolemia primária e de alto risco coronário é uma boa opção terapêutica, podendo atingir benefícios sobre o perfil lipídico semelhantes àqueles obtidos quando esses fármacos são utilizados, isoladamente, ou em associação, e em doses mais elevadas.

  14. In house reverse membrane hybridisation assay versus GenoType MTBDRplus and their performance to detect mutations in the genes rpoB, katG and inhA

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    Sergio Luiz Montego Ferreira Junior

    2014-06-01

    Full Text Available Drug-resistant tuberculosis (TB threatens global TB control and is a major public health concern in several countries. We therefore developed a multiplex assay (LINE-TB/MDR that is able to identify the most frequent mutations related to rifampicin (RMP and isoniazid (INH resistance. The assay is based on multiplex polymerase chain reaction, membrane hybridisation and colorimetric detection targeting of rpoB and katG genes, as well as the inhA promoter, which are all known to carry specific mutations associated with multidrug-resistant TB (MDR-TB. The assay was validated on a reference panel of 108 M. tuberculosis isolates that were characterised by the proportion method and by DNA sequencing of the targets. When comparing the performance of LINE-TB/MDR with DNA sequencing, the sensitivity, specificity and agreement were 100%, 100% and 100%, respectively, for RMP and 77.6%, 90.6% and 88.9%, respectively, for INH. Using drug sensibility testing as a reference standard, the performance of LINE-TB/MDR regarding sensitivity, specificity and agreement was 100%, 100% and 100% (95%, respectively, for RMP and 77%, 100% and 88.7% (82.2-95.1, respectively, for INH. LINE-TB/MDR was compared with GenoType MTBDRplus for 65 isolates, resulting in an agreement of 93.6% (86.7-97.5 for RIF and 87.4% (84.3-96.2 for INH. LINE-TB/MDR warrants further clinical validation and may be an affordable alternative for MDR-TB diagnosis.

  15. Nitrate reductase and glutamine synthetase activity in coffee leaves during fruit development Atividade da redutase do nitrato e glutamina sintetase em folhas de cafeeiro durante o desenvolvimento dos frutos

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    Andre Rodrigues Reis

    2009-04-01

    Full Text Available Nitrate reductase is the first enzyme in the pathway of nitrate reduction by plants, followed by glutamine synthetase, which incorporates ammonia to glutamine. The purpose of this study was to evaluate the nitrate reductase and glutamine synthetase activity, total soluble protein content, N and Ni content in coffee leaves during fruit development under field conditions to establish new informations to help assess the N nutritional status and fertilizer management. The experimental design was in randomized complete blocks, arranged in a 3 x 6 factorial design, with five replications. The treatments consisted of 3 N rates (0 - control, 150 and 300 kg ha-1 and six evaluation periods (January, February, March, April, May, and June in six-year-old coffee (Coffea arabica L. plants of Catuaí Vermelho IAC 44 cv. The nitrate reductase and glutamine synthetase activities, leaf soluble protein, and N concentrations increased linearly with the N rates. During fruit development, the enzyme activity, leaf soluble protein and N content decreased, due to the leaf senescence process caused by nutrient mobilization to other organs, e.g, to the berries. Leaf Ni increased during fruit development. Beans and raisin-fruits of plants well-supplied with N had higher Ni contents. Enzyme activities, total leaf N and leaf soluble protein, evaluated during the green fruit stage in March, were significantly correlated with coffee yield. These variables can therefore be useful for an early assessment of the coffee N nutritional status as well as coffee yield and N fertilization management.A redutase do nitrato (RN é a primeira enzima na via de redução de nitrato pelas plantas, seguida da glutamina sintetase (GS, a qual incorpora amônia à glutamina. O objetivo deste trabalho foi avaliar a atividade dessas enzimas, o teor de proteína solúvel total e a concentração de N e de Ni em folhas e grãos de cafeeiro durante o desenvolvimento dos frutos, em condições de

  16. Nitrate reductase enzyme activity and free proline contents in sugarcane roots under water and acid stress in soilAtividade da enzima redutase do nitrato e teores de prolina livre em raízes de cana-de-açúcar sob os estresses hídrico e ácido no solo

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    Samira Domingues Carlin

    2011-10-01

    Full Text Available The aim of this work was evaluate the physiological indicators of association between water deficiency and soil acidity, by determining the nitrate reductase activity, the levels of free proline and initial growth of the root system of seedlings of sugarcane cv. IAC91-5155. After 30 days, the seedlings were transferred to plastic pots with 12 dm3 of dystrophic alic Red Latosol (Oxisol and submitted to association of three treatments of water availability: no stress (70%, moderate stress (55% and severe stress (40%, in according with field capacity and three acidity treatments: no stress (55%, moderate stress (33% and severe stress (23%, considering the base saturation. The experimental design was that of random blocks under factorial scheme of 3x3, with four replicates. After 60 days under the stress association, the levels of free proline, the nitrate reductase activity and the growth of the sugarcane roots system were evaluated in seedlings of sugarcane. The nitrate reductase enzyme activity can be considered a physiological indicator of the effect of the association of acid and water stress in moderate conditions in soil, while the free proline can be considered physiological indicator to both stress in severe conditions. Water deficiency increasing reduced growth of sugarcane roots.O objetivo deste trabalho foi avaliar indicadores fisiológicos da associação entre deficiência hídrica e acidez do solo, por meio da determinação da atividade da enzima redutase do nitrato, dos teores de prolina livre e do crescimento inicial do sistema radicular de plantas jovens da cultivar IAC91-5155 de cana-de-açúcar. Aos 30 dias, as mudas, originadas da brotação de uma gema, foram transferidas para vasos de plástico de 12 dm3 contendo Latossolo Vermelho distrófico álico e submetidas à associação de três tratamentos de disponibilidade hídrica: sem estresse (70%, estresse moderado (55% e estresse severo (40%, de acordo com a capacidade de

  17. A conservative region of the mercuric reductase gene (merA as a molecular marker of bacterial mercury resistance Região conservada do gene da mercúrio redutase (merA como marcador molecular da resistência bacteriana ao mercúrio

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    Adriana Sotero-Martins

    2008-06-01

    Full Text Available The most common bacterial mercury resistance mechanism is based on the reduction of Hg(II to Hg0, which is dependent of the mercuric reductase enzyme (MerA activity. The use of a 431 bp fragment of a conservative region of the mercuric reductase (merA gene was applied as a molecular marker of this mechanism, allowing the identification of mercury resistant bacterial strains.O mecanismo de resistência bacteriana ao mercúrio mais comum é baseada na redução do Hg(II a Hg0, através da atividade da enzima mercúrio redutase (MerA. O uso do fragmento de 431 pb amplificado de uma região conservada do gene merA, que codifica a enzima MerA,foi utilizado como marcador molecular deste mecanismo, permitindo a identificação de bactérias resistentes ao mercúrio.

  18. Avaliação do teste de nitrato redutase para a detecção rápida de resistência aos medicamentos de primeira linha em cepas de Mycobacterium tuberculosis isoladas de pacientes em um hospital geral Evaluation of the nitrate reductase assay for the rapid detection of resistance to first-line medications in Mycobacterium tuberculosis strains isolated from patients in a general hospital

    Directory of Open Access Journals (Sweden)

    Maria de Fátima Filardi Oliveira Mansur

    2012-04-01

    Full Text Available Comparamos o teste de nitrato redutase com o método de proporções, considerado como padrão ouro, em 57 cepas de Mycobacterium tuberculosis isoladas de pacientes atendidos no Hospital das Clínicas da Universidade Federal de Minas Gerais, em Belo Horizonte (MG. A sensibilidade, a especificidade e a acurácia para rifampicina e isoniazida foram de 100% para todas, enquanto essas foram, respectivamente, de 88,9%, 66,7% e 96,5% para estreptomicina e de 98,0%, 100% e 98,2% para etambutol. A média de tempo para a obtenção dos resultados foi de dez dias. Na amostra estudada, o teste de nitrato redutase mostrou grande acurácia e excelente concordância com o padrão ouro.We compared the nitrate reductase assay with the proportion method, which is considered the gold standard, in 57 Mycobacterium tuberculosis strains isolated from patients treated at the Federal University of Minas Gerais Hospital das Clínicas, located in the city of Belo Horizonte, Brazil. For rifampin and isoniazid, the sensitivity, specificity, and accuracy of the nitrate reductase assay were all 100%, whereas they were 100%, 88.9%, and 66.7%, respectively, for streptomycin and 98.0%, 100%, and 98.2%, respectively, for ethambutol. The mean time to results was ten days. In the study sample, the nitrate reductase assay proved highly accurate and showed excellent concordance with the gold standard.

  19. Avaliação do polimorfismo no gene da metilenotetrahidrofolato redutase e concentração de folato e vitamina B12 em pacientes portadores do HIV-1 em tratamento com anti-retrovirais Evaluation of the polymorphisms in methylenetetrahydrofolate reductase gene and the levels of folate and B12 in HIV-infected patients under antiretroviral therapy

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    Iran Malavazi

    2004-12-01

    Full Text Available Neste trabalho, investigamos concentração da vitamina B12 e folato, considerando-se a influência dos genótipos da metilenotetrahidrofolato redutase, o perfil imunológico e a terapia antiretroviral utilizada na população brasileira portadora do HIV. Um grupo de 86 indivíduos portadores do HIV-1 e 29 doadores de sangue foram recrutados para compor a casuística. Entre os infectados pelo HIV-1, observou-se menor concentração de B12 no grupo com maior número de linfócitos TCD4+. Não encontramos diferença na distribuição genotípica para as mutações MTHFR C677T e A1298C entre infectados e não infectados pelo HIV-1. Indivíduos portadores do HIV, genótipo C677C, apresentaram concentrações menores de B12 em relação ao grupo controle de mesmo genótipo. A terapia antiretroviral não mostrou qualquer influência nos valores de folato e vitamina B12. Estudos adicionais são necessários para reavaliar a prevalência de menores concentrações de B12 e folato e de hiperhomocisteinemia na população portadora do HIV sob a ótica do uso de HAART e da melhoria na sobrevida dos pacientes.In this study we sought to investigate the B12 and folate levels regarding the influence of methylenetetrahydrofolate reductase genotypes, immunological profile and antiretroviral therapy in the Brazilian HIV-infected population. The study group comprised 89 HIV-infected individuals and 29 blood donors. There was a decrease in the B12 levels in the HIV-infected group with higher TCD4+ lymphocyte counts. No differences in the genotype distribution for methylenetetrahydrofolate reductase polymorphisms between the HIV-infected individuals and the controls were found. HIV-infected individuals carrying the C677C genotype presented lower B12 levels (313.91 ± 154.05 than those with the same genotype in the control group (408.27 ± 207.69. Also, the antiretroviral therapy was not a source of variation of the folate and B12 serum levels. Further studies are

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

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    Isaac L. Silva Filho

    2010-02-01

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

  1. Comparative evaluation of the Nitrate Reductase Assay and the Resazurin Microtitre Assay for drug susceptibility testing of Mycobacterium tuberculosis against first line anti-tuberculosis drugs Avaliação comparative dos métodos Nitrato Redutase e Microdiluição com Resazurina para testar a sensibilidade do Mycobacterium tuberculosis frente aos anti-tuberculosos de primeira linha

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    Karine O. Sanchotene

    2008-03-01

    rapid detection of resistance in low-income countries.A tuberculose permanece como uma séria doença infecciosa, com distribuição mundial, alta morbidade e mortalidade, ocorrendo principalmente em paises com baixa condição econômica. O estado de emergência da tuberculose causada por cepas resistentes e multirresistentes tornou-se uma importante ameaça para o tratamento e programas de controle da tuberculose. Uma rápida detecção de cepas resistentes permitirá a implantação de um tratamento adequado e contribuirá para controlar a disseminação destas cepas. Este estudo avaliou a performace do ensaio nitrato redutase em meio sólido (NRA-LJ e meio líquido (NRA-7H9, para determinar a sensibilidade frente aos fármacos antituberculosos de primeira linha: isoniazida (INH, rifampicina (RMP, etambutol (EMB and estreptomicina (SMR. Ambos os métodos, NRA-LJ e NRA-7H9, foram avaliados com 18 cepas com conhecido perfil de sensibilidade. O ensaio de microplaca com resazurina (REMA foi utilizado como método de referência. A concordância observada entre NRA-7H9 and REMA foi de 100% para os quatro fármacos testados. Quando o método NRA-LJ foi comparado com o REMA, a sensibilidade e especificidade para INH RMP e SMR foram de 100%, 100%, 85,7%, 76,9% e 80%, 100%, 75% and 80%, respectivamente. Dos 57 isolados clinicos de M. tuberculosis avaliados por NRA-7H9 e REMA, 56 (98.2% foram sensíveis a todos antibióticos testados (INH, RMP, EMB e SMR pelo método NRA-7H9, enquanto três destas cepas foram resistentes para INH pelo REMA. Uma cepa mostrou resistência para INH e RMP por ambos os métodos, e CMI de 1,0 µg/ml para INH para ambos os métodos, enquanto CMI de 1,0 e 2,0 µg/ml para RMP pelo REMA e NRA-7H9, respectivamente. Os três ensaios mostraram um alto nível ded concordância para uma rápida detecção de resistência a rifampicina e isoniazida. Com relação à rapidez na obtenção dos resultados, a detecção na mudança de cor nos métodos NRA

  2. Adubação do cafeeiro com nitrato de potássio via solo e folha, no outono-inverno e primavera-verão: efeitos na atividade da redutase do nitrato, no crescimento das plantas e na produção Coffee tree fertilization with potassium nitrate via leaf and soil, in autumn-winter and spring-summer: effects on nitrate reductase activity, on plant growth and production

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    Rupert Barros de Freitas

    2007-08-01

    Full Text Available Objetivou-se verificar o efeito da adubação de 100 g de N/ano/planta, em três épocas (outono-inverno, primavera-verão e outono-inverno/primavera-verão e três modos de aplicação (folha, solo e folha/solo, no desenvolvimento, na produção e na atividade da redutase do nitrato (RN da cultivar Rubi-MG com quatro anos de idade. A análise conjunta entre o modo e época de aplicação mostra que o padrão de RN não se altera, sendo no frio maior nas raízes e no calor maior nas folhas. O mesmo aconteceu com o padrão de crescimento, rápido na estação quente/chuvosa e lento no período seco/frio. Em função da época de adubação, concluiu-se que, no outono-inverno, deve-se dar preferência pela aplicação na folha ou folha-solo. Esses modos de adubação foram os mais eficientes na retomada do crescimento. Quando as adubações ocorrerem nos dois períodos, qualquer um dos três modos pode ser utilizado. Apesar da inviabilidade de várias pulverizações, eventualmente, uma ou mais pulverizações com KNO3 podem substituir eficientemente a adubação no solo.The objective of this work was to verify the effect of the fertilization of 100g N/year/plant, in three periods (autumn-winter, spring-summer and autumn-winter/spring-summer and three types of application (leaf, soil and leaf/soil on the development, production and nitrate reductase activity (NR on four years old Rubi-MG cultivar. The analysis of type and time of application showed no alteration on NR activity being higher on roots in cold weather and higher on leaves in warmer weather. The same was observed with growth pattern, fast in hot/rain season and slow in dry/cold period. During autumn-winter, applications may be performed on leaf or leaf-soil. These types of fertilization were the most efficient during growth recovering. When the fertilization occurred in both periods, any type may be used. Although, several pulverizations are nonviable, one or more pulverization with

  3. Biomass accumulation, photochemical efficiency of photosystem II, nutrient contents and nitrate reductase activity in young rosewood plants (Aniba rosaeodora Ducke submitted to different NO3-:NH4+ ratios Acúmulo de biomassa, eficiência fotoquímica do fotossistema II, conteúdo de nutrientes e atividade da redutase do nitrato em plantas jovens de pau-rosa (Aniba rosaeodora Ducke submetidas a diferentes relações NO3-:NH4+

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    Denize Caranhas de Sousa Barreto

    2007-01-01

    Full Text Available The rosewood (Aniba rosaeodora Ducke is a native tree species of Amazon rainforest growing naturally in acidic forest soils with reduced redox potential. However, this species can also been found growing in forest gaps containing oxide soils. Variations in the forms of mineral nitrogen (NO3- or NH4+ may be predicted in these different edaphic conditions. Considering that possibility, an experiment was carried out to analyze the effects of different NO3-:NH4+ ratios on the growth performance, mineral composition, chloroplastid pigment contents, photochemical efficiency photosystem II (PSII, and nitrate redutase activity (RN, E.C.1.6.6.1 on A. rosaeodora seedlings. Nine-month-old seedlings were grown in pots with a washed sand capacity of 7.5 kg and submitted to different NO3-:NH4+ ratios (T1 = 0:100%, T2 = 25:75%, T3 = 50:50%, T4 = 75:25%, and T5 = 100:0%. The lowest relative growth rate was observed when the NO3-:NH4+ ratio was equal to 0:100%. In general, high concentrations of NO3- rather than NH4+ favored a greater nutrient accumulation in different parts of the plant. For the chloroplastid pigment, the highest Chl a, Chl b, Chl tot, Chl a/b and Chl tot/Cx+c contents were found in the treatment with 75:25% of NO3-:NH4+, and for Chl b and Cx+c it was observed no difference. In addition, there was a higher photochemical efficiency of PSII (Fv/Fm when high NO3- concentrations were used. A linear and positive response for the nitrate reductase activity was recorded when the nitrate content increased on the culture substrate. Our results suggest that A. rosaeodora seedlings have a better growth performance when the NO3- concentrations in the culture substrate were higher than the NH4+ concentrations.O pau-rosa (Aniba rosaeodora Ducke habita, naturalmente, solos florestais ácidos com potencial redox reduzido. No entanto, estas espécies têm sido encontradas também em clareiras que, teoricamente, apresentam solos mais oxidados. Nestas diferentes

  4. Papel da redutase do nitrato e da asparagina sintetase em plantas de soja (Glycine max L.) sob condições de estresse de nitrogenio

    OpenAIRE

    Flavia Antunes

    2007-01-01

    Resumo: O sistema radicular da soja (Glycine max L.) é um importante sítio da assimilação do nitrogênio (N), seja pela assimilação do nitrato nas raízes, seja pela fixação simbiótica de N atmosférico nos nódulos. Os principais produtos da assimilação do N inorgânico, os aminoácidos asparagina e glutamina, os ureídeos, a alantoína e o ácido alantóico, são usados no transporte de N para a parte aérea. Assim, esses produtos representam uma fonte de N reduzido tanto para os sítios de consumo, qua...

  5. Comparação de métodos de fingerprints para o rastreio virtual de inibidores de 5α-redutase

    OpenAIRE

    Reis, Pedro Rafael Mendes

    2014-01-01

    Dissertação de mestrado em Química Farmacêutica Industrial, apresentada à Faculdade de Farmácia da Universidade de Coimbra A hiperplasia benigna da próstata e o cancro da próstata são duas doenças proliferativas relacionadas com a idade com morbilidade e mortalidade significativas. O controlo da proliferação e da actividade celular na próstata possibilita minorar os sintomas e o avanço destas patologias, melhorando a qualidade de vida. Este controlo pode ser assegurado pelo blo...

  6. Avaliação do tempo de secagem e da atividade de óxido-redutases de yacon (Smallanthus sonchifolius sob tratamento químico Drying evaluation time and yacon (Smallanthus sonchifolius enzymatic activity inhibition under chemical treatment

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    Vivianne Montarroyos Padilha

    2009-10-01

    Full Text Available Evidências científicas reconhecem o yacon (Smallanthus sonchifolius como fonte promissora de frutanos, tais como inulina e frutooligossacarídeos (FOS. No entanto, a presença de compostos fenólicos torna-o suscetível à reação de escurecimento enzimático. Este trabalho teve como objetivo avaliar o emprego de agentes químicos no processamento de yacon, para obtenção de farinha, a fim de inibir o escurecimento enzimático do produto e favorecer o tempo de secagem. Amostras de yacon sem inibição química, de yacon submetido à solução de cloreto de cálcio 1,0g 100g-1 por 30 minutos e de yacon submetido à solução de metabissulfito de potássio 0,5g 100g-1 por 5 minutos foram secas a 55°C, em estufa ventilada, e o teor de água e curvas de secagem foram determinados. As atividades das enzimas peroxidase (POD e polifenoloxidase (PPO foram determinadas antes e após a secagem como um possível marcador bioquímico do escurecimento enzimático desse tubérculo. No parâmetro umidade, os três tratamentos apresentaram-se semelhantes; porém, o tratamento 2 (cloreto de cálcio reduziu a umidade no menor tempo. Antes e após o tratamento térmico, a atividade enzimática foi maior no tratamento 3 (metabissulfito de potássio. As atividades enzimáticas não foram inativadas completamente pela ação térmica. Para a obtenção da farinha de yacon, o tratamento com cloreto de cálcio 1,0g 100g-1 por 30 minutos foi o que apresentou melhor resultado. Apesar de não inibir totalmente a atividade das enzimas POD e PPO foi o melhor tratamento de inativação, propiciando menor tempo de secagem e melhor firmeza da matéria-prima e facilitando o processamento para obtenção da farinha.Scientific evidences acknowledged the yacon (Smallanthus sonchifolius like a promise source of fructans, as inulin and fructooligosaccharides - FOS, meanwhile, the presence of phenolic coumpounds became this susceptible to enzimatic browning reaction. This project aimed to evaluate the use of chemical agents in yacon processing to obtain flour in a way that inhibits enzymatic darkening of the product besides determining the enzymatic activity in these treatments. Samples of yacon without chemical inhibitions, yacon treated with 1.0g 100g-1 calcium chloride for 30 seconds and yacon treated with 0.5g 100g-1 potassium metabisulfite for 5 minutes were dried at 55oC in a ventilated greenhouse and the proportions of humidity and drying curves were determined. The peroxidase activities and polyphenol oxidase enzymes were checked before and after being dried with an enzymatic darkening possible biochemist marker of this tubercle. Regarding humidity Parameter all the three treatment were equivalent, but treatment 2 (calcium chloride reduced the humidity in lower time. Before and after the thermal treatment the enzymatic activity was higher in treatment 3 (potassium metabisulfite. The thermal action did not inhibit completely polyphenol oxidase and peroxidase. The treatment with calcium chloride at 1.0g 100g-1 for 30 minutes to obtain yacon flour was the one with better result despite the fact that it did not inhibit completely peroxidase and polyphenol oxidase enzymes activity, offering a better drying time better material of row firmness , facilitating the process for obtaining the meal.

  7. Síntese, avaliação biológica e modelagem molecular de arilfuranos como inibidores da enzima tripanotiona redutase Synthesis, biological evaluation and molecular modeling of arylfurans as potential trypanothione reductase inhibitors

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    Renata B. de Oliveira

    2008-01-01

    Full Text Available Trypanosoma cruzi is a protozoan parasite that causes a severe disease (Chagas'disease in Central and South America. The currently available chemotherapeutic agents against this disease are still inadequate. The enzyme trypanothione reductase (TR is considered a validated molecular target for the development of new drugs against this parasite. In this regard, a series of arylfurans based on 2,5-bis-(4-acetamidophenylfuran was synthesized and tested for their in vitro inhibitory activity against TR. Molecular modeling studies of putative enzyme-inhibitor complexes revealed a possible mechanism of interaction. From synthesized compounds, a benzylaminofuran derivative was found to be more active than the lead compound.

  8.  Avaliação da atividade de inibidor da Enzima 3- Metil - Glutaril Coenzimaa Redutase na viabilidade, ciclo celular e síntese de fatores de crescimento em células de meloma múltiplo.

    OpenAIRE

    Paula Josiane Janowski Trojan

    2012-01-01

    The multiple myeloma (MM) reaches plasma cells that are completely different from each other, that produce high amounts of immunoglobulins. The genetic changes are multiple and complexes, affecting the disease course and the sensibility of the tumorous cells to the treatment. The bone marrow constitutes the ideal microenvironment for the disease development, secreting growth factors that are indispensable for the proliferation, survival and migration of tumourous cells. This project is eva...

  9. Inhibin alpha gene expression in human trophoblasts is regulated by interactions between TFAP2 and cAMP signaling pathways.

    Science.gov (United States)

    Depoix, Christophe L; Debiève, Frédéric; Hubinont, Corinne

    2014-11-01

    Inhibin α (Inha) gene expression is regulated, in rat granulosa cells, via a cyclic 3',5'-adenosine monophosphate (AMP)-response element (CRE) found in a region of the promoter that is homologous to the human INHA promoter. We previously found that during in vitro cytotrophoblast differentiation, human INHA gene expression was regulated by TFAP2A via association with an AP-2 site located upstream of this CRE. The aim of this study was to evaluate if the human INHA gene was also regulated by cAMP in trophoblasts, and to investigate the possible crosstalk between TFAP2 and cAMP signaling pathways in the regulation of INHA gene expression. Treatment with cAMP or forskolin increased INHA mRNA expression by 7- and 2-fold in primary cytotrophoblasts and choriocarcinoma-derived BeWo cells, respectively. Treatment with the protein kinase A inhibitor H-89 reduced forskolin-induced luciferase activity by ∼40% in BeWo cells transfected with an INHA promoter-driven luciferase reporter vector. TFAP2 overexpression increased basal luciferase activity, whereas the dominant repressor KCREB abolished it. Surprisingly, mutation of the CRE also eliminated the TFAP2-induced transcription, although TFAP2 overexpression was still able to increase forskolin-induced luciferase activity when the AP-2 binding site, but not the CRE site, was mutated. Thus, INHA gene expression is upregulated by cAMP via CRE in human trophoblasts, and TFAP2 regulates this expression by interacting with CRE.

  10. La parenté hagiographique d’après Jacques de Voragine et les manuscrits enluminés de la Légende dorée (c. 1260-1490, thèse de doctorat sous la direction Jean-Claude Schmitt, soutenue le 6 septembre 2010 (INHA

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    Chloé Maillet

    2011-10-01

    Full Text Available Cette thèse retrace l’histoire d’une parenté déviante et pourtant modélisante, racontée, répétée et représentée inlassablement à la fin du Moyen Âge. Pour tenter de définir la parenté hagiographique médiévale – notion qui s’avère d’emblée paradoxale tant cette l’hagiographie vise à minimiser l’importance des liens de parenté terrestres au profit des liens spirituels – l’enjeu de cette thèse repose sur l’établissement d’une triangulation entre parenté, hagiographie et images. Il y est question...

  11. Caracterização de miopatias mitocondriais através da avaliação das atividades enzimáticas envolvidas no metabolismo energético Characterization of mitochondrial myopathies through the evaluation of the enzymatic activities involved in the energetic metabolism

    OpenAIRE

    Fábio Cesar Pedroso; Annibal P. Campello; Lineu Cesar Werneck; Klüppel,Maria Lúcia W.

    1997-01-01

    Foi determinada a atividade das enzimas NADH desidiogenase, NADH citocromo e redutase, succinato desidiogenase, succinato citocromo e redutase, citocromo e oxidase e citrato sintase em mitocôndrias de músculo esquelético humano normal e doente (suspeito de miopatia mitocondrial). O grupo controle foi constituído de 13 indivíduos normais e que não faziam uso contínuo de fárrnacos. O grupo doente era constituído de 10 pacientes cujo diagnóstico anatomopatológico indicava suspeita de miopatia mi...

  12. Caracterização de miopatias mitocondriais através da avaliação das atividades enzimáticas envolvidas no metabolismo energético

    OpenAIRE

    Pedroso,Fábio Cesar; Annibal P. Campello; Werneck,Lineu Cesar; Klüppel,Maria Lúcia W.

    1997-01-01

    Foi determinada a atividade das enzimas NADH desidiogenase, NADH citocromo e redutase, succinato desidiogenase, succinato citocromo e redutase, citocromo e oxidase e citrato sintase em mitocôndrias de músculo esquelético humano normal e doente (suspeito de miopatia mitocondrial). O grupo controle foi constituído de 13 indivíduos normais e que não faziam uso contínuo de fárrnacos. O grupo doente era constituído de 10 pacientes cujo diagnóstico anatomopatológico indicava suspeita de miopatia mi...

  13. Complex Formation Between Iron(III) and Isonicotinohydroxamic ...

    African Journals Online (AJOL)

    acer

    Nigerian Journal of Basic and Applied Science (2009), 17(2): 181-188 ... ABSTRACT: Complex of Fe(III) with isonicotinohydroxamic acid (INHA) has been .... Matrix rank analysis using nine solution ..... computation of stability constants from ...

  14. Absence of inhibin alpha and retinoblastoma protein leads to early sertoli cell dysfunction.

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    Roopa L Nalam

    Full Text Available Sertoli cells, the support cells of mammalian spermatogenesis, are regulated by a number of nuclear factors and express retinoblastoma (RB tumor suppressor protein. We hypothesized that RB is an important mediator of Sertoli cell tumorigenesis in inhibin alpha knockout (Inha KO mice. In our previous mouse studies, we found that conditional knockout (cKO of Rb in Sertoli cells caused progressive Sertoli cell dysfunction. Initially, loss of RB had no gross effect on Sertoli cell function as the mice were fertile with normal testis weights at 6 weeks of age, but by 10-14 weeks of age, mutant mice demonstrated severe Sertoli cell dysfunction and infertility. Although double knockout (dKO of Rb and Inha did not result in exacerbation of the tumorigenic phenotype of Inha-null mice, we found that the dKO mice demonstrate an acceleration of Sertoli cell dysfunction compared to Rb cKO mice. Specifically, in contrast to Rb cKO mice, Inha/Rb dKO mice showed signs of Sertoli cell dysfunction as early as 4 weeks of age. These results demonstrate that RB is not essential for Sertoli cell tumorigenesis in Inha KO mice but that loss of Inha accelerates the infertility phenotype of Rb cKO mice.

  15. Absence of Inhibin Alpha and Retinoblastoma Protein Leads to Early Sertoli Cell Dysfunction

    Science.gov (United States)

    Nalam, Roopa L.; Andreu-Vieyra, Claudia; Matzuk, Martin M.

    2010-01-01

    Sertoli cells, the support cells of mammalian spermatogenesis, are regulated by a number of nuclear factors and express retinoblastoma (RB) tumor suppressor protein. We hypothesized that RB is an important mediator of Sertoli cell tumorigenesis in inhibin α knockout (Inha KO) mice. In our previous mouse studies, we found that conditional knockout (cKO) of Rb in Sertoli cells caused progressive Sertoli cell dysfunction. Initially, loss of RB had no gross effect on Sertoli cell function as the mice were fertile with normal testis weights at 6 weeks of age, but by 10–14 weeks of age, mutant mice demonstrated severe Sertoli cell dysfunction and infertility. Although double knockout (dKO) of Rb and Inha did not result in exacerbation of the tumorigenic phenotype of Inha-null mice, we found that the dKO mice demonstrate an acceleration of Sertoli cell dysfunction compared to Rb cKO mice. Specifically, in contrast to Rb cKO mice, Inha/Rb dKO mice showed signs of Sertoli cell dysfunction as early as 4 weeks of age. These results demonstrate that RB is not essential for Sertoli cell tumorigenesis in Inha KO mice but that loss of Inha accelerates the infertility phenotype of Rb cKO mice. PMID:20676395

  16. Human immunoglobulin G recognizing fibrinogen-binding surface proteins is protective against both Staphylococcus aureus and Staphylococcus epidermidis infections in vivo.

    Science.gov (United States)

    Vernachio, John H; Bayer, Arnold S; Ames, Brenda; Bryant, Dawn; Prater, Bradley D; Syribeys, Peter J; Gorovits, Elena L; Patti, Joseph M

    2006-02-01

    A human donor-selected immunoglobulin G for intravenous injection (IGIV) product with elevated titers against the staphylococcal fibrinogen-binding MSCRAMM proteins ClfA and SdrG (INH-A21) was tested in vitro and in vivo. INH-A21 contained a significantly increased ability to inhibit the fibrinogen-binding activity of recombinant forms of both ClfA and SdrG. Evaluation of the opsonizing potential of INH-A21 was evaluated using fluorescently labeled bacteria; this assay indicated an increase in phagocytic activity compared to normal IGIV. The prophylactic efficacy of INH-A21 against an intraperitoneal challenge of methicillin-resistant Staphylococcus epidermidis (MRSE) was evaluated in a neonatal rat model. INH-A21 was also evaluated for prophylactic and therapeutic efficacy in a rabbit model of catheter-induced aortic valve infective endocarditis caused by either MRSE or methicillin-resistant Staphylococcus aureus (MRSA). Results from the in vivo models demonstrated potent prophylactic and therapeutic efficacy against both MRSE and MRSA. These data suggest that INH-A21 may be an important tool for the prevention and treatment of staphylococcal infections, especially in high-risk populations.

  17. Epidemiology of isoniazid resistance mutations and their effect on tuberculosis treatment outcomes.

    Science.gov (United States)

    Huyen, Mai N T; Cobelens, Frank G J; Buu, Tran N; Lan, Nguyen T N; Dung, Nguyen H; Kremer, Kristin; Tiemersma, Edine W; van Soolingen, Dick

    2013-08-01

    Isoniazid resistance is highly prevalent in Vietnam. We investigated the molecular and epidemiological characteristics and the association with first-line treatment outcomes of the main isoniazid resistance mutations in Mycobacterium tuberculosis in codon 315 of the katG and in the promoter region of the inhA gene. Mycobacterium tuberculosis strains with phenotypic resistance to isoniazid from consecutively diagnosed smear-positive tuberculosis patients in rural Vietnam were subjected to Genotype MTBDRplus testing to identify katG and inhA mutations. Treatment failure and relapse were determined by sputum culture. In total, 227 of 251 isoniazid-resistant strains (90.4%) had detectable mutations: 75.3% in katG codon 315 (katG315) and 28.2% in the inhA promoter region. katG315 mutations were significantly associated with pretreatment resistance to streptomycin, rifampin, and ethambutol but not with the Beijing genotype and predicted both unfavorable treatment outcome (treatment failure or death) and relapse; inhA promoter region mutations were only associated with resistance to streptomycin and relapse. In tuberculosis patients, M. tuberculosis katG315 mutations but not inhA mutations are associated with unfavorable treatment outcome. inhA mutations do, however, increase the risk of relapse, at least with treatment regimens that contain only isoniazid and ethambutol in the continuation phase.

  18. Action mechanism of inhibin α-subunit on the development of Sertoli cells and first wave of spermatogenesis in mice.

    Directory of Open Access Journals (Sweden)

    Kailai Cai

    Full Text Available Inhibin is an important marker of Sertoli cell (SC activity in animals with impaired spermatogenesis. However, the precise relationship between inhibin and SC activity is unknown. To investigate this relationship, we partially silenced both the transcription and translation of the gene for the α-subunit of inhibin, Inha, using recombinant pshRNA vectors developed with RNAi-Ready pSIREN-RetroQ-ZsGreen Vector (Clontech Laboratories, Mountain View, Calif. We found that Inha silencing suppresses the cell-cycle regulators Cyclin D1 and Cyclin E and up-regulates the cell-cycle inhibitor P21 (as detected by Western blot analysis, thereby increasing the number of SCs in the G1 phase of the cell cycle and decreasing the amount in the S-phase of the cell cycle (as detected by flow cytometry. Inha silencing also suppressed Pdgfa, Igf1, and Kitl mRNA levels and up-regulated Tgfbrs, Inhba, Inhbb, Cyp11a1, Dhh, and Tjp1 mRNA levels (as indicated by real-time polymerase chain reaction [PCR] analysis. These findings indicate that Inha has the potential to influence the availability of the ligand inhibin and its antagonist activin in the SC in an autocrine manner and inhibit the progression of SC from G1 to S. It may also participate in the development of the blood-testis barrier, Leydig cells, and spermatogenesis through its effect on Dhh, Tjp1, Kitl, and Pdgfa. Real-time PCR and Western blot analyses of Inha, Inhba, and Inhbb mRNA and Inha levels over time show that Inha plays an important role in the formation of round spermatid during the first wave of spermatogenesis in mice.

  19. Expression of Wnt and TGF-β pathway components and key adrenal transcription factors in adrenocortical tumors: association to carcinoma aggressiveness.

    Science.gov (United States)

    Parviainen, Helka; Schrade, Anja; Kiiveri, Sanne; Prunskaite-Hyyryläinen, Renata; Haglund, Caj; Vainio, Seppo; Wilson, David B; Arola, Johanna; Heikinheimo, Markku

    2013-08-01

    Factors controlling benign and malignant adrenocortical tumorigenesis are largely unknown, but several mouse models suggest an important role for inhibin-alpha (INHA). To show that findings in the mouse are relevant to human tumors and clinical outcome, we investigated the expression of signaling proteins and transcription factors involved in the regulation of INHA in human tumor samples⋅ Thirty-one adrenocortical tumor samples, including 13 adrenocortical carcinomas (ACCs), were categorized according to Weiss score, hormonal profile, and patient survival data and analyzed using immunohistochemistry and RT-PCR. Expression of the TGF-β signaling mediator SMAD3 varied inversely with Weiss score, so that SMAD3 expression was lowest in the most malignant tumors. By contrast, SMAD2 expression was upregulated in most malignant tumors. Wnt pathway co-receptors LRP5 and LRP6 were predominantly expressed in benign adrenocortical tumors. In ACCs, expression of transcription factors GATA-6 and SF-1 correlated with that of their target gene INHA. Moreover, the diminished expression of GATA-6 and SF-1 in ACCs correlated with poor outcome. We conclude that the factors driving INHA expression are reduced in ACCs with poor outcome, implicating a role for INHA as a tumor suppressor in humans.

  20. Produção, extração e pré-purificação de lovastatina por linhagem de aspergillus terreus utilizando resíduos agroindustriais como meio de cultivo

    OpenAIRE

    Paulo, Anderson José

    2013-01-01

    A lovastatina é um inibidor enzimático comumente chamado de estatina e são de grande importância na medicina para redução dos altos níveis de colesterol. É uma potente droga, produzida através do metabolismo secundário de alguns micro-organismos, e atua inibindo a ação da enzima 3-hidroxi-3-metil-glutaril-CoA redutase (HMG-Coa redutase), que tem papel limitante na biossíntese do colesterol. Este estudo avaliou a produção de lovastatina pela linhagem de Aspergillus terreus URM 4317 através de ...

  1. Validação de métodos para análise de estatinas em medicamentos por cromatografia líquida de alta eficiência e eletroforese capilar

    OpenAIRE

    Karina Litzi González Tejerina

    2011-01-01

    As estatinas são os fármacos mais usados para tratamento das hiperlipidemias em prevenção primária e secundária, com o propósito de diminuir os níveis de lipoproteínas plasmáticas ricas em colesterol e reduzir os riscos de doença arterialcoronária (DAC) (WITZTUM, 2005). Estes efeitos são resultantes da atividade inibidora das estatinas sobre a enzima HMG-CoA redutase (hidroximetilglutaril-CoA redutase), com a propriedade de bloquear a conversão do substrato HMG-CoA em ácido mevalônico, inibin...

  2. Terapêutica médica na hiperplasia benigna da próstata

    OpenAIRE

    Silva, Odila Nunes da

    2009-01-01

    A hiperplasia benigna da próstata (HBP) é um tumor benigno da glândula prostática. A prevalência da HBP aumenta com a idade. A HBP manifesta-se comummente pelos sintomas do tracto urinário baixo (LUTS). O tratamento da HBP mudou drasticamente nos últimos 20 anos. A terapêutica médica clássica para a HBP compreende os antagonistas adrenérgicos alfa-1, os inibidores da enzima 5-alfa redutase, a terapêutica de associação com um antagonista adrenérgico alfa-1 e um inibidor da 5-alfa redutase e a ...

  3. Produção, extração e pré-purificação de lovastatina por linhagem de aspergillus terreus utilizando resíduos agroindustriais como meio de cultivo

    OpenAIRE

    Paulo, Anderson José

    2013-01-01

    A lovastatina é um inibidor enzimático comumente chamado de estatina e são de grande importância na medicina para redução dos altos níveis de colesterol. É uma potente droga, produzida através do metabolismo secundário de alguns micro-organismos, e atua inibindo a ação da enzima 3-hidroxi-3-metil-glutaril-CoA redutase (HMG-Coa redutase), que tem papel limitante na biossíntese do colesterol. Este estudo avaliou a produção de lovastatina pela linhagem de Aspergillus terreus URM 4317 através de ...

  4. Dietary reference intakes for the cuban population, 2008.

    Science.gov (United States)

    Hernández-Triana, Manuel; Porrata, Carmen; Jiménez, Santa; Rodríguez, Armando; Carrillo, Olimpia; García, Alvaro; Valdés, Lourdes; Esquivel, Mercedes

    2009-10-01

    Recommended dietary reference intakes (DRI) for energy and nutrients for the Cuban population were first established by the Nutrition and Food Hygiene Institute (INHA, its Spanish acronym) in 1996.[1] International organizations and Cuban public health research subsequently generated a considerable volume of new information on food-based energy and nutrient requirements,[2-4] resulting in the need for a revision. Updated DRIs were therefore compiled by a multidisciplinary group of specialists and published in 2008 by INHA and the Cuban Ministry of Public Health.[5,6].

  5. Terbutaline accumulates in blood and urine following daily therapeutic inhalation

    DEFF Research Database (Denmark)

    Krogh, Nanna; Rzeppa, Sebastian; Dyreborg, Anders

    2017-01-01

    PURPOSE: This study investigated pharmacokinetics of terbutaline after single and seven consecutive days of inhalation in exercising trained men. METHODS: Twelve healthy young trained men underwent two pharmacokinetic trials comparing single dose (2 mg) and seven consecutive days (2 mg×d) of inha...

  6. Mn and Btex Reference Value Arrays (Final Reports)

    Science.gov (United States)

    These final reports are a summary of reference value arrays with critical supporting documentation for the chemicals manganese, benzene, toluene, ethylbenzene, and xylene. Each chemical is covered in a separate document, and each is a statement of the status of the available inha...

  7. ’Fused-on’ Rotating Bands for Projectiles

    Science.gov (United States)

    1974-12-01

    casting alloys, might be preferable for the procese or as rotating band materials depending on projectile requirements. METHODS AND PROCEDURES In this in...has fractured on impact (left projectile) all parts of the band have remained attached to the steel. Problem Areas One of the problems encountered in

  8. Validation and Application of Pharmacokinetic Models for Interspecies Extrapolations in Toxicity Risk Assessments of Volatile Organics

    Science.gov (United States)

    1991-07-23

    harmaceutics . College of harmacy University of Georgia, Athens. Georgia 30602 1?ereived July 1, 1988&acceptedJanuaryl16,1939 The Utipake and Elimination of l.1...column operation. When using the LCD. gas.- Inhaed as onenirtiu 027 (5 prm eposre) flow ratea of 40 mI/min were employed for nitrogen ( car . (.9111MC_

  9. Expression of Wnt and TGF-β pathway components and key adrenal transcription factors in adrenocortical tumors – association to carcinoma aggressiveness

    Science.gov (United States)

    Parviainen, Helka; Schrade, Anja; Kiiveri, Sanne; Prunskaite-Hyyryläinen, Renata; Haglund, Caj; Vainio, Seppo; Wilson, David B.; Arola, Johanna; Heikinheimo, Markku

    2013-01-01

    Factors controlling benign and malignant adrenocortical tumorigenesis are largely unknown, but several mouse models suggest an important role for inhibin-alpha (INHA). To show that findings in the mouse are relevant to human tumors and clinical outcome, we investigated the expression of signaling proteins and transcription factors involved in the regulation of INHA in human tumor samples. Thirty-one adrenocortical tumor samples, including 13 adrenocortical carcinomas (ACCs), were categorized according to Weiss score, hormonal profile, and patient survival data and analyzed using immunohistochemistry and RT-PCR. Expression of the TGF-β signaling mediator SMAD3 varied inversely with Weiss score, so that SMAD3 expression was lowest in the most malignant tumors. By contrast, SMAD2 expression was upregulated in most malignant tumors. Wnt pathway co-receptors LRP5 and LRP6 were predominantly expressed in benign adrenocortical tumors. In ACCs, expression of transcription factors GATA-6 and SF-1 correlated with that of their target gene INHA. Moreover, the diminished expression of GATA-6 and SF-1 in ACCs correlated with poor outcome. We conclude that the factors driving INHA expression are reduced in ACCs with poor outcome, implicating a role for INHAas a tumor suppressor in humans. PMID:23866946

  10. Development and validation of the inhaled corticosteroid questionnaire : a model for the measurement of patient-perceived side effects by self-report

    NARCIS (Netherlands)

    Foster, Juliet Michelle

    2007-01-01

    Inhaled corticosteroids are effective treatments in asthma and in some cases of chronic obstructive pulmonary disease. Unfortunately these medications are associated with commonly occurring side effects such as dysphonia (problems with the voice) or skin bruising. Some patients will stop taking inha

  11. Pulmonary clearance of 99mTc-DTPA: role of the pulmonary surfactant system.

    NARCIS (Netherlands)

    J.A.H. Bos (Jelle)

    1992-01-01

    textabstractIt is established that in a wide variety of acute and chronic lung diseases the integrity of the alveolar-capillary membrane is altered. Increased permeability of the barrier is thought to cause increased filtration of fluids and proteins into the alveoli and increased absorption of inha

  12. Investigation of isoniazid and ethionamide cross-resistance by whole genome sequencing and association with poor treatment outcomes of multidrug-resistant tuberculosis patients in South Africa

    Directory of Open Access Journals (Sweden)

    L Malinga

    2016-01-01

    Conclusion: Baseline ETH molecular resistance before second-line treatment is a concern. Unfavorable treatment outcomes of patients with ethA, ethR, and inhA mutations highlight the importance of genotypic testing before initiation of treatment containing ETH. The clinical significance of whole genome analysis for early detection of mutations predictive of treatment failure needs further investigation.

  13. Tiotropium in Asthma Poorly Controlled with Standard Combination Therapy

    NARCIS (Netherlands)

    Kerstjens, Huib A. M.; Engel, Michael; Dahl, Ronald; Paggiaro, Pierluigi; Beck, Ekkehard; Vandewalker, Mark; Sigmund, Ralf; Seibold, Wolfgang; Moroni-Zentgraf, Petra; Bateman, Eric D.

    2012-01-01

    BACKGROUND Some patients with asthma have frequent exacerbations and persistent airflow obstruction despite treatment with inhaled glucocorticoids and long-acting beta-agonists (LABAs). METHODS In two replicate, randomized, controlled trials involving 912 patients with asthma who were receiving inha

  14. Inhibition of cell proliferation by a selective inhibitor of the Ca{sup 2+}-activated Cl{sup -} channel, Ano1

    Energy Technology Data Exchange (ETDEWEB)

    Mazzone, Amelia; Eisenman, Seth T.; Strege, Peter R. [Enteric NeuroScience Program, Mayo Clinic, Rochester, MN (United States); Yao, Zhen [Laboratory of Molecular Genetics, UCSF, San Francisco, CA (United States); Ordog, Tamas; Gibbons, Simon J. [Enteric NeuroScience Program, Mayo Clinic, Rochester, MN (United States); Farrugia, Gianrico, E-mail: farrugia.gianrico@mayo.edu [Enteric NeuroScience Program, Mayo Clinic, Rochester, MN (United States)

    2012-10-19

    Highlights: Black-Right-Pointing-Pointer T16A{sub inh}-A01 blocked Ano1 currents in HEK cells expressing Ano1. Black-Right-Pointing-Pointer T16A{sub inh}-A01 reduced proliferation in ICC primary cultures and CFPAC-1 cell line. Black-Right-Pointing-Pointer T16A{sub inh}-A01 reduced proliferation of ICC in intact smooth muscle strips. -- Abstract: Background: Ion channels play important roles in regulation of cellular proliferation. Ano1 (TMEM16A) is a Ca{sup 2+}-activated Cl{sup -} channel expressed in several tumors and cell types. In the muscle layers of the gastrointestinal tract Ano1 is selectively expressed in interstitial cells of Cajal (ICC) and appears to be required for normal gastrointestinal slow wave electrical activity. However, Ano1 is expressed in all classes of ICC, including those that do not generate slow waves suggesting that Ano1 may have other functions. Indeed, a role for Ano1 in regulating proliferation of tumors and ICC has been recently suggested. Recently, a high-throughput screen identified a small molecule, T16A{sub inh}-A01 as a specific inhibitor of Ano1. Aim: To investigate the effect of the T16A{sub inh}-A01 inhibitor on proliferation in ICC and in the Ano1-expressing human pancreatic cancer cell line CFPAC-1. Methods: Inhibition of Ano1 was demonstrated by whole cell voltage clamp recordings of currents in cells transfected with full-length human Ano1. The effect of T16A{sub inh}-A01 on ICC proliferation was examined in situ in organotypic cultures of intact mouse small intestinal smooth muscle strips and in primary cell cultures prepared from these tissues. ICC were identified by Kit immunoreactivity. Proliferating ICC and CFPAC-1 cells were identified by immunoreactivity for the nuclear antigen Ki67 or EdU incorporation, respectively. Results: T16A{sub inh}-A01 inhibited Ca{sup 2+}-activated Cl{sup -} currents by 60% at 10 {mu}M in a voltage-independent fashion. Proliferation of ICC was significantly reduced in primary cultures

  15. Triclosan Derivatives: Towards Potent Inhibitors of Drug-Sensitive and Drug-Resistant Mycobacterium tuberculosis

    Energy Technology Data Exchange (ETDEWEB)

    Freundlich, Joel S.; Wang, Feng; Vilchèze, Catherine; Gulten, Gulcin; Langley, Robert; Schiehser, Guy A.; Jacobus, David P.; Jacobs, Jr., William R.; Sacchettini, James C.; (Einstein); (TAM); (Jacobus)

    2009-06-30

    Isoniazid (INH) is a frontline antitubercular drug that inhibits the enoyl acyl carrier protein reductase InhA. Novel inhibitors of InhA that are not cross-resistant to INH represent a significant goal in antitubercular chemotherapy. The design, synthesis, and biological activity of a series of triclosan-based inhibitors is reported, including their promising efficacy against INH-resistant strains of M. tuberculosis. Triclosan has been previously shown to inhibit InhA, an essential enoyl acyl carrier protein reductase involved in mycolic acid biosynthesis, the inhibition of which leads to the lysis of Mycobacterium tuberculosis. Using a structure-based drug design approach, a series of 5-substituted triclosan derivatives was developed. Two groups of derivatives with alkyl and aryl substituents, respectively, were identified with dramatically enhanced potency against purified InhA. The most efficacious inhibitor displayed an IC{sub 50} value of 21 nM, which was 50-fold more potent than triclosan. X-ray crystal structures of InhA in complex with four triclosan derivatives revealed the structural basis for the inhibitory activity. Six selected triclosan derivatives were tested against isoniazid-sensitive and resistant strains of M. tuberculosis. Among those, the best inhibitor had an MIC value of 4.7 {mu}g mL{sup -1} (13 {mu}M), which represents a tenfold improvement over the bacteriocidal activity of triclosan. A subset of these triclosan analogues was more potent than isoniazid against two isoniazid-resistant M. tuberculosis strains, demonstrating the significant potential for structure-based design in the development of next generation antitubercular drugs.

  16. Efeito da aplicação de nitrogênio e de piraclostrobina em plantas de tomateiro cultivar Micro-Tom

    Directory of Open Access Journals (Sweden)

    Emanuela Garbin Martinazzo

    Full Text Available RESUMO Fungicidas do grupo químico das estrobilurinas ganharam novas perspectivas de uso, em vista das vantagens obtidas pela ação dos efeitos fisiológicos positivos sobre as plantas. Este trabalho teve como objetivo verificar o efeito da piraclostrobina nos teores de pigmentos fotossintéticos e de nitrogênio na atividade da enzima redutase do nitrato, em plantas de tomateiro cv. Micro-Tom, cultivadas com diferentes doses de nitrogênio. Aos 21 dias após o transplante (DAT, as plantas foram submetidas aos tratamentos: solução nutritiva completa, sem piraclostrobina; solução nutritiva completa, com piraclostrobina; solução nutritiva com 1/2 força iônica de nitrogênio, sem piraclostrobina; e solução nutritiva com 1/2 força iônica de nitrogênio, com piraclostrobina. A aplicação da piraclostrobina foi repetida aos 35 DAT. Aos 28 e 42 DAT, determinaram-se o teor de pigmentos fotossintéticos, o nitrogênio total e a atividade da enzima redutase do nitrato. A aplicação de piraclostrobina promoveu aumento dos teores de pigmentos apenas aos 28 DAT, nas plantas com fornecimento reduzido de nitrogênio. Duas aplicações de piraclostrobina não influenciaram no conteúdo de nitrogênio e na atividade da redutase do nitrato de plantas de tomateiro cv. Micro-Tom, cultivadas em solução nutritiva de Hoagland e Arnon.

  17. Metabolismo do nitrogênio e estado nutricional do cafeeiro (Coffea arabica)

    OpenAIRE

    2007-01-01

    Este trabalho foi realizado com a finalidade de avaliar atividade da redutase do nitrato (RN), sintetase da glutamina (GS) e conteúdo de proteína total solúvel (PTS) em folhas de cafeeiro durante o desenvolvimento dos frutos em condições de campo. O experimento foi instalado em blocos ao acaso em esquema fatorial 3x6, constituído pela combinação de 3 doses de N (0, 150 e 350 kg ha-1) e avaliado em seis diferentes fases fenológicas (janeiro, fevereiro, março, abril, maio e junho), com sete rep...

  18. Calcium homeostasis and mitochondrial function during death of prostate cancer cells exposed to statins

    OpenAIRE

    Kivia Aparecida Pontes de Oliveira

    2008-01-01

    Resumo: As estatinas são inibidores da 3-hidroxi-3-metilglutaril CoA (HMG-CoA) redutase usados no tratamento de hipercolesterolemia. Estudos in vitro e in vivo têm demonstrado que as estatinas podem ter efeitos anti-cancerígenos. No presente estudo analisamos os mecanismos de toxicidade de sinvastatina e de lovastatina nas linhagens de câncer de próstata LNCaP e PC-3. Curvas dose-resposta do efeito das estatinas (0,1-100 µM) sobre as células LNCaP e PC-3 mostraram efeitos similares e maior se...

  19. Investigação dos polimorfismos dos genes de polipeptídeos transportadores de ânions orgânicos (OATP) e a resposta ao tratamento com sinvastatina

    OpenAIRE

    Vinicius de Albuquerque Sortica

    2009-01-01

    A prevenção das doenças cardiovasculares é, atualmente, uma das principais metas para o cuidado com a saúde nos países ocidentais. Os inibidores da HMG CoA redutase (estatinas) são os fármacos mais utilizados para a prevenção das doenças cardiovasculares devido a sua eficácia em reduzir os níveis de colesterol e por serem bem tolerados durante o tratamento. Apesar de serem eficazes, existe uma grande variabilidade interindividual à resposta ao tratamento com estatinas. Parte dessa variação po...

  20. Bloqueio do Receptor AT1 da Angiotensina II na atividade basal da AMP- proteína Quinase Hipotalâmica e após estresse de contenção ou cirúrgico

    OpenAIRE

    Mirna Bastos Marques

    2010-01-01

    A 5-monofosfato-adenosina proteína quinase ativada (AMP-Q) foi descoberta há mais de 3 décadas como uma proteína quinase de atividade associada a acetil-CoA carboxilase1 e HMG CoA redutase2, enzimas de papel essencial na síntese de ácidos graxos livres e colesterol. Posteriormente, descobriu-se que AMP-Q é ativada por AMP3 e foi reconhecido que a AMP-Q era, por sua vez, regulada pelo nível de energia celular4 e por fosforilação.5,6

  1. Desenvolvimento e validação de método discriminativo de dissolução para comprimidos revestidos de atorvastatina cálcica associado a dados in vivo

    OpenAIRE

    Jaison Carlosso Machado

    2012-01-01

    A atorvastatina é um dos principais fármacos hipolipemiantes da classe das estatinas, sendo atualmente, prescrita mundialmente para a redução de doenças cardiovasculares. Trata-se de um potente inibidor da enzima HMG – CoA redutase, limitando a síntese de colesterol e triglicerídeos. Mesmo sendo comercializada há praticamente 15 anos, os comprimidos revestidos de atorvastatina não estão inclusos em nenhuma farmacopéia ou compendio oficial, tendo somente uma sugestão de condição para o teste d...

  2. Surto psicótico pela possível interação medicamentosa de sibutramina com finasterida

    OpenAIRE

    Douglas Dogol Sucar; Everton Botelho Sougey; José Brandão Neto

    2002-01-01

    É descrito um caso de interação medicamentosa em um paciente de 30 anos, que fazia com sucesso um tratamento antiobesidade com sibutramina. Esse medicamento é um saciogênico que inibe a recaptação da serotonina e da noradrenalina. Logo após a introdução da finasterida para tratamento da alopécia androgênica, o paciente passou a apresentar surto psicótico. Esse medicamento é um potente inibidor da 5-alfa-redutase do tipo II. A interação medicamentosa demonstra-se por meio de um rigoroso acompa...

  3. Redução de nitrato em plantas jovens de café cultivadas em diferentes níveis de luz e de nitrogênio Nitrate reduction in young coffee trees grown under different levels of light and nitrogen

    Directory of Open Access Journals (Sweden)

    Maria Luiza Carvalho Carelli

    1990-01-01

    Full Text Available Foi estudado o efeito de níveis de luz e de nitrogênio na atividade da enzima redutase de nitrato e nos teores de nitrato e de açúcares nas folhas de plantas jovens de café (Coffea arabica L, assim como as possíveis relações entre a disponibilidade desses compostos e a atividade enzimática. Foram utilizadas plantas de dez meses de idade cultivadas em vasos contendo uma mistura de terra mais composto, e mantidas em condições ambientais em pleno sol e em 50% da luz solar. Metade das plantas de cada tratamento de luz foi suplementada semanalmente com nitrogênio. Os resultados mostraram que a atividade da redutase de nitrato, nos dois tratamentos de luz, foi maior nas plantas suplementadas com nitrogênio. Para um mesmo nível de nitrogênio, as plantas cultivadas em pleno sol apresentaram menor atividade da redutase de nitrato, maiores teores de nitrato e de açúcares e maiores taxas de transpiração, do que as cultivadas na sombra. Tais resultados indicam que a menor atividade da redutase de nitrato nas plantas cultivadas em pleno sol aparentemente não foi devida a limitações na disponibilidade de nitrato e de açúcares para fornecer a energia necessária para a redução de nitrato.The effect of levels of light and nitrogen on the activity of the enzyme nitrate reductase and its relationship with the availability of sugars and nitrate was studied in leaves of coffee plants (Coffea arabica L. cv. Catual. Ten month old plants were grown on pots containing a mixture of soil and compost, and were kept at full or 50% sunlight. Half of the plants of each light treatment received nitrogen supply. The results showed that the activity of nitrate reductase was higher on plants supplied with nitrogen at both light treatments. For the same nitrogen level, plants grown under full sunlight presented lower nitrate reductase activity, higher nitrate and sugars concentrations, and higher transpiration rates than plants kept at 50% sunlight

  4. Characterization of mutations causing rifampicin and isoniazid resistance of Mycobacterium tuberculosis in Syria.

    Science.gov (United States)

    Madania, Ammar; Habous, Maya; Zarzour, Hana; Ghoury, Ifad; Hebbo, Barea

    2012-01-01

    In order to characterize mutations causing rifampicin and isoniazid resistance of M. tuberculosis in Syria, 69 rifampicin resistant (Rif(r)) and 72 isoniazid resistant (Inh(r)) isolates were screened for point mutations in hot spots of the rpoB, katG and inhA genes by DNA sequencing and real time PCR. Of 69 Rif(r) isolates, 62 (90%) had mutations in the rifampin resistance determining region (RRDR) of the rpoB gene, with codons 531 (61%), 526 (13%), and 516 (8.7%) being the most commonly mutated. We found two new mutations (Asp516Thr and Ser531Gly) described for the first time in the rpoB-RRDR in association with rifampicin resistance. Only one mutation (Ile572Phe) was found outside the rpoB-RRDR. Of 72 Inh(r) strains, 30 (41.6%) had a mutation in katGcodon315 (with Ser315Thr being the predominant alteration), and 23 (32%) harbored the inhA(-15C-->T) mutation. While the general pattern of rpoB-RRDR and katG mutations reflected those found worldwide, the prevalence of the inhA(-15C-->T mutation was above the value found in most other countries, emphasizing the great importance of testing the inhA(-15C-->T) mutation for prediction of isoniazid resistance in Syria. Sensitivity of a rapid test using real time PCR and 3'-Minor groove binder (MGB) probes in detecting Rif(r) and Inh(r) isolates was 90% and 69.4%, respectively. This demonstrates that a small set of MGB-probes can be used in real time PCR in order to detect most mutations causing resistance to rifampicin and isoniazid.

  5. 7. The Flemish Health and Environment Study (FLHES): area of residence is associate with significant differences in HPRT mutant frequency in peripheral blood lymphocytes of women aged 50-65

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    @@Both exposure and effect biomarkers were measured in residents of Wilrijk and Hoboken, industrial suburbs of the big city of Antwerp, and in residents of Peer, a rural municipality with intensive agriculture (14,622 inha-bitants, 70 km east of Antwerp). Persons with known occupational exposures, or working in a region with pollution levels clearly different from the area of residence or commuting over long

  6. Polymorphisms in Isoniazid and Prothionamide Resistance Genes of the Mycobacterium tuberculosis Complex

    KAUST Repository

    Projahn, M.

    2011-06-27

    Sequence analyses of 74 strains that encompassed major phylogenetic lineages of the Mycobacterium tuberculosis complex revealed 10 polymorphisms in mshA (Rv0486) and four polymorphisms in inhA (Rv1484) that were not responsible for isoniazid or prothionamide resistance. Instead, some of these mutations were phylogenetically informative. This genetic diversity must be taken into consideration for drug development and for the design of molecular tests for drug resistance.

  7. JPRS Report, Near East & South Asia, Israel

    Science.gov (United States)

    2007-11-02

    nd ser of self - esteem . It entered the jaws of the Palestinian lion in officers met yesterday in Jerusalem with members of Sidon for the first time in...HA’ARETZ 4 Jul] ............................................................ 41 Journalists ’ Ostracized ’ by Egyptian Counterparts [MA’ARIV 5 Jul...Ahime’ir and The Prime Minister has surrounded himself with 13 Posner, define the way they relate to Shamir as esteem . people who bear the title of

  8. Mutaciones asociadas con resistencia a rifampicina o isoniazida en aislamientos clínicos de M. tuberculosis de Sonora, México DNA mutations associated to rifampicin or isoniazid resistance in M. tuberculosis clinical isolates from Sonora, Mexico

    Directory of Open Access Journals (Sweden)

    Enrique Bolado-Martínez

    2012-04-01

    Full Text Available OBJETIVO: Realizar el análisis de regiones específicas de genes asociados con resistencia a isoniazida o rifampicina. MATERIAL Y MÉTODOS: Se estudiaron 22 cepas de M. tuberculosis, aisladas en Sonora, México. Se utilizaron iniciadores para regiones específicas de los genes rpoB, katG e inhA y la región ahpC-oxyR. Los productos de PCR se secuenciaron y analizaron. RESULTADOS: Se identificaron mutaciones en la región promotora del gen inhA, región ahpC-oxyR, codón 315 del gen katG y codones 451 ó 456 del gen rpoB. CONCLUSIONES: La identificación de mutaciones no descritas previamente obliga a continuar el análisis genotípico de cepas aisladas en Sonora.OBJECTIVE: To perform the analysis of specific regions of the major genes associated with resistance to isoniazid or rifampin. MATERIALS AND METHODS: Twenty two M. tuberculosis strains, isolated from human samples obtained in Sonora, Mexico. Specific primers for hotspots of the rpoB, katG, inhA genes and the ahpC-oxyR intergenic region were used. The purified PCR products were sequenced. RESULTS: Mutations in the promoter of inhA, the ahpC-oxyR region, and codon 315 of katG and in 451 or 456 codons of rpoB, were identified. CONCLUSIONS: Detection of mutations not previously reported requires further genotypic analysis of Mycobacterium tuberculosis isolates in Sonora.

  9. On Public Opinion in Time of War

    Science.gov (United States)

    2009-12-01

    5 Stimson, 65. 6 Eric V. Larson, Casualties and Consensus: The Historical Role of Casualties in...has been made. Kennamer boldly states, “Voting is the most visible and concrete sign of public opinion.”11 According to Erikson , Wright, & McIver...en.wikipedia.org/wiki/Opinion_poll. 10 Kennamer, 15. 11 Kennamer, 2. 12 Robert S. Erikson , John P. McIver, and Gerald C. Wright, Statehouse Democracy

  10. A Relative Ranking Approach for Nano-Enabled Applications to Improve Risk-Based Decision Making: A Case Study of Army Materiel

    Science.gov (United States)

    2014-12-24

    in energy and sensor applications , and carbon nanotubes (CNT) also ranked relatively high based on worker inha- lation from CNTs used in research and...accidental expo- sures to carbon nanotubes and copper flakes incorporated into energy and obscurant materiel by Army workers rank highest relative to...that inhalation from accidental exposures to carbon nanotubes and copper flakes incorporated into energy and obscurant materiel by Army workers rank

  11. Evaluation of GenoFlow DR-MTB Array Test for Detection of Rifampin and Isoniazid Resistance in Mycobacterium tuberculosis

    Science.gov (United States)

    Molina-Moya, B.; Kazdaglis, G.; Lacoma, A.; Prat, C.; Gómez, A.; Villar-Hernández, R.; García-García, E.; Haba, L.; Maldonado, J.; Samper, S.; Ruiz-Manzano, J.; Ausina, V.

    2016-01-01

    The aim of this study was to evaluate the GenoFlow DR-MTB array test (DiagCor Bioscience, Hong Kong) on 70 cultured isolates and 50 sputum specimens. The GenoFlow array test showed good sensitivity and specificity compared to the phenotypic Bactec 460TB. This array accurately detected mutations in rpoB, katG, and inhA associated with resistance to rifampin and isoniazid. PMID:26865688

  12. Luteinizing hormone promotes gonadal tumorigenesis in inhibin-deficient mice

    Science.gov (United States)

    Nagaraja, Ankur K.; Agno, Julio E.; Kumar, T. Rajendra; Matzuk, Martin M.

    2009-01-01

    Summary The inhibins are secreted α:β heterodimers of the TGF-β superfamily that are mainly synthesized in Sertoli cells and granulosa cells, and are critical regulators of testicular and ovarian development and function. Mice homozygous for a targeted deletion of the inhibin α subunit gene (Inha-/-) develop sex cord-stromal tumors in a gonadotropin-dependent manner. Here, we determine the contribution of LH to gonadal tumorigenesis by generating mice deficient in both inhibins and LH. Inha-/-Lhb-/- mice have increased survival and delayed tumor progression, and these observations correlate with lower serum FSH and estradiol levels compared to Inha-/- controls. Double mutant testicular tumors demonstrate decreased expression of cyclin D2, while double mutant ovarian tumors have elevated expression of p15INK4b and trend toward higher levels of p27Kip1. We conclude that LH is not required for tumor formation in the absence of inhibins but promotes tumor progression, likely through alterations in serum hormone levels and cell cycle regulators. PMID:18657590

  13. Proteomic Analysis of Bacillus thuringiensis Strain 4.0718 at Different Growth Phases

    Directory of Open Access Journals (Sweden)

    Xiaohui Li

    2012-01-01

    Full Text Available The growth process of Bacillus thuringiensis Bt4.0718 strain was studied using proteomic technologies. The proteins of Bt whole cells at three phases—middle vegetative, early sporulation, and late sporulation—were extracted with lysis buffer, followed with separation by 2-DE and identified by MALDI-TOF/TOF MS. Bioactive factors such as insecticidal crystal proteins (ICPs including Cry1Ac(3, Cry2Aa, and BTRX28, immune inhibitor (InhA, and InhA precursor were identified. InhA started to express at the middle vegetative phase, suggesting its contribution to the survival of Bt in the host body. At the early sporulation phase, ICPs started their expression. CotJC, OppA, ORF1, and SpoIVA related to the formation of crystals and spores were identified, the expression characteristics of which ensured the stable formation of crystals and spores. This study provides an important foundation for further exploration of the stable expression of ICPs, the smooth formation of crystals, and the construction of recombinant strains.

  14. Could the endogenous opioid, morphine, prevent neural stem cell proliferation?

    Science.gov (United States)

    Shoae-Hassani, Alireza; Sharif, Shiva; Tabatabaei, Seyed Abdolreza Mortazavi; Verdi, Javad

    2011-02-01

    In spite of widespread use of morphine to treat pain in patients, little is known about the effects of this opioid on many cells including stem cells. Moreover the studies have been shown controversial results about morphine effects on several kinds of cells. It is well-known that morphine exposure could decrease testosterone levels in brain and spinal cord. Morphine could increase the activity of 5α-redutase, the enzyme that converts testosterone into its respective 5α-redutase derivative dihydrotestosterone (DHT). Also it could increase aromatase activity that converts testosterone to estradiol. Proliferation of neural stem cells was observed in human stem cells after exposure to certain combinations of steroids especially testosterone. On the other hand DHT has negative effect in neural stem cell reproduction. Morphine induces over-expression of p53 gene that could mediate stem cell apoptosis. Therefore we hypothesized that due to reduction in the testosterone levels, elevation in the DHT levels, and over-expression of p53 gene, morphine could prevent neural stem cell proliferation. Copyright © 2010 Elsevier Ltd. All rights reserved.

  15. Atividade enzimática em variedades de cana-de-açúcar cultivadas in vitro sob diferentes níveis de nitrogênio Enzymatic activity in sugar cane varieties cultivated in vitro under nitrogen levels

    Directory of Open Access Journals (Sweden)

    Virgínia Maria Tenório Sabino Donato

    2004-11-01

    Full Text Available O nitrogênio é considerado o elemento mineral mais abundante nas plantas, sendo componente essencial de biomoléculas e inúmeras enzimas. O objetivo deste trabalho foi avaliar a eficiência no processo de assimilação do nitrogênio e encontrar parâmetros indicativos do potencial de fixação biológica em variedades de cana-de-açúcar (Saccharum officinarum cultivadas in vitro. Foram utilizadas as variedades de cana-de-açúcar RB 842021, RB 83102, RB 75126, RB 882980 e Co 997, além da Brachiaria arrecta (testemunha, cultivadas in vitro em diferentes níveis de nitrogênio [M1 (9,83 mM, M2 (2,46 mM, M3 (0,49 mM, M4 (0,0 mM]. Todas as variedades e a cultura testemunha (Brachiaria arrecta apresentaram atividade da nitrato redutase (NR constitutiva e também da glutamina sintetase (GS, mesmo na ausência de amônio e nitrato no meio de cultura. A variedade RB 842021 apresentou a maior atividade da nitrato redutase e o maior conteúdo de clorofilas a e b, e a variedade RB 882980 a maior atividade da glutamina sintetase nas mesmas condições de cultivo, o que pode representar maior potencial de assimilação do nitrogênio.Nitrogen is considered the most abundant mineral element in the plants and an essential component to biomolecules and several enzymes. The objective of this work was to estimate nitrogen assimilation efficiency process and to identify biological fixation potential parameters on sugar cane varieties cultivated in vitro. The sugar cane varieties used were RB 842021, RB 83102, RB 75126, RB 882980, CO 997, besides Brachiaria arrecta (witness, cultivated in vitro in different nitrogen levels [M1 (9.83 mM, M2 (2.46 mM, M3 (0.49 mM, M4 (0.0 mM]. All the varieties and plant witness (Brachiaria arrecta exhibited redutase nitrate activity and glutamina sintetase, even in the ammonium and nitrate absence culture media. The RB 842021 variety showed the highest activity of nitrate reductase and the higher chlorophyll a and b contents

  16. Isoniazid acetylating phenotype in patients with paracoccidioidomycosis and its relationship with serum sulfadoxin levels, glucose-6-phosphate dehydrogenase and glutathione reductase activities

    Directory of Open Access Journals (Sweden)

    Benedito Barraviera

    1991-06-01

    Full Text Available The authors evaluated the isoniazid acetylating phenotype and measured hematocrit, hemoglobin, glucose-6-phosphate dehydrogenase and glutathione reductase activities plus serum sulfadoxin levels in 39 patients with paracoccidioidomycosis (33 males and 6 females aged 17 to 58 years. Twenty one (53.84% of the patients presented a slow acetylatingphenotype and 18(46.16% a fast acetylating phenotype. Glucose-6-phosphate- dehydrogenase (G6PD acti vity was decreased in 5(23.80% slow acetylators and in 4(22.22% fast acetylators. Glutathione reductase activity was decreased in 14 (66.66% slow acetylators and in 12 (66.66% fast acetylators. Serum levels of free and total sulfadoxin Were higher in slow acetylator (p Os autores avaliaram o fenótipo acetilador da isoniazida, hematócrito, hemoglobina, atividade da glicose-6- fosfato desidrogenase, glutationa redutase e os níveis séricos de sulfadoxina de 39 doentes com paracoccidíoidomicose, senão 33 do sexo masculino e 6 do feminino, com idades compreendidas entre 17 e 58 anos. Vinte e um (53,84% doentes apresentaram fenótipo acetilador lento e 18 (46,16% rápido. A atividade da glicose-6-fosfato desidrogenase (G6PD esteve diminuída em 5 (23,80% acetiladores lentos e 4 (22,22% rápidos. A atividade da glutationa redutase esteve diminuída em 14 (66,66% acetiladores lentos e 12 (66,66% rápidos. Os níveis séricos de sulfadoxina livre e total foram maiores nos acetiladores lentos (p < 0,02. A análise dos resultados permite concluir que os níveis séricos de sulfadoxina relaciona-se com o fenótipo acetilador. Além disso, os níveis estiveram sempre acima de 50 µg/ml, níveis estes considerados terapêuticos. Por outro lado, a deficiência de glutationa redutase pode estar relacionada com a má absorção intestinal de nutrientes, entre eles riboflavina, vitamina precursora de FAD.

  17. Study of 5alph-reductase 2 and 3beta-hydroxysteroid dehydrogenase 2enzymes on ambiguous genital and prostate cancer

    OpenAIRE

    Lucio Fabio Caldas Ferraz

    2006-01-01

    O hormônio androgênico di-hidrotestosterona (DHT) possui fundamental importância na diferenciação sexual masculina e no desenvolvimento e manutenção da próstata. Duas enzimas atuam diretamente na concentração deste andrógeno nas células: 1) com uma função anabólica, a enzima 5α-redutase tipo 2 (gene SRD5A2) é responsável pela síntese de DHT ao converter testosterona (T) em 5α-di-hidrotestosterona e 2) com uma função catabólica, a enzima 3β- hidroxi desydrogenase/[...

  18. Inhibition by sivastatin of mitochondrial respiration from skeletal muscle and liver rat biopsies

    OpenAIRE

    Paolo Gadioli La Guardia

    2010-01-01

    Resumo: Inibidores da 3-hidroxi-3-metilglutaril-CoA redutase (estatinas) são fármacos utilizados para diminuir os níveis plasmáticos de colesterol e são, geralmente, seguros e bem tolerados. Ocasionalmente esses fármacos induzem miotoxicidade, como miopatia e rabdomiólise, e hepatotoxicidade. Neste trabalho investigou-se o mecanismo, in vitro e in vivo, pelo qual as estatinas atuam sobre a respiração mitocondrial de biópsias de músculo esquelético e de fígado de ratos. A incubação (1 hora) de...

  19. Avaliação da qualidade do leite cru e resfriado mediante a aplicação de princípios do APPCC Evaluation of the quality of raw and cool milk using HACCP guidelines

    Directory of Open Access Journals (Sweden)

    Leonardo Souza da Rosa

    2007-06-01

    Full Text Available O trabalho teve como objetivo avaliar a qualidade do leite cru e resfriado produzido no município de Canguçu - RS, através da aplicação de princípios do APPCC. Inicialmente, aplicou-se um plano de amostragem simples conforme norma ABC-STD-105, sendo elaboradas cartas controle para os parâmetros redutase, mastite e temperatura. As amostras de leite foram avaliadas quanto aos parâmetros contagem de aeróbios mesófilos, psicrotróficos, redutase, acidez, mastite, índice crioscópico, prova do álcool e resíduos de antibióticos. Os resultados possibilitaram a seleção de quatro rotas de captação do leite e vinte produtores, determinando-se um limite de controle para o parâmetro redutase de 321 minutos, mediante a equação Y = 2,35 x 10(7 - 7,01 x 10(4.X (r = -0,9284. A partir da relação existente entre a contagem de aeróbios mesófilos e psicrotróficos (r = 0,8561, obteve-se, conforme a equação Y = -8,73 x 10(4 + 0,3209 X, um limite de controle para psicrotróficos de 2,33 x 10(5 UFC.mL-1. Em virtude destes resultados, conclui-se que 80,0% das amostras estão acima do limite de controle estabelecido para a redutase, comportamento que se repete em 65,0% das amostras na contagem de psicrotróficos, caracterizando a existência de problemas nas etapas de resfriamento do leite e ordenha, as quais representam dois pontos críticos de controle, contribuindo com a perda de qualidade do leite.The aim of this work was to evaluate the quality of raw and cool milk produced in the municipal district of Canguçu - RS by using guidelines of the HACCP system. Initially, a sampling plan according to norm ABC-STD-105 was set up, and letters for the reductase, mastitis and temperature parameters were developed. The milk samples were evaluated in terms of the parameters total count of aerobious mesophiles, psycotrophic, reductase, acidity, mastitis, cryoscopic index, proof of alcohol and antibiotic residues. The results made it possible to

  20. Cinética do comprometimento do sistema citocromo p-450 microssomal hepático na esquistossomose mansônica murina

    OpenAIRE

    2006-01-01

    Vários são os fatores que afetam o metabolismo hepático normal e o de alguns medicamentos. Dentre eles, podem-se citar as doenças agudas ou crônicas que atingem a arquitetura e/ou função hepáticas, como por exemplo, a esquistossomose. Objetivo: Avaliar as alterações das enzimas citocromo P-450 das subfamílias 1A1, 2A6, 2B1, 2C11, 2E1, 3A2 e redutase, nas fases aguda, crônica e após tratamento com oxamniquine e avaliar a cinética das alterações das enzimas citocromo P-450 das subfamílias 1A...

  1. Açao da dutasterida no tecido prostático humano normal : papel dos receptores hormonais esteroides como possíveis marcadores clínicos

    OpenAIRE

    João Carlos Cardoso Alonso

    2013-01-01

    Introdução: Os andrógenos, além de desempenhar um papel importante no desenvolvimento e no crescimento da próstata, também são responsáveis pelo surgimento e progressão de lesões prostáticas. Portanto, as ações desses hormônios podem ser antagonizadas impedindo a conversão irreversível de testosterona em dihidrotestosterona, por meio da inibição da 5a-redutase. Neste contexto, os objetivos deste estudo foram caracterizar os efeitos clínicos e morfofuncionais da dutasterida, um inibidor de 5a-...

  2. Rapid detection of isoniazid resistance in Mycobacterium tuberculosis isolates by use of real-time-PCR-based melting curve analysis.

    Science.gov (United States)

    Hu, Siyu; Li, Guoli; Li, Hui; Liu, Xiaoli; Niu, Jianjun; Quan, Shengmao; Wang, Feng; Wen, Huixin; Xu, Ye; Li, Qingge

    2014-05-01

    The MeltPro TB/INH assay, recently approved by the Chinese Food and Drug Administration, is a closed-tube, dual-color, melting curve analysis-based, real-time PCR test specially designed to detect 30 isoniazid (INH) resistance mutations in katG position 315 (katG 315), the inhA promoter (positions -17 to -8), inhA position 94, and the ahpC promoter (positions -44 to -30 and -15 to 3) of Mycobacterium tuberculosis. Here we evaluated both the analytical performance and clinical performance of this assay. Analytical studies with corresponding panels demonstrated that the accuracy for detection of different mutation types (10 wild-type samples and 12 mutant type samples), the limit of detection (2×10(3) to 2×10(4) bacilli/ml), reproducibility (standard deviation [SD], real-time PCR machines, with the shortest running time (105 min) obtained with the LightCycler 480 II. Clinical studies enrolled 1,096 clinical isolates collected from three geographically different tuberculosis centers, including 437 INH-resistant isolates and 659 INH-susceptible isolates characterized by traditional drug susceptibility testing on Löwenstein-Jensen solid medium. The clinical sensitivity and specificity of the MeltPro TB/INH assay were 90.8% and 96.4%, respectively. DNA sequencing analysis showed that, except for the 5 mutants outside the detection range of the MeltPro assay, a concordance rate between the two methods of 99.1% (457/461) was obtained. Among the 26 mutation types detected, katG S315T (AGC→ACC), inhA -15C→T, katG S315N (AGC→AAC), and ahpC promoter -10C→T accounted for more than 90%. Overall, the MeltPro TB/INH assay represents a reliable and rapid tool for the detection of INH resistance in clinical isolates.

  3. Drug discovery using chemical systems biology: repositioning the safe medicine Comtan to treat multi-drug and extensively drug resistant tuberculosis.

    Directory of Open Access Journals (Sweden)

    Sarah L Kinnings

    2009-07-01

    Full Text Available The rise of multi-drug resistant (MDR and extensively drug resistant (XDR tuberculosis around the world, including in industrialized nations, poses a great threat to human health and defines a need to develop new, effective and inexpensive anti-tubercular agents. Previously we developed a chemical systems biology approach to identify off-targets of major pharmaceuticals on a proteome-wide scale. In this paper we further demonstrate the value of this approach through the discovery that existing commercially available drugs, prescribed for the treatment of Parkinson's disease, have the potential to treat MDR and XDR tuberculosis. These drugs, entacapone and tolcapone, are predicted to bind to the enzyme InhA and directly inhibit substrate binding. The prediction is validated by in vitro and InhA kinetic assays using tablets of Comtan, whose active component is entacapone. The minimal inhibition concentration (MIC(99 of entacapone for Mycobacterium tuberculosis (M.tuberculosis is approximately 260.0 microM, well below the toxicity concentration determined by an in vitro cytotoxicity model using a human neuroblastoma cell line. Moreover, kinetic assays indicate that Comtan inhibits InhA activity by 47.0% at an entacapone concentration of approximately 80 microM. Thus the active component in Comtan represents a promising lead compound for developing a new class of anti-tubercular therapeutics with excellent safety profiles. More generally, the protocol described in this paper can be included in a drug discovery pipeline in an effort to discover novel drug leads with desired safety profiles, and therefore accelerate the development of new drugs.

  4. Lisotipia Vi e classificação bioquímica de amostras de Salmonella typhi isoladas no Estado da Guanabara

    Directory of Open Access Journals (Sweden)

    Ernesto Hofer

    1972-01-01

    Full Text Available Na presente investigação, os autores estudaram a distribuição dos lisótipos Ví, tipos fermentativos e tetrationato-redutase, em 110 amostras de Salmonella typhi, isoladas no período de 1966-1970, no Estado da Guanabara. Para estabelecer uma melhor configuração epidemiológica dos resultados, fez-se a divisão do Estado em seis regiões, respectivamente as zonas Sul, Centro, Norte, da E. F. Central, da E. F. Leopoldina e rural. Obteve-se no cômputo geral, a prevalência dos lisótipos A (59,09%, E 1a (19,09%, T (7,27% e amostras Vi negativas (4,55%. Verificou-se ainda, que cerca de 70,90% das culturas foram classificadas no tipo fermentativo II, segundo esquema de Kristensen e 95,42% das amostras revelaram-se positivas na prova de detecção da tetrationato-redutase.In the present investigation, the authors have studied the distribution of lysotypes, fermentative types and tetrathionate- reductase in 110 strains of Salmonella typhi, isolated from hemocultures during the period 1966-1970, in the State of Guanabara, Brazil. In order to establish a better epidemiological configuration of the results, this study has been divided and effected in the six following regions: south, middle, north at the Central Railway, the Leopoldina Railway and the rural district. As a general outcome a predomination of the lysotypes A, E 1a, T and negative Vi strains has been obtained. It has stil been verified that 70,90% of the cultures were classified fermentative type Ii and that 95,42% of the strains were tetrathionate-reductase positives.

  5. Antihypercholesterolemic and antioxidant effect of sterol rich methanol extract of stem of Musa sapientum (banana) in cholesterol fed wistar rats.

    Science.gov (United States)

    Dikshit, Piyush; Tyagi, Mool Kumar; Shukla, Kirtikar; Gambhir, Jasvindar K; Shukla, Rimi

    2016-03-01

    Musa sapientum Linn. (English 'Banana' family Musaceae), is a plant with nutritive, as well as medicinal value. Antihypercholesterolemic and antioxidant effect of methanolic extract of stem of this plant was investigated in hypercholesterolemic rats. Rats were made hypercholesterolemic by feeding cholesterol (100 mg/kg/day) suspended in soya oil. Treatment groups received extract at a dose of 10, 20 and 40 mg/kg/day in addition to cholesterol orally once daily. Fasting blood samples were collected before and after 6 weeks treatment. Animals were sacrificed and liver stored at -80 °C. Total cholesterol, HDL-cholesterol and triacylglycerol were estimated in blood. Malondialdehyde, reduced glutathione, superoxide dismutase and catalase were measured in blood and liver. Total lipids, HMG CoA redutase and lipoprotein lipase were investigated in liver. Most effective dose was found to be 20 mg/kg/day. Rise in total cholesterol, LDL + VLDL-cholesterol and triacylglycerol in animals receiving only cholesterol was 179 %, 417 % and 74 % respectively, while in animals receiving 20 mg/kg dose rise in these parameters was restricted to 40 %, 106 % and 24 %. HDL-cholesterol decreased by 12 % in extract treated group, while it decreased to 36 % in untreated hypercholesterolemic rats. Malonaldialdehyde, marker of lipid peroxidation decreased while reduced glutathione and enzymes superoxide dismutase and catalase increased significantly in blood and liver (p HMG CoA redutase and lipoprotein lipase were restored to near normal. Gas chromatography mass spectroscopy indicated high content of sterols in extract. Study demonstrated that methanol extract of stem of Musa sapientum has significant antihypercholesterolemic and antioxidant effects.

  6. Elucidating Drug-Enzyme Interactions and Their Structural Basis for Improving the Affinity and Potency of Isoniazid and Its Derivatives Based on Computer Modeling Approaches

    Directory of Open Access Journals (Sweden)

    Auradee Punkvang

    2010-04-01

    Full Text Available The enoyl-ACP reductase enzyme (InhA from M. tuberculosis is recognized as the primary target of isoniazid (INH, a first-line antibiotic for tuberculosis treatment. To identify the specific interactions of INH-NAD adduct and its derivative adducts in InhA binding pocket, molecular docking calculations and quantum chemical calculations were performed on a set of INH derivative adducts. Reliable binding modes of INH derivative adducts in the InhA pocket were established using the Autodock 3.05 program, which shows a good ability to reproduce the X-ray bound conformation with rmsd of less than 1.0 Å. The interaction energies of the INH-NAD adduct and its derivative adducts with individual amino acids in the InhA binding pocket were computed based on quantum chemical calculations at the MP2/6-31G (d level. The molecular docking and quantum chemical calculation results reveal that hydrogen bond interactions are the main interactions for adduct binding. To clearly delineate the linear relationship between structure and activity of these adducts, CoMFA and CoMSIA models were set up based on molecular docking alignment. The resulting CoMFA and CoMSIA models are in conformity with the best statistical qualities, in which r2cv is 0.67 and 0.74, respectively. Structural requirements of isoniazid derivatives that can be incorporated into the isoniazid framework to improve the activity have been identified through CoMFA and CoMSIA steric and electrostatic contour maps. The integrated results from structure-based, ligand-based design approaches and quantum chemical calculations provide useful structural information facilitating the design of new and more potentially effective antitubercular agents as follow: the R substituents of isoniazid derivatives should contain a large plane and both sides of the plane should contain an electropositive group. Moreover, the steric and electrostatic fields of the 4-pyridyl ring are optimal for greater potency.

  7. Genotypic Analysis of Genes Associated with Independent Resistance and Cross-Resistance to Isoniazid and Ethionamide in Mycobacterium tuberculosis Clinical Isolates.

    Science.gov (United States)

    Rueda, Johana; Realpe, Teresa; Mejia, Gloria Isabel; Zapata, Elsa; Rozo, Juan Carlos; Ferro, Beatriz Eugenia; Robledo, Jaime

    2015-12-01

    Ethionamide (ETH) is an antibiotic used for the treatment of multidrug-resistant (MDR) tuberculosis (TB) (MDR-TB), and its use may be limited with the emergence of resistance in the Mycobacterium tuberculosis population. ETH resistance in M. tuberculosis is phenomenon independent or cross related when accompanied with isoniazid (INH) resistance. In most cases, resistance to INH and ETH is explained by mutations in the inhA promoter and in the following genes: katG, ethA, ethR, mshA, ndh, and inhA. We sequenced the above genes in 64 M. tuberculosis isolates (n = 57 ETH-resistant MDR-TB isolates; n = 3 ETH-susceptible MDR-TB isolates; and n = 4 fully susceptible isolates). Each isolate was tested for susceptibility to first- and second-line drugs using the agar proportion method. Mutations were observed in ETH-resistant MDR-TB isolates at the following rates: 100% in katG, 72% in ethA, 45.6% in mshA, 8.7% in ndh, and 33.3% in inhA or its promoter. Of the three ETH-susceptible MDR-TB isolates, all showed mutations in katG; one had a mutation in ethA, and another, in mshA and inhA. Finally, of the four fully susceptible isolates, two showed no detectable mutation in the studied genes, and two had mutations in mshA gene unrelated to the resistance. Mutations not previously reported were found in the ethA, mshA, katG, and ndh genes. The concordance between the phenotypic susceptibility testing to INH and ETH and the sequencing was 1 and 0.45, respectively. Among isolates exhibiting INH resistance, the high frequency of independent resistance and cross-resistance with ETH in the M. tuberculosis isolates suggests the need to confirm the susceptibility to ETH before considering it in the treatment of patients with MDR-TB.

  8. PRI-Modeler: extracting RNA structural elements from PDB files of protein-RNA complexes.

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    Han, Kyungsook; Nepal, Chirag

    2007-05-01

    A complete understanding of protein and RNA structures and their interactions is important for determining the binding sites in protein-RNA complexes. Computational approaches exist for identifying secondary structural elements in proteins from atomic coordinates. However, similar methods have not been developed for RNA, due in part to the very limited structural data so far available. We have developed a set of algorithms for extracting and visualizing secondary and tertiary structures of RNA and for analyzing protein-RNA complexes. These algorithms have been implemented in a web-based program called PRI-Modeler (protein-RNA interaction modeler). Given one or more protein data bank files of protein-RNA complexes, PRI-Modeler analyzes the conformation of the RNA, calculates the hydrogen bond (H bond) and van der Waals interactions between amino acids and nucleotides, extracts secondary and tertiary RNA structure elements, and identifies the patterns of interactions between the proteins and RNAs. This paper presents PRI-Modeler and its application to the hydrogen bond and van der Waals interactions in the most representative set of protein-RNA complexes. The analysis reveals several interesting interaction patterns at various levels. The information provided by PRI-Modeler should prove useful for determining the binding sites in protein-RNA complexes. PRI-Modeler is accessible at http://wilab.inha.ac.kr/primodeler/, and supplementary materials are available in the analysis results section at http://wilab.inha.ac.kr/primodeler/.

  9. Phenotypic and Genotypic Analysis of Multidrug-Resistant Mycobacterium tuberculosis Isolates from Sudanese Patients

    Science.gov (United States)

    Salih, Mohamed Ahmed; Ali, Manasik; EL-Zaki, Salah-Eldin; Abuzeid, Nadir; Elgadi, Zeinab Abubaker Mohammed; Altayb, Hisham N.; Elegail, Asrar M. A.; Ibrahim, Nuha Y.; Elamin, Bahaeldin K.

    2017-01-01

    Background. Currently, mutations in rpoB, KatG, and rrs genes and inhA promoter were considered to be involved in conferring resistance to rifampicin, isoniazid, and streptomycin in Mycobacterium tuberculosis (MTB). Objective. The aims of this study were to detect the prevalence of first-line tuberculosis (TB) drug resistance among a group of previously treated and newly detected TB patients, to determine the association between prevalence of multidrug resistance (MDR) and demographic information (age and sex), to explain genes correlated with MDR Mycobacterium tuberculosis, and to characterize MTB via 16S ribosomal RNA (16S rRNA) analysis. Methods. A hundred MTB isolates from Sudanese pulmonary TB patients were included in the study. The proportional method of drug susceptibility test was carried out on Löwenstein-Jensen media. Multiplex PCR of rpoB and KatG genes and inhA promoter was conducted; then rrs genes were amplified by conventional PCR and were sequenced. The sequences of the PCR product were compared with known rrs gene sequences in the GenBank database by multiple sequence alignment tools. Result. The prevalence of MDR was 14.7% among old cases and 5.3% among newly diagnosed cases. Conclusion. Mutations in rrs could be considered as a diagnostic marker. PMID:28197340

  10. Aqueous Molecular Dynamics Simulations of the M. tuberculosis Enoyl-ACP Reductase-NADH System and Its Complex with a Substrate Mimic or Diphenyl Ethers Inhibitors

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    Camilo Henrique da Silva Lima

    2015-10-01

    Full Text Available Molecular dynamics (MD simulations of 12 aqueous systems of the NADH-dependent enoyl-ACP reductase from Mycobacterium tuberculosis (InhA were carried out for up to 20–40 ns using the GROMACS 4.5 package. Simulations of the holoenzyme, holoenzyme-substrate, and 10 holoenzyme-inhibitor complexes were conducted in order to gain more insight about the secondary structure motifs of the InhA substrate-binding pocket. We monitored the lifetime of the main intermolecular interactions: hydrogen bonds and hydrophobic contacts. Our MD simulations demonstrate the importance of evaluating the conformational changes that occur close to the active site of the enzyme-cofactor complex before and after binding of the ligand and the influence of the water molecules. Moreover, the protein-inhibitor total steric (ELJ and electrostatic (EC interaction energies, related to Gly96 and Tyr158, are able to explain 80% of the biological response variance according to the best linear equation, pKi = 7.772 − 0.1885 × Gly96 + 0.0517 × Tyr158 (R2 = 0.80; n = 10, where interactions with Gly96, mainly electrostatic, increase the biological response, while those with Tyr158 decrease. These results will help to understand the structure-activity relationships and to design new and more potent anti-TB drugs.

  11. Phenotypic and Genotypic Analysis of Multidrug-Resistant Mycobacterium tuberculosis Isolates from Sudanese Patients

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    Solima M. A. Sabeel

    2017-01-01

    Full Text Available Background. Currently, mutations in rpoB, KatG, and rrs genes and inhA promoter were considered to be involved in conferring resistance to rifampicin, isoniazid, and streptomycin in Mycobacterium tuberculosis (MTB. Objective. The aims of this study were to detect the prevalence of first-line tuberculosis (TB drug resistance among a group of previously treated and newly detected TB patients, to determine the association between prevalence of multidrug resistance (MDR and demographic information (age and sex, to explain genes correlated with MDR Mycobacterium tuberculosis, and to characterize MTB via 16S ribosomal RNA (16S rRNA analysis. Methods. A hundred MTB isolates from Sudanese pulmonary TB patients were included in the study. The proportional method of drug susceptibility test was carried out on Löwenstein-Jensen media. Multiplex PCR of rpoB and KatG genes and inhA promoter was conducted; then rrs genes were amplified by conventional PCR and were sequenced. The sequences of the PCR product were compared with known rrs gene sequences in the GenBank database by multiple sequence alignment tools. Result. The prevalence of MDR was 14.7% among old cases and 5.3% among newly diagnosed cases. Conclusion. Mutations in rrs could be considered as a diagnostic marker.

  12. Genotype MTBDR plus assay for molecular detection of rifampicin and isoniazid resistance in Mycobacterium tuberculosis

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    Soniya Sharma

    2014-01-01

    Full Text Available Aim: This study was performed for the rapid identification of Mycobacterium tuberculosis complex and its resistance to rifampicin and isoniazid, directly from the sputum samples of pulmonary tuberculosis patients. Materials and Methods: A commercially available genotype MTBDR plus assay was used for the identification and detection of mutations in Mycobacterial isolates. A total of 100 sputum samples of pulmonary tuberculosis patients were analyzed by using the genotype MTBDR plus assay. The MTBDR plus assay is designed to detect the mutations in the hotspot region of rpoB gene, katG and regulatory region of inhA gene. Results: The genotype MTBDR plus assay detected 22% multidrug resistant (MDR, 2% rifampicin (RMP monoresistant and 1% isoniazid (INH monoresistant isolates. In 22 MDR isolates, the codons most frequently involved in RMP-associated mutations were codon 531 (54.55%, 516 (31.82% and 526 (13.63%, and 90.90% of MDR isolates showed KatG S315T mutations and 9.1% showed inhA C-15T mutations associated with INH resistance. Conclusion: The new genotype MTBDR plus assay represents a rapid, reliable tool for the detection of MDR-TB, wherein results are obtained in 5 h allowing early and appropriate treatment, which is essential to cut the transmission path and reduce the spread of MDR-TB. The genotype MTBDR plus assay can readily be included in a routine laboratory work for the early diagnosis and control of MDR-TB.

  13. An MspI polymorphism in the inhibin alpha gene and its associations with superovulation traits in Chinese Holstein cows.

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    Tang, Ke-Qiong; Li, Shu-Jing; Yang, Wu-Cai; Yu, Jun-Na; Han, Li; Li, Xiang; Yang, Li-Guo

    2011-01-01

    To identify a predictor to forecast superovulation response on the basis of associations between superovulation performance and gene polymorphism, the PCR-RFLP method was applied to detect an A>G transition determining an MspI polymorphism at position 192 in the exon I of the bovine inhibin alpha (INHA) gene and evaluate its associations with superovulatory response in 118 Chinese Holstein cows treated for superovulation. Association analysis showed that cows with the GG genotype resulted in a significant increase in the number of ova (TNO) than AG and AA genotypes in the first (P=0.023), second (P=0.004) and third (P=0.002) superovulation treatments and produced more transferable embryos (NTE) than that of AG and AA genotypes in the third (P=0.045) superovulation treatment. Moreover, individuals with GG genotype produced more transferable embryos than AA (Psuperovulation treatment and all cows without superovulation response were mutations with genotypes of AA and AG. These results indicate that INHA gene can be used as a predictor for superovulation in Chinese Holstein cows, and imply that cows with AA genotype should be excluded for superovulation practices.

  14. A review of the use of ethionamide and prothionamide in childhood tuberculosis.

    Science.gov (United States)

    Thee, S; Garcia-Prats, A J; Donald, P R; Hesseling, A C; Schaaf, H S

    2016-03-01

    Ethionamide (ETH) and prothionamide (PTH), both thioamides, have proven efficacy in clinical studies and form important components for multidrug-resistant tuberculosis treatment regimens and for treatment of tuberculous meningitis in adults and children. ETH and PTH are pro-drugs that, following enzymatic activation by mycobacterial EthA inhibit InhA, a target shared with isoniazid (INH), and subsequently inhibit mycolic acid synthesis of Mycobacterium tuberculosis. Co-resistance to INH and ETH is conferred by mutations in the mycobacterial inhA promoter region; mutations in the ethA gene often underlie ETH and PTH monoresistance. An oral daily dose of ETH or PTH of 15-20 mg/kg with a maximum daily dose of 1000 mg is recommended in children to achieve adult-equivalent serum concentrations shown to be efficacious in adults, although information on optimal pharmacodynamic targets is still lacking. Gastrointestinal disturbances, and hypothyroidism during long-term therapy, are frequent adverse effects observed in adults and children, but are rarely life-threatening and seldom necessitate cessation of ETH therapy. More thorough investigation of the therapeutic effects and toxicity of ETH and PTH is needed in childhood TB while child-friendly formulations are needed to appropriately dose children.

  15. Use of Genotype MTBDRplus Assay for Diagnosis of Multidrug-Resistant Tuberculosis in Nepal

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    Elina Maharjan

    2017-01-01

    Full Text Available The main aims of this study were to study the patterns of mutations in rpoB, katG, and inhA genes in Mycobacterium tuberculosis strains isolated from patients from Nepal and to evaluate the performance of genotype MTBDRplus assay, taking conventional drug susceptibility testing as gold standard for diagnosis of MDR-TB. A total of 69 Mycobacterium tuberculosis strains isolated from 73 smear positive sputum samples from patients suspected of suffering from multidrug-resistant tuberculosis were used in our study. The drug susceptibility pattern of Mycobacterium tuberculosis isolated from these sputum specimens was determined by using genotype MTBDRplus assay taking conventional drug susceptibility testing as reference. The sensitivity and specificity of the genotype MTBDRplus assay for the detection of MDR-TB were found to be 88.7% and 100%, respectively. 88.7% of the rifampicin resistant isolates had mutations in rpoB gene. Similarly, 79.7% and 9.4% of isoniazid resistant isolates had mutations in katG and inhA genes, respectively. Genotype MTBDRplus assay was found to be very rapid and highly sensitive and specific method for diagnosis of MDR-TB and will be very helpful for early diagnosis of MDR-TB in high tuberculosis burden countries.

  16. Genotypic characterization of multi-drug-resistant Mycobacterium tuberculosis isolates in Myanmar.

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    Aye, Khin Saw; Nakajima, Chie; Yamaguchi, Tomoyuki; Win, Min Min; Shwe, Mu Mu; Win, Aye Aye; Lwin, Thandar; Nyunt, Wint Wint; Ti, Ti; Suzuki, Yasuhiko

    2016-03-01

    The number of multi-drug-resistant tuberculosis (MDR-TB) cases is rising worldwide. As a countermeasure against this situation, the implementation of rapid molecular tests to identify MDR-TB would be effective. To develop such tests, information on the frequency and distribution of mutations associating with phenotypic drug resistance in Mycobacterium tuberculosis is required in each country. During 2010, the common mutations in the rpoB, katG and inhA of 178 phenotypically MDR M. tuberculosis isolates collected by the National Tuberculosis Control Program (NTP) in Myanmar were investigated by DNA sequencing. Mutations affecting the 81-bp rifampicin (RIF) resistance-determining region (RRDR) of the rpoB were identified in 127 of 178 isolates (71.3%). Two of the most frequently affected codons were 531 and 526, with percentages of 48.3% and 14.0% respectively. For isoniazid (INH) resistance, 114 of 178 MDR-TB isolates (64.0%) had mutations in the katG in which a mutation-conferring amino acid substitution at codon 315 from Ser to Thr was the most common. Mutations in the inhA regulatory region were also detected in 20 (11.2%) isolates, with the majority at position -15. Distinct mutation rate and pattern from surrounding countries might suggest that MDR-TB has developed and spread domestically in Myanmar.

  17. Caracterização de miopatias mitocondriais através da avaliação das atividades enzimáticas envolvidas no metabolismo energético Characterization of mitochondrial myopathies through the evaluation of the enzymatic activities involved in the energetic metabolism

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    Fábio Cesar Pedroso

    1997-06-01

    Full Text Available Foi determinada a atividade das enzimas NADH desidiogenase, NADH citocromo e redutase, succinato desidiogenase, succinato citocromo e redutase, citocromo e oxidase e citrato sintase em mitocôndrias de músculo esquelético humano normal e doente (suspeito de miopatia mitocondrial. O grupo controle foi constituído de 13 indivíduos normais e que não faziam uso contínuo de fárrnacos. O grupo doente era constituído de 10 pacientes cujo diagnóstico anatomopatológico indicava suspeita de miopatia mitocondrial. Observou-se redução na atividade das enzimas em todos os pacientes: 7 com anormalidades em todas as enzimas ensaiadas; 2 com deficiências em todas as enzimas exceto na citocromo e oxidase; e 1 paciente com disfunção apenas na atividade da succinato desidiogenase e succinato citocromo e redutase. Este perfil possibilitou caracterizar múltiplas deficiências ou deficiência combinada da cadeia respiratória, além da disfunção na citrato sintase em 9 pacientes. Um dos casos constituiu exceção, sendo a deficiência enzimática restrita ao complexo II. Foi possível concluir que a metodologia usada é adequada e facilmente aplicável aos objetivos clínicos. Os resultados obtidos possibilitam a caracterização dos complexos enzimáticos mitocondriais deficientes, mostrando que tais enfermidades são originadas de disfunção no metabolismo energético.The activities of the enzymes NADH dehydrogenase, NADH cytochrome e reductase, succinate dehydrogenase, succinate cytochrome e reductase, cytochrome e oxidase and citrate synthase in normal and sick human skeletal muscle mitochondria were determined. A control group was formed by 13 normal people and without using continuous medication. The patient group was formed by 10 people whose pathological diagnosis indicated suspicion of mitochondrial myopathy. A decrease in the activity of the enzymes in all patient was observed: 7 with abnormality in all the tested enzymes; 2 with

  18. Structure-activity studies of the inhibition of FabI, the enoyl reductase from Escherichia coli, by triclosan: kinetic analysis of mutant FabIs.

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    Sivaraman, Sharada; Zwahlen, Jacque; Bell, Alasdair F; Hedstrom, Lizbeth; Tonge, Peter J

    2003-04-22

    Triclosan, a common antibacterial additive used in consumer products, is an inhibitor of FabI, the enoyl reductase enzyme from type II bacterial fatty acid biosynthesis. In agreement with previous studies [Ward, W. H., Holdgate, G. A., Rowsell, S., McLean, E. G., Pauptit, R. A., Clayton, E., Nichols, W. W., Colls, J. G., Minshull, C. A., Jude, D. A., Mistry, A., Timms, D., Camble, R., Hales, N. J., Britton, C. J., and Taylor, I. W. (1999) Biochemistry 38, 12514-12525], we report here that triclosan is a slow, reversible, tight binding inhibitor of the FabI from Escherichia coli. Triclosan binds preferentially to the E.NAD(+) form of the wild-type enzyme with a K(1) value of 23 pM. In agreement with genetic selection experiments [McMurry, L. M., Oethinger, M., and Levy, S. B. (1998) Nature 394, 531-532], the affinity of triclosan for the FabI mutants G93V, M159T, and F203L is substantially reduced, binding preferentially to the E.NAD(+) forms of G93V, M159T, and F203L with K(1) values of 0.2 microM, 4 nM, and 0.9 nM, respectively. Triclosan binding to the E.NADH form of F203L can also be detected and is defined by a K(2) value of 51 nM. We have also characterized the Y156F and A197M mutants to compare and contrast the binding of triclosan to InhA, the homologous enoyl reductase from Mycobacterium tuberculosis. As observed for InhA, Y156F FabI has a decreased affinity for triclosan and the inhibitor binds to both E.NAD(+) and E.NADH forms of the enzyme with K(1) and K(2) values of 3 and 30 nM, respectively. The replacement of A197 with Met has no impact on triclosan affinity, indicating that differences in the sequence of the conserved active site loop cannot explain the 10000-fold difference in affinities of FabI and InhA for triclosan.

  19. The genotypic study of Mycobacterium tuberculosis complex resistant to isoniazid: Galicia, Spain (2008-2013).

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    Pérez Del Molino, M L; Barbeito-Castiñeiras, G; Mejuto, B; Alonso, P; Fernández, A; González-Mediero, G

    2016-11-01

    Incorporation of rapid detection systems to identify mutations in M. tuberculosis complex that confer resistance to isoniazid and rifampicin has potentiated the knowledge of their distribution, given the geographical variability. We performed antibiograms of the 2,993 strains isolated in Galicia, Spain (2008-2013). In the strains resistant to isoniazid, a concentration of 0.4 mg/mL and MTBDRplus Genotype test (Hain Lifescience, Germany) were used. We found that 3.64 % of strains were resistant to isoniazid, while 0.43 % were resistant to isoniazid and rifampicin (multidrug resistant, MDR). The MTBDRplus test showed an overall sensitivity of 72.48 %, with 62.5 % sensitivity for non MDR isoniazid-resistant strains and 100 % sensitivity for MDR strains. The katG gene mutation was detected at codon 315 in 38.53 % of strains. The S315T mutation appeared in 61.54 % of MDR strains and 34.38 % of non-MDR strains. The 28.44 % had mutations in inhA, (93.55 % in C15T), and 38.46 % of MDR strains were mutated. In non-MDR strains, 37.50 % were wild-type, 35.42 % and 27.08 % had mutations in katG and inhA, respectively. The most frequent mutation in rpoβ was S531L (46.15 %). The 38.71 % and 41.9 % of strains with resistance to isoniazid and streptomycin had mutations in katG and inhA, respectively (2 strains with mutations in T8C and T8A). The distribution pattern of resistance among strains with high and low concentrations of isoniazid showed statistically significant differences in relation to the mutation in katG and wild-type. The sensitivity of the Genotype MTBDRplus test for non-MDR strains in our area was at the lower threshold described.

  20. Diagnostic moléculaire du complexe Mycobacterium tuberculosis résistant à l'isoniazide et à la rifampicine au Burkina Faso

    Science.gov (United States)

    Désire, Ilboudo; Cyrille, Bisseye; Florencia, Djigma; Souba, Diande; Albert, Yonli; Valerie, Bazie Jean Telesphore; Rebecca, Compaore; Charlemagne, Gnoula; Tamboura, Djibril; Rémy, Moret; Virginio, Pietra; Simplice, Karou Damintoti; Martial, Ouedraogo; Jacques, Simpore

    2015-01-01

    Introduction Cette étude a eu pour objectifs de diagnostiquer la tuberculose pulmonaire par l'examen microscopique et par la PCR des crachats et de déterminer les bases moléculaires de la résistance à la rifampicine et à l'isoniazide. Méthodes Le diagnostic du Complexe Mycobacterium Tuberculosis (CMTB) a été effectué par microscopie après coloration au Ziehl Nielsen et par PCR en temps réel en utilisant le kit d'identification du complexe MTB (Sacace Biotechnologie, Italie). Les résistances à la Rifampicine et à l'Isoniazide ont été étudiées par la technique de la PCR en utilisant le kit MTB résistance 8 (Sacace, Biotechnologie). Résultats Sur les 59 patients diagnostiqués pour la tuberculose pulmonaire, 59,3% étaient positifs en microscopie optique et 44,1% étaient positifs par PCR en Temps réel. Les résistances à la rifampicine (rpoB) et à l'isoniazide (katG et inhA) ont été observées chez 9 patients. La résistance à la rifampicine était due aux mutations (Asp516Val, Ser531Trp, Leu533Pro) et celle à l'isoniazide par les substitutions Ser315Thr du gène katG et C209T du gène inhA. Les multi résistances à la rifampicine et à l'isoniazide ont été observées dans 55,5% des échantillons et concernaient les associations: ropBAsp513Val + inhAC209T et rpoBLeu533Pro + katGSer315Thr. Conclusion La PCR en temps réel qui permet l'identification des allèles mutants rpoB, katG et inhA de M. tuberculosis est un outil de diagnostic épidémiologique de grande importance car elle permet de déterminer le niveau de résistance à la rifampicine et à l'isoniazide. PMID:26491516

  1. Molecular characteristics of rifampin and isoniazid resistant Mycobacterium tuberculosis strains from Beijing, China

    Institute of Scientific and Technical Information of China (English)

    JIAO Wei-wei; Mokrousov Igor; SUN Gui-zhi; LI Mo; LIU Jia-wen; Narvskaya Olga; SHEN A-dong

    2007-01-01

    Background China is one of the high burden countries of Mycobacterium tuberculosis (TB) infection globally, with high incidence and mortality. We studied the molecular characteristics of rifampin (RIF) and isoniazid (INH) resistant Mycobacterium tuberculosis strains from Beijing, China, in order to find out the genetic marker for rapid detection of specific drug resistance.Methods Forty pansusceptible and 81 resistant strains of Mycobacterium tuberculosis isolated from Beijing, China during 2002-2005 were analyzed. The modified rifampin oligonucleotide (RIFO) assay based on reverse line blot hybridization was used to detect mutations in the 81 bp hot-spot region of rpoB gene, which is associated with RIF resistance. The INH resistance associated genes, regulatory region mab-inhA (-15C/T) and structural gene katG S315T were detected by reverse line blot hybridization and PCR-restriction fragment length polymorphism (RFLP) method respectively. All the strains were typed by spoligotying and the Beijing genotype was further subdivided by NTF locus analysis. The distribution of drug resistance associated mutations in the above genes was compared in these groups.Results Sixty-five (91.5%) of 71 RIF resistant and 52 (92.9%) of 56 multidrug-resistant (MDR, Ⅰ.e. Resistant to at least RIF and INH) strains were found to harbor mutations in the rpoB hot-spot region. No mutation was detected in RIF sensitive strains. The specificity and sensitivity of the modified RIFO assay were 100% and 91.5%, respectively. katG315 AGC>ACC and inhA-15C>T mutations were found in 40 (60.6%) and 10 (15.2%) of 66 Ⅰ NH resistant strains, respectively;7.6% of INH-resistant strains had mutations in both of these genes. Therefore, a combined use of both katG315 and inhA-15 identified 68.2% of INH-resistant strains. The Beijing genotype accounted for 91.7% of total strains and was further subdivided into "modern" (76.6%) and "ancestral" (23.4%) group. There is no significant difference between

  2. Comparative evaluation of GenoType MTBDRplus line probe assay with solid culture method in early diagnosis of multidrug resistant tuberculosis (MDR-TB at a tertiary care centre in India.

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    Raj N Yadav

    Full Text Available BACKGROUND: The objectives of the study were to compare the performance of line probe assay (GenoType MTBDRplus with solid culture method for an early diagnosis of multidrug resistant tuberculosis (MDR-TB, and to study the mutation patterns associated with rpoB, katG and inhA genes at a tertiary care centre in north India. METHODS: In this cross-sectional study, 269 previously treated sputum-smear acid-fast bacilli (AFB positive MDR-TB suspects were enrolled from January to September 2012 at the All India Institute of Medical Sciences hospital, New Delhi. Line probe assay (LPA was performed directly on the sputum specimens and the results were compared with that of conventional drug susceptibility testing (DST on solid media [Lowenstein Jensen (LJ method]. RESULTS: DST results by LPA and LJ methods were compared in 242 MDR-TB suspects. The LPA detected rifampicin (RIF resistance in 70 of 71 cases, isoniazid (INH resistance in 86 of 93 cases, and MDR-TB in 66 of 68 cases as compared to the conventional method. Overall (rifampicin, isoniazid and MDR-TB concordance of the LPA with the conventional DST was 96%. Sensitivity and specificity were 98% and 99% respectively for detection of RIF resistance; 92% and 99% respectively for detection of INH resistance; 97% and 100% respectively for detection of MDR-TB. Frequencies of katG gene, inhA gene and combined katG and inhA gene mutations conferring all INH resistance were 72/87 (83%, 10/87 (11% and 5/87 (6% respectively. The turnaround time of the LPA test was 48 hours. CONCLUSION: The LPA test provides an early diagnosis of monoresistance to isoniazid and rifampicin and is highly sensitive and specific for an early diagnosis of MDR-TB. Based on these findings, it is concluded that the LPA test can be useful in early diagnosis of drug resistant TB in high TB burden countries.

  3. Study of rifampicin and isoniazid resistance mutation genes of M. tuberculosis isolates in Nepal.

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    Khadka, J B; Rai, S K; Shrestha, S; Maharjan, B; Bhatta, D R; Ghimire, P

    2011-09-01

    We studied the rifampicin (RIF) and isoniazid (INH) resistant genes in Mycobacterium tuberculosis isolated from sputum samples of re-treated TB cases using phenotypic drug susceptibility test (DST) and rapid molecular method (Genotype; MTBDR plus, Hains Life Science, Nehren, Germany). Of the total 207 isolates included in this study, 90 (42.0%) were RIF resistant, 107 (50.7%) were INH resistant whereas 86 (41.5%) were multidrug resistant (MDR) by DST. According to genotypic analysis, 37.0% (77/207) were RIF resistant, 50.2% (105/207) INH resistant and 70 (33.8%) were MDR. Among the total 90 RIF resistant isolates, 85.5% (77/90) showed mutation in 81 bp of rpoB gene and the mutation locus were distributed as follows; MUT1 Asp516Val (16.7%; 15/90), MUT2A His526Tyr (6.6%; 6/90), MUT2B His526Asp (5.5%; 5/90) and MUT3 Ser531Lue (56.7%; 51/90). Among the total 107 INH resistant isolates, the mutation located in katG gene was 98.1% (105/107) and in inhA MUT1 gene was (16.8%; 18/107). The mutation locus were distributed as follows; MUT1 Ser315Thr (77.6%; 83/107), MUT2 Ser315Ile (3.7%; 4/107) in katG and in inhA MUT1Cys15Thr (16.8%; 18/107). The highest frequency of mutations in rpoB gene was found at the region of codon Ser531Leu (56.7%) while the frequency of mutation in katG and inhA gene were found at the region of codon Ser315Thr (77.6%) and Cyst15Thr (17.1%), respectively. The rapid molecular test kit used in this study was found to be sensitive as well as specific for detection of RIF and INH resistant gene and also helpful in early detection of MDR-TB.

  4. Trocas gasosas e estresse oxidativo em plantas de algodoeiro supridas com silício e infectadas por Ramularia areola

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    Carmen Rosa da Silva Curvêlo

    2013-01-01

    Full Text Available Considerando o efeito positivo do silício (Si na melhoria do desempenho fisiológico de várias espécies de plantas quando infectadas por patógenos, este trabalho teve como meta investigar o efeito desse elemento na capacidade fotossintética e no metabolismo antioxidativo de plantas dos cultivares de algodoeiro NuOpal e BRS Buriti, supridas ou não com Si, durante o processo infeccioso de Ramularia areola. A infecção por R. areola afetou negativamente a taxa líquida de fixação do CO2 (A, a condutância estomática (g s, a transpiração e a razão entre concentração interna e ambiente de CO2 nas plantas dos cultivares NuOpal e BRS Buriti não supridas com Si em relação às plantas supridas com esse elemento. Na presença do Si, houve aumento em A e g s além das concentrações de clorofila a, clorofila b e carotenóides nas plantas dos dois cultivares. A concentração foliar de Si aumentou, o que contribuiu para decrescer em 38% a área abaixo da curva do progresso da mancha de ramulária nas plantas supridas com Si, em relação às plantas não supridas com esse elemento. O fornecimento de Si às plantas de algodoeiro dos dois cultivares aumentou a atividade da dismutase do superóxido (SOD, da peroxidase do ascorbato (APX e da lipoxigenase (LOX, mas houve redução na atividade da catalase (CAT e na concentração de peróxido de hidrogênio (H2O2 e de aldeído malônico (MDA, houve ainda uma maior perda de eletrólitos. A atividade da glutationa redutase (GR foi maior nas plantas do cultivar BRS Buriti supridas com Si, enquanto que nas plantas do cultivar NuOpal houve aumento da atividade da glutationa redutase apenas aos 21 dias após inoculação (dai. Conclui-se que a capacidade fotossintética e o sistema antioxidativo das plantas supridas com Si sofreram menor impacto durante o processo infeccioso de R. areola.

  5. Methemoglobinemia associated with loxoscelism Meta-hemoglobinemia associada ao loxoscelismo

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    Orlando C. de O. Barreto

    1990-02-01

    Full Text Available In twenty five patients who presented the cutaneous form of loxoscelism, serum haptoglobin and lactic dehydrogenase, erythrocyte glucose-6-phosphate dehydrogenase, glutathione reductase, glutathione peroxidase, methemoglobin, bilirubin and reticulocytes were investigated after bite. No hemolysis was detected but an increase in methemoglobin was found in 54% of the cases; in 7% it was between 1.1% and 2%, in 27% it ranged from 2.1% to 4%, and in 20% from 4.1% to 8%. Blood samples of a normal, blood group 0 individual and of a patient who exhibited methemoglobinemia after Loxosceles bite were incubated separately with antisera against Loxosceles gaucho, Crotalus terrificus, Bothrops jararaca, with Loxosceles gaucho venom and 0.3% phenol. No methemoglobin was found after 1, 4,8 and 15 days in both sets of samples. At the 25th day all the samples, including the controls, exhibited similar methemoglobin reductase decrease. The data suggest that the methemoglobinemia which occurs in 50% of the patients probably arises from in vivo venom metabolism, inasmuch as the crude venom does not induce methemoglobinemia.Vinte e cinco pacientes que apresentaram a forma cutânea do loxoscelismo foram estudados após a picada, determinando-se a glicose-6-fosfato desidrogenase, glutationa redutase e glutationa peroxidase eritrocitárias, haptoglobina e latico desidrogenase séricas, bilirrubina, reticulócitos e meta-hemoglobina. Não foi observada hemólise aumentada, mas foi detectado aumento da meta-hemoglobina em 54% dos casos: em 7% entre 1,1% e 2%, em 27% variou de 2,1% a 4%, e em 20% de 4,1 a 8%. Amostras de sangue de um indivíduo normal do grupo 0 de uma paciente que exibiu meta-hemoglobina após picada por Loxosceles foram incubadas separadamente com anti-soros contra Loxosceles gancho, Crotalus terrificus e Bothrops jararaca, com veneno de Loxosceles gaucho e fenol a 3%, e não se detectou aumento de meta-hemoglobina depois de 1, 4, 8 e 15 dias em todas

  6. Alterações no metabolismo de cinco genótipos de trigo com diferentes níveis de resistência ao Soil-borne wheat mosaic virus Alterations in the metabolism of five wheat genotypes with different resistance levels to Soil-borne wheat mosaic virus

    Directory of Open Access Journals (Sweden)

    Rocheli de Souza

    2006-09-01

    Full Text Available Soil-borne wheat mosaic virus - SBWMV, agente causal da virose que se caracteriza, em termos econômicos, como uma das mais importantes enfermidades da cultura de trigo, pode também infectar uma vasta gama de gramíneas. Com o objetivo de conhecer as alterações metabólicas promovidas pelo mosaico do trigo, foram analisados os teores de açúcares totais e a concentração de prolina e determinou-se a atividade da nitrato redutase. O experimento foi conduzido na área experimental da Embrapa Trigo, usando cinco genótipos de trigo (BRS Guabiju, BRS 194, BRS 179, BR 23 e PF 980524 com diferentes níveis de resistência ao SBWMV. As determinações bioquímicas foram realizadas 45 dias após a emergência de plantas. A atividade da nitrato redutase foi mais elevada em plantas sem sintomas, quando comparada às com sintomas. Os níveis de açúcares e de prolina foram mais elevados em plantas com sintomas do que nas sem sintomas. Os resultados encontrados comprovam as alterações metabólicas promovidas pelo SBWMV nos cinco genótipos de trigo testados.Soil-borne wheat mosaic virus - SBWMV causes substantial economic losses to the wheat crop and can also infect a wide range of grass crops. An experiment was conducted in the experimental area of Embrapa Trigo, using five genotypes of wheat (BRS Guabiju, BRS 194, BRS 179, BR 23, and PF 980524 with different resistance levels to SBWMV. Samples were collected 45 days after emergence, and levels of sugars, proline concentration, and nitrate reductase activity were biochemically analyzed to understand the metabolic alterations induced by SBWMV. Nitrate reductase activity was higher in asymptomatic plants, as compared to the level observed in plants with symptoms. Sugar and proline levels were higher in plants with disease symptoms than in asymptomatic plants. The results show that the metabolic changes were caused by the SBWMV in the five different genotypes used in the experiment.

  7. Can statins improve outcome in colorectal surgery?: Part I

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    Júlio César M Santos Jr

    2012-09-01

    Full Text Available Statins are recommended for people who have high serum cholesterol, and this role of statins has been well documented. However, some activities of statins, independent of their lipid-lowering effect, in conditions such as systemic inflammatory response syndrome, nephropathy, and other anti-inflammatory activities that reduce proinflammatory cytokines, are called "pleiotropic" effects of statins. For this reason, many candidates for surgical treatment are users of statins. As a result, benefits are observed in these patients, such as minimized postoperative complications, especially in cardiac or coronary surgery. This study was designed with the purpose of determining the current status of the use of statins as an adjuvant in the prevention of postoperative complications in colorectal surgery. Ongoing studies and future researches will help clarify the potential impact of statins on the prophylaxis of postoperative complications.As estatinas são drogas com o poder de inibir a hidroxi-metil-glutaril coenzima A redutase (HMG-CoA redutase, enzima que age na ativação da cadeia metabólica do colesterol. Portanto, sua principal ação, entre outros efeitos, é diminuir a concentração sérica total desse lipídeo. Por essa razão, muitas pessoas candidatas ao tratamento cirúrgico são pacientes usuários das estatinas. Seus outros efeitos, independente de sua capacidade para baixar os lipídeos circulantes, são denominados "efeitos pleiotrópicos" e estão, principalmente, relacionados à ação de bloqueio das atividades pró-inflamatórias, sobretudo minimizando, nos cardiopatas ou coronariopatas submetidos às operações cardíacas ou coronarianas, a prevalência da síndrome da reação inflamatória sistêmica, inclusive quando desencadeada por infecção. Estudos recentes têm sido elaborados para maiores conhecimentos dos mecanismos de ação das estatinas, especialmente em pacientes cardiopatas submetidos a tratamentos cirúrgicos n

  8. Absorção, fluxo no xilema e assimilação do nitrato em feijão-caupi submetido à salinidade

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    Rafael Magalhães Aragão

    Full Text Available Resumo O presente trabalho foi desenvolvido com o objetivo de avaliar qual etapa da aquisição de nitrato (absorção, fluxo no xilema ou redução assimilatória é mais influenciada pela presença do NaCl em feijão-caupi. Plântulas com 12 dias de idade foram tratadas com NaCl 50 mM em solução nutritiva durante 4 dias e as avaliações realizadas sob duas condições ambientais contrastantes: dia típico (pleno sol e dia completamente nublado (nublado. A salinidade afetou mais intensamente o fluxo da seiva e o fluxo de nitrato no xilema do que a transpiração sob as duas condições ambientais. Plantas tratadas com NaCl mostraram uma intensa redução na taxa de absorção de nitrato e na atividade da redutase de nitrato nas folhas, tanto em pleno sol como em dia nublado, quando comparadas com as plantas não tratadas. A transpiração foi reduzida pela nebulosidade, enquanto que os fluxos da seiva e de nitrato permaneceram inalterados. A absorção de nitrato e a atividade da redutase de nitrato foram menos afetadas pela nebulosidade do que a transpiração, tanto no controle como nas plantas estressadas. A nebulosidade afetou mais intensamente as variáveis de estudo das plantas controle do que de plantas tratadas. Em adição, o NaCl reduziu a concentração de nitrato em raiz, caule e folha nos dois ambientes, enquanto que a nebulosidade afetou somente nas folhas, tanto no controle como nas plantas estressadas. A salinidade afeta mais negativamente o fluxo de nitrato no xilema do que os processos de absorção e assimilação desse íon nas folhas de feijão-caupi.

  9. Eficiência na absorção e utilização de nitrogênio e atividade enzimática em genótipos de milho Nitrogen uptake and utilization efficiency and enzymatic activity in maize genotypes

    Directory of Open Access Journals (Sweden)

    Lucimar Rodrigues de Oliveira

    2013-09-01

    Full Text Available O objetivo desse estudo foi investigar o emprego dos componentes da eficiência no uso de nitrogênio e das atividades das enzimas nitrato redutase e glutamina sintetase na seleção de genótipos de milho eficientes no uso de nitrogênio (N. Foram avaliados 10 genótipos de milho em estádio de V4 em condições de alto e baixo N. O experimento foi composto por um fatorial 2 x 10 (dois níveis de N e 10 genótipos de milho em delineamento inteiramente casualizados com três repetições. Os seguintes caracteres foram avaliados: as eficiências na absorção (EAbN, na utilização (EUtN e no uso de N (EUsN e as atividades das enzimas glutamina sintetase (GS e nitrato redutase (NR. Para os caracteres que o efeito de genótipos foi significativo (pThe aim of this study was to investigate the employment of components of the nitrogen efficiency use (NUE, and of the activities of the nitrate reductase and glutamine synthetase enzymes in the selection of nitrogen use efficient maize genotypes of maize. Ten maize genotypes at V4 stage were evaluated under high and low N. The experiment consisted of a 2 x 10 factorial (two levels of N and ten maize genotypes, in a randomized complete block design with three replicates. The following traits were evaluated: N uptake efficiency (NUpE, N utilization efficiency (NUtE and N use efficiency (NUE, and activities of the glutamine synthetase (GS and nitrate reductase (NR enzymes. For traits which the effect of genotype was significant (p<0.05 in the variance analysis for level of N, the mean were compared using the t test (p<0.05. The indirect gains in NUE and their components with selection over the activities of the enzymes NR and GS were estimated under low N. We conclude that: under high N, NupE is efficient to differentiate nitrogen use efficient maize genotypes; under low N, NupE and NUtE efficient to differentiate nitrogen use efficient maize genotypes, the activity of NR enzyme is not a good physiological

  10. Naissance de la bestialité. Une anthropologie du rapport homme-animal dans les années 1300

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    Pierre-Olivier Dittmar

    2011-05-01

    Full Text Available Composition du juryMonsieur Jérôme Baschet,Monsieur Philippe Descola,Monsieur Michel Pastoureau,Monsieur Jean-Claude Schmitt (dir.,Madame Claudine Vassas,Monsieur Jean-Jacques Vincensini.Thèse soutenue à l’INHA, Salle Walter Benjamin, le 29 juin 2010RésuméLe rapport à l’animal dans le contexte chrétien est paradoxal. Alors qu’il se trouve pour la première fois totalement exclu du sacré légitime (abandon du sacrifice antique, des interdits alimentaires juifs, l’animal est extrêmement présent...

  11. Terra Foundation for American Art Europe

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    Géraldine Chouard

    2009-07-01

    Full Text Available Le 25 novembre 2008, à l'INHA, l'équipe de la Terra Foundation for American Art a annoncé ses nouveaux projets et notamment, à la suite de la fermeture du Musée d’art américain de Giverny, la création d'un centre parisien, situé dans le 2è arrondissement. Ce centre souhaite être, pour tous ceux que l'art américain intéresse, un nouveau lieu de ressources, d'échanges et de contacts.Sophie Lévy, Veerle Thielemans, Katherine Bourguignon, Ewa Bobrowska, Francesca Rose et Véronique Bossard ont tou...

  12. Multidrug-resistant tuberculosis: treatment outcome in Denmark, 1992-2007

    DEFF Research Database (Denmark)

    Bang, Didi; Lillebaek, Troels; Thomsen, Vibeke Østergaard;

    2010-01-01

    A retrospective nationwide study including all culture-verified multidrug-resistant (MDR) tuberculosis (TB) cases was performed in Denmark. The aim was to examine the long-term treatment outcome of MDR-TB, to assess if MDR-TB transmission occurs, and to evaluate a rapid mutation analysis detecting...... rifampin and isoniazid resistance in this cohort. Clinical data were obtained from patient records. A restriction fragment length polymorphism genotype database of all TB cases was compared for identical strains indicating active transmission. Twenty-nine cases of MDR-TB were identified and the incidence...... was low at 0.5%. Acquired MDR-TB and active transmission was rare. Mutations in rifampin (rpoB) and isoniazid (katG, inhA) genes correctly determined resistance in 100% and 82% of all isolates tested, respectively. Initial treatment success was 89% for 27 MDR-TB patients with available outcome data...

  13. Phosphodiesterases in the rat ovary

    DEFF Research Database (Denmark)

    Petersen, Tonny Studsgaard; Stahlhut, Martin; Andersen, Claus Yding

    2015-01-01

    Phosphodiesterases (PDEs) are important regulators of the intracellular cAMP concentration, which is a central second messenger that affects a multitude of intracellular functions. In the ovaries, cAMP exerts diverse functions, including regulation of ovulation and it has been suggested...... that augmented cAMP levels stimulate primordial follicle growth. The present study examined the gene expression, enzyme activity and immunolocalization of the different cAMP hydrolysing PDEs families in the rat ovary. Further, the effect of PDE4 inhibition on primordial follicle activation in cultured neonatal...... and PDE2A in the corpora lutea. Incubating neonatal rat ovaries with PDE4 inhibitors did not increase primordial follicle activation or change the expression of the developing follicle markers Gdf9, Amh, Inha, the proliferation marker Mki67 or the primordial follicle marker Tmeff2. In addition, the cAMP...

  14. Defining multidrug-resistant tuberculosis: correlating GenoType MTBDRplus assay results with minimum inhibitory concentrations.

    Science.gov (United States)

    Kambli, Priti; Ajbani, Kanchan; Sadani, Meeta; Nikam, Chaitali; Shetty, Anjali; Udwadia, Zarir; Georghiou, Sophia B; Rodwell, Timothy C; Catanzaro, Antonino; Rodrigues, Camilla

    2015-05-01

    This study correlates MICs of rifampicin (RIF) and isoniazid (INH) with GenoType MTBDRplus assay results for drug-resistant Mycobacterium tuberculosis (MTB) clinical isolates. MICs of RIF and INH were established for 84 and 90 isolates, respectively, testing 7 concentrations of each drug. Genotypic resistance to each drug was determined by GenoType MTBDRplus assay with 50 representative mutations confirmed by pyrosequencing, with mutations in the rpoB gene associated with RIF resistance and mutations in the katG and/or inhA genes associated with INH resistance. Based upon the correlation of MICs with specific genetic profiles, relative resistance levels were established for each isolate. Results indicate that MTB phenotypic resistance, currently based upon the testing of isolate susceptibility to a single drug concentration, may be more accurately profiled via quantitative MICs, and therefore, the correlation of molecular diagnostic results with specific MICs may allow for more optimal treatment of infections.

  15. Exploration of freely available web-interfaces for comparative homology modelling of microbial proteins.

    Science.gov (United States)

    Nema, Vijay; Pal, Sudhir Kumar

    2013-01-01

    This study was conducted to find the best suited freely available software for modelling of proteins by taking a few sample proteins. The proteins used were small to big in size with available crystal structures for the purpose of benchmarking. Key players like Phyre2, Swiss-Model, CPHmodels-3.0, Homer, (PS)2, (PS)(2)-V(2), Modweb were used for the comparison and model generation. Benchmarking process was done for four proteins, Icl, InhA, and KatG of Mycobacterium tuberculosis and RpoB of Thermus Thermophilus to get the most suited software. Parameters compared during analysis gave relatively better values for Phyre2 and Swiss-Model. This comparative study gave the information that Phyre2 and Swiss-Model make good models of small and large proteins as compared to other screened software. Other software was also good but is often not very efficient in providing full-length and properly folded structure.

  16. Pyrazole clubbed triazolo[1,5-a]pyrimidine hybrids as an anti-tubercular agents: Synthesis, in vitro screening and molecular docking study.

    Science.gov (United States)

    Bhatt, Jaimin D; Chudasama, Chaitanya J; Patel, Kanuprasad D

    2015-12-15

    A series of novel pyrazole linked triazolo-pyrimidine hybrids were synthesized and evaluated for their anti-tuberculosis activity against M.tb H37Rv strain. Some of the screened entities rendered promising anti-tb activity (MIC: 0.39μg/mL) and were found non toxic against Vero cells (IC50: ⩾20μg/mL). Further, the docking study against wild type InhA enzyme of Mycobacterium tuberculosis using Glide reproduced the most active inhibitors (J21 and J27) with lowest binding energies and highest Glide XP scores demonstrating efficient binding to the active pocket. Additionally, the enzyme inhibition assay and ADME prediction of the active proved to be an attest to the possibility of developing compound J27 as a potent anti-tubercular lead.

  17. Phosphodiesterases in the rat ovary: effect of cAMP in primordial follicles.

    Science.gov (United States)

    Petersen, Tonny Studsgaard; Stahlhut, Martin; Andersen, Claus Yding

    2015-07-01

    Phosphodiesterases (PDEs) are important regulators of the intracellular cAMP concentration, which is a central second messenger that affects a multitude of intracellular functions. In the ovaries, cAMP exerts diverse functions, including regulation of ovulation and it has been suggested that augmented cAMP levels stimulate primordial follicle growth. The present study examined the gene expression, enzyme activity and immunolocalization of the different cAMP hydrolysing PDEs families in the rat ovary. Further, the effect of PDE4 inhibition on primordial follicle activation in cultured neonatal rat ovaries was also evaluated. We found varied expression of all eight families in the ovary with Pde7b and Pde8a having the highest expression each accounting for more than 20% of the total PDE mRNA. PDE4 accounted for 15-26% of the total PDE activity. Immunoreactive PDE11A was found in the oocytes and PDE2A in the corpora lutea. Incubating neonatal rat ovaries with PDE4 inhibitors did not increase primordial follicle activation or change the expression of the developing follicle markers Gdf9, Amh, Inha, the proliferation marker Mki67 or the primordial follicle marker Tmeff2. In addition, the cAMP analogue 8-bromo-cAMP did not increase AKT1 or FOXO3A phosphorylation associated with follicle activation or increase the expression of Kitlg known to be associated with follicle differentiation but did increase the Tmeff2, Mki67 and Inha expression in a dose-dependent manner. In conclusion, this study shows that both Pde7b and Pde8a are highly expressed in the rodent ovary and that PDE4 inhibition does not cause an increase in primordial follicle activation. © 2015 Society for Reproduction and Fertility.

  18. Expression of Iron-Related Proteins Differentiate Non-Cancerous and Cancerous Breast Tumors.

    Science.gov (United States)

    Pizzamiglio, Sara; De Bortoli, Maida; Taverna, Elena; Signore, Michele; Veneroni, Silvia; Cho, William Chi-Shing; Orlandi, Rosaria; Verderio, Paolo; Bongarzone, Italia

    2017-02-14

    We have previously reported hepcidin and ferritin increases in the plasma of breast cancer patients, but not in patients with benign breast disease. We hypothesized that these differences in systemic iron homeostasis may reflect alterations in different iron-related proteins also play a key biochemical and regulatory role in breast cancer. Thus, here we explored the expression of a bundle of molecules involved in both iron homeostasis and tumorigenesis in tissue samples. Enzyme-linked immunosorbent assay (ELISA) or reverse-phase protein array (RPPA), were used to measure the expression of 20 proteins linked to iron processes in 24 non-cancerous, and 56 cancerous, breast tumors. We found that cancerous tissues had higher level of hepcidin than benign lesions (p = 0.012). The univariate analysis of RPPA data highlighted the following seven proteins differentially expressed between non-cancerous and cancerous breast tissue: signal transducer and transcriptional activator 5 (STAT5), signal transducer and activator of transcription 3 (STAT3), bone morphogenetic protein 6 (BMP6), cluster of differentiation 74 (CD74), transferrin receptor (TFRC), inhibin alpha (INHA), and STAT5_pY694. These findings were confirmed for STAT5, STAT3, BMP6, CD74 and INHA when adjusting for age. The multivariate statistical analysis indicated an iron-related 10-protein panel effective in separating non-cancerous from cancerous lesions including STAT5, STAT5_pY694, myeloid differentiation factor 88 (MYD88), CD74, iron exporter ferroportin (FPN), high mobility group box 1 (HMGB1), STAT3_pS727, TFRC, ferritin heavy chain (FTH), and ferritin light chain (FTL). Our results showed an association between some iron-related proteins and the type of tumor tissue, which may provide insight in strategies for using iron chelators to treat breast cancer.

  19. Efflux pump gene expression in multidrug-resistant Mycobacterium tuberculosis clinical isolates.

    Science.gov (United States)

    Li, Guilian; Zhang, Jingrui; Guo, Qian; Jiang, Yi; Wei, Jianhao; Zhao, Li-li; Zhao, Xiuqin; Lu, Jianxin; Wan, Kanglin

    2015-01-01

    Isoniazid (INH) and rifampicin (RIF) are the two most effective drugs in tuberculosis therapy. Understanding the molecular mechanisms of resistance to these two drugs is essential to quickly diagnose multidrug-resistant (MDR) tuberculosis and extensive drug-resistant tuberculosis. Nine clinical Mycobacterium tuberculosis isolates resistant to only INH and RIF and 10 clinical pan-sensitive isolates were included to evaluate the expression of 20 putative drug efflux pump genes and sequence mutations in rpoB (RIF), katG (INH), the inhA promoter (INH), and oxyR-ahpC (INH). Nine and three MDR isolates were induced to overexpress efflux pump genes by INH and RIF, respectively. Eight and two efflux pump genes were induced to overexpress by INH and RIF in MDR isolates, respectively. drrA, drrB, efpA, jefA (Rv2459), mmr, Rv0849, Rv1634, and Rv1250 were overexpressed under INH or RIF stress. Most efflux pump genes were overexpressed under INH stress in a MDR isolates that carried the wild-type katG, inhA, and oxyR-ahpC associated with INH resistance than in those that carried mutations. The expression levels of 11 genes (efpA, Rv0849, Rv1250, P55 (Rv1410c), Rv1634, Rv2994, stp, Rv2459, pstB, drrA, and drrB) without drug inducement were significantly higher (P < 0.05) in nine MDR isolates than in 10 pan-sensitive isolates. In conclusion, efflux pumps may play an important role in INH acquired resistance in MDR M. tuberculosis, especially in those strains having no mutations in genes associated with INH resistance; basal expression levels of some efflux pump genes are higher in MDR isolates than in pan-sensitive isolates and the basal expressional differences may be helpful to diagnose and treat resistant tuberculosis.

  20. Efflux pump gene expression in multidrug-resistant Mycobacterium tuberculosis clinical isolates.

    Directory of Open Access Journals (Sweden)

    Guilian Li

    Full Text Available Isoniazid (INH and rifampicin (RIF are the two most effective drugs in tuberculosis therapy. Understanding the molecular mechanisms of resistance to these two drugs is essential to quickly diagnose multidrug-resistant (MDR tuberculosis and extensive drug-resistant tuberculosis. Nine clinical Mycobacterium tuberculosis isolates resistant to only INH and RIF and 10 clinical pan-sensitive isolates were included to evaluate the expression of 20 putative drug efflux pump genes and sequence mutations in rpoB (RIF, katG (INH, the inhA promoter (INH, and oxyR-ahpC (INH. Nine and three MDR isolates were induced to overexpress efflux pump genes by INH and RIF, respectively. Eight and two efflux pump genes were induced to overexpress by INH and RIF in MDR isolates, respectively. drrA, drrB, efpA, jefA (Rv2459, mmr, Rv0849, Rv1634, and Rv1250 were overexpressed under INH or RIF stress. Most efflux pump genes were overexpressed under INH stress in a MDR isolates that carried the wild-type katG, inhA, and oxyR-ahpC associated with INH resistance than in those that carried mutations. The expression levels of 11 genes (efpA, Rv0849, Rv1250, P55 (Rv1410c, Rv1634, Rv2994, stp, Rv2459, pstB, drrA, and drrB without drug inducement were significantly higher (P < 0.05 in nine MDR isolates than in 10 pan-sensitive isolates. In conclusion, efflux pumps may play an important role in INH acquired resistance in MDR M. tuberculosis, especially in those strains having no mutations in genes associated with INH resistance; basal expression levels of some efflux pump genes are higher in MDR isolates than in pan-sensitive isolates and the basal expressional differences may be helpful to diagnose and treat resistant tuberculosis.

  1. Multidrug-Resistant Mycobacterium tuberculosis of the Latin American Mediterranean Lineage, Wrongly Identified as Mycobacterium pinnipedii (Spoligotype International Type 863 [SIT863]), Causing Active Tuberculosis in South Brazil

    KAUST Repository

    Dalla Costa, Elis R.

    2015-09-23

    We recently detected the spoligotype patterns of strains of Mycobacterium pinnipedii, a species of the Mycobacterium tuberculosis complex, in sputum samples from nine cases with pulmonary tuberculosis residing in Porto Alegre, South Brazil. Because this species is rarely encountered in humans, we further characterized these nine isolates by additional genotyping techniques, including 24-locus mycobacterial interspersed repetitive-unit–variable-number tandem-repeat (MIRU-VNTR) typing, verification of the loci TbD1, RD9, pks15/1, RDRio, and fbpC, the insertion of IS6110 at a site specific to the M. tuberculosis Latin American Mediterranean (LAM) lineage, and whole-genome sequencing. The combined analysis of these markers revealed that the isolates are in fact M. tuberculosis and more specifically belong to the LAM genotype. Most of these isolates (n = 8) were shown to be multidrug resistant (MDR), which prompted us to perform partial sequencing of the rpoA, rpoB, rpoC, katG, and inhA genes. Seven isolates (77.8%) carried the S315T mutation in katG, and one of these (11%) also presented the C(−17)T single-nucleotide polymorphism (SNP) in inhA. Interestingly, six of the MDR isolates also presented an undescribed insertion of 12 nucleotides (CCA GAA CAA CCC) in codon 516 of rpoB. No putative compensatory mutation was found in either rpoA or rpoC. This is the first report of an M. tuberculosis LAM family strain with a convergent M. pinnipedii spoligotype. These spoligotypes are observed in genotype databases at a modest frequency, highlighting that care must be taken when identifying isolates in the M. tuberculosis complex on the basis of single genetic markers.

  2. Overview on mechanisms of isoniazid action and resistance in Mycobacterium tuberculosis.

    Science.gov (United States)

    Unissa, Ameeruddin Nusrath; Subbian, Selvakumar; Hanna, Luke Elizabeth; Selvakumar, Nagamiah

    2016-11-01

    Isoniazid (INH) is one of the most active compounds used to treat tuberculosis (TB) worldwide. In addition, INH has been used as a prophylactic drug for individuals with latent Mycobacterium tuberculosis (MTB) infection to prevent reactivation of disease. Importantly, the definition of multidrug resistance (MDR) in TB is based on the resistance of MTB strains to INH and rifampicin (RIF). Despite its simple chemical structure, the mechanism of action of INH is very complex and involves several different concepts. Many pathways pertaining to macromolecular synthesis are affected, notably mycolic acid synthesis. The pro-drug INH is activated by catalase-peroxidase (KatG), and the active INH products are targeted by enzymes namely, enoyl acyl carrier protein (ACP) reductase (InhA) and beta-ketoacyl ACP synthase (KasA). In contrast, INH is inactivated by arylamine N-acetyltransferases (NATs). Consequently, the molecular mechanisms of INH resistance involve several genes in multiple biosynthetic networks and pathways. Mutation in the katG gene is the major cause for INH resistance, followed by inhA, ahpC, kasA, ndh, iniABC,fadE, furA, Rv1592c and Rv1772. The recent association of efflux genes with INH resistance has also gained considerable attention. Interestingly, substitutions have also been observed in nat, fabD, and accD recently in resistant isolates. Understanding the mechanisms operating behind INH action and resistance would enable better detection of INH resistance. This information would aid novel drug design strategies. Herein we review all mechanisms known to potentially contribute to the complexity of INH action and mechanisms of resistance in MTB, with insights into methods for detection of INH resistance as well as their limitations.

  3. Differentiation of rat iPS cells and ES cells into granulosa cell-like cells in vitro

    Institute of Scientific and Technical Information of China (English)

    Juan Zhang; Hui Li; Zhao Wu; XiaoJun Tan; Fengying Liu; Xianghong Huang; Xiaoling Fang

    2013-01-01

    Premature ovarian failure (POF) is an ovarian defect characterized by the premature depletion of ovarian follicles before 40 years of age,representing one major cause of female infertility.Stem cells provide the possibility of a potential treatment for POF.In this study,rat embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) were co-cultured with granulosa cells (GCs) to differentiate to GC-like cells.The level of estradiol (E2) analyzed by radioimmunoassay showed that the E2 concentration of the culture supernatant of co-cultured rat iPSCs and ESCs increased in a time-dependent manner,compared with the GCs group that has an opposite trend.The expression of follicle-stimulating hormone receptor (FSHR) was confirmed by immunostaining.These results indicated that rat iPSCs and ESCs were effectively induced to GC-like cells through indirect cell-to-cell contact.Real-time polymerase chain reaction was performed to analyze the expression level of marker genes in POF,including BMP15,FMR1,FSHR,INHA,AMH,NOBOX,FOXO3,EIF2B,FIGLA,and GDF9.The BMP15,FSHR,INHA,AMH,NOBOX,and GDF9 genes were significantly up-regulated in iPSCs and ESCs cocultured with GCs in comparison with ceils that were not co-cultured.Thus,here we demonstrated an available method to differentiate rat iPSCs and ESCs into GC-like cells in vitro for the possible cell therapy of POF.

  4. Evaluation of GenoType® MTBDRplus assay for rapid detection of drug susceptibility testing of multi-drug resistance tuberculosis in Northern India

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    Anand Kumar Maurya

    2013-01-01

    Full Text Available Background: The problem of multi-drug resistance tuberculosis (MDR-TB is growing in several hotspots throughout the world. Rapid and accurate diagnosis of MDR-TB is crucial to facilitate early treatment and to reduce its spread in the community. The aim of the present study was to evaluate the new, novel GenoType® MTBDRplus assay for rapid detection of drug susceptibility testing (DST of MDR-TB cases in Northern India. Materials and Methods: A total of 550 specimens were collected from highly suspected drug resistant from pulmonary and extra-pulmonary TB cases. All the specimens were processed by Ziehl- Neelsen staining, culture, differentiation by the GenoType® CM assay, first line DST using BacT/ALERT 3D system and GenoType® MTBDRplus assay. The concordance of the GenoType® MTBDRplus assay was calculated in comparison with conventional DST results. Results: Overall the sensitivity for detection of rifampicin, isoniazid and MDR-TB resistance by GenoType® MTBDRplus assay was 98.0%, 98.4% and 98.2% respectively. Out of 55 MDR-TB strains, 45 (81.8%, 52 (94.5% and 17 (30.9% strains showed mutation in rpoB, katG and inhA genes respectively (P < 0.05. The most prominent mutations in rpoB, katG and inhA genes were; 37 (67.3% in S531L, 52 (94.5% in S315T1 and 11 (20% in C15T regions respectively (P < 0.05. Conclusions: Our study demonstrated a high concordance between the GenoType® MTBDRplus assay resistance patterns and those were observed by conventional DST with good sensitivity, specificity with short turnaround times and to control new cases of MDR-TB in countries with a high prevalence of MDR-TB.

  5. [Conventional and molecular diagnosis in a group of patients with drug-resistant tuberculosis].

    Science.gov (United States)

    Mindru, Roxana; Spînu, Victor; Popa, Cristina; Botezatu, Elena; Spătaru, Ruxandra

    2014-01-01

    Worldwide, although the incidence of the sensitive/susceptible tuberculosis diminished, the number of drug resistant tuberculosis is growing. The bacteriological diagnosis, genetic and phenotypic, becomes essential for the epidemic control. The resistance appears as a phenotypic expression of mutations from M. tuberculosis genome. The mutations that appear for Rifampicin are in region rpoB, for Isoniazid in region katG and inhA, for Ethambutol-embB, Quinolone-gyrA, Aminoglicozid and Cyclical Peptides-rrs. To follow the concordance of results of drug sensitivity test (DST) through phenotypic and genetic method, we analyzed a group of 40 patients with TB-DR. We performed drug susceptibility testing on Lowenstein-Jensen medium according to the instructions of the manufacturer. The strains were tested indirect genetic too, Genotype MTBDR plus forl INH and RIF and Genotype MTBDRsl for the second line drugs. The concordance between genetic method and the phenotypic method is 95%, 5% from the patients have different sensitivity to INH and RIF, butphenotypical they are resistant, meaning that they have other mutations undetected by the strip. The most common mutation in region rpoB is MUT3 (52%) associating the absence of band W8. Mutations in the region rpoB MUT1 and MUT2A are 12.5%, and 15% respectively. For high resistance to INH, the mostcommon isMUT1 forkatG 95% and forlow resistance to INHMUTI from region inhA-30%. For the second line drugs, the most frequent concordance between genetic method andphenotypic method is for EMB, of 30%, geneticallyspeaking the strains display no mutation in region embB, but are resistantin phenotypic method. For FQ, KAN, AMKand CAP the concordance between the two methods is of 100% to all tested strains. In conclusion, genetic methods have high sensitivity, they are fast and shorten significantly the diagnosis time.

  6. Performance assessment of the GenoType MTBDRplus test and DNA sequencing in detection of multidrug-resistant Mycobacterium tuberculosis.

    Science.gov (United States)

    Huang, Wei-Lun; Chen, Huang-Yau; Kuo, Yuh-Min; Jou, Ruwen

    2009-08-01

    To facilitate the management of multidrug-resistant (MDR) tuberculosis, two nucleic acid sequence-based methods, the GenoType MTBDRplus test and DNA sequencing, were assessed for the rapid detection of drug-resistant Mycobacterium tuberculosis for the first time in the Asia-Pacific region. The performances of these two assays in detecting the presence of rifampin (rifampicin) (RIF) and isoniazid (INH) resistance-associated mutations in the rpoB, katG, inhA regulatory region, inhA, and oxyR-ahpC genes were compared to that of a conventional agar proportion drug susceptibility test. A total of 242 MDR and 30 pansusceptible M. tuberculosis isolates were evaluated in this study. The sensitivities obtained for RIF-resistant detection by the GenoType MTBDRplus test and by resistance gene sequencing were 95.5% and 97.9%, respectively. The sensitivities for INH resistance detection by the GenoType MTBDRplus test and by resistance gene sequencing were 81.8% and 93.4%, respectively. Together, the sensitivity for MDR tuberculosis detection was 78.5% with the GenoType MTBDRplus test and 91.3% by resistance gene sequencing. The specificity for RIF resistance, INH resistance, and MDR detection was 100% by both methods. The GenoType MTBDRplus test has the advantage of a short turnaround time for drug-resistant M. tuberculosis detection. Overall, the two assays performed equally well in detecting RIF resistance (P = 0.13). However, DNA sequencing demonstrated superior performance in detecting INH resistance (P tuberculosis (P GenoType MTBDRplus test, especially for different geographic areas with genetically diverse M. tuberculosis strains.

  7. Effect of mutation and genetic background on drug resistance in Mycobacterium tuberculosis.

    Science.gov (United States)

    Fenner, Lukas; Egger, Matthias; Bodmer, Thomas; Altpeter, Ekkehardt; Zwahlen, Marcel; Jaton, Katia; Pfyffer, Gaby E; Borrell, Sonia; Dubuis, Olivier; Bruderer, Thomas; Siegrist, Hans H; Furrer, Hansjakob; Calmy, Alexandra; Fehr, Jan; Stalder, Jesica Mazza; Ninet, Béatrice; Böttger, Erik C; Gagneux, Sebastien

    2012-06-01

    Bacterial factors may contribute to the global emergence and spread of drug-resistant tuberculosis (TB). Only a few studies have reported on the interactions between different bacterial factors. We studied drug-resistant Mycobacterium tuberculosis isolates from a nationwide study conducted from 2000 to 2008 in Switzerland. We determined quantitative drug resistance levels of first-line drugs by using Bactec MGIT-960 and drug resistance genotypes by sequencing the hot-spot regions of the relevant genes. We determined recent transmission by molecular methods and collected clinical data. Overall, we analyzed 158 isolates that were resistant to isoniazid, rifampin, or ethambutol, 48 (30.4%) of which were multidrug resistant. Among 154 isoniazid-resistant strains, katG mutations were associated with high-level and inhA promoter mutations with low-level drug resistance. Only katG(S315T) (65.6% of all isoniazid-resistant strains) and inhA promoter -15C/T (22.7%) were found in molecular clusters. M. tuberculosis lineage 2 (includes Beijing genotype) was associated with any drug resistance (adjusted odds ratio [OR], 3.0; 95% confidence interval [CI], 1.7 to 5.6; P mutations (OR, 6.4; 95% CI, 2.0 to 20.7; P = 0.002). We found that the genetic strain background influences the level of isoniazid resistance conveyed by particular mutations (interaction tests of drug resistance mutations across all lineages; P tuberculosis drug resistance mutations were associated with various levels of drug resistance and transmission, and M. tuberculosis lineages were associated with particular drug resistance-conferring mutations and phenotypic drug resistance. Our study also supports a role for epistatic interactions between different drug resistance mutations and strain genetic backgrounds in M. tuberculosis drug resistance.

  8. Genotypic assessment of drug-resistant tuberculosis in Baghdad and other Iraqi provinces using low-cost and low-density DNA microarrays.

    Science.gov (United States)

    Al-Rubaye, Dalal Saleh; Henihan, Grace; Al-Abasly, Ameraa K Abas; Seagar, Amie-Louise; Al-Attraqchi, Azhar Ab F; Schulze, Holger; Hashim, Dhafer Salman; Kamil, Jinan Khalid; Laurenson, Ian F; Bachmann, Till T

    2016-02-01

    We report on a molecular investigation carried out to ascertain the prevalence of drug-resistant tuberculosis (TB) and the specific gene mutations responsible for resistance to rifampicin (RIF) and/or isoniazid (INH) in Iraq. In total, 110 clinical isolates from category II TB cases from Baghdad (58%) and several Iraqi provinces (42%) were analysed using colorimetric, low-cost and low-density (LCD) microarrays (MYCO-Direct and MYCO-Resist LCD array kits) to identify the point mutations responsible for resistance in Mycobacterium tuberculosis isolates. We found 76 patients (69.1%) had resistant strains, of which 40 (36%) were multidrug-resistant (MDR)-TB. Where mono-resistance was identified, it was found to be predominantly to RIF (83%). The most common mutations were rpoB S531L (50%), inhA C15T (25%) and katG S315T (15%). The most common MDR-TB genotypes were rpoB S531L with inhA C15T (60%) and rpoB S531L with katG S315T (20%). Where phenotypic analysis of clinical isolates was also performed, genotypic data were found to show excellent correlation with phenotypic results. Correlation was found between the MYCO-Resist LCD array and GenoType MTBDRplus for detection of resistance to RIF. Our study shows MDR-TB in 36% of category II TB cases in Baghdad and surrounding Iraqi provinces, which reflects the World Health Organization findings based on phenotypic studies. Diagnosis of TB and MDR-TB using culture-based tests is a significant impediment to global TB control. The LCD arrays investigated herein are easy to use, sensitive and specific molecular tools for TB resistance profiling in resource-limited laboratory settings.

  9. Atividade alelopática dos lixiviados de Asemeia extraaxillaris (Polygalaceae sobre o crescimento de Ipomoea cordifolia

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    C.B. SILVA

    2016-01-01

    Full Text Available RESUMO O objetivo desse trabalho foi avaliar a atividade alelopática dos lixiviados das raízes e folhas secas de Asemeia extraaxillaris sobre o crescimento de corda de viola Ipomoea cordifolia em casa de vegetação, para avaliação dos efeitos sobre o estresse oxidativo. Nos bioensaios foram utilizadas três concentrações (0,25 - 5,00 e 10,00 g e um controle, com quatro repetições de 10 plantas, cultivadas em solo durante 30 dias. Os lixiviados possuem ação alelopática e o maior efeito foi verificado para o pó das raizes. O efeito inibitório foi dose-dependente e o aumento do estresse oxidativo foi observado pelos níveis da catalase, peroxidase, superóxido dismutase, glutationa redutase, peroxidação lipídica e síntese de formazan. Concluiu-se que A. extraaxillaris possui substâncias capazes de afetar a emergência, o crescimento e o estresse oxidativo de I. cordifolia.

  10. Fisiologia e produtividade do algodoeiro em solo encharcado na fase de plântula

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    Souza José Gomes de

    2001-01-01

    Full Text Available O algodoeiro herbáceo (Gossypium hirsutum L. r. latifolium Hutch. é uma planta considerada sensível à deficiência de oxigênio do solo. O objetivo deste trabalho foi avaliar a fisiologia e produtividade do algodoeiro cultivar CNPA 7H, em casa de vegetação, submetido à anoxia, por encharcamento do solo, na fase de plântula. Foram conduzidos dois experimentos em blocos ao acaso, com sete períodos de encharcamento e seis repetições. No encharcamento de quatro dias ocorreu uma redução na atividade da invertase de 76,69%, da beta-amilase de 77,37% e da redutase do nitrato de 51,10%. A fotossíntese foi afetada a partir do primeiro dia e alcançou decréscimo de 58,63% no décimo quarto dia de encharcamento; os carboidratos foram acumulando-se nas folhas, caule e raízes. O rendimento de algodão em caroço foi reduzido em 35,76% no décimo dia do estresse anoxítico.

  11. Increment of antioxidase activity of transgenic tobacco with betaine aldehyde dehydrogenase

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    Superoxide dismutase (SOD) activity in the leaves of transgenic tobacco plants with betaine aldehyde dehydrogenase (BADH) gene was about 36% higher than that in the control plants (parent plants),activities of peroxi-dase (POD) and catalase (Cat) increased by about 62% and 88% respectively. Activities of ascorbate peroxidase (AsSPOD),dehydroascorbate redutase (DAsAR) and gluta-thione reductase (GR) in ascorbate-glutothion pathway lo-cated at chloroplasts increased by 67.7%,47.9% and 38.8% respectively. These results indicated that the H2O2 produced by SOD catalyzing superoxide anion radicals (O2- ) could be fully decomposed,and could not derive to form the strongest toxicant radicals ·OH. This is the first report to elucidate quantitatively that the activities of two kinds of antioxidative enzymes decomposed radicals and active oxygen were matched. Photoinhibition tolerant capacity of the transgenic tobacco plants was 35% higher than that in the parent plants. Increment of photoinhibition tolerant capacity in the trans-genic tobacco plants might be due to increment of antioxida-tive enzymes activities,in turn being able to more effectively scavenge active oxygen and radicals,protect organization and function of chloroplasts. These results showed that the increment of antioxidative enzymes activities in the trans-genic tobacco might be one of the reasons for the increment of resistance in the transgenic tobacco.

  12. Doença de Lafora: diagnóstico pela biopsia de músculo esquelético (relato de caso Lafora's disease: diagnosis by muscle biopsy (case report

    Directory of Open Access Journals (Sweden)

    ALZIRA ALVES DE SIQUEIRA CARVALHO

    2000-12-01

    Full Text Available Uma paciente de 16 anos apresentava epilepsia mioclônica causada pela doença de Lafora. A biopsia muscular mostrou padrão vacuolar nas fibras musculares com as reações nicotinamida adenina tetrazolium redutase desidrogenase, hematoxilina-eosina e PAS. O aspecto morfológico permite o diagnóstico através da biopsia muscular. Este é um procedimento menos agressivo que a biopsia de fígado e de cérebro. A microscopia eletrônica deve ser reservada para casos nos quais as alterações musculares à microscopia óptica são muito discretas deixando dúvidas quanto ao diagnóstico.A 16-year-old female patient had myoclonic epilepsy caused by Lafora's disease. Muscle biopsy showed a prominent splitting pattern in muscle fibers with the nicotinamide adenine nucleotide dehydrogenase-tetrazolium reductase reaction, hematoxylin-eosin, and PAS stains. This morphologic appearance of the tissue permits diagnosis using the benign technique of muscle biopsy. The ultrastructural examination of muscle may be necessary to confirm the diagnosis of Lafora myoclonus epilepsy if light microscopical findings are equivocal.

  13. Effect of pravastatin on survival in patients with advanced hepatocellular carcinoma. A randomized controlled trial.

    Science.gov (United States)

    Kawata, S; Yamasaki, E; Nagase, T; Inui, Y; Ito, N; Matsuda, Y; Inada, M; Tamura, S; Noda, S; Imai, Y; Matsuzawa, Y

    2001-04-06

    Chemotherapy is not effective for hepatocellular carcinoma (HCC). HMG-CoA redutase inhibitors have cytostatic activity for cancer cells, but their clinical usefulness is unknown. To investigate whether pravastatin, a potent HMG-CoA reductase inhibitor, prolongs survival in patients with advanced HCC, this randomized controlled trial was conducted between February 1990 and February 1998 at Osaka University Hospital. 91 consecutive patients <71 years old (mean age 62) with unresectable HCC were enroled in this study. 8 patients were withdrawn because of progressive liver dysfunction; 83 patients were randomized to standard treatment with or without pravastatin. All patients underwent transcatheter arterial embolization (TAE) followed by oral 5-FU 200 mg(-1)d for 2 months. Patients were then randomly assigned to control (n = 42) and pravastatin (n = 41) groups. Pravastatin was administered at a daily dose of 40 mg. The effect of pravastatin on tumour growth was assessed by ultrasonography. Primary endpoint was death due to progression of HCC. The duration of pravastatin administration was 16.5 +/- 9.8 months (mean +/- SD). No patients in either group were lost to follow-up. Median survival was 18 months in the pravastatin group versus 9 months in controls (P = 0.006). The Cox proportional hazards model showed that pravastatin was a significant factor contributing to survival. Pravastatin prolonged the survival of patients with advanced HCC, suggesting its value for adjuvant treatment.

  14. [Establishment of a model for evaluating hypolipidemic effect in HepG2 cells].

    Science.gov (United States)

    Niu, Yucun; Lü, Na; Li, Ying; Zhao, Dan; Sun, Changhao

    2010-03-01

    To establish a model of evaluating hypolipidemic effect in vitro. Adding cholesterol to the culture medium for HepG2 cells to induce a hypercholesterolemia model. The content of cellular cholesterol and the expression of protein regulating cholesterol metabolism in HepG2 cells were determined. The validation of the model was identified by lovastatin, a widely used cholesterol-lowering drug. Free fatty acid was added to the culture medium for HepG2 cells to induce a hypertriglyceridemia model. The content of cellular triglyceride and the absorption rate of free fatty acid were determined. The validation of the model was identified by fenofibrate, a triglyceride-lowering drug. Cellular cholesterol content was increased and the expression of HMG-CoA redutase, SREBP-2 and LDLR were decreased after adding cholesterol and 25-hydrocholesterol to the culture medium. Cellular cholesterol was decreased and the expression of SREBP-2 and LDLR were up-regulated by Lovastatin. The absorption of oleic acid in cells was up to 40% after adding oleic acid (50 micromol) to the culture medium for 6 h. The absorption of free fatty acid was increased but the content of cellular triglyceride was not increased in cells by Fenofibrate. This model might be an effective method for screening and assessing functional factors for lowing plasma lipids.

  15. Fatores de risco e medidas de prevenção do câncer de próstata: subsídios para a enfermagem

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    Adriane Pinto de Medeiros

    2011-04-01

    Full Text Available Apresenta-se um artigo de reflexão sobre fatores de risco e medidas de prevenção do câncer de próstata, visando contribuir para a abordagem de homens, especialmente durante a consulta de enfermagem. Idade avançada, origem étnica e hereditariedade são fatores de risco potenciais. Dieta pobre em gorduras, rica em frutas, verduras, legumes, grãos e outros é citada como minimizadora. A finasterida, droga inibidora da 5 alpha-redutase tem sido estudada como redutor de risco. O rastreamento da doença pela dosagem do Antígeno Prostático Específico (PSA e toque retal é também medida recomendada. Entretanto, maiores evidências científicas ainda estão sendo buscadas sobre esses fatores. A abordagem dos homens na consulta de enfermagem pode contribuir para a identificação de fatores de risco, sinais e sintomas de possíveis alterações.

  16. Surto psicótico pela possível interação medicamentosa de sibutramina com finasterida

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    Sucar Douglas Dogol

    2002-01-01

    Full Text Available É descrito um caso de interação medicamentosa em um paciente de 30 anos, que fazia com sucesso um tratamento antiobesidade com sibutramina. Esse medicamento é um saciogênico que inibe a recaptação da serotonina e da noradrenalina. Logo após a introdução da finasterida para tratamento da alopécia androgênica, o paciente passou a apresentar surto psicótico. Esse medicamento é um potente inibidor da 5-alfa-redutase do tipo II. A interação medicamentosa demonstra-se por meio de um rigoroso acompanhamento clínico e de uma aplicação do algoritmo de Naranjo. Também descrevem-se as condições clínicas gerais do paciente e sua evolução. Discutem-se os mecanismos prováveis que levaram à interação medicamentosa, com a conclusão de que a finasterida inibiu o metabolismo hepático da sibutramina. Esta deslocou a finasterida de seus pontos de ligações com as proteínas, aumentando sua fração livre plasmática e inibindo ainda mais o metabolismo da sibutramina. Com aumento de serotonina, noradrenalina e dopamina na fenda sináptica, desencadeia-se o surto psicótico.

  17. Impact of the processes of testicular regression and recrudescence in the prostatic complex of the bat Myotis nigricans (Chiroptera: Vespertilionidae).

    Science.gov (United States)

    Beguelini, Mateus R; Góes, Rejane M; Rahal, Paula; Morielle-Versute, Eliana; Taboga, Sebastião R

    2015-07-01

    Myotis nigricans is a species of vespertilionid bat, whose males show two periods of total testicular regression during the annual reproductive cycle in the northwest São Paulo State, Brazil. Thus, the aim of this study was to investigate the impact of total testicular regression on the prostatic morphophisyology and its regulation. The prostatic complex (PC) of animals from the four periods of the reproductive cycle (active, regressing, regressed, and recrudescence) was analyzed by different histological, morphometric, and immunohistochemical procedures to characterize its variations, analyze its hormonal regulation and evaluate whether the prostate is affected by the processes of testicular regression and recrudescence. The results indicated a decrease in the prostatic parameters from the active to regressed periods, which are related to decreases in the testicular production of testosterone and in the prostatic expression of androgen receptor (AR), estrogen receptor α (ERα) and aromatase. However, in regressed-recrudescence periods, the prostatic expression of AR, ERα and aromatase increased, indicating the reactivation of the PC. Despite this, the PC appears to have a slower reactivation and seems not to follow the testicular recrudescence in morphological and morphometric terms. With these data, we demonstrate that the prostatic physiology is directly affected by total testicular regression and conclude that it is regulated by testosterone and estrogen, via the production of testosterone by the testes, its conversion to dihydrotestosterone by 5α-redutase and to estrogen by aromatase, and the activation/deactivation of AR and ERα in epithelial cells, which regulate cell expression and proliferation.

  18. Direct sequencing for rapid detection of multidrug resistant Mycobacterium tuberculosis strains in Morocco

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    Zakham F

    2013-11-01

    Full Text Available Fathiah Zakham,1,4 Imane Chaoui,1 Amina Hadbae Echchaoui,2 Fouad Chetioui,3 My Driss Elmessaoudi,3 My Mustapha Ennaji,4 Mohammed Abid,2 Mohammed El Mzibri11Unité de Biologie et Recherché Médicale, Centre National de l'Energie, des Sciences et des Techniques Nucléaires (CNESTEN, Rabat, 2Laboratoire de Génétique Mycobacterienne, Institut Pasteur, Tangier, 3Laboratoire de Tuberculose Institut Pasteur, Casablanca, 4Laboratoire de Microbiologie, Hygiène et Virologie, Faculté des Sciences et Techniques, Mohammedia, MoroccoBackground: Tuberculosis (TB is a major public health problem with high mortality and morbidity rates, especially in low-income countries. Disturbingly, the emergence of multidrug resistant (MDR and extensively drug resistant (XDR TB cases has worsened the situation, raising concerns of a future epidemic of virtually untreatable TB. Indeed, the rapid diagnosis of MDR TB is a critical issue for TB management. This study is an attempt to establish a rapid diagnosis of MDR TB by sequencing the target fragments of the rpoB gene which linked to resistance against rifampicin and the katG gene and inhA promoter region, which are associated with resistance to isoniazid.Methods: For this purpose, 133 sputum samples of TB patients from Morocco were enrolled in this study. One hundred samples were collected from new cases, and the remaining 33 were from previously treated patients (drug relapse or failure, chronic cases and did not respond to anti-TB drugs after a sufficient duration of treatment. All samples were subjected to rpoB, katG and pinhA mutation analysis by polymerase chain reaction and DNA sequencing.Results: Molecular analysis showed that seven strains were isoniazid-monoresistant and 17 were rifampicin-monoresistant. MDR TB strains were identified in nine cases (6.8%. Among them, eight were traditionally diagnosed as critical cases, comprising four chronic and four drug-relapse cases. The last strain was isolated from a

  19. Encurtamento pelo frio de fibras musculares oxidativas de bovinos pela técnica de NADH-TR Cold shortening on bovine oxidative muscle fibers by NADH-TR

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    Juliana Maria Pereira Felício Gonfiantini Fernandes

    2006-12-01

    Full Text Available Carcaças bovinas resfriadas rapidamente podem apresentar uma contração muscular conhecida como encurtamento pelo frio. Esse fenômeno, prejudicial à textura da carne, ocorre principalmente nas fibras musculares oxidativas. O objetivo deste trabalho foi disponibilizar uma ferramenta analítica para distinguir essas fibras e determinar com maior precisão a contração do tecido muscular pela mensuração do comprimento dos sarcômeros. Amostras do músculo Longissimus dorsi de 12 novilhas nelore foram coletadas. As amostras obtidas de uma das meias-carcaças foram submetidas a uma refrigeração rápida, e as amostras provenientes da outra meia-carcaça à refrigeração lenta. Foi testado um método analítico, baseado na técnica de coloração por Nicotinamida Adenina Dinucleotídeo - Tetrazolium Redutase (NADH-TR, para mensurar, por microscopia, o comprimento dos sarcômeros das fibras vermelhas. Foram determinadas as velocidades de queda de pH e temperatura, a área do olho de lombo (AOL e a força de cisalhamento. Os resultados demonstraram que a temperatura é o principal fator responsável pelo comprimento do sarcômero quando a velocidade de resfriamento é rápida, sendo essa influência menor quando a queda de temperatura é mais lenta. Desta forma, demonstrou-se que a técnica de coloração com NADH-TR é capaz de detectar a ocorrência do encurtamento pelo frio nos músculos esqueléticos de bovinos.Bovine carcasses quickly cooled may develop a muscle contraction known as cold shortening. This process, harmful to meat texture, occurs mainly in the oxidative muscle fibers. This work was aimed at studing an analytical tool to distinguish these fibers and to evaluate, more precisely, the muscle contraction through the measurement of sarcomeres length. Longissimus dorsi muscles of 12 heifers were used as samples. Samples from one of the half carcasses were fast cooled and the samples from the others half carcasses were slowly cooled

  20. Deficiências de macronutrientes no estado nutricional da mamoneira cultivar Iris Deficiencies of macronutrients on nutritional status of castor bean cultivar Iris

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    José Lavres Junior

    2005-02-01

    Full Text Available Os objetivos deste trabalho foram avaliar o efeito da deficiência dos macronutrientes no crescimento e estado nutricional da mamoneira (Ricinus communis L. e a obtenção do quadro sintomatológico das deficiências de N, P, K, Ca, Mg e S. Cultivou-se Ricinus communis L. cultivar Iris em solução nutritiva, tendo como tratamentos soluções completa e deficientes em N, P, K, Ca, Mg e S. Foram determinadas a concentração indireta de clorofila, a atividade da redutase do nitrato, os teores solúveis de N (NO3-, P (H2PO4- e K+ e realizados testes rápidos para N, P e K. As deficiências de N, Ca, S e Mg foram as que mais restringiram a produção de massa de matéria seca, na ordem decrescente: N>Ca>S>Mg>K>P. Os maiores acúmulos de macronutrientes, no tratamento completo, foram observados nas raízes e no limbo das folhas inferiores. As concentrações críticas para N e Ca, ambas no limbo das folhas superiores e, Mg, no pecíolo das inferiores foram, respectivamente, 46,7, 13,8 e 6,5 g kg-1. As deficiências influenciaram as leituras do clorofilômetro. A atividade da redutase do nitrato guardou relação direta com a determinação da clorofila. Os teores de N (NO3-, P (H2PO4- e K+ foram menores nas folhas deficientes em N, P e K, respectivamente, o mesmo acontecendo quando avaliadas pela análise de toque.The objectives of this work were to evaluate the effect of macronutrient deficiencies on growth and on nutritional status of castor beans (Ricinus communis L., and to obtain the symptoms of the lack of N, P, K, Ca, Mg and S. Nutrient solutions containing all macronutrients and those from which one of these elements was omitted were the treatments used. The following determinations were made: chlorophyll (indirectly, nitrate reductase activity, NO3--N, H2PO4--P and K+, both on soluble fractions, and by spot tests. Deficiencies of N, Ca, S and Mg were the most limiting for dry matter production, followed in decreasing order by those of K and

  1. Nitrate levels and stages of growth in hypernodulating mutants of Lupinus albus. II. Enzymatic activity and transport of N in the xylem sap Diferentes níveis de nitrato e estágio de crescimento em mutantes hipernodulantes de Lupinus albus II. Atividade enzimática e transporte de N na seiva do xilema

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    Hélio Almeida Burity

    1999-04-01

    decisiva em evitar produtos inibidores da nitrogenase pelo amônio, porém poucas são as informações deste mecanismo com o qual o xilema carrega estes produtos, em compensação, este processo, pode ser a chave da função em evitar acumulação de aminoácidos nas células dos nódulos infectados. O comportamento das enzimas nitrato redutase (NR, fosfoenolpiruvato carboxilase (PEPC, glutamina sintetase (GS e dos compostos nitrogenados derivados da fixação como: N-a-amino, N-ureídos e N-amida em genótipos mutados foi observado. A atividade da enzima nitrato redutase foi altamente inflenciada pela aplicação de nitrato apresentando valores bastante superiores em relação a não aplicação do nitrato, independentemente dos genótipos, sendo que o melhor período de avaliação foi na décima semana. O genótipo L-62, com caracteríticas de resistência ao nitrato, mostrou de forma bem evidenciada que a enzima PEPC é inibida pela presença de nitrato. O genótipo L-135 (nod- fix- apresentou atividade da GS extremamente baixo, mostrando desta forma que a GS é uma enzima altamente correlacionada com a fixação. Em relação ao melhor estágio de crescimento para a GS, o Lupinus albus, deve ser avaliado na sétima semana.

  2. Farmacocinética das estatinas Pharmacokinetics of statins

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    Francisco Antonio Helfenstein Fonseca

    2005-10-01

    Full Text Available As estatinas são agentes hipolipemiantes que exercem os seus efeitos através da inibição da HMG-CoA redutase, enzima fundamental na síntese do colesterol, levando a uma redução do colesterol tecidual e um conseqüente aumento na expressão dos receptores de LDL. Existem consideráveis diferenças entre as estatinas, no que tange às propriedades farmacocinéticas, bem como ao coeficiente de hidrofilicidade, via hepática de metabolização (especialmente, do citocromo P450 e isoenzimas, meia-vida plasmática e eficácia na redução lipídica. As estatinas também podem diferir na capacidade de interação com outras drogas que utilizam a mesma via de metabolização. Recentemente, muitos efeitos pleiotrópicos têm sido relatados com estas drogas, bem como propriedades antiinflamatórias, melhora na função endotelial e benefícios na hemostasia.Statins are lipid lowering agents that promote their effects on plasma lipids through the inhibition of HMG-CoA reductase, a crucial enzyme in the cascade of cholesterol synthesis, leading to reduction of tissue cholesterol pool and consequently, to an upregulation of the LDL receptor expression. There are considerable differences among statins regarding some pharmacokinetic properties, such as the coefficient of hydrophilicity, via liver metabolism (especially regarding P450 cytochrome and isoenzymes, plasma half-life and efficacy of serum lipid changes. They may also differ regarding interactions with other drugs that share the same pathway of metabolism. Recently, many pleiotropic effects have been reported with these drugs, such as anti-inflammatory properties, improvement in endothelial function and benefits on hemostasis.

  3. Carvedilol atenua o estresse oxidativo na cardiopatia chagásica crônica

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    Patrícia Budni

    2012-03-01

    Full Text Available FUNDAMENTO: Há cada vez mais evidências sugerindo que doença de Chagas envolve dano oxidativo e contribui para a progressão da doença cardíaca. OBJETIVO: Avaliar o efeito do carvedilol sobre marcadores de estresse oxidativo na doença de Chagas crônica. MÉTODOS: A população de estudo incluiu 42 pacientes com cardiopatia chagásica e os biomarcadores de estresse oxidativo foram medidos antes e após um período de seis meses de tratamento com carvedilol (37,5 mg/dia. Os pacientes foram considerados de acordo com a classificação de Los Andes, e a atividade da superóxido dismutase, catalase, glutationa peroxidase, S-transferase e redutase, mieloperoxidase e adenosina deaminase; e os níveis de glutationa reduzida, de espécies reativas do ácido tiobarbitúrico, proteína carbonil, vitamina E e óxido nítrico foram medidos no sangue. RESULTADOS: Após o tratamento com carvedilol, todos os grupos apresentaram reduções significativas nos níveis de proteína carbonil e glutationa reduzida, enquanto os níveis de óxido nítrico e atividade da adenosina aumentaram significativamente somente no grupo IA. Além disso, a maioria das enzimas antioxidantes apresentou diminuição de suas atividades, nos grupos IA e IB. CONCLUSÃO: Os dados sugerem que o tratamento com carvedilol foi eficaz na atenuação do dano oxidativo, um efeito que pode ser particularmente importante em doença de Chagas crônica com cardiopatia.

  4. Hypolipidemic activity of Moringa oleifera Lam., Moringaceae, on high fat diet induced hyperlipidemia in albino rats Atividade hipolipidemica de Moringa oleifera Lam., Moringaceae, na hiperlipidemia induzida por dieta rica em gordura em ratos albinos

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    Pankaj G. Jain

    2010-12-01

    Full Text Available The leaves of Moringa oleifera Lam., Moringaceae, are used by the Indians in their herbal medicine as a hypolipidemic agent in obese patients. Albino Wistar rats were fed with methanolic extract of M. oleifera (150, 300 and 600 mg/kg, p.o. and simvastatin (4 mg/kg, p.o. along with hyperlipidemic diet for 30 days. Moringa oleifera and simvastatin were found to lower the serum cholesterol, triacylglyceride, VLDL, LDL, and atherogenic index, but were found to increase the HDL as compared to the corresponding high fed cholesterol diet group (control. The Moringa oleifera methanolic extract was also investigated for its mechanism of action by estimating HMG CO-A reductase activity. Moringa oleifera was found to increase the excretion of fecal cholesterol. Thus, the study demonstrates that M. oleifera possesses a hypolipidemic effect.As folhas de Moringa oleifera Lam., Moringaceae, são usados na medicina natural da Índia como um agente hipolipemiante em pacientes obesos. Ratos albinos Wistar foram alimentados com extrato metanólico de M. oleifera (150, 300 e 600 mg/kg, p.o. e sinvastatina (4 mg/kg, p.o., juntamente com dieta hiperlipídica por 30 dias. Moringa oleifera e sinvastatina reduziram o colesterol, triacilglicerídeoss, VLDL, LDL e índice aterogênico, mas não aumentaram o HDL em comparação com o grupo controle, com dieta rica em colesterol. O mecanismo de ação do extrato metanólico de Moringa oleifera foi também investigado estimando atividade de HMG CO-A redutase. Moringa oleifera aumentou a excreção fecal de colesterol. Assim, o estudo demonstra que a M. oleifera parece ter efeito hipolipemiante.

  5. Doses de nitrogênio e potássio no crescimento do trigo

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    Eloise Mello Viana

    2010-12-01

    Full Text Available O fornecimento do potássio para as plantas estimula o aproveitamento do nitrogênio, possibilitando que sua absorção, assimilação, nutrição e, consequentemente, a produtividade, sejam aumentadas. Objetivou-se com este trabalho avaliar o efeito de combinações de doses de nitrogênio e potássio no crescimento e assimilação do nitrogênio por plantas de trigo. O experimento foi realizado em casa de vegetação, utilizando-se vasos com capacidade para 3 kg onde foram acondicionadas amostras da camada 0-0,2 m de um Argissolo Vermelho Amarelo Abrúptico. Utilizou-se um esquema fatorial 5² fracionado, com combinações de cinco doses de nitrogênio (0; 70; 140; 210 e 280 mg dm-3 e de cinco doses de potássio (0; 50; 100; 150 e 200 mg dm-3, as quais foram distribuídas segundo o delineamento experimental de blocos aleatorizados, com quatro repetições. O fornecimento combinado das maiores doses de nitrogênio e potássio incrementou a produção de massa seca da parte aérea e teor de clorofila nas folhas. O acúmulo de nitrogênio e potássio nas folhas foi aumentado apenas pelas doses desses nutrientes separadamente. O potássio não influenciou a atividade da redutase do nitrato e o acúmulo de nitrogênio no tecido vegetal.

  6. Metabolismo de nitrogênio em dois sistemas de cultivo de café sob veranico da estação úmida

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    Carmem Lacerda Lemos

    2010-02-01

    Full Text Available Com o objetivo de avaliar o efeito do veranico ocorrido na estação úmida sobre o metabolismo de nitrogênio em cafeeiros em sistema a pleno sol e associados com abacateiro (Persea americana Mill. e ingazeiro (Inga edulis Mart., foi realizado este experimento. O estudo foi conduzido em propriedade situada no município de Barra do Choça, BA, composto por plantas de café (Coffea arabica L., variedade Catuaí, sendo caracterizados dois campos experimentais (sistema sombreado x sistema a pleno sol. As avaliações foram realizadas em janeiro (período de veranico e março (final da estação úmida, em cinco repetições por campo experimental. Os dados foram submetidos ao teste t por meio do programa SAEG, versão 9.1. Maior teor de NO3 - foi verificado no sistema a pleno sol, tanto no período de veranico como no final da estação úmida. Em março foi observado maior acúmulo de N-orgânico no terço superior do cafeeiro a pleno sol e no terço médio do cafeeiro sombreado. Maiores teores de nitrogênio total e a tendência de maior atividade enzimática da redutase do nitrato foram verificados nos sistemas arborizados, quando avaliados dentro do mesmo terço do cafeeiro nas duas estações.

  7. Lactobacillus plantarum strains isolated from naturally fermented sausages and their technological properties for application as starter cultures Lactobacillus plantarum isolados de salames artesanais naturalmente fermentados e suas propriedades tecnológicas como culturas iniciadoras

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    Maristela Cortez Sawitzki

    2009-06-01

    Full Text Available In the present study, technological properties of L. plantarum strains isolated from naturally fermented sausages manufactured in the South region of Brazil were investigated in order to obtain a starter culture. The technological properties evaluated were the following: ability to growth at different pH values, at different temperatures, in different salt concentrations and in the presence of commercial curing salt, fast production of acid, determination of D - and L - lactic acid; nitrate reductase activity; antagonistic activity and stability of the isolated cultures after fermentation, concentration, and freeze-drying process. The isolated strains showed effectiveness to improve technological properties as starter cultures.No presente estudo foram investigadas as propriedades tecnológicas de culturas de L. plantarum, isoladas de salames artesanais, naturalmente fermentados, manufaturados na região Sul do Brasil, a fim de obter um cultivo iniciador. As propriedades tecnológicas investigadas foram as seguintes: habilidade das culturas para crescer em diferentes valores de pH, em diferentes concentrações de sal e na presença de sal de cura comercial; rápida produção de ácido, produção do isômero D - ou L - ácido láctico, atividade nitrato redutase, atividade antagonística e estabilidade das culturas após processo de fermentação, concentração e liofilização. Todas as culturas apresentaram eficiência quanto às propriedades tecnológicas investigadas.

  8. Assimilação metabólica de nitrogênio em plântulas de seringueira cultivadas com nitrato ou amônio

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    Andrea Yu Kwan Villar Shan

    2012-06-01

    Full Text Available O objetivo deste trabalho foi avaliar a assimilação metabólica de nitrogênio em plântulas de seringueira cultivadas na presença de nitrato ou de amônio, por meio da quantificação da atividade das enzimas redutase do nitrato (RN, glutamina sintetase (GS, glutamato sintase (GOGAT e glutamato desidrogenase (GDH. Os seguintes tratamentos foram avaliados: fontes de N - controle, sem aplicação de N; amônio a 8 mmol L-1; e nitrato a 8 mmol L-1 - e tempos de exposição - 0 e 27 dias -, em arranjo fatorial 3x2, no delineamento inteiramente casualizado, com quatro repetições. A forma amoniacal promoveu maior assimilação metabólica de N, o que resultou em maior acúmulo de aminoácidos. A assimilação, tanto da forma nítrica quanto da amoniacal, aconteceu principalmente via GS/GOGAT; parte do N nítrico é assimilado nas raízes, com pequena participação da GDH, e parte é translocada para outros órgãos. O N amoniacal é totalmente assimilado nas raízes, com significativa participação da GDH. Nas folhas, a GS participa tanto da assimilação primária como da reassimilação do amônio fotorrespirado, independentemente da fonte nitrogenada, enquanto a enzima GDH tem papel secundário, independentemente da fonte ou do tempo de exposição. A atividade da RN in vitro é detectada somente em raízes e caules.

  9. METABOLISMO DO NITROGÊNIO ASSOCIADO À DEFICIÊNCIA HÍDRICA E SUA RECUPERAÇÃO EM GENÓTIPOS DE MILHO

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    Ferreira Vilma Marques

    2002-01-01

    Full Text Available O estresse hídrico afeta profundamente o metabolismo celular vegetal. Neste trabalho, objetivou-se quantificar os efeitos da deficiência hídrica e sua recuperação sobre a atividade das enzimas do metabolismo do nitrogênio: redutase do nitrato (RN, glutamina sintetase (GS e glutamato sintase (GOGAT e sobre o acúmulo de prolina em plantas dos genótipos de milho BR 2121 e BR 205. O experimento foi conduzido em casa de vegetação, sob o delineamento inteiramente casualizado, com quatro repetições, utilizando-se vasos que continham 14,3kg de solo. Os tratamentos consistiram da combinação dos dois genótipos e quatro intervalos entre irrigações (1, 3, 5 e 7 dias. No dia da avaliação (49 dias após emergência, os tratamentos com intervalos entre 3 e 7 dias, haviam sido irrigados no dia anterior, caracterizando-se portanto como recuperação da deficiência hídrica leve e severa, respectivamente. As extrações e análises foram realizadas utilizando-se a terceira folha basípeta completamente expandida. As atividades das enzimas estudadas não diferiram entre os tratamentos de estresse hídrico, controle e recuperação do estresse moderado, entretanto as plantas sob recuperação do estresse severo apresentaram atividade enzimática superior à das plantas controle. O acúmulo de prolina livre nas folhas aumentou com o estresse hídrico e respondeu à recuperação do estresse apresentando redução. De modo geral, a atividade enzimática e o acúmulo de prolina apresentaram respostas inversas dentro dos tratamentos.

  10. Frequency and Distribution of Tuberculosis Resistance-Associated Mutations between Mumbai, Moldova, and Eastern Cape.

    Science.gov (United States)

    Georghiou, S B; Seifert, M; Catanzaro, D; Garfein, R S; Valafar, F; Crudu, V; Rodrigues, C; Victor, T C; Catanzaro, A; Rodwell, T C

    2016-07-01

    Molecular diagnostic assays, with their ability to rapidly detect resistance-associated mutations in bacterial genes, are promising technologies to control the spread of drug-resistant tuberculosis (DR-TB). Sequencing assays provide detailed information for specific gene regions and can help diagnostic assay developers prioritize mutations for inclusion in their assays. We performed pyrosequencing of seven Mycobacterium tuberculosis gene regions (katG, inhA, ahpC, rpoB, gyrA, rrs, and eis) for 1,128 clinical specimens from India, Moldova, and South Africa. We determined the frequencies of each mutation among drug-resistant and -susceptible specimens based on phenotypic drug susceptibility testing results and examined mutation distributions by country. The most common mutation among isoniazid-resistant (INH(r)) specimens was the katG 315ACC mutation (87%). However, in the Eastern Cape, INH(r) specimens had a lower frequency of katG mutations (44%) and higher frequencies of inhA (47%) and ahpC (10%) promoter mutations. The most common mutation among rifampin-resistant (RIF(r)) specimens was the rpoB 531TTG mutation (80%). The mutation was common in RIF(r) specimens in Mumbai (83%) and Moldova (84%) but not the Eastern Cape (17%), where the 516GTC mutation appeared more frequently (57%). The most common mutation among fluoroquinolone-resistant specimens was the gyrA 94GGC mutation (44%). The rrs 1401G mutation was found in 84%, 84%, and 50% of amikacin-resistant, capreomycin-resistant, and kanamycin (KAN)-resistant (KAN(r)) specimens, respectively. The eis promoter mutation -12T was found in 26% of KAN(r) and 4% of KAN-susceptible (KAN(s)) specimens. Inclusion of the ahpC and eis promoter gene regions was critical for optimal test sensitivity for the detection of INH resistance in the Eastern Cape and KAN resistance in Moldova. (This study has been registered at ClinicalTrials.gov under registration number NCT02170441.).

  11. Whole Genome Sequencing Based Characterization of Extensively Drug-Resistant Mycobacterium tuberculosis Isolates from Pakistan

    KAUST Repository

    Ali, Asho

    2015-02-26

    Improved molecular diagnostic methods for detection drug resistance in Mycobacterium tuberculosis (MTB) strains are required. Resistance to first- and second- line anti-tuberculous drugs has been associated with single nucleotide polymorphisms (SNPs) in particular genes. However, these SNPs can vary between MTB lineages therefore local data is required to describe different strain populations. We used whole genome sequencing (WGS) to characterize 37 extensively drug-resistant (XDR) MTB isolates from Pakistan and investigated 40 genes associated with drug resistance. Rifampicin resistance was attributable to SNPs in the rpoB hot-spot region. Isoniazid resistance was most commonly associated with the katG codon 315 (92%) mutation followed by inhA S94A (8%) however, one strain did not have SNPs in katG, inhA or oxyR-ahpC. All strains were pyrazimamide resistant but only 43% had pncA SNPs. Ethambutol resistant strains predominantly had embB codon 306 (62%) mutations, but additional SNPs at embB codons 406, 378 and 328 were also present. Fluoroquinolone resistance was associated with gyrA 91-94 codons in 81% of strains; four strains had only gyr B mutations, while others did not have SNPs in either gyrA or gyrB. Streptomycin resistant strains had mutations in ribosomal RNA genes; rpsL codon 43 (42%); rrs 500 region (16%), and gidB (34%) while six strains did not have mutations in any of these genes. Amikacin/kanamycin/capreomycin resistance was associated with SNPs in rrs at nt1401 (78%) and nt1484 (3%), except in seven (19%) strains. We estimate that if only the common hot-spot region targets of current commercial assays were used, the concordance between phenotypic and genotypic testing for these XDR strains would vary between rifampicin (100%), isoniazid (92%), flouroquinolones (81%), aminoglycoside (78%) and ethambutol (62%); while pncA sequencing would provide genotypic resistance in less than half the isolates. This work highlights the importance of expanded

  12. Assessment of a 96-Well Plate Assay of Quantitative Drug Susceptibility Testing for Mycobacterium Tuberculosis Complex in China

    Science.gov (United States)

    Xia, Hui; Zheng, Yang; Zhao, Bing; van den Hof, Susan; Cobelens, Frank; Zhao, YanLin

    2017-01-01

    Objective To evaluate the performance of the Sensitire MYCOTB MIC Plate (MYCOTB) which could measure the twelve anti-tuberculosis drugs susceptibility on one 96-wells plate. Methods A total of 140 MDR-TB strains and 60 non-MDR strains were sub-cultured and 193 strains were finally tested for drug resistance using MYCOTB and agar proportion method (APM) and another 7 strains failed of subculture. The drugs included ofloxacin (Ofx), moxifloxacin (Mfx), rifampin (RFP), amikacin (Am), rifabutin (Rfb), para-aminosalicylic acid (PAS), ethionamide (Eth), isoniazid (INH), kanamycin (Km), ethambutol (EMB), streptomycin (Sm), and cycloserine(Cs). The categorical agreement, conditional agreement, sensitivity and specificity of MYCOTB were assessed in comparison with APM. For strains with inconsistent results between MYCOTB and APM, the drug resistance related gene fragments were amplified and sequenced: gyrA for Ofx and Mfx; rpoB for RFP and Rfb; embB for EMB; rpsl for Sm; katG and the promoter region of inhA for INH, ethA and the promoter region of inhA for Eth. The sequence results were compared with results of MYCOTB and APM to analyze the consistency between sequence results and MYCOTB or APM. Results The categorical agreement between two methods for each drug ranged from 88.6% to 100%. It was the lowest for INH (88.6%). The sensitivity and specificity of MYCOTB ranged from 71.4% to 100% and 84.3% to 100%, respectively. The sensitivity was lowest for Cs(71.4%), EMB at 10μg/ml (80.0%) and INH at 10.0μg/ml (84.6%). The specificity was lowest for Rfb (84.3%). Overall discordance between the two phenotypic methods was observed for 96 strains, of which 63 (65.6%) were found susceptible with APM and resistant with MYCOTB and the remaining 33(34.4%) strains were resistant by APM and susceptible with MYCOTB. 34/52 (65.4%) sequenced APM susceptible and MYCOTB resistant(APM-S/MYCOTB-R) strains had mutations or insertions in the amplified regions. 20/30 (66.7%) sequenced APM

  13. [Evaluation of the genotype MTBDR assay for rapid detection of rifampin and isoniazid resistance in clinical Mycobacterium tuberculosis complex clinical isolates].

    Science.gov (United States)

    Aslan, Gönül; Tezcan, Seda; Emekdaş, Gürol

    2009-04-01

    Rapid identification of resistant Mycobacterium tuberculosis complex isolates is quite important for the establishment of early and appropriate therapy. The Genotype MTBDR (Hain Lifescience, Nehren, Germany) is a commercially available DNA strip assay designed for the rapid detection of rpoB and katG gene mutations in clinical isolates. This study was conducted to determine the mutation types of phenotypically drug resistant 26 M. tuberculosis complex clinical isolates [15 isoniazid (INH), 1 rifampin (RMP) and 10 INH and RMP resistant] by Genotype MTBDR (G-MTBDR) DNA strip assay and to compare the diagnostic performance of this test. Sixteen of 25 (64%) INH-resistant and 9 of 11 (81.8%) RMP-resistant clinical isolates were correctly identified with the presence of hybridization in mutation probe or lack of hybridization at least by one of the wild type probes, by G-MTBDR assay. Hybridization with mutation probes was detected in only 5 of the RMP resistant isolates. We observed rpoB MUT3 (S531L, Ser-->4Leu) mutation in 4 and rpoB MUT1 (D516V) in one of these isolates. In 56% (14/25) of the INH-resistant isolates, katG T1 (S315T1) hybridization pattern was observed at katG mutation probe. G-MTBDR assay couldn't identify two of the 11 (18.2%) RMP-resistant isolates and one of these iSolates was shown to have a mutation at codon 531 (TCG-GCG) and the other at codon 545 (CTG-->ATG), possibly not associated with resistance, by sequence analysis. In four of the eight (8/25; 32%) INH-resistant isolates not identified by G-MTBDR assay, DNA cycle sequencing revealed different nucleotide changes outside the most common mutation zone. One of these were at codon 293 (GCT-->ACT) in katG, one with dual mutation at 279 (GGC-->ACC) in katG and at 15th C-->T in inhA gene, one at 15th C-->T in inhA gene and one at 279 (GGC-->ACC) in katG gene region. DNA membrane strip assay can be a use ful tool for the rapid detection of resistant M. tuberculosis complex isolates and therefore

  14. Interferon Gamma Release Assays in active Tuberculosis: new medical insights

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    Sandro Pierdomenico

    2011-09-01

    Full Text Available Since first presentation, Interferon γ Release Assays (IGRAs have had basic and wide application to LTBI, in accordance with international consensus and CDC recommendations, leaving their use in active TB to the field of study and research.We reviewed the results of 633 patients investigated from 2004 to 2008 targeting active TB, with the objective to highlight immunological data supporting test performances.We evaluated Quantiferon TB Gold (1st generation IGRA kit in association to Culture (MGIT 960 and Lowenstein Jensen and PCR (Probetec-ET having the positivity of culture plus clinical diagnosis as the standard true value to compare. QTB Gold was studied in 69 TB positive patients (42 pulmonary and 27 extra-pulmonary, with Sensitivity, Specificity, PPV and NPV average to 61.8%, 94.5%, 54.3% and 95.9% respectively, after indeterminate results discharging. Significant statistical differences didn’t emerge between pulmonary and extra-pulmonary infections (CI 95%.The overall indeterminate ratio arose up to 20.3% in patients with active TB vs 2.7% of global population (p<0.001. In 22% of patients with active pulmonary disease, IGRA conversed to positivity after 15 days in replicated tests, in spite of current treatment. 4 patients, with pulmonary TB and Quantiferon persistent negativities, underwent 18 months follow-up as not respondent although SIRE phenotypic susceptibilities and enough DOT compliance. Molecular DST documented hetero resistance for rpoB (MUT 1, MUT 3 plus wild lines and katG (MUT 1 plus wild in association to lack of inhA wild lines (Genotype MTBDR plus, Hain Lifescience. These reports suggest a mutational relationship between Rv3874 – 3875 cassette, encoding ESAT-6 / CFP-10, and rpoB, katG, inhA genes plausibly implying weak or absent selective clonal Th 1 activation to IGRA antigens. Our data seem to point out: 1 positive results are able to match true active TB in less than 50% of patients; 2 negative results could leave

  15. Discovery of novel phthalimide analogs: Synthesis, antimicrobial and antitubercular screening with molecular docking studies.

    Science.gov (United States)

    Rateb, Heba S; Ahmed, Hany E A; Ahmed, Sahar; Ihmaid, Saleh; Afifi, Tarek H

    2016-01-01

    In continuation of our endeavor towards the design and development of potent and effective antimicrobial agents, three series of phthalimide derivatives (4a-i, 5a-f, and 6a-c) were synthesized, fully characterized and evaluated for their potential antibacterial, antifungal and antimycobacterial activities. These efforts led to the discovery of nine compounds 4c, 4f, 4g, 4h, 4i, 5c, 5d, 5e, and 6c (MIC range from 0.49 to 31.5 μg/mL) with potent antibacterial, antifungal, and antimycobacterial activities. Ampicillin, ciprofloxacin, amphotericin B were used as references for antibacterial and antifungal screening respectively, while isoniazid was used as a reference for antimycobacterial testing. Furthermore, molecular modeling studies were done to explore the binding mode of the most active derivatives to M. tuberculosis enoyl reductase (InhA) and DNA gyrase B. Our study showed the importance of both hydrogen bonding and hydrophobic interactions as a key interaction with the target enzymes.

  16. CodY orchestrates the expression of virulence determinants in emetic Bacillus cereus by impacting key regulatory circuits.

    Science.gov (United States)

    Frenzel, Elrike; Doll, Viktoria; Pauthner, Matthias; Lücking, Genia; Scherer, Siegfried; Ehling-Schulz, Monika

    2012-07-01

    Bacillus cereus causes gastrointestinal diseases and local and systemic infections elicited by the depsipeptide cereulide, enterotoxins, phospholipases, cytolysins and proteases. The PlcR-PapR quorum sensing system activates the expression of several virulence factors, whereas the Spo0A-AbrB regulatory circuit partially controls the plasmid-borne cereulide synthetase (ces) operon. Here, we show that CodY, a nutrient-responsive regulator of Gram-positive bacteria, has a profound effect on both regulatory systems, which have been assumed to operate independently of each other. Deletion of codY resulted in downregulation of virulence genes belonging to the PlcR regulon and a concomitant upregulation of the ces genes. CodY was found to be a repressor of the ces operon, but did not interact with the promoter regions of PlcR-dependent virulence genes in vitro, suggesting an indirect regulation of the latter. Furthermore, CodY binds to the promoter of the immune inhibitor metalloprotease InhA1, demonstrating that CodY directly links B. cereus metabolism to virulence. In vivo studies using a Galleria mellonella infection model, showed that the codY mutant was substantially attenuated, highlighting the importance of CodY as a key regulator of pathogenicity. Our results demonstrate that CodY profoundly modulates the virulence of B. cereus, possibly controlling the development of pathogenic traits in suitable host environments.

  17. Detection of Mycobacterium tuberculosis resistance mutations to rifampin and isoniazid by real-time PCR

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    Hristea A

    2010-01-01

    Full Text Available Objective: The objective of our study was to evaluate the use of a real-time polymerase chain reaction (PCR-based technique for the prediction of phenotypic resistance of Mycobacterium tuberculosis. Materials and Methods: We tested 67 M tuberculosis strains (26 drug resistant and 41 drug susceptible using a method recommended for the LightCycler platform. The susceptibility testing was performed by the absolute concentration method. For rifampin resistance, two regions of the rpoB gene were targeted, while for identification of isoniazid resistance, we searched for mutations in katG and inhA genes. Results: The sensitivity and specificity of this method for rapid detection of mutations for isoniazid resistance were 96% (95% CI: 88% to 100% and 95% (95% CI: 89% to 100%, respectively. For detection of rifampin resistance, the sensitivity and specificity were 92% (95% CI: 81% to 100% and 74% (95% CI: 61% to 87%, respectively. The main isoniazid resistance mechanism identified in our isolates is related to changes in the katG gene that encodes catalase. We found that for rifampin resistance the concordance between the predicted and observed phenotype was less than satisfactory. Conclusions: Using this method, the best accuracy for genotyping compared with phenotypic resistance testing was obtained for detecting isoniazid resistance mutations. Although real-time PCR assay may be a valuable diagnostic tool, it is not yet completely satisfactory for detection of drug resistance mutations in M tuberculosis.

  18. Synthesis of Novel Pyrazinamide Derivatives Based on 3-Chloropyrazine-2-carboxamide and Their Antimicrobial Evaluation

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    Ondrej Jandourek

    2017-02-01

    Full Text Available Aminodehalogenation of 3-chloropyrazine-2-carboxamide with variously substituted benzylamines yielded a series of fifteen 3-benzylaminopyrazine-2-carboxamides. Four compounds possessed in vitro whole cell activity against Mycobacterium tuberculosis H37Rv that was at least equivalent to that of the standard pyrazinamide. MIC values ranged from 6 to 42 μM. The best MIC (6 μM was displayed by 3-[(4-methylbenzylamino]pyrazine-2-carboxamide (8 that also showed low cytotoxicity in the HepG2 cell line (IC50 ≥ 250 μM. Only moderate activity against Enterococcus faecalis and Staphylococcus aureus was observed. No activity was detected against any of tested fungal strains. Molecular docking with mycobacterial enoyl-ACP reductase (InhA was performed to investigate the possible target of the prepared compounds. Active compounds shared common binding interactions of known InhAinhibitors. Antimycobacterial activity of the title compounds was compared to the previously published benzylamino-substituted pyrazines with differing substitution on the pyrazine core (carbonitrile moiety. The title series possessed comparable activity and lower cytotoxicity than molecules containing a carbonitrile group on the pyrazine ring.

  19. Topological analysis of electron density and the electrostatic properties of isoniazid: an experimental and theoretical study.

    Science.gov (United States)

    Rajalakshmi, Gnanasekaran; Hathwar, Venkatesha R; Kumaradhas, Poomani

    2014-04-01

    Isoniazid (isonicotinohydrazide) is an important first-line antitubercular drug that targets the InhA enzyme which synthesizes the critical component of the mycobacterial cell wall. An experimental charge-density analysis of isoniazid has been performed to understand its structural and electronic properties in the solid state. A high-resolution single-crystal X-ray intensity data has been collected at 90 K. An aspherical multipole refinement was carried out to explore the topological and electrostatic properties of the isoniazid molecule. The experimental results were compared with the theoretical charge-density calculations performed using CRYSTAL09 with the B3LYP/6-31G** method. A topological analysis of the electron density reveals that the Laplacian of electron density of the N-N bond is significantly less negative, which indicates that the charges at the b.c.p. (bond-critical point) of the bond are least accumulated, and so the bond is considered to be weak. As expected, a strong negative electrostatic potential region is present in the vicinity of the O1, N1 and N3 atoms, which are the reactive locations of the molecule. The C-H···N, C-H···O and N-H···N types of intermolecular hydrogen-bonding interactions stabilize the crystal structure. The topological analysis of the electron density on hydrogen bonding shows the strength of intermolecular interactions.

  20. Defining Multidrug-Resistant Tuberculosis: Correlating GenoType MTBDRplus Assay Results with Minimum Inhibitory Concentrations

    Science.gov (United States)

    Kambli, Priti; Ajbani, Kanchan; Sadani, Meeta; Nikam, Chaitali; Shetty, Anjali; Udwadia, Zarir; Georghiou, Sophia B; Rodwell, Timothy C; Catanzaro, Antonino; Rodrigues, Camilla

    2015-01-01

    This study correlates Minimum Inhibitory Concentrations (MICs) of rifampicin (RIF) and isoniazid (INH) with GenoType MTBDRplus assay results for drug-resistant Mycobacterium tuberculosis (MTB) clinical isolates. MICs of RIF and INH were established for 84 and 90 isolates, respectively, testing six concentrations of each drug. Genotypic resistance to each drug was determined by GenoType MTBDRplus assay with 50 representative mutations confirmed by pyrosequencing, with mutations in the rpoB gene associated with RIF-resistance and mutations in the katG and/or inhA genes associated with INH-resistance. Based upon the correlation of MICs with specific genetic profiles, relative resistance levels were established for each isolate. Results indicate that MTB phenotypic resistance, currently based upon the testing of isolate susceptibility to a single drug concentration, may be more accurately profiled via quantitative MICs, and therefore the correlation of molecular diagnostic results with specific MICs may allow for more optimal treatment of infections. PMID:25749461

  1. Clinical implications of molecular drug resistance testing for Mycobacterium tuberculosis: a TBNET/RESIST-TB consensus statement.

    Science.gov (United States)

    Domínguez, J; Boettger, E C; Cirillo, D; Cobelens, F; Eisenach, K D; Gagneux, S; Hillemann, D; Horsburgh, R; Molina-Moya, B; Niemann, S; Tortoli, E; Whitelaw, A; Lange, C

    2016-01-01

    The emergence of drug-resistant strains of Mycobacterium tuberculosis is a challenge to global tuberculosis (TB) control. Although culture-based methods have been regarded as the gold standard for drug susceptibility testing (DST), molecular methods provide rapid information on mutations in the M. tuberculosis genome associated with resistance to anti-tuberculosis drugs. We ascertained consensus on the use of the results of molecular DST for clinical treatment decisions in TB patients. This document has been developed by TBNET and RESIST-TB groups to reach a consensus about reporting standards in the clinical use of molecular DST results. Review of the available literature and the search for evidence included hand-searching journals and searching electronic databases. The panel identified single nucleotide mutations in genomic regions of M. tuberculosis coding for katG, inhA, rpoB, embB, rrs, rpsL and gyrA that are likely related to drug resistance in vivo. Identification of any of these mutations in clinical isolates of M. tuberculosis has implications for the management of TB patients, pending the results of in vitro DST. However, false-positive and false-negative results in detecting resistance-associated mutations in drugs for which there is poor or unproven correlation between phenotypic and clinical drug resistance complicate the interpretation. Reports of molecular DST results should therefore include specific information on the mutations identified and provide guidance for clinicians on interpretation and on the choice of the appropriate initial drug regimen.

  2. A user-friendly web portal for analyzing conformational changes in structures of Mycobacterium tuberculosis.

    Science.gov (United States)

    Hassan, Sameer; Thangam, Manonanthini; Vasudevan, Praveen; Kumar, G Ramesh; Unni, Rahul; Devi, P K Gayathri; Hanna, Luke Elizabeth

    2015-10-01

    Initiation of the Tuberculosis Structural Consortium has resulted in the expansion of the Mycobacterium tuberculosis (MTB) protein structural database. Currently, 969 experimentally solved structures are available for 354 MTB proteins. This includes multiple crystal structures for a given protein under different functional conditions, such as the presence of different ligands or mutations. In depth analysis of the multiple structures reveal that subtle differences exist in conformations of a given protein under varied conditions. Therefore, it is immensely important to understand the conformational differences between the multiple structures of a given protein in order to select the most suitable structure for molecular docking and structure-based drug designing. Here, we introduce a web portal ( http://bmi.icmr.org.in/mtbsd/torsion.php ) that we developed to provide comparative data on the ensemble of available structures of MTB proteins, such as Cα root means square deviation (RMSD), sequence identity, presence of mutations and torsion angles. Additionally, torsion angles were used to perform principal component analysis (PCA) to identify the conformational differences between the structures. Additionally, we present a few case studies to demonstrate this database. Graphical Abstract Conformational changes seen in the structures of the enoyl-ACP reductase protein encoded by the Mycobacterial gene inhA.

  3. The Genotype MTBDRplus ver. 2.0 test as a quick indicator of resistance to rifampicin and isoniazid in Mycobacterium tuberculosis strains

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    Salvatore Nisticò

    2013-08-01

    Full Text Available Tuberculosis is still a global emergency and a major public health problem, in some cases related to the appearance of strains of multi drug resistance (MDR and extensive drug resistance (XDR Mycobacterium tuberculosis complex.The correct determination of antibiotic sensitivity profiles is therefore crucial to carry out appropriate treatment aimed to decrease the infectivity of each patient and to reduce mortality. The poor adherence to treatment by the patient or the use of therapies based on a single drug, as a result of incorrect requirements, promote the development of drug-resistance. Have some time on the market of molecular diagnostic tests that allow, quickly and directly from biological sample to search for resistance genes some key drugs of anti-TB therapy (Rifampicin and Isoniazid. One of the tests in question is the Genotype MTBDRplus ver 2.0 which can reveal the presence of genes for resistance to Isoniazid (INH and Rifampin (RMP.The loci analyzed are those corresponding to the rpoB gene for rifampicin, katG and inhA for isoniazid. Our study is based on the analysis of 83 strains of tubercular Mycobacteria identified and isolated from patients with tuberculosis disease and subjected to the tests sensitivity, searching for mutations and phenotypic susceptibility testing for Rifampicin and Isoniazid.The comparison of the results has shown that the results obtained using the Genotype MTBDRplus ver 2.0 test, were similar to the results obtained by the traditional susceptibility testing.

  4. Atherinella blackburni (Schultz, 1949 at Itamambuca Beach, Ubatuba, SP: ecological characterization and distribution on the Brazilian coast (Teleostei: Atheriniformes: Atherinopsidae

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    GMT. Mattox

    Full Text Available Atherinella blackburni is a silverside species whose occurrence on the Brazilian coast was not properly registered until recently. So far, records of its distribution along the Brazilian shore were limited to Itaparica Island, Bahia State, and Porto Inhaúma, Rio de Janeiro State. In a recent survey of the ichthyofauna of Itamambuca Beach, Ubatuba, São Paulo State, 100 specimens of this species were collected, yielding a considerable source of information regarding its ecology and a new southern limit of its distribution. A detailed survey of an ichthyological collection revealed lots of this species from regions of the Brazilian Northeast, resulting in a northward expansion of the occurrence of A. blackburni in Brazilian waters. Besides the populations found on the Brazilian coast, the species also occurs discontinuously in the Atlantic coasts of Costa Rica, Panama and Venezuela. Meristic and morphometric analysis showed overlapped values between Brazilian populations and the ones closer to the Caribbean. An ecological study of A. blackburni at Itamambuca Beach revealed that it is a predominantly diurnal species with preference for warm and salty waters, often occurring in sandy-bottom environments. It was more abundant in Summer and Winter, when individuals with smaller values of length and weight also occurred.

  5. Evaluation of efflux pump gene expression among drug susceptible and drug resistant strains of Mycobacterium tuberculosis from Iran.

    Science.gov (United States)

    Kardan Yamchi, Jalil; Haeili, Mehri; Gizaw Feyisa, Seifu; Kazemian, Hossein; Hashemi Shahraki, Abdolrazagh; Zahednamazi, Fatemeh; Imani Fooladi, Abbas Ali; Feizabadi, Mohammad Mehdi

    2015-12-01

    Absence of mutations within the genes encoding drug targets in some phenotypically drug resistant strains of Mycobacterium tuberculosis suggests possible involvement of alternative mechanisms such as over-expression of efflux pumps. We investigated the expression level of Rv1410c, Rv2459, Rv1218c and Rv1273c efflux pumps gene by real-time quantitative reverse transcription PCR (qRT-PCR) in 31 clinical isolates of M. tuberculosis. Susceptibility to first-line drugs was performed using the proportion method. Twenty one isolates were characterized with drug resistance (DR), and among them 12 showed a significantly elevated level of expression (>4 fold) for at least one of the studied genes encoding for efflux pumps. Point mutations in the katG (codons 315 or 335) and rpoB (codons 456 and 441) genes were found in 42.85% and 66.6% of drug resistant isolates, respectively. Only one isolate showed mutation at position -15 of the inhA promoter region. Among the 7 isolates (33.33%) which had no mutation in the studied regions of drug target genes, 5 isolates showed over-expression for efflux pumps. Our results demonstrated that over-expression of efflux pumps can contribute to drug resistance in M. tuberculosis.

  6. Design of new phenothiazine-thiadiazole hybrids via molecular hybridization approach for the development of potent antitubercular agents.

    Science.gov (United States)

    Ramprasad, Jurupula; Nayak, Nagabhushana; Dalimba, Udayakumar

    2015-12-01

    A new library of phenothiazine and 1,3,4-thiadiazole hybrid derivatives (5a-u) was designed based on the molecular hybridization approach and the molecules were synthesized in excellent yields using a facile single-step chloro-amine coupling reaction between 2-chloro-1-(10H-phenothiazin-10-yl)ethanones and 2-amino-5-subsituted-1,3,4-thiadiazoles. The compounds were evaluated for their in vitro inhibition activity against Mycobacterium tuberculosis H37Rv (MTB). Compounds 5 g and 5 n were emerged as the most active compounds of the series with MIC of 0.8 μg/mL (∼ 1.9 μM). Also, compounds 5a, 5b, 5c, 5e, 5l and 5m (MIC = 1.6 μg/mL), and compounds 5j, 5k and 5o (MIC = 3.125 μg/mL) showed significant inhibition activity. The structure-activity relationship demonstrated that an alkyl (methyl/n-propyl) or substituted (4-methyl/4-Cl/4-F) phenyl groups on the 1,3,4-thiadiazole ring enhance the inhibition activity of the compounds. The cytotoxicity study revealed that none of the active molecules are toxic to a normal Vero cell line thus proving the lack of general cellular toxicity. Further, the active molecules were subjected to molecular docking studies with target enzymes InhA and CYP121.

  7. Antimicrobial susceptibility determined by the E test, Löwenstein-Jensen proportion, and DNA sequencing methods among Mycobacterium tuberculosis isolates discrepancies, preliminary results

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    Maria Inês Moura Freixo

    2004-02-01

    Full Text Available Mycobacterium tuberculosis strains resistant to streptomycin (SM, isoniazid (INH, and/or rifampin (RIF as determined by the conventional Löwenstein-Jensen proportion method (LJPM were compared with the E test, a minimum inhibitory concentration susceptibility method. Discrepant isolates were further evaluated by BACTEC and by DNA sequence analyses for mutations in genes most often associated with resistance to these drugs (rpsL, katG, inhA, and rpoB. Preliminary discordant E test results were seen in 75% of isolates resistant to SM and in 11% to INH. Discordance improved for these two drugs (63% for SM and none for INH when isolates were re-tested but worsened for RIF (30%. Despite good agreement between phenotypic results and sequencing analyses, wild type profiles were detected on resistant strains mainly for SM and INH. It should be aware that susceptible isolates according to molecular methods might contain other mechanisms of resistance. Although reproducibility of the LJPM susceptibility method has been established, variable E test results for some M. tuberculosis isolates poses questions regarding its reproducibility particularly the impact of E test performance which may vary among laboratories despite adherence to recommended protocols. Further studies must be done to enlarge the evaluated samples and looked possible mutations outside of the hot spot sequenced gene among discrepant strains.

  8. Numeri caratteristici dei flocculi d'idrogeno e di calcio c dei filamenti d'idrogeno per l'anno 1958

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    g. godoli

    1959-06-01

    Full Text Available Proseguendo la pubblicazione annuale dei numeri caratteristici deiflocculi d'idrogeno e di calcio e dei filamenti d'idrogeno (', vengonoriportati nelle seguenti tre tabelle di immediata interpretazione i valorigiornalieri per il 1958.I numeri caratteristici sono stati stimati sugli spettroeliogrammieseguiti alla torre solare di Arcetri con l'immagine di 63 mm; le caratteristichestrumentali e la tecnica delle osservazioni sono state descrittein altra sede (2.Durante tutto il 1958 sono stati eseguiti quotidianamente tre spettroeliogrammisuccessivi nella riga Ea3 dell'idrogeno e tre spettroeliogrammisuccessivi nella riga K2i 3_ 2 del calcio ionizzato. L'esecuzionedi spettroeliogrammi successivi ha lo scopo di mettere in evidenza levariazioni di intensità delle zone più attive. Tali variazioni di intensità,interessantissime nello studio dell'evoluzione dei fenomeni, non hannoperò alcuna influenza nella stima dei numeri caratteristici. Per questoabbiamo continuato a stimare i numeri caratteristici su di un solo spettroeliogrammain lì«;l ed uno solo in K2 3> 2. Gli spettroeliogrammi inHa3 esaminati erano generalmente i primi eseguiti. Solo in qualche rarocaso, quando il primo risultava difettoso, veniva esaminato il secondood il, terzo spettroeliogramma. Gli spettroeliogrammi in K2i 3> 2 esaminatierano quelli ridotti in sede di programma AGI.

  9. Comparative proteomic analysis of experimental evolution of the Bacillus cereus-Ketogulonicigenium vulgare co-culture.

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    Qian Ma

    Full Text Available The microbial co-culture system composing of Ketogulonicigenium vulgare and Bacillus cereus was widely adopted in industry for the production of 2-keto-gulonic acid (2-KGA, the precursor of vitamin C. We found serial subcultivation of the co-culture could enhance the yield of 2-KGA by 16% in comparison to that of the ancestral co-culture. To elucidate the evolutionary dynamics and interaction mechanisms of the two microbes, we performed iTRAQ-based quantitative proteomic analyses of the pure cultures of K. vulgare, B. cereus and their co-culture during serial subcultivation. Hierarchy cluster analyses of the proteomic data showed that the expression level of a number of crucial proteins associated with sorbose conversion and oligopeptide transport was significantly enhanced by the experimental evolution. In particular, the expression level of sorbose/sorbosone dehydrogenase was enhanced in the evolved K. vulgare, while the expression level of InhA and the transport efficiency of oligopeptides were increased in the evolved B. cereus. The decreased sporulating protein expression and increased peptide transporter expression observed in evolved B. cereus, together with the increased amino acids synthesis in evolved K. vulgare suggested that serial subcultivation result in enhanced synergistic cooperation between K. vulgare and B. cereus, enabling an increased production of 2-KGA.

  10. Testicular activin and follistatin levels are elevated during the course of experimental autoimmune epididymo–orchitis in mice

    Science.gov (United States)

    Nicolas, Nour; Michel, Vera; Bhushan, Sudhanshu; Wahle, Eva; Hayward, Susan; Ludlow, Helen; de Kretser, David M.; Loveland, Kate L.; Schuppe, Hans-Christian; Meinhardt, Andreas; Hedger, Mark P.; Fijak, Monika

    2017-01-01

    Experimental autoimmune epididymo-orchitis (EAEO) is a model of chronic inflammation, induced by immunisation with testicular antigens, which reproduces the pathology of some types of human infertility. Activins A and B regulate spermatogenesis and steroidogenesis, but are also pro-inflammatory, pro-fibrotic cytokines. Expression of the activins and their endogenous antagonists, inhibin and follistatin, was examined in murine EAEO. Adult untreated and adjuvant-treated control mice showed no pathology. All mice immunised with testis antigens developed EAEO by 50 days, characterised by loss of germ cells, immune cell infiltration and fibrosis in the testis, similar to biopsies from human inflamed testis. An increase of total CD45+ leukocytes, comprising CD3+ T cells, CD4 + CD8− and CD4 + CD25+ T cells, and a novel population of CD4 + CD8+ double positive T cells was also detected in EAEO testes. This was accompanied by increased expression of TNF, MCP-1 and IL-10. Activin A and B and follistatin protein levels were elevated in EAEO testes, with peak activin expression during the active phase of the disease, whereas mRNA expression of the inhibin B subunits (Inha and Inhbb) and activin receptor subunits (Acvr1b and Acvr2b) were downregulated. These data suggest that activin–follistatin regulation may play a role during the development of EAEO. PMID:28205525

  11. [Drug sensitivity in Mycobacterium tuberculosis versus its viability, cytotoxicity, genotype, and the course of the process in patients with pulmonary tuberculosis].

    Science.gov (United States)

    Manicheva, O A; Lasunskaia, E B; Zhuravlev, V Iu; Otten, T F; Barnaulov, A O; Mokrousov, I V; Pavlova, M V; Vishnevskiĭ, B I; Narvskaia, O V

    2008-01-01

    The authors studied drug sensitivity, mutations in the katG, in-hA, alpC, rpoB genes, virulence via the cytotoxicity test on THP-1 cells, and the viability and genetic affiliation of 53 clinical M. tuberculosis isolates versus data on the form and dynamics of a process. Sensitive and resistant strains did not significantly differ in viability and cytotoxicity. The highest death of infected macrophages was observed was seen with infection of M. tuberculosis of the Beijing B0 genotype, the least one seen with that of LAM with the similar rate of multiple drug resistance. There was a correlation of the changes in the count of lymphocytes in patients with the genetic affiliation of a causative agent. The severest course of the tuberculous process was observed in baseline lymphopenia (before treatment) in combination with multidrug resistance of mycobacteria, high and moderate cytotoxicity and high viability. Ser-Leu 531 mutation resulted in cross resistance to rifampicin and mycobutin in most cases.

  12. Discovery of novel phthalimide analogs: Synthesis, antimicrobial and antitubercular screening with molecular docking studies

    Science.gov (United States)

    Rateb, Heba S.; Ahmed, Hany E. A.; Ahmed, Sahar; Ihmaid, Saleh; Afifi, Tarek H

    2016-01-01

    In continuation of our endeavor towards the design and development of potent and effective antimicrobial agents, three series of phthalimide derivatives (4a-i, 5a-f, and 6a-c) were synthesized, fully characterized and evaluated for their potential antibacterial, antifungal and antimycobacterial activities. These efforts led to the discovery of nine compounds 4c, 4f, 4g, 4h, 4i, 5c, 5d, 5e, and 6c (MIC range from 0.49 to 31.5 μg/mL) with potent antibacterial, antifungal, and antimycobacterial activities. Ampicillin, ciprofloxacin, amphotericin B were used as references for antibacterial and antifungal screening respectively, while isoniazid was used as a reference for antimycobacterial testing. Furthermore, molecular modeling studies were done to explore the binding mode of the most active derivatives to M. tuberculosis enoyl reductase (InhA) and DNA gyrase B. Our study showed the importance of both hydrogen bonding and hydrophobic interactions as a key interaction with the target enzymes.

  13. Analysis of facial bone fractures: An 11-year study of 2,094 patients

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    Hwang Kun

    2010-01-01

    Full Text Available Purpose: The medical records of these patients were reviewed and analysed to determine the clinical characteristics and treatment of facial bone fractures. Patients and Methods: This is a retrospective study of 2,094 patients with facial bone fractures from various accidents that were treated at the Inha University Hospital from 1996 to 2007. Results: The most common age group was the third decade of life (29%. Males were more common than females (3.98:1. The most common aetiology was violent assault or nonviolent traumatic injury (49.4%. The most common isolated fracture site was the nasal bone (37.7%, followed by the mandible (30%, orbital bones (7.6%, zygoma (5.7%, maxilla (1.3% and the frontal bone (0.3%. The largest group with complex fractures included the inferior region of the orbital floor and zygomaticomaxilla (14%. Closed reduction was performed in 46.3% of the cases while 39.7% of the cases required open reduction. For open reductions, the most commonly used soft-tissue approach was the intraoral approach (32.3%. The complication rate was 6.4% and the most common complication was hypoesthesia (68.4% followed by diplopia (25.6%. Conclusion: Long-term collection of epidemiological data regarding facial fractures and concomitant injuries is important for the evaluation of existing preventive measures and useful in the development of new methods of injury prevention and treatment.

  14. Efeito da coenzima Q10 nos danos oxidativos induzidos pela L-tiroxina no músculo sóleo de ratos

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    Fabiana Barreiro de Freitas Silva

    2015-04-01

    Full Text Available INTRODUÇÃO: os músculoesqueléticos são tecidos dinâmicos que podem alterar suas características fenotípicas proporcionando melhor adaptação funcional com estímulos variados. A L-tiroxina é um hormônio produzido pela glândula tireoide e tem sido utilizada como modelo experimental para estimulação de estresse oxidativo no músculo esquelético. A coenzima Q10 é uma provitamina lipossolúvel sintetizada endogenamente e naturalmente encontrada em alimentos como carne vermelha, peixes, cereais, brócolis e espinafre. Apresenta propriedade antioxidante e tem potencial no tratamento de doenças degenerativas e neuromusculares. OBJETIVO: avaliar o efeito protetor da coenzima Q10 no músculo sóleo de ratos frente aos danos oxidativos provocados pela L-tiroxina. MÉTODOS: os ratos foram distribuídos em quatro grupos de seis animais cada: Grupo 1 controle; Grupo 2 coenzima Q10; Grupo 3 L-tiroxina e Grupo 4 coenzima Q10 e L-tiroxina. Após a eutanásia, o sangue dos animais foi colhido e foi analisada a atividade sérica das enzimas creatina quinase CK e aspartato aminotransferase AST. No homogenato do músculo sóleo foram avaliados fatores relacionados ao estresse oxidativo. RESULTADOS: a coenzima Q10 protegeu o músculo sóleo dos danos provocados pela L-tiroxina e favoreceu a manutenção da atividade das enzimas antioxidantes glutationa redutase e glutationa peroxidase, da concentração de glutationa reduzida e oxidada, além de evitar a lipoperoxidação. CONCLUSÃO: os resultados indicam que a coenzima Q10 protege o músculo sóleo de ratos dos danos oxidativos provocados pela L-tiroxina.

  15. Efeitos de tratamentos térmicos aplicados sobre frutas cítricas armazenadas sob refrigeração

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    Kluge Ricardo Alfredo

    2006-01-01

    Full Text Available Foram aplicados tratamentos térmicos em laranja "Valência", tangor "Murcott" e lima ácida "Tahiti" armazenadas em baixa temperatura e avaliados os efeitos na redução de injúrias pelo frio e sobre outras características bioquímicas. Os seguintes tratamentos foram aplicados: T1: frutas armazenadas a 1degreesC (Controle; T2: aquecimento rápido das frutas em água quente a 53degreesC, durante 3 minutos, sendo, em seguida, armazenadas a 1degreesC; T3: aquecimento lento das frutas em câmara regulada para 37degreesC, por 2 dias, com posterior armazenamento a 1degreesC; e T4: aquecimento intermitente em ciclos de 6 dias a 1degreesC + 1 dia a 25degreesC. As frutas foram armazenadas durante 90 dias a 1degreesC e 90-95% de Umidade Relativa (UR, sendo avaliadas a cada 15 dias. Além da incidência dos danos, foram avaliadas também as atividades das enzimas antioxidativas (catalase, glutationa redutase e ascorbato peroxidase. A lima ácida "Tahiti" e o tangor "Murcott" suportaram até 90 dias de armazenamento a 1degreesC com aquecimento intermitente, não apresentando danos pelo frio. No tratamento controle (armazenamento contínuo a 1degreesC, os danos pelo frio surgiram aos 30 dias de armazenamento para a lima "Tahiti" e aos 45 dias para o tangor "Murcott". Em laranjas "Valência", as injúrias pelo frio surgiram aos 45 dias de armazenamento, sendo significativamente menores no condicionamento térmico. Os efeitos dos tratamentos térmicos no aumento da resistência das frutas ao frio podem estar relacionados com a atividade das enzimas antioxidativas. Para a laranja "Valência", os tratamentos térmicos, aplicados na forma de condicionamento térmico ou aquecimento intermitente, reduzem as injúrias pelo frio e podem prolongar a conservação das frutas a 1degreesC e 90-95% UR. Para tangor "Murcott" e lima "Tahiti", o aquecimento intermitente é o tratamento mais eficiente para a redução de injúrias pelo frio e para o aumento na capacidade

  16. Efeito do exercício físico e estatinas na função muscular em animais com dislipidemia

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    Marilita Falângola Accioly

    2012-06-01

    Full Text Available As estatinas são utilizadas no tratamento das dislipidemias, com grande tolerância; no entanto, vários efeitos colaterais podem surgir, destacando-se miopatia. A prática regular do exercício físico (EF produz modificações favoráveis no perfil lipídico; entretanto, pode gerar lesões musculares. OBJETIVO: Avaliar o efeito da associação entre exercício físico e estatinas na função muscular, pela análise histológica, em modelo experimental animal com dislipidemia. MÉTODOS: Foram utilizados 80 ratos machos Wistar, distribuídos em oito grupos, incluindo animais submetidos à dieta hipercolesterolêmica (DH, sinvastatina com (G1 e sem (G2 EF; DH e fluvastatina, com (G3 e sem EF (G4; alimentados com ração comercial (RC na presença (G5 e ausência de (G6 EF; DH submetidos (G7 ou não (G8 a EF. A DH foi administrada por 90 dias, as estatinas e prática de EF em esteira rolante por oito semanas. Os animais foram sacrificados, e o músculo sóleo retirado para análise histológica. Aplicaram-se os testes t de Student pareado e análise multivariada, com nível significante para p < 0,05. RESULTADOS: As principais alterações histológicas encontradas foram fibras de diferentes diâmetros, atróficas, em degeneração, splitting, edema, infiltrado inflamatório. Essas alterações foram observadas em 90% dos animais do grupo G1, 80% do G2, 70% do G3, 30% do G4, 40% do G5 e 30% do G7. Nos grupos G6 e G8 identificaram-se fibras musculares com morfologia preservada. CONCLUSÕES: Na avaliação histológica muscular, a associação entre fluvastatina, sinvastatina e exercício físico acarreta alterações morfológicas com predomínio no uso da sinvastatina, variando de grau leve a grave, no músculo sóleo de ratos, induzidos pelos inibidores da HMG-CoA redutase.

  17. Physiological responses in the banana plantlets treateds with strobilurinsRespostas fisiológicas em mudas de banananeira tratadas com estrobilurinas

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    Sivia Helena Modenese-Gorla da Silva

    2012-04-01

    âmetro do pseudocaule e a massa da matéria seca da parte aérea de plantas tratadas com estrobilurinas foram superiores às de plantas não tratadas. O efeito do tratamento com fungicidas foi diferenciado, sendo mais pronunciado nas plantas tratadas com piraclostrobina do que nas plantas tratadas com azoxistrobina. Plantas tratadas com piraclostrobina apresentaram área foliar, atividade da redutase do nitrato e teor de clorofila a e de nitrogênio total foliar superiores às plantas tratadas com azoxistrobina e água, que não diferiram entre si. As estrobilurinas afetaram a fisiologia das mudas de bananeira, com destaque para a piraclostrobina.

  18. Modulation of Kupffer cells on hepatic drug metabolism

    Institute of Scientific and Technical Information of China (English)

    Hong Ding; Jing Tong; Shi-Cheng Wu; Deng-Ke Yin; Xian-Fen Yuan; Jian-Yuan Wu; Jun Chen; Gang-Gang Shi

    2004-01-01

    AIM: To observe the effects of Kupffer cells on hepatic drug metabolic enzymes.METHODS: Kunming mice were ip injected with GdCl310,20, 40 mg/kg to decrease the number and block the function of kupffer cells selectively. The contents of drug metabolic enzymes, cytochrome P450, NADPH-cytochrom C redutase (NADPH-C), aniline hydroxylase (ANH), aminopyrine Ndemethylase (AMD), erythromycin N-demethylase (EMD),and glutathione s-transferase (mGST) in hepatic microsome and S9-GSTpi, S9-GST in supernatant of 9 000 g were accessed 1 d after the injection. The time course of alteration of drug metabolic enzymes was observed on d 1, 3, and 6 treated with a single dose GdCl3. Mice were treated with Angelica sinensis polysaccharides (ASP) of 30, 60, 120 mg/kg, ig, qd ×6 d, respectively and the same assays were performed.RESULTS: P450 content and NADPH-C, ANH, AMD, and END activities were obviously reduced 1 d after Kupffer cell blockade. However, mGST and S9-GST activities were significantly increased. But no relationship was observed between GdCl3 dosage and enzyme activities. With single dose GdCl3 treatment, P450 content, NADPH-C, and ANH activities were further decreased following Kupffer cell blockade lasted for 6 d, by 35.7%, 50.3%, 36.5% after 3 d, and 57.9%, 57.9%, 63.2% after 6 d, respectively. On the contrary, AMD, EMD, mGST, and Sg-GST activities were raised by 36.5%, 71.9%, 23.1%, 35.7% after 3 d,and 155%, 182%, 21.5%, 33.7% after 6 d, respectively.Furthermore, the activities of drug metabolic enzymes were markedly increased after 30 mg/kg ASP treatment,and decreased significantly after 120 mg/kg ASP treatment.No change in activity of Sg-GSTpi was observed in the present study.CONCLUSION: Kupffer cells play an important role in the modulation of drug metabolic enzymes. The changes of drug metabolic enzyme activities depend on the time of kupffer cell blockade and on the degree of Kupffer cells activated. A low concentration of ASP increases the activities of drug

  19. Finasteride-associated male infertility Infertilidade masculina associada ao uso de finasterida

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    Sidney Glina

    2004-01-01

    Full Text Available Finasteride is a potent and specific inhibitor of the 5alpha-reductase enzyme in men. Clinical studies have shown that finasteride 1mg/day is effective for promoting hair growth in men with male pattern hair loss. However, there is a concern about the use of finasteride, especially in young fertile patients, because of its action on testosterone metabolism. This paper describes 3 cases of young patients who had very poor seminal quality during finasteride treatment (1 mg/day, and their seminal quality greatly improved after cessation of finasteride treatment. Two of them presented with a left varicocele and the other was obese. We hypothesize that finasteride may not dramatically change the spermatogenesis process in healthy men, but in patients with conditions related to infertility, an amplification of the negative influence of finasteride could occur. Future studies should be done to clarify the extent of the effect of finasteride in patients fertility problems.A finasterida é um potente e específico inibidor da enzima 5alfa-redutase em homens. Estudos clínicos demonstraram que finasterida 1mg/dia diminui a progressão da queda e aumenta o crescimento do cabelo em homens que sofrem de queda de cabelo hereditária. Por sua influência no metabolismo dos andrógenos existe uma preocupação a respeito do seu uso, principalmente em pacientes em idade fértil. Neste trabalho são descritos 3 casos de pacientes jovens, que apresentaram piora do espermograma durante o uso continuado de finasterida 1mg revertida após a suspensão do mesmo. Dois deles tinham varicocele unilateral e o terceiro era obeso. Aparentemente o tratamento com finasterida promoveu alteração significativa na qualidade seminal. Pode-se especular que talvez a finasterida por si só não traga alteração para a espermatogênese como reportado por Overstreet et al. (1999, mas que em pacientes de risco com possíveis causas de infertilidade associadas, possa ocorrer a

  20. Influence of a native strain of Staphylococcus xylosus on the microbiological, physicochemical and sensorial characteristics on milano salami type

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    Ângela Maria Fiorentini

    2010-08-01

    Full Text Available In this work, the influence of native starter cultures on the microbiological, physicochemical and sensorial characteristics of Milano salami type was studied. Two batches of Milano salami type were produced: Batch A, with the addition of Staphylococcus xylosus U5 and Batch B (control without the starter culture. The Milano salami type was characterized by an important microbial activity of coagulase-negative staphylococci (CNS that resulted in substantial growth in Batch A during the ripening with an initial count of 7.60 log cfu.g-1 and reached 9.84 log CFU.g-1 after 14 days. Bacterial enzymes that showed efficient activity under the conditions found in Milano salami type were catalase, nitrite and nitrate reductase, contributing for sensory and physicochemical properties of the product. There were no significant differences in general free fatty acids composition among the batches, while the color parameters (L *, a * and b * in the Batch A presented significantly higher values in relation to Batch B. Moreover, batch A had the higher preference in sensorial analysis.A análise da influência de culturas iniciadoras nativas nas características microbiológicas, físicoquímicas e sensoriais de salame tipo Milano foi o objeto deste estudo. Foram produzidos dois grupos de salame tipo Milano: Grupo A - com aplicação de linhagem Staphylococcus xylosus U5 enquanto o controle, Grupo B, foi produzido sem culturas iniciadoras. O salame tipo Milano foi caracterizado pela importante atividade microbiana de estafilococos coagulase negativo (SCN, que resultou significativo crescimento no Grupo A durante a maturação, com contagem inicial de 7,60 ufc.g-1 e alcançando um crescimento de 9,84 cfu.g-1 depois de 14 dias. As enzimas bacterianas que mostraram eficiente atividade sob as condições encontradas no salame tipo Milano foram catalase, nitrito e nitrato redutase, contribuindo para as propriedades físicoquímicas e sensoriais do produto. N

  1. Muscle biopsy in Pompe disease Biópsia muscular na doença de Pompe

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    Lineu Cesar Werneck

    2013-05-01

    Full Text Available Pompe disease (PD can be diagnosed by measuring alpha-glucosidase levels or by identifying mutations in the gene enzyme. Muscle biopsies can aid diagnosis in doubtful cases. Methods: A review of muscle biopsy from 19 cases of PD (infantile, 6 cases; childhood, 4 cases; and juvenile/adult, 9 cases. Results: Vacuoles with or without glycogen storage were found in 18 cases. All cases had increased acid phosphatase activity. The vacuole frequency varied (almost all fibers in the infantile form to only a few in the juvenile/adult form. Atrophy of type 1 and 2 fibers was frequent in all forms. Atrophic angular fibers in the NADH-tetrazolium reductase and nonspecific esterase activity were observed in 4/9 of the juvenile/adult cases. Conclusion: Increased acid phosphatase activity and vacuoles were the primary findings. Most vacuoles were filled with glycogen, and the adult form of the disease had fewer fibers with vacuoles than the infantile or childhood forms.O diagnóstico da doença de Pompe (PD pode ser feito pela dosagem da enzima alfa-glicosidase ou pela mutação do seu gene codificador. A biópsia muscular pode ajudar em casos duvidosos. Métodos: Revisão das biópsias musculares de 19 casos de PD (forma infantil, 6 casos; infantil tardia, 4; e juvenil/adulto, 9. Resultados: Encontrados vacúolos em 18 casos, com ou sem depósito de glicogênio. Todos mostraram aumento da fosfatase ácida. Os vacúolos estavam presentes na maioria das fibras nas formas infantis, menos frequentes nas formas juvenil e mais raros nas formas do adulto. A atrofia de fibras dos tipos 1 e 2 ocorreram em todas as formas. Fibras atróficas na NADH-tetrazolium redutase e esterase não específica foram observadas em 4/9 das formas infantil tardia/adulta. Conclusões: Os dados mais frequentes foram vacúolos, preenchidos por glicogênio com atividade aumentada da fosfatase ácida. A forma adulta apresenta menor número de vacúolos que as formas infantil e infantil

  2. Resistência à sulfadoxina-pirimetamina em Maputo, Moçambique: presença de mutações nos genes dhfr e dhps do Plasmodium falciparum Sulfadoxine-pyrimethamine resistance in Maputo, Mozambique: presence of mutations in the dhfr and dhps genes of Plasmodium falciparum

    Directory of Open Access Journals (Sweden)

    Natércia Emília Pedro Fernandes

    2007-08-01

    Full Text Available Foram analisadas a freqüência e distribuição de mutações nos genes dihidrofolato redutase e dihidropteroato sintetase do Plasmodium falciparum, usando a metodologia de reação em cadeia da polimerase e polimorfismos de hidrólise por enzimas de restrição, em amostras de sangue infectado proveniente de crianças moçambicanas, residentes em Maputo. A análise foi feita antes e 7 dias após o tratamento com sulfadoxina-pirimetamina (S/P. Os resultados mostraram a ocorrência de mutações pontuais nos genes estudados e a presença de combinações de três alelos em dhfr (51Ile, 59Arg e 108Asn e do quintúplo mutante (dhfr 51Ile, 59Arg, 108Asn e dhps 437Gly, 540Glu, ambas situações associadas à falha terapêutica no sétimo dia após tratamento com S/P. Esses achados mostram a importância de se estudar a resistência à S/P em Moçambique, e como os marcadores moleculares de resistência aos antimaláricos podem fornecer dados importantes para a política nacional de controlo da malária.The frequency and distribution of mutations in Plasmodium falciparum, dihydrofolate reductase and dihydropteroate synthase genes were analyzed, using the polymerase chain reaction and restriction fragment length polymorphism methodology, in infected blood samples from Mozambican children living in Maputo, before and seven days after treatment with sulfadoxine/pyrimethamine (S/P. The results showed the occurrence of point mutations in the genes studied and the presence of combinations of three alleles in dhfr (51Ile, 59Arg and 108Asn and "quintuple" mutant (dhfr 51Ile, 59Arg, 108Asn and dhps 437Gly, 540Glu. Both of these situations were associated with seven-day therapeutic failure, following treatment with S/P. These findings show the importance of studying S/P resistance in Mozambique, and how molecular markers for antimalarial resistance can provide important data for national malaria control policy.

  3. Pharmacokinetic comparison of sustained- and immediate-release formulations of cilostazol after multiple oral doses in fed healthy male Korean volunteers

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    Kim YH

    2015-07-01

    Full Text Available Yo Han Kim,1 Jong-Lyul Ghim,2 Jin Ah Jung,2 Sang-Heon Cho,3 Sangmin Choe,4 Hee Youn Choi,1 Kyun-Seop Bae,1 Hyeong-Seok Lim1 1Department of Clinical Pharmacology and Therapeutics, College of Medicine, University of Ulsan, Asan Medical Center, Seoul, 2Department of Clinical Pharmacology, Inje University, Busan Paik Hospital, Busan, 3Department of Clinical Pharmacology, Inha University Hospital, Inha University School of Medicine, Incheon, 4Clinical Trials Center, Pusan National University Hospital, Busan, Republic of Korea Background: A new extended-release form of cilostazol has recently been developed. This study was conducted to compare the pharmacokinetic characteristics of sustained-release (SR and immediate-release (IR formulations of cilostazol after multiple oral doses in healthy male Korean volunteers.Methods: This was an open-label, randomized, multiple-dose, crossover study conducted in 30 healthy Korean subjects. In each treatment period, subjects received oral doses of 200 mg SR formulation every 24 hours or 100 mg IR formulation every 12 hours for 5 consecutive days in a fed state, with a washout period of 9 days. The plasma concentrations of cilostazol and its metabolites were determined using a validated liquid chromatography-tandem mass spectrometry method. The area under the plasma concentration–time curve within a dosing interval (AUCT, the measured peak plasma concentration at steady state (Cmax,ss, and the time to reach Cmax,ss (tmax,ss were analyzed using a noncompartmental method. Results: A total of 24 healthy male subjects completed the study. The mean (standard deviation [SD] AUCT (96–120 hours values for SR and IR were 27,378.0 (10,301.6 ng·h/mL and 27,860.3 (7,152.3 ng·h/mL, respectively. The mean (SD Cmax,ss values were 2,741.4 (836.0 ng/mL and 2,051.0 (433.2 ng/mL, respectively. The median tmax,ss values were 8.0 hours and 4.0 hours, respectively. The geometric mean ratios (90% confidence intervals of

  4. GenoType MTBDRplus assay for rapid detection of rifampin and isoniazid resistance,in Mycobacterium tuberculosis in Sichuan%分子线性探针技术分析四川地区结核分枝杆菌耐药情况

    Institute of Scientific and Technical Information of China (English)

    多丽娜; 王婷婷; 宋兴勃; 谢轶; 陆小军; 范红; 应斌武; 王兰兰; 张磊

    2011-01-01

    Objective To explore the molecular and epidemic characteristics of rifampin (RFP) and isoniazid (INH) resistance of mycobacterium tuberculosis (MTB) in Sichuan. Methods GenoType · MTBDRplus Assay Gtplus was used to examine 68 clinical isolates of MTB and 105 clinical specimens for mutations in rpoB, katG and inhA genes related to RFP and INH resistance. Results Of the 151 valid tests obtained, 44 (29.14%) and 26 (17.22%) showed drug resistance and multidrug resistance, respectively. Resistance to RFP and INH was found in 21.85% (33/151) and 24.50% (37/151) of the samples, respectively. The most prevalent mutations were rpoB S531L, katG S315T1 and inhA C-15T. The multidrug resistance rate in the sputum specimens was significantly higher than that in the non-respiratory samples (19.35% vs 7.41%). Conclusion Drug-resistant, especially multidrug-resistant tuberculosis is highly prevalent in Sichuan. The multidrug-resistant bacteria most frequently show rpoB S531L combined with katG S315T1 mutations, suggesting the necessity of developing rapid clinical identification methods for drug-resistant MTB to control the spread of the resistant strains.%目的 应用分子线性探针Geno Type(R)MTBDRplus Assay(Gtplus)试剂盒检测四川地区耐多药结核分枝杆菌(MTB)株的基因型特征,并初步了解耐药结核病的流行情况.方法 选取结核确诊患者105份临床标本和68株临床分离株,应用Gtplus试剂盒检测MTB耐利福平和异烟肼相关的rpoB、katG、inhA基因的突变,推测其对利福平和异烟肼的耐药性.结果 151例样本的Gtplus结果有效,总耐药率和耐多药率分别为29.14%(44/151)和17.22%(26/151);耐利福平和高、低水平耐异烟肼的突变菌株分别为21.85%(33/151)、21.19%(32/151)和3.31%(5/151);突变型以rpoB S531L、katGS315T1和inhA C-15T为主.痰标本组的耐多药菌株比率明显高于肺外标本组.结论 四川地区MTB耐药形势严峻,尤以耐多药结核病的情

  5. Rapid detection of multidrug-resistant Mycobacterium tuberculosis by use of real-time PCR and high-resolution melt analysis.

    Science.gov (United States)

    Ramirez, Melissa V; Cowart, Kelley C; Campbell, Patricia J; Morlock, Glenn P; Sikes, David; Winchell, Jonas M; Posey, James E

    2010-11-01

    The current study describes the development of a unique real-time PCR assay for the detection of mutations conferring drug resistance in Mycobacterium tuberculosis. The rifampicin resistance determinant region (RRDR) of rpoB and specific regions of katG and the inhA promoter were targeted for the detection of rifampin (RIF) and isoniazid (INH) resistance, respectively. Additionally, this assay was multiplexed to discriminate Mycobacterium tuberculosis complex (MTC) strains from nontuberculous Mycobacteria (NTM) strains by targeting the IS6110 insertion element. High-resolution melting (HRM) analysis following real-time PCR was used to identify M. tuberculosis strains containing mutations at the targeted loci, and locked nucleic acid (LNA) probes were used to enhance the detection of strains containing specific single-nucleotide polymorphism (SNP) transversion mutations. This method was used to screen 252 M. tuberculosis clinical isolates, including 154 RIF-resistant strains and 174 INH-resistant strains based on the agar proportion method of drug susceptibility testing (DST). Of the 154 RIF-resistant strains, 148 were also resistant to INH and therefore classified as multidrug resistant (MDR). The assay demonstrated sensitivity and specificity of 91% and 98%, respectively, for the detection of RIF resistance and 87% and 100% for the detection of INH resistance. Overall, this assay showed a sensitivity of 85% and a specificity of 98% for the detection of MDR strains. This method provides a rapid, robust, and inexpensive way to detect the dominant mutations known to confer MDR in M. tuberculosis strains and offers several advantages over current molecular and culture-based techniques.

  6. Multiplex real-time PCR melting curve assay to detect drug-resistant mutations of Mycobacterium tuberculosis.

    Science.gov (United States)

    Luo, Tao; Jiang, Lili; Sun, Weiming; Fu, G; Mei, Jian; Gao, Qian

    2011-09-01

    Early diagnosis of drug-resistant Mycobacterium tuberculosis is urgently needed to optimize treatment regimens and to prevent the transmission of resistant strains. Real-time PCR assays have been developed to detect drug resistance rapidly, but none of them have been widely applied due to their complexity, high cost, or requirement for advanced instruments. In this study, we developed a real-time PCR method based on melting curve analysis of dually labeled probes. Six probes targeting the rpoB 81-bp core region, katG315, the inhA promoter, the ahpC promoter, and embB306 were designed and validated with clinical isolates. First, 10 multidrug-resistant (MDR) strains with a wide mutation spectrum were used to analyze the melting temperature (T(m)) deviations of different mutations by single real-time PCR. All mutations can be detected by significant T(m) reductions compared to the wild type. Then, three duplex real-time PCRs, with two probes in each, were developed to detect mutations in 158 MDR isolates. Comparison of the results with the sequencing data showed that all mutations covered by the six probes were detected with 100% sensitivity and 100% specificity. Our method provided a new way to rapidly detect drug-resistant mutations in M. tuberculosis. Compared to other real-time PCR methods, we use fewer probes, which are labeled with the same fluorophore, guaranteeing that this assay can be used for detection in a single fluorescent channel or can be run on single-channel instruments. In conclusion, we have developed a widely applicable real-time PCR assay to detect drug-resistant mutations in M. tuberculosis.

  7. RP-HPLC法分离异黄酮中移动相条件的优化%Optimum of Mobile Phase Condition for Resolving Isoflavones in RP-HPLC

    Institute of Scientific and Technical Information of China (English)

    郑金珠; 盧敬昊

    2007-01-01

    In reversed-phase high performance liquid chromatography (RP-HPLC),the mobile phase condition for separating eight isoflavones (daidzin,glycitin,genistin,6'-o-acetyl daidzin,6'-o-malonyl genistin,daidzein,glycitein and genistein) was optimized using the HCI (High-Purity Separation Laboratory,Department of Chemical Engineering,Inha University) program software.The optimum composition of mobile phase for the separation of the eight isoflavones was obtained.The elution profiles were calculated by the plate theory based on the equations of retention factor,In k=A+BF+CF2,where F Was the volume percentage of acetonitrile with 0.1% acetic acid (AA).The first mobile phase composition was water with 0.1% AA/acetonitrile with 0.1% AA (88%/12%,by volume),followed at 9min later by the second composition of mobile phase which was step-changed to 85%/15%,at 19min by the third composition which was step-changed to 73%/27%,at 30min when it was changed to 65%/35% and finally it was maintained in isocratic mode to the end of the run time at 50min.Although,using step gradient mode to separate the isoflavones,the calculated and experimented data were not achieved very good agwement,we could estimate the closed retention time before experiment.And the agreement between the experimental data and the calculated values was relatively good using isocratic separation for eight isoflavones,but the retention time is very long.

  8. Effects of carbon nanofiber on physiology of Drosophila

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    Lee SH

    2015-05-01

    Full Text Available Shin-Hae Lee,1,* Hye-Yeon Lee,1,* Eun-Ji Lee,1,* Dongwoo Khang,2 Kyung-Jin Min11Department of Biological Sciences, Inha University, Incheon, Republic of Korea; 2Department of Molecular Medicine, Graduate School of Medicine, Gachon University, Incheon, Republic of Korea*These authors contributed equally to this workAbstract: As nanomaterials are now widely utilized in a wide range of fields for both medical and industrial applications, concerns over their potential toxicity to human health and the environment have increased. To evaluate the toxicity of long-term exposure to carbon nanofibers (CNFs in an in vivo system, we selected Drosophila melanogaster as a model organism. Oral administration of CNFs at a concentration of 1,000 µg/mL had adverse effects on fly physiology. Long-term administration of a high dose of CNFs (1,000 µg/mL reduced larval viability based on the pupa:egg ratio, adult fly lifespan, reproductive activity, climbing activity, and survival rate in response to starvation stress. However, CNFs at a low concentration (100 µg/mL did not show any significant deleterious effect on developmental rate or fecundity. Furthermore, long-term administration of a low dose of CNFs (100 µg/mL increased lifespan and climbing ability, coincident with mild reactive oxygen species generation and stimulation of the antioxidant system. Taken together, our data suggest that a high dose of CNFs has obvious physiological toxicity, whereas low-dose chronic exposure to CNFs can actually have beneficial effects via stimulation of the antioxidant defense system.Keywords: toxicity, Drosophila melanogaster, lifespan, reactive oxygen species

  9. The Characteristics of a Research Network for Radiation Oncology in Korea

    Energy Technology Data Exchange (ETDEWEB)

    Choi, Jin Hyun; Park, Seo Hyun; Kang, Jin Oh [Kyung Hee University School of Medicine, Seoul (Korea, Republic of)

    2010-11-15

    To evaluate the structural characteristics of a scientific network of radiation oncology society A total of 1,512 articles published from 1986 to April 2010 with the terms 'radiation oncology' or 'therapeutic radiology' were obtained in the Korea Med database. The co-authors were analyzed according to their affiliation, and their relationship was used to build a matrix. With the matrix, centralization indices and the Key Player index were analyzed. We used UCINET 6.0 for the network analysis, Netdraw for determining a sociogram and Key Player 1.44 for the key player analysis. The centralization of the radiation oncology field decreased from 8.29% for the period from 1986-1990 to 1.84% from 2006-2010. However, when the Korean Journal of Medical Physics was excluded, centralization increased from 2.32% for the period from 2001-2005 to 3.80% from 2006-2010. This suggested that the communication in the clinical research field of radiation oncology is decreasing. In a node centralization analysis, Seoul National University was found to be the highest at 7.9%. Seoul National University showed the highest indices in the Outdegree (6.50%) and Indegree (8.54%), in addition to Betweenness (14.94%) and Eigenvector (135.234%). The Key Player analysis indicated that Inha University had the highest index at 0.491, but when the Korean Journal of Medical Physics was excluded, Yonsei University had the highest Key Player index at 0.584. The degree centrality in the network of radiation oncology decreased in the most recent period as more institutions are participating in network. However, the Betweenness centrality is still increasing, suggesting that the communications among research groups (clique) in radiation oncology is warranted.

  10. Cognitive function correlations with apolipoprotein Eε4 single nucleotide polymorphism in 1 000 elderly patients following general anesthesia A randomized controlled study

    Institute of Scientific and Technical Information of China (English)

    Yingmin Cai; Haitao Hu; Rongliang Xue; Pengbin Liu; Gaifeng Feng; Weijiang Dong; Bin Yu; Minggang Zhao

    2009-01-01

    BACKGROUND: Cognitive dysfunction occurs in elderly patients following general anesthesia, and this might be associated with genetics. Studies have shown that theε4 allele gene is closely associated with senile dementia.OBJECTIVE: To compare and analyze the correlations between cognitive dysfunction and single nucleotide polymorphism of apolipoprotein E (ApoE) following inhaJation or intravenous anesthesia.DESIGN, TIME AND SETTING: A randomized, controlled study was performed. The patients were recruited from the Department of Anesthesia, Second Affiliated Hospital, Medical College, Xi'an Jiaotong University, China between May 2005 and December 2008. Genetic analyses were conducted at the Departments of Neuroanatomy and Forensic Medicine, Medical College, Xi'an Jiaotong University, China.PARTICIPANTS: A total of 1 000 patients of ASA Ⅰ-Ⅱ grade, without genetic connection, were enrolled in this study, comprising 520 males and 480 females, aged (70.1±4.6) years and weighing (57.3±7.5) kg. No patients suffered from cognitive dysfunction.METHODS: The patients were equally and randomly divided into intravenous anesthesia and gas anesthesia groups. Total intravenous anesthesia and inhaled anesthesia were used. Genomic DNA from whole blood was extracted. The ApoE gene was amplified by PCR. Restriction fragment length polymorphism of ApoE gene was analyzed. Cognitive function was evaluated by Mini-Mental State Examination (MMSE). Patients scoring 0.05).CONCLUSION: Results demonstrated a correlation between cognitive dysfunction and ApoE single nucleotide polymorphism in elderly patients after gas anesthesia. However, no relationship between cognitive dysfunction and ApoE single nucleotide polymorphism was determined in elderly patients following intravenous anesthesia. Therefore, elderly patients, especially those expressing the ApoEε4 gene, should be cautiously exposed to gas anesthesia.

  11. Conditional Deletion of the Retinoblastoma (Rb) Gene in Ovarian Granulosa Cells Leads to Premature Ovarian Failure

    Science.gov (United States)

    Andreu-Vieyra, Claudia; Chen, Ruihong; Matzuk, Martin M.

    2008-01-01

    The retinoblastoma protein (RB) regulates cell proliferation and survival by binding to the E2F family of transcription factors. Recent studies suggest that RB also regulates differentiation in a variety of cell types, including myocytes, neurons, adipocytes, and chondrocytes. Rb mutations have been found in ovarian cancer; however, the role of RB in normal and abnormal ovarian function remains unclear. To test the hypothesis that loss of Rb induces ovarian tumorigenesis, we generated an ovarian granulosa cell conditional knockout of Rb (Rb cKO) using the Cre/lox recombination system. Rb cKO females showed 100% survival and no ovarian tumor formation through 9 months of age, but they developed progressive infertility. Prepubertal Rb cKO females showed increased ovulation rates compared with controls, correlating with increased follicle recruitment, higher Fshr and Kitl mRNA levels, and lower anti-Müllerian hormone levels. In contrast, the ovulation rate of 6-wk-old females was similar to that of controls. Morphometric analysis of Rb cKO ovaries from 6-wk-old and older females showed increased follicular atresia and apoptosis. Rb cKO ovaries and preantral follicles had abnormal levels of known direct and indirect target genes of RB, including Rbl2/p130, E2f1, Ccne2, Myc, Fos, and Tgfb2. In addition, preantral follicles showed increased expression of the granulosa cell differentiation marker Inha, decreased levels of Foxl2 and Cyp19a1 aromatase, and abnormal expression of the nuclear receptors Nr5a1, Nr5a2, and Nr0b1. Taken together, our results suggest that RB is required for the temporal-specific pattern of expression of key genes involved in follicular development. PMID:18599617

  12. Harvesting energy wood in connection to young stand treatment; Energiapuun korjuu taimikon harvennuksen yhteydessae

    Energy Technology Data Exchange (ETDEWEB)

    Haemaelaeinen, J.; Lilleberg, R.; Poikela, A.; Rieppo, K. [Metsaeteho Oy, Helsinki (Finland)

    1997-12-01

    In the study felling-bunching and the Chipset chipharvester was tried in late thinning of a young stand. Also the impact of energy wood recovery in the young stand stage on the profitability of first thinning was monitored. On the study site the amount of trees to be cut was 5 300 - 22 000 trees/ha, mean height 2,7 - 5,0 m and the total biomass amounted to 26 - 75 m{sup 3}/ha. The felling-bunching was made with a chain-saw equipped with felling handles. The work consisted of opening of the strip roads, of thinning the stand between strip roads and of bunching the trees for the chip-harvester. The effective time expenditure of felling-bunching was on the different sites 8,5 - 27 in/ha or 0,24 - 0,64 h/m{sup 3}. The calculated costs of the work were 900 - 2 800 FIM/ha or 26 - 67 FIM/m{sup 3}. The density and volume of the trees strongly influenced work productivity. The Chipset chipping productivity was 5,7 - 8,7 m{sup 3} per gross effective hour. The chipping costs, when hauling distance was 250 meters, were 44 - 81 FIM/m{sup 3} or 21 - 43 FIM/MVVh. The total costs of energy wood procurement at the plant were 112 - 144 FIM/m{sup 3} or 56 - 73 FIM/MWh. The opening of strip roads during young stand treatment decreases net volume later in the first thinning. According to calculations made for the example sites the net volume decreased by approx. 9 m{sup 3}/ha. The first thinning costs thus increased by 4 FIM/m{sup 3} or ca 5 %. In the extension project the technical, economical and productional preconditions for combining young stand treatment, energy wood harvesting and first thinning will be monitored

  13. Differentially expressed miRNAs in trisomy 21 placentas.

    Science.gov (United States)

    Svobodová, Iveta; Korabečná, Marie; Calda, Pavel; Břešťák, Miroslav; Pazourková, Eva; Pospíšilová, Šárka; Krkavcová, Miroslava; Novotná, Michaela; Hořínek, Aleš

    2016-08-01

    Molecular pathogenesis of Down syndrome (DS) is still incompletely understood. Epigenetic mechanisms, including miRNAs gene expression regulation, belong to potential influencing factors. The aims of this study were to compare miRNAs expressions in placentas with normal and trisomic karyotype and to associate differentially expressed miRNAs with concrete biological pathways. A total of 80 CVS samples - 41 with trisomy 21 and 39 with normal karyotype - were included in our study. Results obtained in the pilot study using real-time PCR technology and TaqMan Human miRNA Array Cards were subsequently validated on different samples using individual TaqMan miRNA Assays. Seven miRNAs were verified as upregulated in DS placentas (miR-99a, miR-542-5p, miR-10b, miR-125b, miR-615, let-7c and miR-654); three of these miRNAs are located on chromosome 21 (miR-99a, miR-125b and let-7c). Many essential biological processes, transcriptional regulation or apoptosis, were identified as being potentially influenced by altered miRNA levels. Moreover, miRNAs overexpressed in DS placenta apparently regulate genes involved in placenta development (GJA1, CDH11, EGF, ERVW-1, ERVFRD-1, LEP or INHA). These findings suggest the possible participation of miRNAs in Down syndrome impaired placentation and connected pregnancy pathologies. © 2016 John Wiley & Sons, Ltd. © 2016 John Wiley & Sons, Ltd.

  14. Co-expression network analysis and genetic algorithms for gene prioritization in preeclampsia.

    Science.gov (United States)

    Tejera, Eduardo; Bernardes, João; Rebelo, Irene

    2013-11-12

    In this study, we explored the gene prioritization in preeclampsia, combining co-expression network analysis and genetic algorithms optimization approaches. We analysed five public projects obtaining 1,146 significant genes after cross-platform and processing of 81 and 149 microarrays in preeclamptic and normal conditions, respectively. After co-expression network construction, modular and node analysis were performed using several approaches. Moreover, genetic algorithms were also applied in combination with the nearest neighbour and discriminant analysis classification methods. Significant differences were found in the genes connectivity distribution, both in normal and preeclampsia conditions pointing to the need and importance of examining connectivity alongside expression for prioritization. We discuss the global as well as intra-modular connectivity for hubs detection and also the utility of genetic algorithms in combination with the network information. FLT1, LEP, INHA and ENG genes were identified according to the literature, however, we also found other genes as FLNB, INHBA, NDRG1 and LYN highly significant but underexplored during normal pregnancy or preeclampsia. Weighted genes co-expression network analysis reveals a similar distribution along the modules detected both in normal and preeclampsia conditions. However, major differences were obtained by analysing the nodes connectivity. All models obtained by genetic algorithm procedures were consistent with a correct classification, higher than 90%, restricting to 30 variables in both classification methods applied.Combining the two methods we identified well known genes related to preeclampsia, but also lead us to propose new candidates poorly explored or completely unknown in the pathogenesis of preeclampsia, which may have to be validated experimentally.

  15. A prospective comparison of acute intestinal toxicity following whole pelvic versus small field intensity-modulated radiotherapy for prostate cancer

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    Kim YJ

    2016-03-01

    Full Text Available Yeon Joo Kim, Jin-hong Park, In-Ha Yun, Young Seok KimDepartment of Radiation Oncology, Asan Medical Center, College of Medicine, University of Ulsan, Seoul, Republic of KoreaPurpose: To compare the acute intestinal toxicity of whole pelvic (WP and small field (SF intensity-modulated radiotherapy (IMRT for prostate cancer using dosimetric and metabolic parameters as well as clinical findings.Methods: Patients who received IMRT in either a definitive or postoperative setting were prospectively enrolled. Target volume and organs at risk including intestinal cavity (IC were delineated in every patient by a single physician. The IC volume that received a 10–50 Gy dose at 5-Gy intervals (V10–V50 and the percentage of irradiated volume as a fraction of total IC volume were calculated. Plasma citrulline levels, as an objective biological marker, were checked at three time points: baseline and after exposure to 30 Gy and 60 Gy.Results: Of the 41 patients, only six experienced grade 1 acute intestinal toxicity. Although all dose–volume parameters were significantly worse following WP than SF IMRT, there was no statistically significant relationship between these dosimetric parameters and clinical symptoms. Plasma citrulline levels did not show a serial decrease by radiotherapy volume difference (WP versus SF and were not relevant to the irradiated doses.Conclusion: Given that WP had comparable acute intestinal toxicities to those associated with SF, WP IMRT appears to be a feasible approach for the treatment of prostate cancer despite dosimetric disadvantages.Keywords: prostate cancer, intensity-modulated radiotherapy, intestinal toxicity, citrulline

  16. Quadruple-first line drug resistance in Mycobacterium tuberculosis in Vietnam: What can we learn from genes?

    Science.gov (United States)

    Nguyen, Huy Quang; Nguyen, Nhung Viet; Contamin, Lucie; Tran, Thanh Hoa Thi; Vu, Thuong Thi; Nguyen, Hung Van; Nguyen, Ngoc Lan Thi; Nguyen, Son Thai; Dang, Anh Duc; Bañuls, Anne-Laure; Nguyen, Van Anh Thi

    2017-02-21

    In Vietnam, a country with high tuberculosis (137/100.000 population) and multidrug-resistant (MDR)-TB burdens (7.8/100.000 population), little is known about the molecular signatures of drug resistance in general and more particularly of second line drug (SLD) resistance. This study is specifically focused on Mycobacterium tuberculosis isolates resistant to four first-line drugs (FLDs) that make TB much more difficult to treat. The aim is to determine the proportion of SLD resistance in these quadruple drug resistant isolates and the genetic determinants linked to drug resistance to better understand the genetic processes leading to quadruple and extremely drug resistance (XDR). 91 quadruple (rifampicin, isoniazid, ethambutol and streptomycin) FLD resistant and 55 susceptible isolates were included. Spoligotyping and 24-locus MIRU-VNTR techniques were performed and 9 genes and promoters linked to FLD and SLD resistance were sequenced. SLD susceptibility testing was carried out on a subsample of isolates. High proportion of quadruple-FLD resistant isolates was resistant to fluoroquinolones (27%) and second-line injectable drugs (30.2%) by drug susceptibility testing. The sequencing revealed high mutation diversity with prevailing mutations at positions katG315, inhA-15, rpoB531, embB306, rrs1401, rpsL43 and gyrA94. The sensitivity and specificity were high for most drug resistances (>86%), but the sensitivity was lower for injectable drug resistances (<69%). The mutation patterns revealed 23.1% of pre-XDR and 7.7% of XDR isolates, mostly belonging to Beijing family. The genotypic diversity and the variety of mutations reflect the existence of various evolutionary paths leading to FLD and SLD resistance. Nevertheless, particular mutation patterns linked to high-level resistance and low fitness costs seem to be favored.

  17. Predicting protein-binding RNA nucleotides using the feature-based removal of data redundancy and the interaction propensity of nucleotide triplets.

    Science.gov (United States)

    Choi, Sungwook; Han, Kyungsook

    2013-11-01

    Several learning approaches have been used to predict RNA-binding amino acids in a protein sequence, but there has been little attempt to predict protein-binding nucleotides in an RNA sequence. One of the reasons is that the differences between nucleotides in their interaction propensity are much smaller than those between amino acids. Another reason is that RNA exhibits less diverse sequence patterns than protein. Therefore, predicting protein-binding RNA nucleotides is much harder than predicting RNA-binding amino acids. We developed a new method that removes data redundancy in a training set of sequences based on their features. The new method constructs a larger and more informative training set than the standard redundancy removal method based on sequence similarity, and the constructed dataset is guaranteed to be redundancy-free. We computed the interaction propensity (IP) of nucleotide triplets by applying a new definition of IP to an extensive dataset of protein-RNA complexes, and developed a support vector machine (SVM) model to predict protein binding sites in RNA sequences. In a 5-fold cross-validation with 812 RNA sequences, the SVM model predicted protein-binding nucleotides with an accuracy of 86.4%, an F-measure of 84.8%, and a Matthews correlation coefficient of 0.66. With an independent dataset of 56 RNA sequences that were not used in training, the resulting accuracy was 68.1% with an F-measure of 71.7% and a Matthews correlation coefficient of 0.35. To the best of our knowledge, this is the first attempt to predict protein-binding RNA nucleotides in a given RNA sequence from the sequence data alone. The SVM model and datasets are freely available for academics at http://bclab.inha.ac.kr/primer.

  18. Use of the GenoType® MTBDRplus assay to assess drug resistance of Mycobacterium tuberculosis isolates from patients in rural Uganda

    Directory of Open Access Journals (Sweden)

    Bwanga Fred

    2010-08-01

    Full Text Available Abstract Background Drug resistance levels and patterns among Mycobacterium tuberculosis isolates from newly diagnosed and previously treated tuberculosis patients in Mbarara Uganda were investigated. Methods We enrolled, consecutively, all newly diagnosed and previously treated smear-positive TB patients aged ≥ 18 years. Isolates were tested for drug resistance against rifampicin (RIF and isoniazid (INH using the Genotype® MDRTBplus assay and results were compared with those obtained by the indirect proportion method on Lowenstein-Jensen media. HIV testing was performed using two rapid HIV tests. Results A total of 125 isolates from 167 TB suspects with a mean age 33.7 years and HIV prevalence of 67.9% (55/81 were analysed. A majority (92.8% of the participants were newly presenting while only 7.2% were retreatment cases. Resistance mutations to either RIF or INH were detected in 6.4% of the total isolates. Multidrug resistance, INH and RIF resistance was 1.6%, 3.2% and 4.8%, respectively. The rpoβ gene mutations seen in the sample were D516V, S531L, H526Y H526 D and D516V, while one strain had a Δ1 mutation in the wild type probes. There were three strains with katG (codon 315 gene mutations while only one strain showed the inhA promoter region gene mutation. Conclusion The TB resistance rate in Mbarara is relatively low. The GenoType® MTBDRplus assay can be used for rapid screening of MDR-TB in this setting.

  19. MOLECULAR CHARACTERIZATION OF MYCOBACTERIUM TUBERCULOSIS STRAINS CIRCULATING IN THE URAL REGION, RUSSIA

    Directory of Open Access Journals (Sweden)

    T. V. Umpeleva

    2013-01-01

    Full Text Available Abstract. Overall 178 Mycobacterium tuberculosis isolates recovered in 2009–2011 from newly diagnosed epidemiologically unlinked to TB patients in the Ural region of Russia have been studied. The absolute concentration method was used for drug susceptibility testing. Mutations katG, inhA and rpoB associated with resistance to isoniazid and rifampicin were detected by microchip technology («TB-Biochip». The isolates were genotyped by real-time PCR for the detection of Beijing/non-Beijing genotypes and 15-locus MIRU-VNTR typing according to «MIRU-VNTRplus» (http://www.miru-vntrplus.org. More than half (55.1% of 178 isolates belonged to the Beijing family, 58.7% of them were multiple drug resistant (MDR mostly due to rpoBSer531→Leu and katGSer315→Thr1 substitutions. Fifty VNTR profiles were found in 98 Beijing isolates; 57 of them grouped into 9 clusters. The largest VNTR cluster included 23 (23.5% Beijing isolates and 21 of them were MDR. The 80 non-Beijing isolates showed 64 distinct VNTR patterns which belonged to 6 genetic families: LAM, Ural, Haarlem, etc. Among LAM and Ural isolates 30.4% and 28.6% were MDR, respectively. The 5 of 7 MDR LAM isolates had specific mutation profile:  rpoBAsp516→Val substitution and mutations katGSer315→Thr1 and inhA_T15. The MDR Ural isolates showed the heterogeneity of mutations in rpoB gene compared to other genotypes. Taken together, these findings suggest the emergence and spread of MDR-TB in the Ural region which is characterized by circulation of MDR strains of different genotypes with the Beijing family genotype to be predominant.

  20. Detection of multi-drug resistance & characterization of mutations in Mycobacterium tuberculosis isolates from North- Eastern States of India using GenoType MTBDRplus assay

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    Ritu Singhal

    2014-01-01

    Full Text Available Background & objectives: Information on drug resistance tuberculosis is sparse from North-East (N-E States of India. We undertook this study to detect multi-drug resistant tuberculosis (MDR-TB among MDR-TB suspects, and common mutations among MDR-TB cases using GenoType MTBDRplus. Methods: All MDR suspect patients deposited sputum samples to peripheral designated microscopy centres (DMC in North-East States. The district TB officers (DTOs facilitated the transport of samples collected during January 2012 to August 2012 to our laboratory. The line probe assay to detect common mutations in the rpoB gene for rifampicin (RIF and katG and inhA genes for isoniazid (INH, respectively was performed on 339 samples or cultures. Results: A total of 553 sputum samples from MDR suspects were received of which, 181 (32.7% isolates were found to be multi-drug resistant. Missing WT8 along with mutation in codon S531L was commonest pattern for rifampicin resistant isolates (65.1% and missing WT along with mutations in codon S315T1 of katG gene was commonest pattern for isoniazid resistant isolates (86.2%. Average turn-around time for dispatch of LPA result to these States from cultures and samples was 23.4 and 5.2 days, respectively. Interpretations & conclusions: The MDR-TB among MDR-TB suspects in North-Eastern States of India was found to be 32.7 per cent. The common mutations obtained for RIF and INH in the region were mostly similar to those reported earlier.

  1. Molecular approaches for detection of the multi-drug resistant tuberculosis (MDR-TB in Bangladesh.

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    Tafsina Haque Aurin

    Full Text Available The principal obstacles in the treatment of tuberculosis (TB are delayed and inaccurate diagnosis which often leads to the onset of the drug resistant TB cases. To avail the appropriate treatment of the patients and to hinder the transmission of drug-resistant TB, accurate and rapid detection of resistant isolates is critical. Present study was designed to demonstrate the efficacy of molecular techniques inclusive of line probe assay (LPA and GeneXpert MTB/RIF methods for the detection of multi-drug resistant (MDR TB. Sputum samples from 300 different categories of treated and new TB cases were tested for the detection of possible mutation in the resistance specific genes (rpoB, inhA and katG through Genotype MTBDRplus assay or LPA and GeneXpert MTB/RIF tests. Culture based conventional drug susceptibility test (DST was also carried out to measure the efficacy of the molecular methods employed. Among 300 samples, 191 (63.7% and 193 (64.3% cases were found to be resistant against rifampicin in LPA and GeneXpert methods, respectively; while 189 (63% cases of rifampicin resistance were detected by conventional DST methods. On the other hand, 196 (65.3% and 191 (63.7% isolates showed isoniazid resistance as detected by LPA and conventional drug susceptibility test (DST, respectively. Among the drug resistant isolates (collectively 198 in LPA and 193 in conventional DST, 189 (95.6% and 187 (96.9% were considered to be MDR as examined by LPA and conventional DST, respectively. Category-II and -IV patients encountered higher frequency of drug resistance compared to those from category-I and new cases. Considering the higher sensitivity, specificity and accuracy along with the required time to results significantly shorter, our study supports the adoption of LPA and GeneXpert assay as efficient tools in detecting drug resistant TB in Bangladesh.

  2. Early detection of multi-drug resistance and common mutations in Mycobacterium tuberculosis isolates from Delhi using GenoType MTBDRplus assay

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    R Singhal

    2015-01-01

    Full Text Available Purpose: There is scarcity of prevalence data of multi-drug-resistant tuberculosis (MDR-TB data and common mutations responsible in North India. This study aimed to detect MDR-TB among MDR-TB suspects from Delhi and mutation patterns using GenoType MTBDRplus assay. Materials and Methods: All MDR suspects in five districts of New Delhi were referred to the laboratory from 1 st October 2011 to 31 st December 2012 as per criterion defined by Programmatic Management of Drug Resistant Tuberculosis (PMDT. GenoType MTBDRplus assay was performed on 2182 samples or cultures and mutations in the rpoB gene for rifampicin (RIF and katG and inhA genes for isoniazid (INH were analyzed. Results: A total of 366 (16.8% MDR-TB cases were diagnosed. MDR rate was found to be 32%, 16.6% and 10.2% during criterion A, B and C respectively. The most common mutation detected for RIF was S531L (59.0% and for INH was S315T1 (88.3%. Mutations S531L and S315T1 occurred significantly higher in MDR strains as compared to RIF mono-resistant and INH mono-resistant strains, respectively. Average laboratory turn-around time (TAT for dispatch of result to districts for test conducted on samples was 4.4 days. Conclusion: GenoType MTBDRplus is a useful assay for rapid detection of MDR-TB. The common mutations for RIF and INH were similar to those seen in other regions. However, mutations determining MDR strains and mono-resistant strains differed significantly for both RIF and INH.

  3. [Comparing performance of "TB-BIOCHIP", "Xpert MTB/RIF" and "genotype MTBDRplus" assays for fast identification of mutations in the Mycobacterium tuberculosis complex in sputum from TB patients].

    Science.gov (United States)

    Nosova, E Iu; Krasnova, M A; Galkina, K Iu; Makarova, M V; Litvinov, V I; Moroz, A M

    2013-01-01

    The frequency of mutations causing drug resistance in MTB isolates were studied in the respiratory material obtained from TB-patients in the Moscow Region. In izoniazid-resistant isolates, the most prevalent mutation was found to be the Ser315Thr substitution in the katG gene (15.8%) whereas the most frequent mutations in multidrug-resistant isolates were Ser531Leu and Ser315Thr in the rpoB and katG genes (26.3%), or a combination of these two substitutions with a T15 mutation in the inhA gene (5.3%). We compared performance of three molecular assays--"TB-BIOCHIP" ("BIOCHIP-IMB", Ltd, Russia), Xpert MTB/RIF ("Cepheid", USA) and GenoType MTBDRplus ("Hain Life-science", Germany), with the efficiency of luminescent microscopy, and phenotypic drug-suscepibility testing in an automated system BACTEC MGIT 960 (Becton, Disckinson and Company, USA). Xpert MTB/RIF, TB-BIOCHIP and GenoType MTBDRplus detected MTB in sputum in 92, 78 and 49% of all culture-positive cases, respectively. The agreement between standard cultural data and molecular DST results for Xpert MTB/RIF (resistance towards rifampicin), for TB-BIOCHIP and GenoType MTBDRplus (resistance towards rifampicin and izoniazid) amounted to 100, 97 and 100% respectively. Summing up, Xpert MTB/RIF was concluded to be the most efficient assay for primary detection of MTB, whereas the TB-BIOCHIP was shown to be the only molecular assay sensitive enough for simultaneous detection of MTB DNA and for revealing multidrug resistance in sputum (i.e. resistance to both first-line anti-TB drugs, rifampicin and izoniazid).

  4. Drug resistance of Mycobacterium tuberculosis strains in southern Brazil

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    Laynara Katize Grutzmacher

    2012-02-01

    Full Text Available INTRODUCTION: The aim of this work was to evaluate the prevalence of Mycobacterium tuberculosis (MT strains with mutations that could result in resistance to the main drugs used in treatment in a region with one of the highest numbers of tuberculosis (TB cases in southern Brazil. METHODS: Deoxyribonucleic acid (DNA from 120 sputum samples from different patients suspicious of pulmonary tuberculosis who attended the Municipal Public Laboratory for Mycobacterium sp. diagnosis was directly amplified and analyzed by PCR-SSCP. The DNA was amplified in known hotspot mutation regions of the genes rpoB, ahpC, embB, katG, inhA, and pncA. RESULTS: The percentage of samples positive by culture was 9.2% (11/120; 5% (6/120 were positive by bacilloscopy and MT-PCR, and DNA fragments of the aforementioned resistance genes could be amplified from seven (7 of the eleven (11 samples with positive results, either by culture or PCR/bacilloscopy. All presented a SSCP pattern similar to a native, nonresistant genotype, with the ATCC strain 25177 as control, except for one sample (0.01%, which presented a SSCP profile demonstrating mutation at the embB gene. CONCLUSIONS: These results are consistent with the empirical observations by physicians treating TB patients in our region of a low occurrence of cases that are refractory to conventional treatment schemes, in contrast to other parts of the country. Continued surveillance, especially molecular, is essential to detect and monitor the outbreak of MT-resistant strains.

  5. "Vibrational spectroscopic analysis and molecular docking studies of (E)-4-methoxy-N‧-(4-methylbenzylidene) benzohydrazide by DFT"

    Science.gov (United States)

    Maheswari, R.; Manjula, J.

    2016-07-01

    (E)-4-methoxy-N‧-(4-methylbenzylidene)benzohydrazide (4MN'MBH) a novel, organic, hydrazone Schiff base compound was synthesized and its structure was characterized by Fourier Transform Infrared (4000-400 cm-1), Fourier Transform Raman (3500-50 cm-1), Ultraviolet-Visible (200-800 nm) and 1H and 13C NMR spectroscopic analysis. Optimized molecular structure, vibrational frequencies and corresponding vibrational assignments regarding 4MN'MBH has become screened tentatively as well as hypothetically utilizing Gaussian09Wprogram package. Potential energy distributions of the normal modes of vibrations connected with vibrations are generally accomplished by applying VEDA program. Natural Bonding Orbital (NBO) assessment was completed with a reason to clarify charge transfer or conjugative interaction, the intra-molecular-hybridization and delocalization of electron density within the molecule. Electronic transitions were studied employing UV-Visible spectrum and the observed values were compared with theoretical values. 1H and13C NMR spectral assessment had been made with choosing structure property relationship by chemical shifts along with magnetic shielding effects of title compound. The first order hyperpolarizability (β0) and related properties (β, α0 and Δα) of 4MN'MBH were calculated. The computed first order hyperpolarizability commensurate with the documented worth of very similar structure and could be an interesting thing for more experiments on non linear optics. Molecular docking study has been performed by in silico method to analysis their antituberculosis aspects against Enoyl acyl carrier protein reductase (Mycobacterium tuberculosis InhA) protein.

  6. Effect of a Caregiver's Education Program on Stroke Rehabilitation

    Science.gov (United States)

    2017-01-01

    Objective To evaluate effects of caregiver's education program on their satisfaction, as well as patient functional recovery, performed in addition to daily conventional rehabilitation treatment. Methods Three hundred eleven subjects diagnosed with first-onset stroke and transferred to the Department of Physical Medicine and Rehabilitation of Inha University Hospital were surveyed. In 2015, caregivers attended an education program for acute and subacute stroke patients. Patients who received an additional rehabilitation therapy were assigned to the experimental group (n=81), whereas the control group (n=100) consisted of transfer cases in 2014 with only conventional treatment. The experimental group was classified by severity using the Korean version of the National Institutes of Health Stroke Scale (K-NIHSS), which was administered to all 181 subjects, in addition to, the Korean version of the Mini Mental Status Examination (K-MMSE), a Modified Barthel Index (K-MBI), and the Berg Balance Scale (K-BBS). Caregiver satisfaction and burden before and after education programs were assessed using the Canadian Occupational Performance Measure (COPM), as well as family burden and caregiver burnout scales. Results No significant intergroup difference was observed between initial K-NIHSS, K-MMSE, K-BBS, K-MBI scores, and times from admission to transfer. Those with moderate or severe strokes under the experimental condition showed a more significant improvement than the control group as determined by the K-NIHSS and K-BBS, as well as tendential K-MMSE and K-MBI score increases. Satisfaction was significantly greater for family members and formal caregivers of patients with strokes of moderate severity in the experimental group. Conclusion The caregiver's education program for stroke subjects had a positive outcome on patients' functional improvement and caregiver satisfaction. The authors believe that the additional rehabilitation therapy with the education program aids

  7. Phenotypic and genotypic analysis of anti-tuberculosis drug resistance in Mycobacterium tuberculosis isolates in Myanmar.

    Science.gov (United States)

    Aung, Wah Wah; Ei, Phyu Win; Nyunt, Wint Wint; Swe, Thyn Lei; Lwin, Thandar; Htwe, Mi Mi; Kim, Kyung Jun; Lee, Jong Seok; Kim, Chang Ki; Cho, Sang Nae; Song, Sun Dae; Chang, Chulhun L

    2015-09-01

    Tuberculosis (TB) is one of the most serious health problems in Myanmar. Because TB drug resistance is associated with genetic mutation(s) relevant to responses to each drug, genotypic methods for detecting these mutations have been proposed to overcome the limitations of classic phenotypic drug susceptibility testing (DST). We explored the current estimates of drug-resistant TB and evaluated the usefulness of genotypic DST in Myanmar. We determined the drug susceptibility of Mycobacterium tuberculosis isolated from sputum smear-positive patients with newly diagnosed pulmonary TB at two main TB centers in Myanmar during 2013 by using conventional phenotypic DST and the GenoType MTBDRplus assay (Hain Lifescience, Germany). Discrepant results were confirmed by sequencing the genes relevant to each type of resistance (rpoB for rifampicin; katG and inhA for isoniazid). Of 191 isolates, phenotypic DST showed that 27.7% (n=53) were resistant to at least one first-line drug and 20.9% (n=40) were resistant to two or more, including 18.3% (n=35) multidrug-resistant TB (MDR-TB) strains. Monoresistant strains accounted for 6.8% (n=13) of the samples. Genotypic assay of 189 isolates showed 17.5% (n=33) MDR-TB and 5.3% (n=10) isoniazid-monoresistant strains. Genotypic susceptibility results were 99.5% (n=188) concordant and agreed almost perfectly with phenotypic DST (kappa=0.99; 95% confidence interval 0.96-1.01). The results highlight the burden of TB drug resistance and prove the usefulness of the genotypic DST in Myanmar.

  8. Computer-Aided Design of Orally Bioavailable Pyrrolidine Carboxamide Inhibitors of Enoyl-Acyl Carrier Protein Reductase of Mycobacterium tuberculosis with Favorable Pharmacokinetic Profiles

    Science.gov (United States)

    Kouassi, Affiba Florance; Kone, Mawa; Keita, Melalie; Esmel, Akori; Megnassan, Eugene; N’Guessan, Yao Thomas; Frecer, Vladimir; Miertus, Stanislav

    2015-01-01

    We have carried out a computational structure-based design of new potent pyrrolidine carboxamide (PCAMs) inhibitors of enoyl-acyl carrier protein reductase (InhA) of Mycobacterium tuberculosis (MTb). Three-dimensional (3D) models of InhA-PCAMx complexes were prepared by in situ modification of the crystal structure of InhA-PCAM1 (Protein Data Bank (PDB) entry code: 4U0J), the reference compound of a training set of 20 PCAMs with known experimental inhibitory potencies (IC50exp). First, we built a gas phase quantitative structure-activity relationships (QSAR) model, linearly correlating the computed enthalpy of the InhA-PCAM complex formation and the IC50exp. Further, taking into account the solvent effect and loss of inhibitor entropy upon enzyme binding led to a QSAR model with a superior linear correlation between computed Gibbs free energies (ΔΔGcom) of InhA-PCAM complex formation and IC50exp (pIC50exp = −0.1552·ΔΔGcom + 5.0448, R2 = 0.94), which was further validated with a 3D-QSAR pharmacophore model generation (PH4). Structural information from the models guided us in designing of a virtual combinatorial library (VL) of more than 17 million PCAMs. The VL was adsorption, distribution, metabolism and excretion (ADME) focused and reduced down to 1.6 million drug like orally bioavailable analogues and PH4 in silico screened to identify new potent PCAMs with predicted IC50pre reaching up to 5 nM. Combining molecular modeling and PH4 in silico screening of the VL resulted in the proposed novel potent antituberculotic agent candidates with favorable pharmacokinetic profiles. PMID:26703572

  9. Computer-Aided Design of Orally Bioavailable Pyrrolidine Carboxamide Inhibitors of Enoyl-Acyl Carrier Protein Reductase of Mycobacterium tuberculosis with Favorable Pharmacokinetic Profiles

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    Affiba Florance Kouassi

    2015-12-01

    Full Text Available We have carried out a computational structure-based design of new potent pyrrolidine carboxamide (PCAMs inhibitors of enoyl-acyl carrier protein reductase (InhA of Mycobacterium tuberculosis (MTb. Three-dimensional (3D models of InhA-PCAMx complexes were prepared by in situ modification of the crystal structure of InhA-PCAM1 (Protein Data Bank (PDB entry code: 4U0J, the reference compound of a training set of 20 PCAMs with known experimental inhibitory potencies (IC50exp. First, we built a gas phase quantitative structure-activity relationships (QSAR model, linearly correlating the computed enthalpy of the InhA-PCAM complex formation and the IC50exp. Further, taking into account the solvent effect and loss of inhibitor entropy upon enzyme binding led to a QSAR model with a superior linear correlation between computed Gibbs free energies (ΔΔGcom of InhA-PCAM complex formation and IC50exp (pIC50exp = −0.1552·ΔΔGcom + 5.0448, R2 = 0.94, which was further validated with a 3D-QSAR pharmacophore model generation (PH4. Structural information from the models guided us in designing of a virtual combinatorial library (VL of more than 17 million PCAMs. The VL was adsorption, distribution, metabolism and excretion (ADME focused and reduced down to 1.6 million drug like orally bioavailable analogues and PH4 in silico screened to identify new potent PCAMs with predicted IC50pre reaching up to 5 nM. Combining molecular modeling and PH4 in silico screening of the VL resulted in the proposed novel potent antituberculotic agent candidates with favorable pharmacokinetic profiles.

  10. Fsh controls gene expression in fish both independently of and through steroid mediation.

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    Elisabeth Sambroni

    Full Text Available The mechanisms and the mediators relaying Fsh action on testicular functions are poorly understood. Unlike in mammals, in fish both gonadotropins (Fsh and Lh are able to efficiently stimulate steroidogenesis, likely through a direct interaction with their cognate receptors present on the Leydig cells. In this context, it is crucial to understand if Fsh effects are mediated through the production of steroids. To address this issue we performed transcriptome studies after in vitro incubations of rainbow trout testis explants in the presence of Fsh alone or in combination with trilostane, an inhibitor of Δ4- steroidogenesis. Trilostane significantly reduced or suppressed the response of many genes to Fsh (like wisp1, testis gapdhs, cldn11, inha, vt1 or dmrt1 showing that, in fish, important aspects of Fsh action follow indirect pathways and require the production of Δ4-steroids. What is more, most of the genes regulated by Fsh through steroid mediation were similarly regulated by Lh (and/or androgens. In contrast, the response to Fsh of other genes was not suppressed in the presence of trilostane. These latter included genes encoding for anti-mullerian hormone, midkine a (pleiotrophin related, angiopoietine-related protein, cyclins E1 and G1, hepatocyte growth factor activator, insulin-like growth factor 1b/3. A majority of those genes were preferentially regulated by Fsh, when compared to Lh, suggesting that specific regulatory effects of Fsh did not depend on steroid production. Finally, antagonistic effects between Fsh and steroids were found, in particular for genes encoding key factors of steroidogenesis (star, hsd3b1, cyp11b2-2 or for genes of the Igf system (igf1b/3. Our study provides the first clear evidence that, in fish, Fsh exerts Δ4-steroid-independent regulatory functions on many genes which are highly relevant for the onset of spermatogenesis.

  11. Dual channel real-time PCR melting curve analysis-based assay for detecting of anti-tuberculosis drug-resistant mutations in Mycobacterium tuberculosis%双通道实时荧光PCR熔解曲线法检测结核分枝杆菌药物耐药相关基因突变

    Institute of Scientific and Technical Information of China (English)

    柳清云; 罗涛; 李静; 梅建; 高谦

    2013-01-01

    目的 基于双标记荧光探针熔解曲线分析技术,建立一种利用实时荧光PCR快速检测结核分枝杆菌耐药突变的方法.方法 根据结核分枝杆菌一线药物常见耐药突变位点(包括rpoB 81 bp耐药决定区、inhA启动子、katG315、ahpC启动子以及embB306)设计6条荧光双标记探针和对应引物,通过PCR扩增耐药突变位点所在基因片段,在扩增完成后通过熔解曲线检测分析实现对耐药突变的快速检测.通过对2008年上海市疾控中心收集的76株临床耐多药(MDR)菌株进行检测,验证本方法的敏感度和特异度.结果 本方法成功从76株MDR菌株中检测出相关耐药突变,各种突变对应ATm值范围为1.8~14.4℃.将检测结果和测序结果对比表明该方法检测敏感度和特异度都为100%(rpoB,80/80:inhA,7/7;katG315,59/59;ahpC,8/8;embB306,27/27).本方法可以成功从最低浓度为100拷贝/μl的结核分枝杆菌DNA样本中准确地检测耐药突变.结论 双通道实时荧光PCR熔解曲线法可以快速灵敏地检测结核分枝杆菌常见耐药突变.该方法具有检测迅速准确、结果易判读、交叉污染概率低等特点,可用于快速检测临床结核耐药相关的基因突变,并对结核耐药情况进行评估.%Objective Based on dual channel melting curve analysis-based assay,we developed a method to rapidly detect the drug-resistant mutations in Mycobacterium tuberculosis through real-time PCR.Methods According to the common first-line drug-resistant mutations of Mycobacterium tuberculosis,we designed six dual-labeled fluorescence probes to rapidly detect the drug-resistant mutations through realtime PCR melting curve after amplifications of drug-resistant related gene region of DNA.The targets include rpoB 81 bp core region,katG315,inhA promoter,ahpC promoter and embB306.To validate the sensitivity and specificity of our method,we performed real-time PCR assays to detect drug-resistant mutations in 76

  12. Seeds with high molybdenum concentration improved growth and nitrogen acquisition of rhizobium-inoculated and nitrogen-fertilized common bean plants Sementes com alta concentração de molibdênio aumentaram o crescimento e a aquisição de nitrogênio de plantas de feijoeiro inoculadas com rizóbio ou sob fertilização nitrogenada

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    Fernanda Fátima Delgado Almeida

    2013-04-01

    crescimento e na aquisição de N de plantas de feijoeiro, sob N simbiótico ou N mineral, pela mensuração das atividades da nitrogenase e da nitrato redutase e da contribuição da fixação biológica de N2, em diferentes estádios de crescimento. Sementes enriquecidas ou não com Mo foram semeadas sob duas fontes de N, inoculadas com rizóbio ou fertilização nitrogenada, em vasos com 10 kg de solo. No experimento 1, um tratamento adicional consistiu de sementes enriquecidas com Mo e aplicação de Mo ao solo. No experimento 2, a contribuição da fixação biológica de N2 foi estimada pela diluição isotópica de 15N. Plantas de feijoeiro originadas de sementes com alto teor de Mo floresceram um dia antes. Sementes com alto teor de Mo aumentaram a área foliar, a massa de parte aérea e a acumulação de N, nas duas fontes de N. A ausência de efeitos do Mo adicional aplicado ao solo indicou que as sementes enriquecidas com esse nutriente foram suficientes para o crescimento vegetal. Sementes enriquecidas com Mo elevaram a atividade da nitrogenase, no estádio vegetativo de plantas inoculadas, e da nitrato redutase, em estádios tardios nas duas fontes de N. A contribuição da fixação biológica de N2 foi de 17 e 61 %, em plantas oriundas de sementes com baixo e alto teor de Mo, respectivamente. Os resultados evidenciaram os benefícios das sementes enriquecidas com Mo para o crescimento e a nutrição nitrogenada do feijoeiro, quando inoculado com rizóbio ou sob adubação nitrogenada.

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

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    Patrícia Budni

    2013-10-01

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

  14. Prevalência dos fatores trombofílicos em mulheres com infertilidade Prevalence of thrombophilic factors in infertile women

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    Adriana de Góes e Silva Soligo

    2007-05-01

    Full Text Available OBJETIVO: determinar a prevalência dos fatores trombofílicos em mulheres inférteis. MÉTODOS: estudo de corte transversal, no qual foram admitidas mulheres inférteis atendidas em clínica privada e submetidas à investigação de trombofilia, conforme protocolo da referida clínica, no período de março de 2003 a março de 2005, após aprovação do Comitê de Ética e Pesquisa da Universidade Estadual de Campinas (UNICAMP. Foram incluídas mulheres com história de infertilidade, definida como um ano de coito desprotegido sem concepção. Foram excluídas mulheres com hepatopatia e dados incompletos em prontuário, obtendo-se uma amostra de 144 mulheres. Os fatores trombofílicos avaliados foram: o anticorpo anticardiolipina (ACL, o anticoagulante lúpico (ACGL, a deficiência de proteína C (DPC, a deficiência de proteína S (DPS, a deficiência de antitrombina III (DAT, a presença do fator V de Leiden, uma mutação no gene da protrombina e a mutação do metileno tetrahidrofolato redutase (MTHFR. Resultados: os valores de prevalência obtidos para ACL e ACGL foram de 2%. A prevalência dos fatores trombofílicos hereditários foram: DPC=4%, DPS=6%, DAT=5%, fator V de Leiden=3%, mutação da protrombina=3% e mutação MTHFR=57%. CONCLUSÕES: das 144 pacientes selecionadas, 105 mulheres, ou seja, 72,9% apresentavam pelo menos um fator trombofílico presente. Isto reforça a importância e justifica a necessidade da investigação destes fatores neste grupo de mulheres.PURPOSE: to establish the prevalence of thrombophilic factors in infertile women. METHODS: a cross-sectional study was performed, in which infertile women, seen in a private clinic with investigation for thrombophilia were included, according to the protocol of the clinic, between March 2003 and March 2005, after the approval of the Research Ethics Committee of the Universidade Estadual de Campinas (UNICAMP. One hundred and forty-four infertile women without any liver

  15. O papel das selenoproteínas no câncer The role of selenoproteins in cancer

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    Kaluce Gonçalves de Sousa Almondes

    2010-01-01

    Full Text Available Evidências têm demonstrado que distúrbios do metabolismo são comuns em células tumorais, levando ao aumento do estresse oxidativo. A elevação na produção de espécies reativas de oxigênio (EROs associada à baixa atividade antioxidante tem sido relacionada a vários tipos de câncer. O selênio, micronutriente antioxidante, pode funcionar como um agente antimutagênico, prevenindo transformações malignas de células normais. Realizou-se um levantamento bibliográfico no período 2000 a 2009 mediante consulta à base de dados PubMed (National Library of Medicine´s Medline Biomedical Literature, USA, selecionando-se 39 artigos que avaliaram a relação entre câncer, estresse oxidativo e suplementação com selênio. O efeito protetor desse mineral é especialmente associado à sua presença na glutationa peroxidase e na tioredoxina redutase, enzimas protetoras do DNA e outros componentes celulares contra o dano oxidativo causado pelas EROs. Vários estudos têm demonstrado a expressão reduzida destas enzimas em diversos tipos de câncer, principalmente quando associados a uma baixa ingestão de selênio, que pode acentuar os danos causados. A suplementação de selênio parece ocasionar redução do risco de alguns tipos de câncer diminuindo o estresse oxidativo e o dano ao DNA. No entanto, mais estudos são necessários para esclarecer as doses de selênio adequadas para cada situação (sexo, localização geográfica e tipo de câncer.There are evidences that metabolic disorders are common in tumoral cells, leading to increased oxidative stress. The rising in the production of reactive oxygen species associated to low antioxidant activity have been associated to different types of cancer. Selenium, an antioxidant micronutrient can work as an anti-cancer agent preventing malignant modification in healthy cells. A literature review was carried out in the period 2000-2009 in the database PubMed selecting 39 articles which assessed

  16. The pentose phosphate pathway in Trypanosoma cruzi: a potential target for the chemotherapy of Chagas disease

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    Mariana Igoillo-Esteve

    2007-12-01

    Full Text Available Trypanosoma cruzi is highly sensitive to oxidative stress caused by reactive oxygen species. Trypanothione, the parasite's major protection against oxidative stress, is kept reduced by trypanothione reductase, using NADPH; the major source of the reduced coenzyme seems to be the pentose phosphate pathway. Its seven enzymes are present in the four major stages in the parasite's biological cycle; we have cloned and expressed them in Escherichia coli as active proteins. Glucose 6-phosphate dehydrogenase, which controls glucose flux through the pathway by its response to the NADP/NADPH ratio, is encoded by a number of genes per haploid genome, and is induced up to 46-fold by hydrogen peroxide in metacyclic trypomastigotes. The genes encoding 6-phosphogluconolactonase, 6-phosphogluconate dehydrogenase, transaldolase and transketolase are present in the CL Brener clone as a single copy per haploid genome. 6-phosphogluconate dehydrogenase is very unstable, but was stabilized introducing two salt bridges by site-directed mutagenesis. Ribose-5-phosphate isomerase belongs to Type B; genes encoding Type A enzymes, present in mammals, are absent. Ribulose-5-phosphate epimerase is encoded by two genes. The enzymes of the pathway have a major cytosolic component, although several of them have a secondary glycosomal localization, and also minor localizations in other organelles.Trypanosoma cruzi é altamente sensível ao estresse oxidativo causado por espécies reativas do oxigênio. Tripanotiona, o principal protetor do parasita contra o estresse oxidativo, é mantido reduzido pela tripanotiona redutase, pela presença deNADPH; a principal fonte da coenzima reduzida parece ser a via da pentose fosfato. As sete enzimas dessa via estão presentes nos quatro principais estágios do ciclo biológico do parasita; nós clonamos e expressamos as enzimas em Escherichia coli como proteínas ativas. Glucose 6-fosfato desidrogenase, que controla o fluxo da glucose da

  17. Potencial de Cajanus cajan e Crotalaria spectabilis parafitorremediação: absorção de arsênio e respostas antioxidativas Potential of Cajanus cajan and Crotalaria spectabilis for phytoremediation: arsenic absorption and antioxidative responses

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    Rafaella Teles Arantes Felipe

    2009-04-01

    Full Text Available Neste estudo, avaliaram-se os efeitos tóxicos do arsênio (As sobre o crescimento e atividade de enzimas antioxidativas dismutase do superóxido (SOD, catalase (CAT, peroxidases (POX, peroxidase do ascorbato peroxidase (APX e glutationa redutase (GR em plantas de Cajanus cajan e Crotalaria spectabili. Plantas das duas espécies foram expostas a diferentes concentrações de As, em solução nutritiva de Hoagland, pH 6,5. A taxa de crescimento relativo (TCR das duas espécies reduziu-se com o aumento da concentração de As na solução nutritiva, sendo de forma mais intensa em C. cajan. A concentração de As na solução capaz de reduzir em 50% a TCR de C. cajan foi de 0,93 mg L-1, enquanto em C. spectabilis foi de 4,80 mg L-1. As duas espécies apresentaram reduzida translocação de As para a parte aérea, sendo em raízes de C. spectabilis observada a maior concentração desse elemento. C. cajan exposto ao As teve as atividades das enzimas CAT, POX e APX reduzidas, ao contrário do que ocorreu em C. spectabilis. Esta espécie, quando submetida ao As, apresentou incremento na atividade de todas as enzimas avaliadas. A maior tolerância ao As observada em C. spectabilis pode estar relacionada à maior capacidade em reter o As nas raízes e à indução das enzimas SOD, CAT, POX, APX e GR. Dessa forma, C. spectabilis é uma espécie com potencial para utilização em programas de revegetação de áreas contaminadas com As.The toxic effects of Arsenic (As on the growth and activities of the antioxidative enzymes superoxide dismutase (SOD, catalase (CAT, peroxidase (POX, ascorbate peroxidase (APX and glutathione reductase (GR of Cajanus cajan and Crotalaria spectabilsi were studied. AQUI Plants of the two species were exposed to different As concentrations in Hoagland's nutrient solution, pH 6.5. The relative growth rate (RGR of the two species reduced with the increase in As concentration in the nutrient solution, especially in C. cajan

  18. Redução assimilatória de NO-3 em plantas de cajueiros cultivados em meio salinizado NO-3 assimilatory reduction in cashew plants grown in salinized medium

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    Ricardo A. Viégas

    2004-12-01

    Full Text Available Com o objetivo de se avaliar os efeitos e mecanismos iniciais da ação da salinidade sobre a redução de NO-3 e a acumulação de solutos nitrogenados, plantas de cajueiro (Anacardium occidentale L., CCP - 1001, com 42 dias de idade foram cultivadas em solução nutritiva contendo 100 mmol L-1 de NaCl, durante 8 dias, em casa de vegetação. A transpiração foi o principal fator de regulação da atividade de redutase de NO-3 (RN que foi fortemente reduzida nas folhas. Contrariamente, a atividade de glutamina sintetase (GS aumentou nas folhas em resposta à salinidade. Esta tendência de aumento da atividade de GS nas folhas foi acompanhada pelas concentrações de aminoácidos solúveis totais, particularmente prolina e proteínas solúveis totais. Por outro lado, o aumento nas concentrações das frações orgânicas de N, nas folhas, foi seguido por decréscimos nas concentrações de clorofilas e por acréscimos nas concentrações de NH+4 . É provável que o aumento da atividade de GS nas folhas de plantas de cajueiro, tenha sido induzido por um adicional suprimento de NH+4 e glutamato, a partir do catabolismo de aminoácidos e de proteínas e por aumentos na atividade fotorrespiratória.To evaluate the effects and initial-action mechanism of salinity on NO-3 reduction and N compounds accumulation, 42-day-old Anacardium occidentale (CCP-1001 plants were hydroponically grown in 100 mmol L-1 NaCl for 8 days (short-term experiment, in greenhouse. The results show that reduction in transpiration rates due to salinity had a relevant role in the regulation of NO-3 reductase (NR activity, which decreased considerably in leaf. On the contrary, glutamine sintetase (GS activity increased in leaf due to salinity. The increase in activity of GS in leaf was followed by the soluble amino acids concentration, in particular proline, and by the total soluble proteins content as well. On the other hand, increase in concentration of organic N fractions in

  19. Efeito da disponibilidade de nitrato em solução nutritiva sobre a absorção de nitrogênio e atividade enzimática de duas cultivares de arroz Effect of nitrate availability in nutrient solution on nitrogen uptake and enzimatic activity of two rice cultivars

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    André Marques dos Santos

    2009-01-01

    Full Text Available Duas cultivares de arroz, Piauí (tradicional e IAC-47 (melhorada, foram colocadas em solução nutritiva contendo 20 mg de N-NO3- L-1 até 32 dias após a germinação (DAG. Depois disso, um grupo de plantas passou a receber 200 mg de N-NO3- L-1, enquanto o outro continuou recebendo 20 mg de N-NO3- L-1 até os 42 DAG. De 42 até os 56 DAG, todas as plantas receberam 5 mg de N-NO3- L-1 até o fim do ciclo. Esses tratamentos foram empregados visando simular o fluxo sazonal de nitrato existente nos solos de regiões tropicais. As plantas foram coletadas aos 133 DAG e separadas em folha bandeira, folha 2, colmos e raízes. Foram determinados os teores de N na forma de aminoácidos, nitrato e amônio, açúcares solúveis, atividades da nitrato redutase (NR e glutamina sintetase (GS. O fluxo sazonal de nitrato influenciou significativamente a absorção e o uso de N. As variedades estudadas foram responsivas de maneira diferenciada em relação ao acúmulo e à translocação de N, quando submetidas ao fluxo sazonal de nitrato. A cultivar Piauí parece acumular mais NO3- em seus tecidos na fase inicial do seu ciclo, podendo permitir sua posterior translocação. A folha bandeira e folha 2 parecem atuar como local preferencial de acúmulo de N, sendo este acúmulo mais intenso na cultivar Piauí.Two rice cultivars, Piaui (a landrace and IAC-47 (an improved variety, were grown in nutrient solution containing 20 mg nitrate (N-NO3- L-1 up to 32 days after germination (DAG. After this, a group of plants received 200 mg N-NO3- L-1, while the other was kept at 20 mg N-NO3- L-1 up to 42 DAG. From 42 until 56 DAG, all plants received 5 mg N-NO3- L-1 until the end of the cycle. These treatments were applied with the objective to simulate nitrate flush in tropical regions. Plants were collected at 133 DAG and separated in to leaf flag, leaf 2, stem and roots. N contents in amino acids, nitrate and ammonium forms, soluble sugars, nitrate reductase (NR and

  20. Regulação da absorção e assimilação do nitrogênio nas plantas Regulation of nitrogen absortion and assimilation in plants

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    Christian Bredemeier

    2000-04-01

    Full Text Available A presente revisão tem por objetivo discutir os mecanismos de regulação da absorção e assimilação do nitrogênio nas plantas, de modo que a eficiência de uso desse nutriente seja aumentada. O nitrogênio é absorvido nas raízes sob a forma de NO3- ou NH4+, sendo então incorporado em aminoácidos na própria raiz ou na parte aérea. A taxa e a quantidade de nitrogênio absorvido e assimilado durante o ciclo da planta dependem da presença de carregadores específicos na membrana plasmática, da atividade das enzimas envolvidas no seu ciclo, da disponibilidade de energia necessária para os processos de absorção e assimilação e do estádio de desenvolvimento da planta. Inúmeros grupos de pesquisa têm centrado esforços na identificação dos pontos mais limitantes do metabolismo do nitrogênio nas plantas. Os limitadores têm sido relacionados com: a a afinidade dos carregadores pelo nitrato e amônio; b o suprimento de carboidratos às raízes; c o nível de aminoácidos no floema radicular; d a atividade das enzimas redutase do nitrato (RN, sintetase da glutamina (GS e sintase do glutamato (GOGAT; e a fonte de N suprida às plantas (NO3- ou NH4+; f o local de assimilação do N (raiz ou parte aérea. Esses estudos têm mostrado que o metabolismo do N é multiregulado e integrado ao metabolismo geral da planta. A identificação de pontos metabólicos específicos que são mais limitantes para o incremento da produtividade é complexa. A perspectiva é que a clonagem dos transportadores de N poderá auxiliar os programas de melhoramento genético na obtenção de plantas mais eficientes na absorção do nitrogênio.Nitrogen is absorbed as NO3- ou NH4+ and assimilated into aminoacids both in roots and shoots. Root absortion (rate and total amount at each plant developmental stage depends on specific plasma membrane carriers, enzyme activity for reduction and assimilation and energy availability for both processes. Several research

  1. Thrombophilic mutations and risk of retinal vein occlusion Mutações trombofílicas e risco de oclusão venosa retiniana

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    Ana Luiza Biancardi

    2007-12-01

    Full Text Available PURPOSE: The association of retinal vein occlusion and hereditary thrombophilia abnormalities is not established, with controversial results in the literature. This study investigates the association between retinal vein occlusion and three thrombophilic mutations: factor V 1691A (factor V Leiden, prothrombin 20210A (PT 20210A and homozygous methylenetetrahydrofolate reductase 677T (MTHFR 677TT. METHODS: 55 consecutive retinal vein occlusion patients and 55 controls matched by age, gender and race, were tested for the presence of the following mutations: factor V Leiden, PT 20210A and MTHFR 677TT. The frequencies of the three mutations in cases and controls were compared. RESULTS: Factor V Leiden was found in 3.6% of patients and in 0% of controls; PT 20210A was found in 1.8% of patients and 3.6% of controls, (matched-pair odds ratio, 0.5; 95% confidence interval, 0.04 to 5.51; MTHFR 677TT was found in 9% of patients and 9% of controls (matched-pair odds ratio, 1; 95% confidence interval, 0.92 to 3.45. Arterial hypertension was more frequent in patients than controls (matched-pair odds ratio, 3.4; 95% confidence interval, 1.25 to 9.21. CONCLUSIONS: This study suggests that thrombophilic mutations are not risk factors for RVO. Routine investigation of hereditary thrombophilia in these patients is not justified.OBJETIVOS: A associação entre oclusão venosa retiniana e trombofilias hereditárias não está estabelecida, com resultados controversos na literatura. O presente estudo investiga a associação entre a oclusão venosa retiniana e três mutações trombofílicas: fator V 1691A (fator V Leiden, protrombina 20210A (PT 20210A e mutação C677T do gene da metileno-tetra-hidro-folato redutase (MTHFR 677TT. MÉTODOS: Cinquenta e cinco pacientes portadores de oclusão venosa retiniana e 55 controles pareados por idade, sexo e raça foram testados para a presença das seguintes mutações: fator V Leiden, PT 20210A e MTHFR 677TT. As freq

  2. Protective and super-imposing effects of Estradiol , Breviscapini, Lumbrokinase In focal cerebral ischemic-reperfusion injury on rats

    Institute of Scientific and Technical Information of China (English)

    Shang huiFang; Luo ZuMing

    2000-01-01

    Baekgrouad: Postmenopausal estrogen replacement therapy is always associated with a decrease in risk of stroke incidence and mortality. The breviscapini and lumbrokinase can protect brain from ischemic injury..Experiments in animal models suggest estradiol can protect brain from ischemic injury. But we know little about the mechanism of brain protection role of estradiol. Espectically we don't know if estrodiol plus other brain protective drugs such as breviscapini and lumbrokinase have superimposing protection. Objective: To study protective function and its mechanism of estradio breviscapini and lumbrokinase after cerebral ischemic reperfusion tn overiectimiaed rat. Methods: We made the local ischemic repenfusion model with thread emgolism of right middle cerebral artery (MCA) of young female SD rars which werc bilaterally ovariectomized two weeks ago were used. These rats were randomized into sham-operated group, control group, estrodiol group, brevisacapini group, 10 SD rats each group. We admistrated each group with reseponsed treatment when rats were subjected to half an hour of MCAO. MCA were oecluded 2 hours and followed by 70 hours of reperfusion. Sham-poerated group were exposed cervical common artery and weren't occluded MCA. The rats were killed at 72 hours. The observed patameters each group listed: 1) The neurological deficit scores were made after MCAO 2 hours and bcforc killed. 2) Each group excluded sham operaated group were caculaled cerebral infarct volume ratio and edema volume with stereological microscoppe. 3) The level of Nitrc oxide (NO) in serum were determined with nitrate redutase method. 4) The level of Interlukin-I (IL-1) and tumor necrosis factot (TNF) were determined by biological activity method. 5) Ultrastructure of neuron were observed with electron microscope. All the rcsults from different groups were compared with each other. The drugs identified effictive in formcr study plus estradiot wcre admistratcd to rats to obscrvcd

  3. Metabolic pathway of oligosaccharides in sorghum honeydew caused by Claviceps africana Rota metabólica de oligossacarídeos em exsudato da "doença açucarada" do sorgo causado por Claviceps africana

    Directory of Open Access Journals (Sweden)

    Amauri Bogo

    2008-08-01

    nutritiva. Parte da frutose não utilizada da sacarose foi reduzida a mannitol por enzimas do patógeno. As enzimas envolvidas também atuaram no exsudato da "doença açucarada", ativando a ação da redutase pela incorporação do 14C manitol. O manitol foi ligado na posição 2 da fructose, formando o dissacarídeo 1-beta-D-fructofuranosyl-D-mannitol e, posteriormente, adicionada mais uma fructose ao mannitol do dissacarídeo, formando o trisacarídeo 1,6-di-beta-D-fructofuranosyl-D-manitol, que se tornou predominate. A formação direta de oligossacarídeos contendo alditol a partir de monossacarídeos parece ser exclusiva em C. africana, contrastando com a ação enzimática da frutose transferase a partir de sucrose para sucrose, a qual é muito comum em outros patógenos tipo "ergot".

  4. Absorção e redução de nitrato em duas cultivares de arroz na presença de alumínio Uptake and reduction of nitrate in two rice cultivars in the presence of aluminum

    Directory of Open Access Journals (Sweden)

    Gilberto Costa Justino

    2006-08-01

    Full Text Available O objetivo deste trabalho foi estudar os efeitos do alumínio (Al sobre a absorção e a redução de nitrato, em duas cultivares de arroz: Fernandes (tolerante e Maravilha (sensível, expostas a 0 e 500 µM de Al. Depois de 21 dias de crescimento, foram determinados o comprimento, a produção de matéria seca, os teores de Al e de nitrato e a atividade in vitro da redutase do nitrato (RN, nas raízes e na parte aérea, bem como as constantes cinéticas de absorção de nitrato pelas raízes. O Al reduziu o crescimento em comprimento e a produção de matéria seca, nas duas partes das plantas, apenas da cultivar Maravilha. Os teores de Al aumentaram nas raízes e parte aérea das plantas nas duas cultivares, enquanto o teor de nitrato sofreu redução apenas nas raízes da cultivar Maravilha. A Vmax não se modificou, enquanto o Km da absorção de nitrato diminuiu cerca de 11% na cultivar Fernandes e aumentou 310% na Maravilha. Em presença de Al, houve redução na atividade da RN nas raízes das duas cultivares, e na parte aérea apenas da cultivar Maravilha. A cultivar tolerante foi mais eficiente na absorção e na redução de nitrato, o que indica que esses processos são importantes componentes da tolerância ao Al em arroz.The objective of this work was to evaluate the effects of Al on the nitrate uptake and reduction in two rice cultivars, Fernandes (Al tolerant and Maravilha (Al sensitive, exposed to 0 and 500 µM Al. After 21 days of Al treatment, growth in size and dry matter yield, Al and nitrate contents and in vitro nitrate reductase activity (NR in the tops and roots, and the kinetic constants of root nitrate uptake were determined. Aluminum reduced the growth and dry matter yield of both plant parts only in the Maravilha cultivar. Aluminum content increased in the tops and roots of both cultivars, while nitrate content decreased only in the roots of Maravilha cultivar. Vmax did not change, while Km of the nitrate uptake

  5. Is MTHFR polymorphism a risk factor for Alzheimer's disease like APOE? Polimorfismo da MTHFR é um fator de risco para demência de Alzheimer como APOE?

    Directory of Open Access Journals (Sweden)

    Liana Lisboa Fernandez

    2005-03-01

    Full Text Available BACKGROUND: The role of methylenetetrahydrofolate reductase (MTHFR gene polymorphisms as risk factors for the occurence of Alzheimer's disease (AD is still controversial: OBJECTIVE: To verify the association between MTHFR and apolipoprotein E (APOE polymorphisms and Alzheimer's disease. METHOD: This work was conducted as a case-control study. Cases included thirty patients with probable AD. Controls were constituted by 29 individuals without dementia according to neuropsychological tests paired to age, sex, race and educational level. DNA was isolated from peripheral leukocytes of anticoagulated venous blood. Genotyping of APOE and MTHFR were performed by DNA amplification and digestion. The frequences of APOE and MTHFR genotypes were submitted by chi-square test corrected by Fisher test; the APOE genotypes, to chi-square linear tendency test and the frequences of MTHFR mutant and AD, by stratificated anlysis adjust by Mantel-Haenszel method. RESULTS: There was significant difference about APOE4 and APOE2 in the groups. (p=0.002 The odds ratio increased exponentially with the increased number of E4 allele (chi2 linear tendency test. No significant difference was detected on MTHFR genotypes in both case and control groups. CONCLUSION: The APOE4 is a risk factor and demonstrated a dose-depenent effect while APOE2 allele conferred a protection to AD. The MTHFR mutation had no correlation with AD.INTRODUÇÃO: O papel do polimorfismo do gene da metilenotetrahidrofolato redutase (MTHFR como um fator de risco para demência de Alzheimer (DA é controverso ainda. OBJETIVO: Verificar a associação entre os polimorfismos da MTHFR e apolipoproteína E (APOE e DA. MÉTODO: O trabalho foi conduzido como um estudo caso-controle. Trinta pacientes com DA provável foram incluídos no grupo caso. Vinte e nove indivíduos sem demência comprovadas por testes neuropsicológicos, emparelhados pela idade, sexo, cor e nível educacional constituíram o grupo

  6. Biochemical and genetic alterations in the freshwater neotropical fish Prochilodus lineatus after acute exposure to Microcystis aeruginosa

    Directory of Open Access Journals (Sweden)

    Cylene Zambrozi Garcia

    2012-09-01

    E. Os resultados mostraram ativação da via de biotransformação devido ao aumento da atividade da CYP1A hepática e da atividade da glutationa S-transferase (GST hepática e branquial. As defesas antioxidantes foram muito afetadas pela exposição a MA levando a alterações, tanto no fígado como nas brânquias, na atividade da superóxido dismutase (SOD, da catalase (CAT, da glutationa peroxidase (GPx, glutationa redutase (GR e no conteúdo do tripeptídeo glutationa (GSH. Apesar dessas alterações a peroxidação lipídica não foi detectada em nenhum dos tecidos, mas danos na molécula de DNA foram observados nas células do sangue. Em conclusão, pode-se afirmar que o liofilizado de MA é capaz de promover alterações em parâmetros bioquímicos e genéticos de P. lineatus

  7. Biochemical and histological characterization of tomato mutants

    Directory of Open Access Journals (Sweden)

    Carolina C. Monteiro

    2012-06-01

    guaiacol peroxidase (GPOX aumentou em ambos os mutantes sitiens (sit e notabilis (not, enquanto no mutante not houve aumento da atividade de ascorbato peroxidase (APX. Baseado nas análises de PAGE, as isoformas III, IV, V e VI de glutationa redutase (GR aumentaram nas folhas do mutante l, enquanto a isoforma III de superóxido dismutase (SOD reduziu nas folhas dos mutantes sit, epi, Never ripe (Nr e green flesh (gf. Análises por microscopia revelaram que os mutantes hp1 e au tiveram um aumento nos espaços intercelulares de folhas, enquanto o mutante sit mostrou um decréscimo. Micrografias eletrônicas revelaram que a superfície foliar dos mutantes au e hp1 apresenta reduzida distribuição de tricomas. A caracterização destes mutantes é essencial para que possam ser utilizados em estudos de desenvolvimento e ecofisiologia, tais como estresses abióticos e bióticos no metabolismo oxidativo.

  8. Protective and super-imposing effects of Estradiol , Breviscapini, Lumbrokinase In focal cerebral ischemic-reperfusion injury on rats

    Institute of Scientific and Technical Information of China (English)

    Shang huiFang; Luo ZuMing

    2000-01-01

    Baekgrouad: Postmenopausal estrogen replacement therapy is always associated with a decrease in risk of stroke incidence and mortality. The breviscapini and lumbrokinase can protect brain from ischemic injury..Experiments in animal models suggest estradiol can protect brain from ischemic injury. But we know little about the mechanism of brain protection role of estradiol. Espectically we don't know if estrodiol plus other brain protective drugs such as breviscapini and lumbrokinase have superimposing protection. Objective: To study protective function and its mechanism of estradio breviscapini and lumbrokinase after cerebral ischemic reperfusion tn overiectimiaed rat. Methods: We made the local ischemic repenfusion model with thread emgolism of right middle cerebral artery (MCA) of young female SD rars which werc bilaterally ovariectomized two weeks ago were used. These rats were randomized into sham-operated group, control group, estrodiol group, brevisacapini group, 10 SD rats each group. We admistrated each group with reseponsed treatment when rats were subjected to half an hour of MCAO. MCA were oecluded 2 hours and followed by 70 hours of reperfusion. Sham-poerated group were exposed cervical common artery and weren't occluded MCA. The rats were killed at 72 hours. The observed patameters each group listed: 1) The neurological deficit scores were made after MCAO 2 hours and bcforc killed. 2) Each group excluded sham operaated group were caculaled cerebral infarct volume ratio and edema volume with stereological microscoppe. 3) The level of Nitrc oxide (NO) in serum were determined with nitrate redutase method. 4) The level of Interlukin-I (IL-1) and tumor necrosis factot (TNF) were determined by biological activity method. 5) Ultrastructure of neuron were observed with electron microscope. All the rcsults from different groups were compared with each other. The drugs identified effictive in formcr study plus estradiot wcre admistratcd to rats to obscrvcd

  9. Brazilian blood donation eligibility criteria for dermatologic patients Critérios brasileiros de elegibilidade à doação de sangue para pacientes dermatológicos

    Directory of Open Access Journals (Sweden)

    Carlos Gustavo Wambier

    2012-08-01

    determinado pela Portaria Ministerial no 1353/2011 do Ministério da Saúde e atuais normas internas do Hemocentro de Ribeirão Preto, um centro de referência regional em procedimentos hemoterápicos. Critérios para inaptidão definitiva: doenças autoimunes (>1 órgão comprometido, antecedente pessoal de câncer diferente de carcinoma basocelular, dermatite atópica ou psoríase graves, pênfigo foliáceo, porfirias, filariose, hanseníase, tuberculose ou paracoccidioidomicose extrapulmonares e uso prévio de etretinato. São condições de inelegibilidade temporária: uso de outros retinóides sistêmicos, corticoides sistêmicos, inibidores da 5-alfa-redutase, vacinas, metotrexato, betabloqueadores, minoxidil, anticonvulsivantes e antipsicóticos. Outras condições responsáveis por inaptidão temporária: acidente ocupacional com material biológico, "piercing", tatuagem, doenças sexualmente transmissíveis, herpes, infecções bacterianas, entre outras. Discussão: Talidomida atualmente não consta na lista de medicações teratogênicas. Apesar do uso da finasterida já ter sido considerada como critério para inaptidão definitiva, de acordo com sua meia-vida curta a restrição atual de 1 mês ainda é demasiadamente longa. Dermatologistas devem ser capazes de aconselhar seus pacientes sobre o momento adequado para doar sangue e discutir o impacto da suspensão de medicações nos resultados do tratamento de forma a respeitar os períodos de restrição designados.

  10. Efeitos neuromusculares in vitro e in vivo do atracúrio e do rocurônio em ratos submetidos a tratamento de sete dias com carbamazepina Efectos neuromusculares in vitro e in vivo del atracurio y del rocuronio en ratones sometidos a tratamiento de siete días con carbamazepina In vitro and in vivo neuromuscular effects of atracurium and rocuronium in rats treated with carbamazepine for seven days

    Directory of Open Access Journals (Sweden)

    Caroline Coutinho de Barcelos

    2008-04-01

    Full Text Available JUSTIFICATIVA E OBJETIVOS: Trata-se de um estudo experimental que investigou in vitro e in vivo o bloqueio neuromuscular produzido pelo rocurônio e atracúrio em ratos tratados com carbamazepina e determinou as concentrações de citocromo P450 e b5 redutase em microssomos hepáticos. MÉTODO: Ratos foram tratados por sete dias com carbamazepina (CBZ - 40 mg.kg-1 pelo método de gavagem e sacrificados no oitavo dia sob anestesia com uretana. As preparações in vitro e in vivo foram montadas de acordo com as técnicas de Bulbring e de Leeuwin e Wolters, respectivamente. As concentrações e doses utilizadas dos bloqueadores nas preparações in vitro e in vivo foram, respectivamente, 20 µg.mL-1 e 0,5 mg.kg-1 para atracúrio (ATC; 4 µg.mL-1 e 0,6 mg.kg-1 para rocurônio (ROC. Cada protocolo teve um n = 5 e as respostas foram observadas por 60 minutos. Os efeitos do ATC e ROC foram avaliados nas preparações de ratos tratados (Cbz t e comparados com os observados nas de ratos não-tratados (CBZst. As concentrações de citocromo P450 e b5 redutase foram determinadas em microssomos isolados de fígados de ratos tratados (CBZt e comparadas com as obtidas em ratos não tratados (CBZst. RESULTADOS: A carbamazepina não alterou a amplitude das respostas musculares; in vitro e in vivo, não houve diferença entre o bloqueio neuromuscular produzido pelo atracúrio nas preparações CBZt versus CBZst; o bloqueio neuromuscular produzido pelo rocurônio nas preparações CBZt foi potencializado in vitro. A carbamazepina não alterou as concentrações de citocromo P450 e b5. CONCLUSÕES: O tratamento por sete dias com carbamazepina não influenciou no bloqueio produzido pelo atracúrio, e alterou in vitro os efeitos do rocurônio. O tempo de tratamento não foi suficiente para causar indução enzimática e diminuir a sensibilidade ao rocurônio.JUSTIFICATIVA Y OBJETIVOS: Se trata de un estudio experimental que investigó in vitro e in vivo el bloqueo

  11. PSEN1 L226F mutation in a patient with early-onset Alzheimer’s disease in Korea

    Directory of Open Access Journals (Sweden)

    Bagyinszky E

    2016-10-01

    Full Text Available Eva Bagyinszky,1,* Sun Ah Park,2,* Hyung Jun Kim,2 Seong Hye Choi,3 Seong Soo A An,1 SangYun Kim4 1Department of BioNano Technology, Gachon University, Seongnam-si, 2Department of Neurology, Soonchunhyang University Bucheon Hospital, Bucheon, 3Department of Neurology, Inha University School of Medicine, Incheon, 4Department of Neurology, Seoul National University College of Medicine & Neurocognitive Behavior Center, Seoul National University Bundang Hospital, Seongnam-si, Republic of Korea *These authors contributed equally to this work Abstract: In this study, we report a first 226leucine (Leu mutation to phenylalanine (Phe in (PSEN1, CTC>TTC, L226F in Asia from a Korean early-onset Alzheimer’s disease (EOAD patient. Polymerase chain reaction (PCR–single strand conformation polymorphism, sequencing, and in silico predictions were performed. Previously, L226F was reported in EOAD patients by Zekanowski et al and Gómez-Tortosa et al. Disease phenotypes appeared in their thirties, and family history was positive in both cases. In our patient, age of onset was similar (37 years of age, but the mutation seemed to be de novo, since no affected family member was found. This leucine to phenylalanine substitution may cause additional stresses inside the transmembrane region due to large aromatic side chain and increased hydrophobic interactions with hydrocarbon chains in the membrane and its binding partners. Clinical phenotype of the mutation was aggressive progression into neurodegeneration, resulting in rapid cognitive decline. One of the patients was initially diagnosed with frontotemporal dementia, but the diagnosis was revised to AD upon postmortem studies in which Aβ plaques were seen. A second mutation, L226R, was found for the L226 residue. Similar to L226F, the patient with L226R also developed the first symptoms in his 30s, but EOAD was diagnosed in his 40s. These findings suggested that L226 might be an important residue in PSEN1

  12. Analysis of gene mutations associated with isoniazid, rifampicin and ethambutol resistance among Mycobacterium tuberculosis isolates from Ethiopia

    Science.gov (United States)

    2012-01-01

    Background The emergence of drug resistance is one of the most important threats to tuberculosis control programs. This study was aimed to analyze the frequency of gene mutations associated with resistance to isoniazid (INH), rifampicin (RMP) and ethambutol (EMB) among Mycobacterium tuberculosis isolates from Northwest Ethiopia, and to assess the performance of the GenoType® MTBDRplus and GenoType® MTBDRsl assays as compared to the BacT/ALERT 3D system. Methods Two hundred sixty Mycobacterium tuberculosis isolates from smear positive tuberculosis patients diagnosed between March 2009 and July 2009 were included in this study. Drug susceptibility tests were performed in the Institute of Medical Microbiology and Epidemiology of Infectious Diseases, University Hospital of Leipzig, Germany. Results Of 260 isolates, mutations conferring resistance to INH, RMP, or EMB were detected in 35, 15, and 8 isolates, respectively, while multidrug resistance (MDR) was present in 13 of the isolates. Of 35 INH resistant strains, 33 had mutations in the katG gene at Ser315Thr 1 and two strains had mutation in the inhA gene at C15T. Among 15 RMP resistant isolates, 11 had rpoB gene mutation at Ser531Leu, one at His526Asp, and three strains had mutations only at the wild type probes. Of 8 EMB resistant strains, two had mutations in the embB gene at Met306Ile, one at Met306Val, and five strains had mutations only at the wild type probes. The GenoType® MTBDRplus assay had a sensitivity of 92% and specificity of 99% for INH resistance, and 100% sensitivity and specificity to detect RMP resistance and MDR. The GenoType® MTBDRsl assay had a sensitivity of 42% and specificity of 100% for EMB resistance. Conclusion The dominance of single gene mutations associated with the resistance to INH and RMP was observed in the codon 315 of the katG gene and codon 531 of the rpoB gene, respectively. The GenoType® MTBDRplus assay is a sensitive and specific tool for diagnosis of resistance to INH

  13. Rapid Screening of MDR-TB in Cases of Extra Pulmonary Tuberculosis Using Geno Type MTBDRplus.

    Directory of Open Access Journals (Sweden)

    Richa Kumari

    Full Text Available Drug resistance in tuberculosis is a major public health challenge in developing countries. The limited data available on drug resistance in extra pulmonary tuberculosis stimulated us to design our study on anti-tuberculosis drug resistance pattern in cases of extra pulmonary tuberculosis in a tertiary referral hospital of North India. We performed Geno Type MTBDRplus assay in comparison with conventional drug susceptibility testing by proportion method to study the mutation patterns in rpoB, katG and inhA genes.A total of 510 extra pulmonary samples were included in this study. After the smear microscopy, all the specimens were subjected for culture on Lowenstein Jensen (LJ media. Phenotypic drug susceptibility testing (DST was performed on LJ media for all the MTB isolates and compared with the results of Geno Type MTBDRplus assay which was performed with the DNA isolated from the culture by conventional method.Of 510 specimens cultured, the total culture positivity obtained was 11.8% (60 encompassing 54 (10.6% Mycobacterium tuberculosis and 6 (1.2% non-tubercular mycobacteria (NTM. DST results by Geno Type MTBDRplus assay and solid culture methods were compared in 51 MTB isolates excluding the two Rif indeterminate and one invalid test. Geno Type MTBDRplus accurately identified 13 of 14 rifampicin-resistant strains, 14 of 15 isoniazid-resistant strains and 13 of 14 as multi drug resistant tuberculosis (MDR-TB in comparison with conventional method. Sensitivity and specificity were 92.86% and 97.30% respectively for detection of RIF resistance, 93.33% and 94.44% respectively for detection of INH resistance, 92.86% and 97.30% respectively for detection of MDR-TB, while the overall concordance of Geno Type MTBDRplus assay with conventional DST was 94.11%. The turn-around time for performing Geno Type MTBDRplus assay test was 48 hours.The problem of MDR in extra pulmonary tuberculosis (EPTB cannot be overlooked and due attention on patients

  14. Estudos aplicados à recuperação biológica da Baía de Guanabara

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    Lejeune P. H. de Oliveira

    1976-01-01

    Full Text Available Continuamos observações sobre as biocenoses litorâneas, na parte oeste do Rio de Janeiro, na Enseada mais poluída, onde está a ilha do Pinheiro. Chegamos à conclusão que há graus de estragos acima dos verificados em publicações anteriores, os 7º, 8º, 9º e 10º graus de estragos, com desaparecimento de todas cinturas vegetais. Anotamos os assoreamentos; devido a aterros artificiais na vizinhança, a lama subiu muito, meio metro em 20 anos, ou seja 2,5 centímetros por ano. Apresentamos as alterações ecológicas resultantes do assoreamento, mostramos a mudança de vários nichos de alguns componentes do manguezal. Fizemos 4 mapas ecológicos, em local que constitui a média representativa do que se passou na Enseada de Inhaúma nesses últimos 5 anos. Chegamos a índices de recuperação de zonações do manguezal, como 60/1 para o Avicennietum; e nenhum para o Laguncularietum, comunidade de "mangues mansos" que está em regressão; Houve regeneração das populações de caranguejos Ucides e Cardisoma que voltaram a habitar a Ilha do Pinheiro. Mostramos uma correlação entre o guaiamu e a aroeira da praia, Schinus. Concluímos que: podem ser usadas as técnicas de recuperação de poluição para esses caranguejos e guaiamus, para a alimentação humana, assim como para peixes apanhados em alagados de manguezais poluídos. O material pescado no Rio de Janeiro, tendo algas do gênero Oscillatoria, apresenta um cheiro de urina pútrida, bem como outros cheiros, mas pode ser recuperado para a alimentação humana. Damos sugestões para se utilizar cercados de vegetação de manguezal contra a poluição em locais apropriados, alagados com camarões e peixes, como usam-se na Ásia. As plantas utilizadas baseiam-se no estudo ecológico que estabelece o nicho apropriado a cada uma delas, resistente á poluição, e que pode combater alguns estragos; entre elas as Iresine e as do gênero Avicennia, pois elas promoveram uma pequena

  15. Analysis of gene mutations associated with isoniazid, rifampicin and ethambutol resistance among Mycobacterium tuberculosis isolates from Ethiopia

    Directory of Open Access Journals (Sweden)

    Tessema Belay

    2012-02-01

    Full Text Available Abstract Background The emergence of drug resistance is one of the most important threats to tuberculosis control programs. This study was aimed to analyze the frequency of gene mutations associated with resistance to isoniazid (INH, rifampicin (RMP and ethambutol (EMB among Mycobacterium tuberculosis isolates from Northwest Ethiopia, and to assess the performance of the GenoType® MTBDRplus and GenoType® MTBDRsl assays as compared to the BacT/ALERT 3D system. Methods Two hundred sixty Mycobacterium tuberculosis isolates from smear positive tuberculosis patients diagnosed between March 2009 and July 2009 were included in this study. Drug susceptibility tests were performed in the Institute of Medical Microbiology and Epidemiology of Infectious Diseases, University Hospital of Leipzig, Germany. Results Of 260 isolates, mutations conferring resistance to INH, RMP, or EMB were detected in 35, 15, and 8 isolates, respectively, while multidrug resistance (MDR was present in 13 of the isolates. Of 35 INH resistant strains, 33 had mutations in the katG gene at Ser315Thr 1 and two strains had mutation in the inhA gene at C15T. Among 15 RMP resistant isolates, 11 had rpoB gene mutation at Ser531Leu, one at His526Asp, and three strains had mutations only at the wild type probes. Of 8 EMB resistant strains, two had mutations in the embB gene at Met306Ile, one at Met306Val, and five strains had mutations only at the wild type probes. The GenoType® MTBDRplus assay had a sensitivity of 92% and specificity of 99% for INH resistance, and 100% sensitivity and specificity to detect RMP resistance and MDR. The GenoType® MTBDRsl assay had a sensitivity of 42% and specificity of 100% for EMB resistance. Conclusion The dominance of single gene mutations associated with the resistance to INH and RMP was observed in the codon 315 of the katG gene and codon 531 of the rpoB gene, respectively. The GenoType® MTBDRplus assay is a sensitive and specific tool for

  16. Whole genome sequencing-based characterization of extensively drug resistant (XDR) strains of Mycobacterium tuberculosis from Pakistan

    KAUST Repository

    Hasan, Zahra

    2015-03-01

    Objectives: The global increase in drug resistance in Mycobacterium tuberculosis (MTB) strains increases the focus on improved molecular diagnostics for MTB. Extensively drug-resistant (XDR) - TB is caused by MTB strains resistant to rifampicin, isoniazid, fluoroquinolone and aminoglycoside antibiotics. Resistance to anti-tuberculous drugs has been associated with single nucleotide polymorphisms (SNPs), in particular MTB genes. However, there is regional variation between MTB lineages and the SNPs associated with resistance. Therefore, there is a need to identify common resistance conferring SNPs so that effective molecular-based diagnostic tests for MTB can be developed. This study investigated used whole genome sequencing (WGS) to characterize 37 XDR MTB isolates from Pakistan and investigated SNPs related to drug resistance. Methods: XDR-TB strains were selected. DNA was extracted from MTB strains, and samples underwent WGS with 76-base-paired end fragment sizes using Illumina paired end HiSeq2000 technology. Raw sequence data were mapped uniquely to H37Rv reference genome. The mappings allowed SNPs and small indels to be called using SAMtools/BCFtools. Results: This study found that in all XDR strains, rifampicin resistance was attributable to SNPs in the rpoB RDR region. Isoniazid resistance-associated mutations were primarily related to katG codon 315 followed by inhA S94A. Fluoroquinolone resistance was attributable to gyrA 91-94 codons in most strains, while one did not have SNPs in either gyrA or gyrB. Aminoglycoside resistance was mostly associated with SNPs in rrs, except in 6 strains. Ethambutol resistant strains had embB codon 306 mutations, but many strains did not have this present. The SNPs were compared with those present in commercial assays such as LiPA Hain MDRTBsl, and the sensitivity of the assays for these strains was evaluated. Conclusions: If common drug resistance associated with SNPs evaluated the concordance between phenotypic and

  17. Molecular detection of multidrug-resistant tuberculosis among smear-positive pulmonary tuberculosis patients in Jigjiga town, Ethiopia

    Science.gov (United States)

    Brhane, Mussie; Kebede, Ameha; Petros, Yohannes

    2017-01-01

    Background Molecular methods that target drug resistance mutations are suitable approaches for rapid drug susceptibility testing to detect multidrug-resistant tuberculosis (MDR-TB). The aim of the study was to determine MDR-TB cases and to analyze the frequency of gene mutations associated with rifampicin (RIF) and/or isoniazid (INH) resistance of Mycobacterium tuberculosis among smear-positive pulmonary tuberculosis patients. Methods Institution-based cross-sectional study design was employed. Sputum specimens were collected, and using a pretested questionnaire, data for associated risk factors for drug resistance were collected from 105 consecutive smear-positive pulmonary tuberculosis patients in Karamara General Hospital. Specimens were transported to Harar Health Research and Regional Laboratory, Harar, where molecular drug susceptibility testing was performed using GenoType® MTBDRplus assay. Results Of the total 105 sputum specimens, 98 (93.3%) gave interpretable results, in which 67 (68.4%) were new cases and 31 (31.6%) were previously treated cases. Of these, 80 (81.6%) were sensitive to both drugs and 18 (18.4%) were resistant to RIF and/or INH. The prevalences of MDR-TB in total cases, new, and previously treated cases were 10 (10.2%), 3 (4.5%), and 7 (22.6%), respectively. Among the ten total RIF-resistant specimens, eight (80%) had resulted because of absence of rpoB WT8 and presence of MUT3 and in all specimens, the amino acids changed were Ser531Lue. Of the 18 total INH-resistant specimens, 15 (83.3%) had mutations in the katG gene (katG MUT1, Ser315Thr1), indicating high-level resistance, while 3 (14.7%) had mutations in the inhA promoter gene (Cys15Thr), indicating low-level resistance. Conclusion Among the mutations associated with resistance to RIF and INH, the majority were in codon 531 of the rpoB gene and codon 315 of the katG gene. Relatively high prevalence of MDR-TB was observed in the study.

  18. Family quality of life among families of children with atopic dermatitis

    Science.gov (United States)

    Jang, Hae Ji; Hwang, Seonyeong; Ahn, Youngmee; Lim, Dae Hyun

    2016-01-01

    Background Atopic dermatitis (AD) may cause emotional distress and impairs the quality of life (QoL) in children and their families. Objective We examined family QoL of children with AD and explored associated factors such as disease severity and psychosocial factors among parents of children with AD. Methods Study participants were 78 children (1 month to 16 years old) diagnosed with AD and their parents visiting an outpatient clinic of the Department of Pediatrics in Inha University Hospital. Data were collected using structured questionnaires and medical record review. Parents completed the Dermatitis Family Impact questionnaire (DFI), the Positive Affect and Negative Affect Schedule, the Satisfaction with Life Scale, and the Korean Parenting Stress Index. For children aged below 6-year-old, parents were asked to complete the Infants' Dermatologic Quality of Life. SCOring Atopic Dermatitis (SCORAD), Children's Dermatology Life Quality Index, and the Pediatric Quality of Life Inventory version 4.0 Generic Core Scale were also completed. Results The mean age of parents and children were 37.4 ± 5.3 years and 65.1 ± 45.7 months, respectively. Among them, 87.2% of parents were mothers and 60.3% of children were boys. The mean score of DFI was 11.2 ± 6.0. The mean SCORAD score was 28.3 ± 16.1. Family who experienced strong negative emotionality had a 3.8 times higher probability of experiencing a lower QoL than parents who did not (odds ratio [OR], 3.82; p = 0.041). Family of children with higher severity of AD had a 6.6 times (OR, 6.55; p = 0.018) higher probability of experiencing a low family QoL than their less-severe counterparts. Families of girls with AD had a lower QoL (OR, 8.40; p = 0.003) than families of boys. Conclusion Family QoL among parents of children with AD was low and associated with parent’s psychosocial characteristics as well as disease severity of the children. Considering parental involvement in AD management for children, emotional

  19. Investigation of the influence factors of oxygen inhalation through nasal tube and the intervention effect of health education on patients in CCU ward%冠心病重症监护病房患者拒绝鼻导管吸氧的影响因素及健康教育效果

    Institute of Scientific and Technical Information of China (English)

    刘文静; 王阳

    2015-01-01

    目的对影响冠心病重症监护病房(CCU)患者拒绝鼻导管吸氧的影响因素进行调查分析,并探讨健康信念模式的健康教育方式对患者鼻导管吸氧的影响。方法选取80例 CCU 鼻导管吸氧患者为研究对象,自身前后对照,记录患者1天吸氧情况;第2天对患者相关情况进行调查分析后实施健康教育,并再次观察吸氧情况。比较教育前后患者拒绝吸氧的时间。结果不同年龄、性别、文化程度、手术史、心功能等级患者的吸氧情况差异有统计学意义(P <0.05),实施健康教育后患者拒绝吸氧的时间少于实施前(Z =-6.029,P <0.01)。结论健康信念模式的健康教育方式可减少 CCU 鼻导管吸氧患者拒绝吸氧的时间,改善患者吸氧的效果。%Objective To investigate the influence factors of oxygen inhalation through nasal tube and tinter-vention effect of health education.Method The investigation was conducted in the patients undergoing oxygen inhala-tion through nasal tube in CCU ward.The time of daily oxygen inhalation was recorded.After the health education,The before -after study wa performed and the rejection time of oxygen inhalation was compared.Results The oxygen inha-lation was significantly different in age,sex,education state,surgeon history and the level of cardiac function(P <0.05).The rejection time of Oxygen inhalation was significantly reduced after health education(Z =-6.029,P <0.01).Conclusion Health education could reduce the rejection time of oxygen inhalation and improve the effects of oxygen inhalation.

  20. Health Risk Assessment of the Arsenic in Soil and Groundwater around A Gold Ore%某金矿周边土壤和地下水中砷的健康风险评价

    Institute of Scientific and Technical Information of China (English)

    鲁敏; 李荟敏; 贺小敏; 李爱民; 范俊楠

    2015-01-01

    以湖北某含砷金矿污染场地为例,依据《污染场地风险评估技术导则》(HJ 25.3—2014),对其周边土壤和地下水中砷元素进行健康风险评价,结果表明:该金矿周边土壤和地下水中砷分别通过经口摄入土壤、皮肤接触土壤和吸入土壤颗粒物、饮用地下水4种暴露途径的致癌风险均超过可接受风险水平(10-6),对周边居民造成潜在危害。计算得到经口摄入、皮肤接触和吸入途径基于致癌效应的土壤风险控制值分别为1.6 mg /kg、9.3 mg /kg 和12.1 mg /kg,应予以重视,并建议对矿山进行综合治理和目标值修复。%The health risks of arsenic in soil and groundwater near an arsenic-bearing gold ore in Hubei province was investigated in this work,which was carried out according to the state environmental protection de-partment of contaminated sites risk assessment technology guideline(HJ 25.3—201 4).The results indicates that the carcinogenic risk of the four paths including oral intake of soil and underground water,skin contact and inha-lation of soil particles exceed the acceptable risk level(1 0 -6 ),which could cause potential damage to the people. The calculated risk control value for the oral intake of soil,skin contact and inhalation of soil particles are 1 .6 mg /kg、9.3 mg /kg and 1 2.1 mg /kg,respectively.The results indicated that attention should be paid to the mine comprehensive treatment and target remediation.

  1. FIND Tuberculosis Strain Bank: a Resource for Researchers and Developers Working on Tests To Detect Mycobacterium tuberculosis and Related Drug Resistance.

    Science.gov (United States)

    Tessema, Belay; Nabeta, Pamela; Valli, Eloise; Albertini, Audrey; Collantes, Jimena; Lan, Nguyen Huu; Romancenco, Elena; Tukavdze, Nestani; Denkinger, Claudia M; Dolinger, David L

    2017-04-01

    The spread of multidrug-resistant (MDR) tuberculosis (TB) and extensively drug-resistant (XDR) TB hampers global efforts in the fight against tuberculosis. To enhance the development and evaluation of diagnostic tests quickly and efficiently, well-characterized strains and samples from drug-resistant tuberculosis patients are necessary. In this project, the Foundation for Innovative New Diagnostics (FIND) has focused on the collection, characterization, and storage of such well-characterized reference materials and making them available to researchers and developers. The collection is being conducted at multiple centers in Southeast Asia, South America, Eastern Europe, and soon the sub-Saharan Africa regions. Strains are characterized for their phenotypic resistances and MICs to first-line drugs (FLDs) and second-line drugs (SLDs) using the automated MGIT 960 system following validated procedures and WHO criteria. Analysis of resistance-associated mutations is done by whole-genome sequencing (WGS) using the Illumina NextSeq system. Mycobacterial interspersed repetitive-unit-variable-number tandem-repeat analysis and WGS are used to determine strain lineages. All strains are maintained frozen at -80°C ± 10°C as distinct mother and daughter lots. All strains are extensively quality assured. The data presented here represent an analysis of the initial part of the collection. Currently, the bank contains 118 unique strains with extracted genomic DNA and matched sputum, serum, and plasma samples and will be expanded to a minimum of 1,000 unique strains over the next 3 years. Analysis of the current strains by phenotypic resistance testing shows 102 (86.4%), 10 (8.5%), and 6 (5.1%) MDR, XDR, and mono/poly resistant strains, respectively. Two of the strains are resistant to all 11 drugs that were phenotypically tested. WGS mutation analysis revealed FLD resistance-associated mutations in the rpoB, katG, inhA, embB, embA, and pncA genes; SLD resistance in the gyrA, gyr

  2. Molecular detection of rifampin and isoniazid resistance to guide chronic TB patient management in Burkina Faso

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    Pinsi Gabriele

    2009-08-01

    were the H526D (15.6%, the H526Y (15.6%, and the S531L (9.4%. All isoniazid-resistant cases (n 36 identified by the molecular assay were carrying a S315T substitution in the katG gene. In 41.7% of cases, a mutation affecting the promoter region of the inhA gene was also detected. Conclusion The GenoType® MTBDRplus assay performed directly on sputum specimens improves the management of chronic TB cases allowing more appropriate anti-TB regimens.

  3. 液相芯片快速检测结核耐药基因方法建立%Rapid detection of drug-resistant genes of clinical isolates in Mycobatterium tuberculosis by Multi-analyte Suspension Array

    Institute of Scientific and Technical Information of China (English)

    王清; 丁显平; 李天俊; 罗涛; 韩婷婷

    2012-01-01

    基于Luminex100TM技术平台,结合多重PCR技术,以5’生物素标记引物为信号指示,在H37 RV结核标准株做对照下,对104株结核分枝杆菌分离株的LFP耐药rpoB基因和INH耐药的katG,inhA基因进行检测,并与药敏试验绝对浓度法和测序结果比较.结果表明:液相芯片检出结果与药敏法比较,104株分离株中LFP、INH、多重耐药的检出结果分别为:液态芯片检测73株、50株、48株;药敏检测结果为74株、58株、53株;对比检出率98.6%,86.2%,90.6%.液相基因芯片技术对耐药,耐多药结核的检测,具有特异性高,敏感性强的特点,是检测结核分枝杆菌耐药基因的有效方法,可用于临床检测.%104 Mycobacterium tuberculosis strains and H37 Rv strain as control were genotyped by muti-PCR based on Luminex 100?platform for detecting the mutant sequences in rpoB, katG and inhA gene to identify the result of the drug resistance. Of 104 strains, Mutations were found in 73 strains (98. 6%) of 74 randomly selected rifampin-resistant MTB , 50 strains (86. 2%) of 58 randomly selected isoniazid-resistant MTB and 48 strains (90. 6%) of 53 randomly selected both rifampin and isoniazid-resistant MTB by MASA. Multi-analyte Suspension Array technology has high sensitivity and specificity in detection of drug-resistant of MTB and may be applied in clinical diagnosis.

  4. Detecting mutation pattern of drug-resistant Mycobacterium tuberculosis isolates in Himachal Pradesh using GenoType® MTBDRplus assay

    Directory of Open Access Journals (Sweden)

    C Thakur

    2015-01-01

    Full Text Available Context: Tuberculosis (TB is a major public health problem in India and a principal cause of death in adults, especially among the economically productive age group. India accounts for one-fifth of the global burden of TB. It is estimated that about 40% of Indian population is infected with TB bacillus. The GenoType® MTBDRplus molecular method allows rapid diagnosis of the clinical samples and detection of the most common mutations in the genes associated with rifampicin (R and isoniazid (H resistance. Aims: To study the drug resistance and mutational patterns in multidrug-resistant (MDR suspects clinical strains using GenoType® MTBDRplus assay. Subjects and Methods: A total of 770 sputum samples of the MDR-TB suspects were included in this study, which were received at Intermediate Reference Laboratory, Government TB Sanatorium, Dharampur, Solan, Himachal Pradesh from the Designated Microscopy Centres of Himachal Pradesh for the culture and susceptibility testing. All the 521 Mycobacterium tuberculosis complex (MTBC strains were subjected to GenoType® MTBDRplus (HAIN Lifescience assay to detect molecular resistance pattern to first line anti-tubercular drugs (isoniazid and rifampicin. Results: Of 770 samples, 556 (72.20% were from male and 214 (27.80% were from female. Among the 521 MTBC strains, 19.76% were found to be MDR and mono-resistance to isoniazid and rifampicin was detected in 8.63% and 6.14% strains respectively. About 74.81%, 76.35% and 5.40% strains harboured known mutation in rpoB, katG and inhA genes respectively. Conclusions: In rpoB gene, the most common mutation is associated with S531 L region. The GenoType® MTBDRplus assay is a rapid test for the detection of the most common mutations in MDR-TB strains. In our study, unknown rpoB gene mutations were found in 25.18% strains that may further be detected by gene sequencing.

  5. Mycobacterium tuberculosis spoligotypes and drug susceptibility pattern of isolates from tuberculosis patients in South-Western Uganda

    Directory of Open Access Journals (Sweden)

    Bwanga Freddie

    2011-03-01

    showed the inhA promoter gene mutation. Conclusion The present study shows that the TB epidemic in Mbarara is caused by modern M. tuberculosis strains mainly belonging to the Uganda genotype and anti-TB drug resistance rate in the region is low.

  6. Activin A balances Sertoli and germ cell proliferation in the fetal mouse testis.

    Science.gov (United States)

    Mendis, Sirisha H S; Meachem, Sarah J; Sarraj, Mai A; Loveland, Kate L

    2011-02-01

    Activin affects many aspects of cellular development, including those essential for reproductive fitness. This study examined the contribution of activin A to murine fetal testicular development, revealing contrasting outcomes of activin actions on Sertoli cells and gonocytes. Shortly after sex determination, from Embryonic Day 12.5 (E12.5) through to birth (0 dpp), the activin A subunit transcript (Inhba) level rises in testis but not ovary, followed closely by the Inha transcript (encoding the inhibitory inhibin alpha subunit). Activin receptor transcript levels also change, with Acvr1 (encoding ALK2) and Acvr2b (ActRIIB) significantly higher and lower, respectively, at 0 dpp compared with E13.5 and E15.5. Transcripts encoding the signaling mediators Smad1, Smad3, and Smad4 were higher at 0 dpp compared with E13.5 and E15.5, whereas Smad2, Smad5, and Smad7 were lower. Detection of phosphorylated (P-)SMAD2/3 in nearly all testis cell nuclei indicated widespread transforming growth factor beta (TGFB) and/or activin ligand signaling activity. In contrast to wild-type littermates, activin betaA subunit knockout (Inhba(-/-)) mice have significantly smaller testes at birth, attributable to a 50% lower Sertoli cell number and decreased Sertoli cell proliferation from E13.5. Inhba(-/-) testes contained twice the normal gonocyte number at birth, with some appearing to bypass quiescence. Persistence of widespread P-SMAD2/3 in Inhba(-/-) cells indicates other TGFB superfamily ligands are active in fetal testes. Significant differences in Smad and cell cycle regulator transcript levels correlating to Inhba gene dosage correspond to differences in Sertoli and germ cell numbers. In Inhba(-/-) testes, Cdkn1a (encoding p21(cip1)), identified previously in fetal gonocytes, was lower at E13.5, whereas Cdkn1b (encoding p(27kip1) in somatic cells) was lower at birth, and cyclin D2 mRNA and protein were lower at E15.5 and 0 dpp. Thus, activin A dosage contributes to establishing the

  7. Evaluation of usefulness of portal image using Electronic Portal Imaging Device (EPID) in the patients who received pelvic radiation therapy

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Woo Chul; Kim, Heon Jong; Park, Seong Young; Cho, Young Kap; Loh, John J. K. [College of Medicine, Inha Univ., Pusan (Korea, Republic of); Park, Won; Suh, Chang Ok; Kim, Gwi Eon [College of Medicine, Yonsei Univ., Seoul (Korea, Republic of)

    1998-12-01

    To evaluate the usefulness of electronic portal imaging device through objective compare of the images acquired using an EPID and a conventional port film. From Apr. to Oct. 1997, a total of 150 sets of images from 20 patients who received radiation therapy in the pelvis area were evaluated in the Inha University Hospital and Severance Hospital. A dual image recording technique was devised to obtain both electronic portal images and port film images simultaneously with one treatment course. We did not perform double exposure. Five to ten images were acquired from each patient. All images were acquired from posteroanterior (PA) view except images from two patients. A dose rate of 100-300 MU/min and a 10-MV X-ray beam were used and 2-10 MUs were required to produce a verification image during treatment. Kodak diagnostic film with metal/film imaging cassette which was located on the top of the EPID detector was used for the port film. The source to detector distance was 140 cm. Eight anatomical landmarks (pelvic brim, sacrum, acetabulum, iliopectineal line, symphysis, ischium, obturator foramen, sacroiliac joint) were assessed. Four radiation oncologist joined to evaluate each image. The individual landmarks in the port film or in the EPID were rated-very clear (1), clear (2), visible (3), notclear (4), not visible (5). Using an video camera based EPID system, there was no difference of image quality between no enhanced EPID images and port film images. However, when we provided some change with window level for the portal image, the visibility of the sacrum and obturator foramen was improved in the portal images than in the port film images. All anatomical landmarks were more visible in the portal images than in the port film when we applied the CLAHE mode enhancement. The images acquired using an matrix ion chamber type EPID were also improved image quality after window level adjustment. The quality of image acquired using an electronic portal imaging device was

  8. 实时聚合酶链反应熔解曲线法快速检测耐多药结核分枝杆菌%Rapid detection of rifampin- and isoniazid-resistant Mycobacterium tuberculosis using real-time PCR and melting curve analysis

    Institute of Scientific and Technical Information of China (English)

    王峰; 崔运勇; 胡思玉; 桂静; 杨慧; 李庆阁; 刘小立

    2011-01-01

    Objective To evaluate the application of a real-time PCR and melting curve analysis assay for rapid detection of rifampin and isoniazid resistance in Mycobacterium tuberculosis ( M.tuberculosis).Methods A total of 311 clinical isolates of M.tuberculosis obtained from Shenzhen Center for Chronic Disease Control (SZCCC) were included in the study.These isolates were collected originally from national baseline survey on drug-resistant tuberculosis,project for drug resistance surveillance in Shenzhen and clinical patients in SZCCC between 2007 and 2009.rpoB gene resistance-determining region,ahpC promoter (-44 to -30 and -15 to -3),inhA promoter (-17 to -8),inhA 94 and katG 315 were detected by melting curve analysis after real-time PCR,and the results were compared with that of proportion method and DNA sequencing.The performances of the assay in detecting the resistance of rifampin and isoniazid were compared to that of reference proportion method drug susceptibility test.Results Real-time PCR and melting analysis was a closed-tube assay that could be completed within 2 -3 h.Compared to the results of the proportion method,the sensitivity,specificity and accuracy of the assay for rifampin resistance were 97.8%,97.1% and 97.4% respectively,and for isoniazid resistance were 86.6%,98.7% and 92.6% respectively.Conclusions Real-time PCR and melting analysis is a rapid,accurate and closed-tube method that can be used as a screening test for rapid identification of Multidrug-resistant tuberculosis.%目的 应用实时PCR熔解曲线法快速检测MTB对利福平和异烟肼的耐药性,并与比例法检测结果进行比较.方法 收集深圳市2007-2009年耐药监测项目、全国耐药基线调查项目和深圳市慢性病防治中心门诊患者临床分离株311株,采用实时PCR熔解曲线法检测rpoB基因耐药突变热点区、ahpC启动子区(-44 -30及-15~3位点)、inhA启动子区(-17~8位点)、inhA 94和katG 315位耐药突变,并与

  9. EDITORIAL: Welcome to the 2012 volume Welcome to the 2012 volume

    Science.gov (United States)

    Garcia, Ephrahim

    2012-01-01

    articles last year, as well as our authors and contributors for submitting their high quality research. We very much look forward to your further involvement with the journal in 2012. Finally, I would like to extend a huge thank you to our diverse Board of Associate Editors who have devoted their time and energy giving invaluable advice on each submitted article, and have been dedicated ambassadors for SMS. A special welcome to Professor Kwang J Kim who joined the Board a few months ago. Professor D Abbott, University of Adelaide, Australia Professor G Akhras, Royal Military College of Canada, Ontario, Canada Professor C Boller, University of Saarland, Saarbrücken, Germany; Fraunhofer-Institut für Zerstörungsfreie Prüfverfahren, Dresden, Germany Professor Cagnol, École Centrale Paris, France Professor G Carman, University of California-Los Angeles, USA Professor S-B Choi, Inha University, Incheon, Korea Dr S H Choi, NASA Langley Research Center, Hampton, VA, USA Professor A Del Grosso, Università degli Studi di Genova, Italy Professor D Erickson, Cornell University, Ithaca, NY, USA Professor A Flatau, University of Maryland, College Park, USA Professor U Gabbert, Universität Magdeburg, Germany Professor A Güemes, Universidad Politecnica de Madrid, Spain Professor S Gopalakrishnan, Indian Institute of Science, Bangalore, India Professor J Kim, Inha University, Incheon, Korea Professor K J Kim, University of Nevada, Reno, USA Professor S J Kim, Seoul National University, Korea Professor D Lagoudas, Texas A&M University, College Station, USA Professor R Lammering, Helmut-Schmidt-Universität/Universität der Bundeswehr Hamburg, Hamburg, Germany Professor C K Lee, National Taiwan University, Taiwan Professor J Leng, Harbin Institute of Technology, China Professor W Li, University of Wollongong, Australia Professor W H Liao, Chinese University of Hong Kong, China Professor C S Lynch, University of California-Los Angeles, USA Dr S Masri, University of Southern California

  10. 基因芯片法用于检测长沙地区结核分枝杆菌耐药性研究%Identification of drug-resistance gene type in Mycobacterium tuberculosis by gene chip in Hunan province

    Institute of Scientific and Technical Information of China (English)

    邓叶华; 向延根; 马小华; 石国民; 喻容; 彭雪峰

    2015-01-01

    Objective To investigate the drug resistance of 1 031 Mycobacterium tuberculosis to rifampicin and isoniazide in the Center Hospital of Changsha from January 1 ,2013 to September 30 ,2014 .Methods A total of 1 031 strains with positive culture result and identified as strains of Mycobacterium tuberculosis were used absolute concentration method to do the conventional drug susceptibility ,and detected rifampicin and isoniazide resistance gene including rpoB ,katG and inhA gene locus mutation by chip technology ,the results of two methods were compared using card square test statistics .Results By gene chip method ,the sensitive strain of rifampicin was 896 ,the drug-resistant strains was 135 ,the sensitive strains of isoniazide was 901 strains ,130 drug-resistant strains .Compared with the absolute concentration method ,resistance chip detection results were consistent with rifampicin resistant strains 1 011 strains(including 894 drug-resistant strains ,and 117 sensitive strains) ,the coincidence rate was 98 .00% ,consistent with isoniazideresistant strains 1 005 strains(including 890 drug-resistant strains ,115 sensitive strains) ,the coincidence rate was 97 .48% .The most common spot of rifampin resistance related mutations of rpoB gene was 531TCG to TTG ,accounted for 51 .11% ,followed by 526CAC→TAC ,accounted for 10 .37% ,11 strains with 526TCG to TTG ,accounted for 8 .15% .Isoniazid re-sistance was caused by mutations in katG315AGC→ACC resistant strains ,accounted for 83 .85% ,inhA-15C→T mutations accoun-ted for only 12 .30% .Conclusion The results of gene chip method is highly consistent with that of absolute concentration method , could be a fast and effective method for screening rifampicin and isoniazide ,the resistant gene of Mycobacterium tuberculosis to rif-ampicin and isoniazide almost mutate in rpoB531 ,526 and katG315 in Changsha .%目的:了解长沙市中心医院2013年1月1日至2014年9月30日临床分离的1031株结核分枝杆菌对

  11. First evaluation in Argentina of the GenoType® MTBDRplus assay for multidrug-resistant Mycobacterium tuberculosis detection from clinical isolates and specimens Primera evaluación en Argentina de GenoType® MTBDRplus para la detección de Mycobacterium tuberculosis multidrogo-resistente desde aislamientos y especímenes clínicos

    Directory of Open Access Journals (Sweden)

    Belén R Imperiale

    2012-12-01

    Full Text Available Tuberculosis (TB and multidrug and extensively drug-resistant (DR TB are important public health problems that are spreading worldwide. The aims of this study were to determine the sensitivity and specificity of the GenoType® MT BDRplus assay from smear-positive clinical specimens and isolates and to explore its possible application in routine work. Clinical samples were previously decontaminated using NaOH-N-acetyl-L-cystein or NaOH-ClNa hypertonic solution for Ziehl-Neelsen staining and cultures. The leftover sediments of smear-positive samples were stored at -20 ºC, 70 of which were selected to be included in this study according to their DR profile. Thirty DR Mycobacterium tuberculosis isolates were also assessed. Sequencing was used as gold standard to detect mutations conferring isoniazid (INH and rifampicin (RIF resistance. Valid results were obtained in 94.0 % of the samples and 85.5 % (53/62 of the INH-R samples were properly identified. Mutations in the katGS315T gene and inhA C-15T gene promoter region were present in 59.7 % (37/62 and 25.8% (16/62 of the INH-R samples, respectively. The system could also identify 97.7 % (41/42 of the RIF-R samples; the mutations found were rpoBS531L (66.7 %, 28/42, D516V (19.0 %, 8/42, H526Y and S531P/W (4.8 %, 2/42 each one, and S522L/Q (2.4 %, 1/42. A 98.8 % concordance between the GenoType assay and sequencing was obtained. GenoType® MT BDRplus has demonstrated to be easy to implement and to perform in clinical laboratories and useful for a rapid detection of DR M. tuberculosis from decontaminated sputa and clinical isolates. Therefore, this assay could be applied as a rapid tool to predict INH-R and/or RIF-R in DR risk cases.La tuberculosis (TBC, y la TBC multi y extensivamente drogo-resistentes (DR son importantes problemas de salud pública mundial. El objetivo de este estudio fue determinar la sensibilidad y especificidad del sistema GenoType® MT BDRplus a partir de esputos (baciloscop

  12. First line antituberculosis drugs and the molecular mechanism of drug resistance in Mycobacterium tuberculosis%抗结核一线药物及结核分枝杆菌的耐药分子机制

    Institute of Scientific and Technical Information of China (English)

    贾平平; 余利岩; 岑山

    2011-01-01

    近年来涌现出了耐多药结核病(Multidrug-resistanttuberculosis,MDR-TB)/广泛耐药结核病(Extensively drug-resistant tuberculosis,XDR-TB)。MDR-TB是指由耐异烟肼和利福平两种或以上抗结核药物的结核分枝杆菌引起的结核病。XDR-TB是指不仅对一线抗结核药物耐药,而且对主要的二线抗结核药物中的至少3种也耐药的结核分枝杆菌引起的结核病。抗结核的一线药物分别为异烟肼、利福平、吡嗪酰胺、链霉素和乙胺丁醇。与异烟肼的耐药相关的基因主要有katG,inhA等。它们的突变会引起结核分枝杆菌对异烟肼的耐药。rpoB、pncA及embB基因的突变分别是导致结核分枝杆菌对利福平、吡嗪酰胺、乙胺丁醇产生耐药性的主要原因。耐链霉素的结核菌主要突变基因rpsL基因,其次是rrS基因,它们的突变也会导致耐药性的产生。对药物耐药机制的深入研究极大地推动了抗结核药物的研发及化学治疗方案的发展。%In recent years, Emergence of multidrug-resistant tuberculosis (MDR-TB) and extensively drug-resistant tuberculosis (XDR-TB) are reported world wide. MDR-TB is defined as TB caused by a Mycobacterium tuberculosis strain(MTB) that is resistant to at least the two best first-line medications, isoniazid and rifampicin. XDR-TB is TB showing resistance to at least rifampicin and isoniazid, which is the definition of MDR-TB, in addition to resistance to at least three of the six main classes of second-line drugs. There are five first-line antituberculosis drugs: isoniazid (INH), rifampicin (RIF), pyrazinamide (PZA), ethambutol (EMB) and streptomycin (SM). Resistance to first line anti-TB drugs has been linked to mutations in at least 7 main genes: katG, inhA for INH resistance, rpoB for RIF resistance, embB for EMB resistance, pncA for PZA resistance and rpsL and rrs for STR resistance. These genes encode either drug-targeting molecule or enzyme involving in

  13. A genetic screen of the mutations in the Korean patients with early-onset Alzheimer’s disease

    Directory of Open Access Journals (Sweden)

    An SS

    2016-12-01

    Full Text Available Seong Soo An,1,* Sun Ah Park,2,* Eva Bagyinszky,1 Sun Oh Bae,1 Yoon-Jeong Kim,2 Ji Young Im,2 Kyung Won Park,3 Kee Hyung Park,4 Eun-Joo Kim,5 Jee Hyang Jeong,6 Jong Hun Kim,7 Hyun Jeong Han,8 Seong Hye Choi,9 SangYun Kim10 1Department of Bionano Technology, Gachon University, Seongnam-si, 2Department of Neurology, Soonchunhyang University Bucheon Hospital, Bucheon, 3Department of Neurology, Dong-A University College of Medicine and Institute of Convergence Bio-Health, Busan, 4Department of Neurology, Gachon University Gil Medical Center, Incheon, 5Department of Neurology, Pusan National University Hospital, Busan, 6Department of Neurology, Ewha Womans University Mokdong Hospital, Seoul, 7Department of Neurology, Ilsan Hospital, National Health Insurance Corporation, 8Department of Neurology, Myongii Hospital, Goyang, 9Department of Neurology, Inha University School of Medicine, Incheon, 10Department of Neurology, Seoul National University College of Medicine & Neurocognitive Behavior Center, Seoul National University Bundang Hospital, Seongnam-si, Republic of Korea *These authors contributed equally to this work Abstract: Early-onset Alzheimer’s disease (EOAD has distinct clinical characteristics in comparison to late-onset Alzheimer’s disease (LOAD. The genetic contribution is suggested to be more potent in EOAD. However, the frequency of causative mutations in EOAD could be variable depending on studies. Moreover, no mutation screening study has been performed yet employing large population in Korea. Previously, we reported that the rate of family history of dementia in EOAD patients was 18.7% in a nationwide hospital-based cohort study, the Clinical Research Center for Dementia of South Korea (CREDOS study. This rate is much lower than in other countries and is even comparable to the frequency of LOAD patients in our country. To understand the genetic characteristics of EOAD in Korea, we screened the common Alzheimer’s disease (AD

  14. 利用数字基因表达谱对小尾寒羊高繁性状相关基因的筛选研究%The Study on Related Genes of High Fecundity Using Digital Gene Expression Tag Profiling in Small-Tail-Han-Sheep

    Institute of Scientific and Technical Information of China (English)

    罗庆苗; 秦晓玉; 苗向阳

    2012-01-01

    pathway and pyrimidine metabolism. According to published papers, the genes including STAR, FLCN, TGFI1, INHA, INHBA and the Notch signaling Pathway were considered as the most important genes and Pathway involved in fecundity differences between FecBBFecBB and FecB+FecB+ewes of small-tail-han-sheep.

  15. PREFACE: 13th International Conference on Electrorheological Fluids and Magnetorheological Suspensions (ERMR2012)

    Science.gov (United States)

    Unal, Halil Ibrahim

    2013-02-01

    China) gave plenary talks on novel MR shear thickening fluids, MR rubber composites and MR plastomers, respectively. H J Choi (INHA University, Korea), X Zhao (Northwestern Polytechnical University, China) and R Tao (Temple University, USA) delivered plenary lectures on issues relating to the area of electrorheological (ER) fluids. In their talks, Choi and Zhao covered novel ER materials with core-shell structured microspheres and micro/nano hierarchical structured titania particles, respectively, and Tao focused on the reduction of the viscosity of liquid suspensions for energy applications in transporting crude oil via pipelines. Attendance of the presentations was exceptionally high. Poster presentations were divided into two sessions and held in the afternoons after the close of the sessions, which were filled with stimulating discussions. The award for the best student research in the area of electrorheological fluids, sponsored by the Winslow family, was given to Y D Liu of INHA University, Korea on research entitled 'Copolyaniline coated monodisperse polystyrene microparticles and their electrorheological response'. The award for the best student research on magnetorheological suspensions, sponsored by the LORD Corporation, was given to S Kaneko of Keio University, Japan on research entitled 'Effect of a magnetic field on sloshing pressure in a magnetic fluid'. Besides the purely scientific program during the five days, some special events were also organized. A guided tour of the old city and some important landmarks of the capital city of Ankara were organized before the Gala Dinner. Special samples of Turkish classical music and folk dancing were performed by a group from the Republic of Turkey Ministry of Culture and Tourism to exhibit Turkish culture to the delegates at Gala Night, which was truly appreciated. Following the conclusion of this successful meeting, the next conference is expected to be organized at the University of Granada, Spain by Professor

  16. EDITORIAL: Welcome to the 2013 volume Welcome to the 2013 volume

    Science.gov (United States)

    Garcia, Ephrahim

    2013-01-01

    Professor Derek Abbott, Professor Alison Flatau, Professor Jinsong Leng and Professor Yuji Matsuzaki who have retired from the Board. Professor Matsuzaki worked on the journal for almost two decades. Associate Editors Professor G Akhras, Royal Military College of Canada, Ontario, Canada Professor C Boller, University of Saarland, Saarbrücken, Germany; Fraunhofer-Institut für Zerstörungsfreie Prüfverfahren, Dresden, Germany Professor Cagnol, École Centrale Paris, France Professor G Carman, University of California-Los Angeles, USA Professor S-B Choi, Inha University, Incheon, Korea Professor S H Choi, NASA Langley Research Center, Hampton, VA, USA Professor A Del Grosso, Università degli Studi di Genova, Italy Professor D Erickson, Cornell University, Ithaca, NY, USA Professor U Gabbert, Universität Magdeburg, Germany Professor A Güemes, Universidad Politecnica de Madrid, Spain Professor S Gopalakrishnan, Indian Institute of Science, Bangalore, India Professor J Kim, Inha University, Incheon, Korea Professor K J Kim, University of Nevada, Reno, USA Professor S J Kim, Seoul National University, Korea Professor D Lagoudas, Texas A&M University, College Station, USA Professor R Lammering, Helmut-Schmidt-Universität/Universität der Bundeswehr Hamburg, Hamburg, Germany Professor C K Lee, National Taiwan University, Taiwan Professor W Li, University of Wollongong, Australia Professor W H Liao, Chinese University of Hong Kong, China Professor Y Liu, Harbin Institute of Technology, China Professor C S Lynch, University of California-Los Angeles, USA Professor S Masri, University of Southern California, Los Angeles, USA Professor W M Ostachowicz, Polish Academy of Sciences, Gdansk, Poland Professor K Peters, North Carolina State University, Raleigh, USA Professor M Shahinpoor, University of New Mexico, Albuquerque, USA Professor H Sodano, Arizona State University, Tempe, USA Professor G Song, University of Houston, TX, USA Professor W J Staszewski, AGH University of Science

  17. Welcome to the 2014 volume of Smart Materials and Structures

    Science.gov (United States)

    Garcia, Ephrahim

    2014-01-01

    last year after many years in service as Associate Editors. Associate Editors in 2013: Professor G Akhras, Royal Military College of Canada, Ontario, Canada Professor C Boller, University of Saarland, Saarbrücken, Germany Fraunhofer-Institut für Zerstörungsfreie Prüfverfahren, Dresden, Germany Professor J Cagnol, École Centrale Paris, France Professor G Carman, University of California-Los Angeles, USA Professor S-B Choi, Inha University, Incheon, Korea Professor S H Choi, NASA Langley Research Center, Hampton, VA, USA Professor A Del Grosso, Università degli Studi di Genova, Italy Professor A Erturk, Georgia Institute of Technology, GA, USA Professor U Gabbert, Universität Magdeburg, Germany Professor A Güemes, Universidad Politecnica de Madrid, Spain Professor S Gopalakrishnan, Indian Institute of Science, Bangalore, India Professor J Kim, Inha University, Incheon, Korea Professor K J Kim, University of Nevada, Reno, USA Professor S J Kim, Seoul National University, Korea Professor D Lagoudas, Texas A&M University, College Station, USA Professor R Lammering, Helmut-Schmidt-Universität/Universität der Bundeswehr Hamburg, Hamburg, Germany Professor C K Lee, National Taiwan University, Taiwan Professor W Li, University of Wollongong, Australia Professor W H Liao, Chinese University of Hong Kong, China Professor Y Liu, Harbin Institute of Technology, China Professor C S Lynch, University of California-Los Angeles, USA Professor S Masri, University of Southern California, Los Angeles, USA Professor W M Ostachowicz, Polish Academy of Sciences, Gdansk, Poland Professor K Peters, North Carolina State University, Raleigh, USA Professor M Shahinpoor, University of Maine, Orono, USA Professor H Sodano, University of Florida, Gainsville, USA Professor G Song, University of Houston, TX, USA Professor W J Staszewski, AGH University of Science and Technology, Kraków, Poland Professor N Takeda, University of Tokyo, Japan Professor D-H Wang, Chongqing University, China

  18. Carvedilol atenua o estresse oxidativo na cardiopatia chagásica crônica Carvedilol atenúa el estrés oxidativo en la cardiopatía chagásica crónica Carvedilol attenuates oxidative stress in chronic chagasic cardiomyopathy

    Directory of Open Access Journals (Sweden)

    Patrícia Budni

    2012-03-01

    Full Text Available FUNDAMENTO: Há cada vez mais evidências sugerindo que doença de Chagas envolve dano oxidativo e contribui para a progressão da doença cardíaca. OBJETIVO: Avaliar o efeito do carvedilol sobre marcadores de estresse oxidativo na doença de Chagas crônica. MÉTODOS: A população de estudo incluiu 42 pacientes com cardiopatia chagásica e os biomarcadores de estresse oxidativo foram medidos antes e após um período de seis meses de tratamento com carvedilol (37,5 mg/dia. Os pacientes foram considerados de acordo com a classificação de Los Andes, e a atividade da superóxido dismutase, catalase, glutationa peroxidase, S-transferase e redutase, mieloperoxidase e adenosina deaminase; e os níveis de glutationa reduzida, de espécies reativas do ácido tiobarbitúrico, proteína carbonil, vitamina E e óxido nítrico foram medidos no sangue. RESULTADOS: Após o tratamento com carvedilol, todos os grupos apresentaram reduções significativas nos níveis de proteína carbonil e glutationa reduzida, enquanto os níveis de óxido nítrico e atividade da adenosina aumentaram significativamente somente no grupo IA. Além disso, a maioria das enzimas antioxidantes apresentou diminuição de suas atividades, nos grupos IA e IB. CONCLUSÃO: Os dados sugerem que o tratamento com carvedilol foi eficaz na atenuação do dano oxidativo, um efeito que pode ser particularmente importante em doença de Chagas crônica com cardiopatia.FUNDAMENTO: Hay cada vez más evidencias sugiriendo que la enfermedad de Chagas envuelve daño oxidativo y contribuye a la progresión de la enfermedad cardíaca. OBJETIVO: Evaluar el efecto del carvedilol sobre marcadores de estrés oxidativo en la enfermedad de Chagas crónica. MÉTODOS: La población de estudio incluyó 42 pacientes con cardiopatía chagásica y los biomarcadores de estrés oxidativo fueron medidos antes y después de un período de seis meses de tratamiento con carvedilol (37,5 mg/día. Los pacientes fueron

  19. Light exercise causes adaptive changes in muscles of young Brasileiro de Hipismo horses Atividade física leve causa mudanças adaptativas musculares em eqüinos jovens da raça Brasileiro de Hipismo

    Directory of Open Access Journals (Sweden)

    Flora Helena de Freitas D'Angelis

    2008-08-01

    policiamento desenvolvido pela PM-SP por 10 meses, foram retirados fragmentos do músculo Gluteus medius nas profundidades de 20mm e 60mm. Para a determinação dos tipos de fibra muscular, foram feitas análises histoquímicas por meio das técnicas de adenosina trifosfatase miofibrilar (mATPase e nicotinamida adenina dinucleotídeo tetrazólio redutase (NADH-TR. Na análise morfométrica das fibras, calculou-se a área de secção transversal (AST, a freqüência média (F e a área de secção transversal relativa (ASTR. Após o período de atividade física, as fibras tipo IIA apresentaram aumento da F e da ASTR nas duas profundidades. As fibras tipo I apresentaram diminuição da F na profundidade de 60mm e as fibras tipo IIX não sofreram alteração da F nas duas profundidades. A ASTR das fibras tipo I e IIX não apresentaram alteração após os 10 meses. A atividade física de baixa intensidade produziu adaptações significativas no músculo Gluteus medius de cavalos da raça BH, incluindo mudança nas propriedades metabólicas e contráteis evidenciadas pelo aumento da área ocupada pelas fibras tipo IIA, nas duas profundidades de biópsia.

  20. O pediatra frente a uma criança com ambigüidade genital The role of the pediatrician in the management of children with genital ambiguities

    Directory of Open Access Journals (Sweden)

    Gil Guerra-Júnior

    2007-11-01

    Full Text Available OBJETIVO: Apresentar os critérios diagnósticos de ambigüidade genital, a conduta médica inicial e a postura esperada do pediatra. FONTES DOS DADOS: Revisão de literatura científica por meio de artigos publicados no MEDLINE nos idiomas inglês e português, no período de 1990 a 2007 e na faixa etária pediátrica. SÍNTESE DOS DADOS: O pediatra tem papel fundamental na avaliação da ambigüidade genital, cujo objetivo é obter diagnóstico etiológico preciso no menor tempo possível para definição do sexo e estabelecimento dos procedimentos terapêuticos. Há critérios diagnósticos específicos, porém, de modo geral, uma genitália é ambígua sempre que houver dificuldade para se atribuir o sexo à criança. O pediatra deve informar à família que a definição do sexo dependerá de investigação laboratorial minuciosa, feita preferencialmente por equipe interdisciplinar em serviço terciário. O cariótipo 46,XX ou 46,XY não é suficiente para definir o sexo de criação, porém esse exame é fundamental para direcionar a investigação. Quando não houver gônadas palpáveis, a primeira hipótese deve ser hiperplasia adrenal congênita. Entre as outras causas, estão insensibilidade parcial a andrógenos, deficiência da enzima 5alfa-redutase, disgenesia gonadal parcial e hermafroditismo. A família deve receber apoio e informações durante todo o processo de avaliação, e sua participação é fundamental na decisão sobre o sexo de criação. CONCLUSÕES: Embora casos de ambigüidade genital sejam relativamente raros para o pediatra, este deve estar informado sobre o tema e a conduta adequada a tomar, pois freqüentemente será o responsável pela orientação inicial da família e pela ligação entre esta e a equipe interdisciplinar.OBJECTIVE: To present the diagnostic criteria of genital ambiguity, the initial medical management and the attitude expected of pediatricians. SOURCES: Review of the scientific literature in

  1. Comportamento celular e resposta antioxidante diferenciados de Saccharomyces cerevisiae e de Saccharomyces chevalieri ao metavanadato de amónio Different cellular behaviour and antioxidant response of Saccharomyces cerevisiae and Saccharomyces chevalieri growing in presence of ammonium metavanadate

    Directory of Open Access Journals (Sweden)

    R. Ferreira

    2007-01-01

    Full Text Available A fermentação do vinho é um processo microbiológico complexo que requere a presença de leveduras adaptadas a condições de stresse. No ambiente celular de organismos aeróbios ocorrem naturalmente espécies reactivas de oxigénio (ROS como subprodutos da respiração mitocondrial. A elevada reactividade destas espécies químicas pode gerar danos moleculares que, em alguns casos, levam à morte celular. Em condições fisiológicas normais ou como resposta ao stresse oxidativo, a célula pode desencadear respostas adaptativas que envolvem mecanismos antioxidantes como os enzimas glutationo redutase (GR; EC 1.6.4.2 e catalases T (CAT T; EC 1.11.1.6 e A (CAT A; EC 1.11.1.6. O vanádio, um metal pesado presente em alguns fitofármacos, pode também com portar-se como um gerador de ROS, alterando o estado redox intracelular e exercendo efeitos nocivos em leveduras expostas a quantidade excessiva deste elemento. O principal objectivo deste trabalho foi comparar o efeito do metavanadato de amónio (NH4VO3, um sal pentavalente de vanádio, na viabilidade celular e nas actividades enzimáticas GR, CAT T e CAT A das leveduras vínicas Saccharomyces cerevisiae UE-ME3 e Saccharomyces chevalieri UE-ME1. Os resultados obtidos mostram que S. chevalieri UE-ME1 revelou menor tolerância ao NH4VO3 do que S. cerevisiae UE-ME3, uma vez que culturas de S. chevalieri não sobreviveram para valores de concentração do sal de vanádio superiores a 7,5 mM enquanto que células de S. cerevisiae mantiveram-se viáveis em presença de metavanadato de amónio 75 mM. As actividades enzimáticas estudadas apresentaram em S. chevalieri valores muito inferiores aos que foram determinados em S. cerevisiae embora em ambas as espécies de levedura o NH4VO3 pareça comportarse como um indutor de stresse oxidativo ao provocar um decréscimo significativo da actividade GR (PThe fermentation of wine is a complex microbiological process which requires yeast adaptation to stress

  2. Biosynthesis of secondary metabolites in sugarcane

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    S.C. França

    2001-12-01

    relacionados ao metabolismo secundário da cana de açúcar e investigar o padrão de expressão gênica de enzimas chaves reguladoras das principais vias biossintéticas ativas nos diferentes tipos de tecidos e situações de estresse físico-químico e biológico a que estão submetidas plantas cultivadas em casas de vegetação, campo ou in vitro. A estratégia de mineração dos dados da database de sequências expressas de cana de açúcar, SUCEST, usando ferramentas de bioinformática, focalizou classes de compostos como isoprenóides e fenilpropanóides que comprovadamente desempenham um papel na resposta de plantas a variações ambientais. Foram identificados e caracterizados genes que codificam enzimas chaves para a síntese de terpenóides, como a sesquiterpeno ciclase (SC; (CHS para síntese de flavonóides; isoflavona sintase (IFS envolvida na biossíntese de isoflavonóides que desempenharm importante papel na defesa de plantas e nodulação de raízes; isoflavona redutases (IFR enzimas chaves para a síntese de fenilpropanóide fitoalexinas, bem como enzimas relacionadas à síntese de precursores de lignina, como a enzima ácido caféico- O- metiltransferase. O efeito do estresse causado por bactérias como Herbaspirillum rubri e Gluconacetobacter diazotroficans também foi avaliado tendo sido constatada a indução da expressão de chalcona sintase (CHS em plântulas infectadas com esses agentes, sugerindo a ativação da via de flavonóides em resposta a este estresse biológico. Esses resultados apontam para o fato de que as vias do metabolismo de isopropanóides e de fenilpropanóides em cana de açúcar são ativadas de acordo com o estágio de desenvolvimento, especificidade de tecidos e em resposta a situações de estresse. Essas observações deverão ser confirmadas por meio de experimentação genética e bioquímica.

  3. Efeito da desnutrição protéica pré e pós-natal sobre a morfologia, a diferenciação e o metabolismo do tecido muscular estriado esquelético em ratos The effects of prenatal and postnatal malnutrition on the morphology, differentiation, and metabolism of skeletal striated muscle tissue in rats

    Directory of Open Access Journals (Sweden)

    Alessandra P. Alves

    2008-06-01

    Full Text Available OBJETIVO: Estudar características morfológicas, metabolismo e habilidades contráteis do músculo submetido a desnutrição protéica pré e pós-natal. MÉTODOS: Distribuição dos animais em dois grupos: controle, dieta normoprotéica (GC; n = 15; 5/5/5 e desnutrido, dieta hipoprotéica (GD; n = 15; 5/5/5, observados respectivamente no sétimo, 14º e 28º dia do período experimental. Foram avaliados massa corporal total, peso, habilidades contráteis e a morfologia do músculo tibial anterior. Amostras de tecidos com 8 m de espessura de ratos com idades de 7, 14 e 28 dias, corados por hematoxilina e eosina, e outros submetidos aos métodos histoquímicos nicotinamida adenina tetrazólio redutase (NADH-TR e miofibrilar (m-ATPase (pH = 4,4. RESULTADOS: Os pesos corporal e muscular apresentaram-se menores nos grupos desnutridos. Aos 7 dias de desnutrição, o músculo apresentou fibras com menor diâmetro, maior polimorfismo e maior teor de tecido conjuntivo endomisial. Nas histoquímicas, tipos de fibras sem delimitação segura. Aos 14 dias de desnutrição, fibras menores, mais polimórficas, muitas com núcleos centrais e moderado teor de tecido conjuntivo endomisial. Quanto à contração, a reação m-ATPase evidenciou fibras lentas e rápidas. A reação NADH-TR revelou os tipos de fibras slow oxidative, fast oxidative glycolytic e fast glycolytic. Aos 28 dias de desnutrição, fibras menores, agrupadas com contornos variáveis. Quanto ao tipo de contração e metabolismo, os três tipos de fibras apresentaram limites de reconhecimento indistinto. CONCLUSÃO: Os resultados experimentais sugerem que, além da redução no número de fibras, a desnutrição promove um retardamento na diferenciação das características morfológicas, metabólicas e contráteis dos tipos de fibras musculares esqueléticas em ratos na fase de crescimento.OBJECTIVE: To study the contractile properties, metabolism and morphological characteristics of

  4. Distúrbios trombofílicos em crianças e adolescentes com trombose da veia porta Thrombophilic disorders in children and adolescents with portal vein thrombosis

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    Raquel B. Pinto

    2003-04-01

    Full Text Available OBJETIVO: determinar a freqüência de deficiência da proteína C (PC, proteína S (PS e antitrombina (AT e das mutações fator V Leiden (FVL, G20210A, no gene da protrombina (PTR, e C677T da metileno-tetraidrofolato redutase (MTFR em crianças e adolescentes com trombose da veia porta (TVP, e definir o padrão hereditário de uma eventual deficiência. MÉTODOS: durante o período de dois anos, foi investigada a presença de distúrbios trombofílicos em 14 crianças e adolescentes (grupo 1 com TVP, seus pais (grupo 2, n = 25 e dois grupos controles constituídos, o primeiro, por crianças e adolescentes sem hepatopatia, pareados por idade com as crianças do grupo 1 (n = 28, e o outro, por pacientes com cirrose (n = 24. Os pacientes com TVP foram investigados do ponto de vista clínico, laboratorial, endoscópico e com biópsia de fígado. O diagnóstico da trombose foi realizado por ultra-sonografia abdominal com Doppler e/ou estudo angiográfico. RESULTADOS: a freqüência da deficiência de PC, PS e AT nos pacientes com TVP foi de 6/14 (42,9% (p 0,05 e 1/14 (7,1% (p > 0,05, respectivamente. Nos pacientes com cirrose, a freqüência da deficiência de PC, PS e AT foi de 14/24 (58,3%, 7/24 (29,2% e 11/24 (45,8%, respectivamente (p OBJECTIVE: to determine the frequency of protein C, protein S and antithrombin deficiency, and factor V Leiden, prothrombin G20210A, and methylenetetrahydrofolate reductase C677T mutations in children and adolescents with portal vein thrombosis, as well as assessing the hereditary character of this disorders. METHODS: a two-year study was carried out to determine the frequency of thrombophilic disorders in children and adolescents with portal vein thrombosis (n = 14, their parents (n = 24, and two control groups, one age-matched children and adolescents free of liver disease (n = 28 and another group with cirrhosis (n = 24. The portal vein thrombosis patients were investigated by clinical and laboratory means

  5. Efeitos do nível de inóculo de Meloidogyne incognita em algodoeiro Effects of low inoculum level of Meloidogyne incognita on cotton plants

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    MARLUCI MUNDIN ABRÃO

    2001-01-01

    Full Text Available Pesquisas acerca da fisiologia de plantas em resposta ao ataque de nematóides normalmente são feitas com doses altas de inóculo, quer sejam ovos ou juvenis de segundo estágio. Diferentemente, as avaliações da resistência de plantas a nematóides em programas de melhoramento são realizadas com doses bastante inferiores. Neste estudo acompanhou-se a variação de alguns parâmetros fisiológicos em algodão, em resposta à aplicação de baixas doses de inóculo de Meloidogyne incognita raça 3. Duas cultivares foram utilizadas, Acala e IAC-20, respectivamente suscetível e moderadamente resistente. Plântulas com duas folhas receberam 500 ou 5.000 ovos e, 90 dias depois, foram comparadas a plantas que não receberam o inóculo, quanto à massa da parte aérea e raízes, fotossíntese, atividade da enzima redutase do nitrato e teores de clorofila e açúcares solúveis. Aos 90 dias algumas plantas também foram submetidas à deficiência hídrica e o potencial da água determinado nas folhas. O maior nível de inóculo levou à redução da parte aérea (massa de folhas em 'Acala'. Com 500 ovos parece ter ocorrido um estímulo em ambas as cultivares, uma vez que houve tendência de maior crescimento da parte aérea. Em 'Acala' houve maior crescimento de raiz com o aumento do inóculo, provavelmente devido à emissão de raízes secundárias nos pontos de penetração do nematóide e também pela formação de galhas. Houve aumento dos teores de clorofila e de açúcares solúveis, bem como da fotossíntese, com o aumento do inóculo. Discute-se se tal ocorrência deve-se à maior demanda de fotoassimilados para suportar o crescimento de raízes, em 'Acala'. Em IAC-20 esse fato estaria ligado aos mecanismos de defesa induzidos pelo ataque do nematóide. Após o estresse hídrico o potencial da água diminuiu em 'IAC-20' e aumentou na 'Acala', com o aumento do inóculo. A elevação do potencial da água, nesta última cultivar, poderia

  6. Amenorréia primária e cariótipo XY: identificando pacientes em risco Primary amenorrhea and XY karyotype: identifying patients in risk

    Directory of Open Access Journals (Sweden)

    Rafael Fabiano Machado Rosa

    2008-11-01

    Full Text Available OBJETIVO: verificar a prevalência e as características clínicas de pacientes com amenorréia primária e cariótipo XY avaliadas em nosso Serviço com o intuito de identificar achados que possam auxiliar em seu reconhecimento. MÉTODOS: no período de Janeiro de 1975 a Novembro de 2007, foram avaliadas 104 pacientes com amenorréia primária. Para todos os casos foi realizada a análise pelo cariótipo por bandas GTG. Destas, 21 (20,2% apresentavam uma constituição 46,XY. Contudo, duas foram excluídas do estudo por terem prontuários incompletos. Das 19 pacientes que compuseram a amostra, a maior parte veio encaminhada pela ginecologia (63,2%. Suas idades variaram entre 16 e 41 anos (média de 22,1 anos. Realizou-se uma coleta de dados sobre sua história familiar e pregressa, exame físico e resultados de exames complementares. Para determinação dos seus diagnósticos levaram-se em consideração essas informações. RESULTADOS: a síndrome de resistência aos androgênios foi o diagnóstico predominante (n=12; 63,2%. Cinco pacientes (26,3% apresentavam disgenesia gonadal pura XY (DGP XY, uma (5,3% deficiência de 17-alfa hidroxilase e uma (5,3% deficiência de 5-alfa redutase. Achados clínicos freqüentemente observados nessas pacientes incluíram desenvolvimento anormal dos caracteres sexuais secundários (n=19, agenesia uterina com vagina em fundo de saco (n=14, história familiar de amenorréia (n=8 e gônadas palpáveis no canal inguinal (n=5. Duas delas apresentavam história de hérnia inguinal. Hipertensão arterial sistêmica foi diagnosticada somente na paciente com deficiência de 17-alfa hidroxilase, e malignização gonadal, naquela com DGP XY. CONCLUSÕES: a freqüência de pacientes com cariótipo XY (20% foi superior à usualmente descrita na literatura (3 a 11%. Acreditamos que isso tenha relação com a forma de encaminhamento das pacientes ao Serviço. Certos achados da história clínica e do exame físico deveriam

  7. Efeito renoprotetor da estatina: modelo animal de isquemia-reperfusão Renoprotective effect of statin: a ischemia-reperfusion animal model

    Directory of Open Access Journals (Sweden)

    Claudia Akemi Shibuya Teshima

    2010-09-01

    Full Text Available OBJETIVO: A lesão renal aguda isquêmica, de causa multifatorial, apresenta morbidade e mortalidade alarmantes. A estatina, inibidor de HMG-CoA redutase, tem demonstrado papel renoprotetor, com componente antioxidante, antiinflamatório e vascular. A atividade de heme oxigenase-1 pode ser mediadora desses efeitos pleitrópicos da estatina sobre o rim, ou seja, independente da ação de redução de lipídio. Esse estudo visou avaliar se o efeito renoprotetor da estatina pode ter mecanismo heme de proteção em ratos. MÉTODOS: O modelo isquêmico foi obtido por meio do clampeamento dos pedículos renais bilaterais por 30 minutos, seguido de reperfusão. Foram utilizados ratos Wistar, machos, pesando entre 250-300g, distribuídos nos seguintes grupos: SHAM (controle, sem clampeamento renal; Isquemia; Iquemia+Estatina (sinvastatina 0,5 mg/kg, via oral por 3 dias; Isquemia+Hemin (indutor de HO-1, 1 mg/100g, intraperitoneal 24h antes da cirurgia; Isquemia+SnPP (inibidor de HO-1, 2μmol/kg intraperitoneal 24h antes da cirurgia; Isquemia+Estatina+Hemin e Isquemia+Estatina+SnPP. Foram avaliados a função renal (clearance de creatinina, Jaffé, osmolalidade urinária, peróxidos urinários e a imunohistoquímica para ED-1. RESULTADOS: Os resultados mostraram que a estatina melhorou a função renal, a osmolalidade urinária, reduziu a excreção de peróxidos urinários e a infiltração de macrófagos em rins de animais submetidos à isquemia renal. O indutor da heme oxigenase-1 e a sua associação com sinvastatina reproduziram o padrão de melhora determinado pela sinvastatina. CONCLUSÃO: O estudo confirmou o efeito renoprotetor da estatina, com ação antioxidante e antiinflamatória, e sugere que esse efeito tenha interface com o sistema heme de proteção renal.OBJECTIVE: Ischemic acute kidney injury (iLRA, with multifatorial cause, presents alarming morbidity and mortality. Statin, HMG-CoA inhibition reductase has shown a renoprotective effect

  8. Inoculação de Azospirillum amazonense em dois genótipos de milho sob diferentes regimes de nitrogênio Inoculation of Azospirillum amazonense in two maize genotypes under different N treatments

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    Fábio Bueno dos Reis Junior

    2008-06-01

    Full Text Available A adaptabilidade de genótipos de milho a ambientes onde os nutrientes não estejam prontamente disponíveis pode ser relacionada com a adaptação à predominância, nos solos, das formas de N nítrica e amoniacal e à associação com microrganismos benéficos como as bactérias diazotróficas e, ou, promotoras de crescimento de plantas. O objetivo deste trabalho foi verificar o comportamento de dois híbridos intervarietais de milho sob diferentes doses e formas de N, assim como o efeito da inoculação com Azospirillum amazonense, em um ensaio em casa de vegetação, utilizando vasos preenchidos com vermiculita e solução nutritiva de Hoagland como fonte de nutrientes. O delineamento experimental foi de blocos ao acaso em esquema fatorial, constituindo os tratamentos em: dois híbridos intervarietais, H1 (Carioca x Eldorado e H2 (Palha Roxa ES x Sol da Manhã; plantas inoculadas ou não inoculadas com A. amazonense, e três regimes de N, conforme doses e proporções seguintes: 126 mg semana-1 de N (75 % NH4+ : 25 % NO3-; 126 mg semana-1 de N (25 % NH4+ : 75 % NO3-, e 12,6 mg semana-1 de N (50 % NH4+ : 50 % NO3-. Vinte e cinco dias após o plantio, as plantas foram separadas em raiz e parte aérea para determinação da matéria seca, conteúdos totais de N e P, atividade das enzimas nitrato redutase e glutamina sintetase e açúcares solúveis totais. O híbrido H1, mais eficiente em avaliações preliminares em campo, produziu mais matéria seca e apresentou maior eficiência na utilização de N e P. Plantas que receberam a maior dose de N, independentemente do predomínio da forma nítrica ou amoniacal, apresentaram maior acúmulo de N (nas raízes e parte aérea e P (na parte aérea e maior índice de utilização de P. Com o predomínio da forma amoniacal, observou-se incremento na produção de matéria seca da parte aérea e na atividade da glutamina sintetase, bem como decréscimo na concentração de açúcares solúveis totais

  9. Committees

    Science.gov (United States)

    2012-11-01

    Quebec IREQ, Varennes Canada Etienne PARKINSON Andritz Hydro Ltd. Switzerland B V S S S PRASAD Indian Institute of Technology Madras India Stefan RIEDELBAUCH Stuttgart University Germany Michel SABOURIN Alstom Hydro Canada Inc Canada Bruno SCHIAVELLO Flowserve Corporation USA Katsumasa SHIMMEI Hitachi Ltd Japan Christoph SINGRTüN VDMA Germany Ale? SKOTAK CKD Blansko Engineering, a s Czech Republic Toshiaki SUZUKI Toshiba Corporation Japan Andy C C TAN Queensland University of Technology Australia Geraldo TIAGO FILHO Universidade Federal de Itajuba Brazi Thi C VU Andritz Hydro Ltd Canada Satoshi WATANABE Kyushu University Japan S H WINOTO National University of Singapore Singapore Woo-Seong WOO STX Institute of Technology Korea International Technical Committee François AVELLAN (principal) EPFL-LMH Switzerland Xingqi LUO (principal) Xi'an University of Technology China Martin BÖHLE Kaiserslautern University Germany Gerard BOIS ENSAM France Young-Seok CHOI KITECH Korea Luca d'AGOSTINO University of Pisa Italy Eduard EGUSQUIZA Polytechnical University Catalonia Spain Arpad FAY University of Miskolcz Hungary Richard FISHER Voith Hydro Inc USA Regiane FORTES-PATELLA Institute Polytechnique de Grenoble France Aleksandar GAJIC University of Belgrade Serbia José GONZÁLEZ Universidad de Oviedo Spain François GUIBAULT Ecole Polytechnique de Montreal Canada Toshiaki IKOHAGI Tohoku University Japan Chisachi KATO University of Tokyo Japan Kwang-Yong KIM Inha University Korea Youn-Jea KIM Sungkyunkwan University Korea Smaine KOUIDRI Université Pierre et Marie Curie (Paris 6) France Shengcai LI Warwick University UK Adrian LUNGU Dunarea de Jos University of Galati Romania Torbjøm K NIELSEN NTNU Norway Michihiro NISHI Tsinghua University China Peter PELZ Darmstadt University Germany Frantisek POCHYLY Brno University Czech Republic Albert RUPRECHT University of Stuttgart Germany Rudolf SCHILLING Technische University München Germany Wei SHYY HKUST Hong Kong,China Romeo SUSAN

  10. Prospecção hidrobiológica da Baía de Sepetiba Color and transparence of waters of Sepetiba Bay, in Rio de Janeiro State

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    Lejeune P. H. de Oliveira

    1971-01-01

    green in the localities with blooms of the diatom Coscinodiscus. The central portion of the bay, 150 Km[squared] hole-green, full of larvae of shrimps Penaeus schmidti; some shoals of shrimps moves its pleopods and by displacement of botton with diatoms and reddish sediment produces a rust color spot in surface of waters seen by fishermen. The estuaries are Isabell color, and caramel. That is a preliminar survey, to future comparison, because to day Sepetiba Bay is not polluted. But futurelly, it will be polluted by an harbour for 3.000,000 tons of iron ore, and by a quay for cargo boats, and siderurgy mills, airports, and other industries. Several polluted waters of Guanabara Bay are black and asphalt color, and cub color. That colors are not found in Sepetiba Bay to day. In 1965, we do not find the "red waters" and "red tides" with blooms of dinoflagellates in Sepetiba Bay, that they are present sometimes in Guanabara Bay with mortalities of fishes. Sepetiba Bay is not biologic problem, to day, is not polluted. The fig. 3 shows a map of transparence of waters by the disc of Secchi and a relation is given of color and transparence, in Sepetiba: Forel color 5: transparence 2.8 m to 2.9 m; Forel n. 6: 2.0 m; Forel color n. 7: 1.0 m to 1.5 m. Moss green waters: 0.6 to 1.5 m; hellebore green waters: 0.9 to 1.5 m; emerald green waters: 2.0 m to 2.2 m; In Guanabara Bay, into the polluted Enseada de Inhaúma: asphalt color waters: 0.02 - 0.05 m; and in black waters: trasnparence of 1 cm. The extintion koeficient of light is indicated in explanation of the fig. 3.

  11. Meta-analysis of pyrosequencing for the rapid detection of isoniazid-resistance in Mycobacterium tuberculosis%焦磷酸测序技术检测结核分枝杆菌异烟肼耐药性的Meta分析

    Institute of Scientific and Technical Information of China (English)

    郭琪; 李觉; 胡忠义; 郑瑞娟

    2013-01-01

    research papers were collected and 9 articles were selected.The accordance between pyrosequencing and conventional sequencing result was 100%.Eight studies were involved including 945 specimens when katG gene was detected.The overall sensitivity and specificity were 0.77 (0.73,0.80) and 1.00 (0.99,1.00),respectively.The area under the SROC was 0.9882.As inhA gene was detected,the overall sensitivity and specificity were 0.19 (0.15,0.24) and 1.00 (0.98,1.00).The test was stable.Conclusions Our meta-analysis reveals that pyrosequencing is a highly specific tool for detection mutation of katG gene of isoniazid resistance.This result suggests that it is useful for screening of isoniazid resistance in diagnostic test.(Chin J Lab Med,2013,36:329-332)

  12. 结核杆菌耐药基因芯片在结核性胸膜炎中的应用%Application of drug-resistance mycobacterium tuberculosis gene chip in tuberculous pleuritis

    Institute of Scientific and Technical Information of China (English)

    陈小可; 周一平; 高秋霞; 刘键; 刘慧; 夏利萍; 李艾芬

    2014-01-01

    目的:探讨结核性胸膜炎患者结核杆菌的耐药基因突变情况及其对疗效的影响。方法结核性胸膜炎患者行纤支镜代胸腔镜检查,胸膜活检标本行结核分枝杆菌耐药基因芯片检测,留置胸腔引流管,常规抗痨治疗。结果46例患者,共检出野生型32例,结核耐药基因10例(21.7%),其中利福平rpoB基因突变5例,异烟肼katG基因突变6例,inhA 基因突变2例,乙胺丁醇embB基因突变1例,链霉素rpsL基因突变1例,多耐药基因突变5例。耐药基因组住院时间12.1±3.1d,显著长于非耐药基因组的8.2±2.5 d,P<0.01;耐药基因组胸管留置时间7.0±2.6d,显著长于非耐药基因组的4.0±1.8 d,P<0.01;耐药基因组抗痨后热退时间6.8±1.6 d,显著长于非耐药基因组的4.2±1.4 d,P<0.01。结论结核性胸膜炎存在一定的耐药率,耐药基因的存在显著影响其疗效,结核杆菌耐药基因芯片快捷简便,对早期发现耐药结核性胸膜炎,作针对性抗痨治疗有一定临床意义。%Objective To study the drug-resistance gene mutations in patients with mycobacterium tubercu-lous pleuritis and their influence on the therapeutic effect. Methods 46 patients with mycobacterium tuberculous pleuritis were given fiber bronchoscopy instead of thoracoscopy, and pleural biopsy specimens were detected by myco-bacterium tuberculous gene chip. All cases were treated with chest tube drainage and anti-tuberculous therapy. Re-sults There were 32 cases of wild-type gene and 10 cases of drug-resistance gene mutations, including 5 cases of mutations of rpoB gene against RFP, 6 cases of mutations of katG gene against INH, 2 cases of mutations of inhA gene against INH, 1 case of mutations of embB gene against EMB, 1 case of mutations of rpsL gene against SM, and 6 cases of multidrug resistance gene. The duration of hospital stay was obviously longer in the group with drug-resist-ance gene mutations (12. 1 ± 3. 1 d

  13. 药物外排泵基因表达与结核分枝杆菌耐药关系的探讨%Association of drug efflux pump gene expression with Mycobacterium tuberculosis drug resistance

    Institute of Scientific and Technical Information of China (English)

    李桂莲; 王撷秀; 谢彤; 巨韩芳; 赵慧; 穆成; 赵德福

    2011-01-01

    isolates resistant to one or more of drugs including isoniazid, rifampicin, streptomycin and ethambutol, and 26 isolates all sensitive to the above four drugs from Tianjin Tuberculosis Control Institute in 2007 were involved in this study. Direct sequencing was applied to detect the mutations in the corresponding resistance genes(isoniazid:katG, inhA, oxyR-ahpC, ndh, rifampicin:rpoB, streptomycin:rpsL, rrs, and ethambutol:embB, embC and embA). After RNA extration and reverse transcription, real-time PCR was conducted to assess the expressions of putative drug efflux pump genes Rv1410c, Rv2136c, Rv0783c and Rv2136c, and Students' t test and ANOVA analysis were used to analyze the expression differences in Mycobacterium tuberculosis with different phenotypic drug resistance and drug resistance related gene mutation patterns.Results Compared to pan-sensitive isolates[(5.67±3.29)×10-5], Rv1410c showed higher expression in streptomycin[(8.48±6.33)×10-5, t'=2.18, P<0.05], isoniazid[(8.43±6.38)×10-5, t'=2.20, P<0.05], rifampicin[(9.59±7.27)×10-5, t'=2.29, P<0.05], multi-drug[(10.37±7.86)×10-5, t'=2.34, P<0.05] resistant isolates, and in isoniazid + streptomycin resistant isolates[(9.39±6.81)×10-5, t'=2.43, P<0.05];Rv2136c showed higher expression in isoniazid resistant[(3.51±2.43)×10-5, t'=2.03, P<0.05], multidrug-resistant isolates[(4.21±2.94)×10-5, t'=2.22, P<0.05] and resistant to isoniazid+streptomycin[(3.81±2.46)×10-5, t'=2.28, P<0.05] isolates . The expression of Rv0783c in rifampicin resistant isolates with rpoB 531 mutations [(5.41±3.03)×10-6] was higher than those with wild type of rpoB 531[(2.29±1.62)×10-6, t=2.81, P<0.05].Conclusions The expression of Rv1410c and Rv2136c are associated with mutiple-drug resistance of Mycobacterium tuberculosis.The expression of Rv0783c in rifampicin resistant isolates is associated with mutation in rpoB 531.

  14. PREFACE: International Conference on Inverse Problems 2010

    Science.gov (United States)

    Hon, Yiu-Chung; Ling, Leevan

    2011-03-01

    Conference 2010, we wish to express our cordial thanks to all the keynote, plenary, and invited speakers and members of the Advisory Board. We would like to thank the City University of Hong Kong, the Hong Kong Mathematical Society, the K C Wong Education Foundation, the Lee Hysan Foundation, and the Wei Lun Foundation for their generous financial support. Particularly, we wish to thank Miss Lonn Chan for her excellent administrative work in making all the arrangements for the conference. March 2011Yiu-Chung Hon, City University of Hong Kong, HKSARLeevan Ling, Hong Kong Baptist University, HKSAR Invited Keynote SpeakerH Thomas Banks, North Carolina State University, USA Invited Plenary SpeakersRyuichi Ashino, Osaka Kyoiku University, JapanAndrea Caponnetto, City University of Hong Kong, HKSARJeng-Tzong Chen, National Taiwan Ocean University, TaiwanHiromichi Itou, Gunma University, JapanHyeonbae Kang, Inha University, South KoreaMichael Klibanov, University of North Carolina at Charlotte, USAKim Knudsen, Technical University of Denmark, DenmarkRainer Kress, Georg-August-Universität Göttingen, GermanyJérôme Le Rousseau, University of Orleans, FranceChang-Ock Lee, Korea Advanced Institute of Science and Technology (KAIST), South KoreaGui-Rong Liu, University of Cincinnati, USAShuai Lu, Fudan University, ChinaFadil Santosa, University of Minnesota, USATomoya Takeuchi, North Carolina State University, USAXiang Xu, Zhejiang University, ChinaJun Zou, The Chinese University of Hong Kong, HKSAR Invited SpeakersMohamed Abdel-Mooty, The American University in Cairo, EgyptKrishna Agarwal, National University of Singapore, SingaporeHui Cao, Sun Yat-sen University, ChinaWen Chen, Hohai University, ChinaJin Cheng, Fudan University, ChinaLian Duan, University of Oxford, UKMeibao Ge, Zhejiang Sci-Tech University, ChinaRalf Hielscher, TU Chemnitz, GermanyGuanghui Hu, Weierstrass Institute for Applied Analysis and Stochastics (WIAS), GermanyKiwan Jeon, National Institute for Mathematical

  15. Evaluation of the laryngeal mask airway for spontaneous breathing in short pediatric operation under Sevoflurane inhalation anesthesia%经喉罩自主吸入七氟醚用于小儿短小手术麻醉的可行性研究

    Institute of Scientific and Technical Information of China (English)

    汪芳俊; 胡建华; 万勇; 孙桂英; 蔡蓉

    2015-01-01

    宜适当手控辅助呼吸,以减轻二氧化碳潴留。%Objective To evaluate the effects of the laryngeal mask airway for spontaneous breath-ing in short pediatric operation under Sevoflurane inhalation anesthesia.Methods Eighty children undergo-ing hernia surgery, of whom ASA Ⅰ or Ⅱ, in accordance with the random number table, were randomly divided into two groups( each 40 cases):the laryngeal mask airway for spontaneous breathing with Sevoflu-rane inhalation anesthesia in group A,and tracheal intubation for controlled ventilation with Sevoflurane inha-lationa nesthesia in group B.Children in the two groups were fasting for 6 hours and without drinking for 4 hours before anesthse ia.Anestheis a was induced with Sevoflurane inhalation fro all patients.The laryngela mask airway was insertde soon afet r induction of anesthesia with Sevoflurane inhalation,and anse thesia main-tained with Sevoflurane in halta ion in gor up A.P atients in gruo p B wereg iven cisatracurium 0.15 mg/kg intravenous injectio n before tracheal intubation, and anesthesia maintained with Sevoflurane inhalation, 40 minutes before the end of the surgeryt o sot p using muscle relaxant.Ten minutes before the end of the operation to stop Sevoflurane inhalation,in two groups.The end-tidal carbon dioxide partial pressure(PETCO2 ), mean blood pressure,heart rate,respiratory rate,pulse oxygen saturation and end-tidal sevoflurane partial pres-sure(PETSev) were recorded one minute before the induction of anesthesia,15 min,30 min,45 min,and 60 min after the induction of anesthesia.Both the time to regain consciousness and the time to remove laryngeal mask airway or tracheal catheter were recorded in two groups.Results The mean blood pressure, heart rate decreased in two groups after the induction of anesthesia(P<0.05).The value of PaCO2 increased and that of base excess decreased in 30 min after the induction of anesthesia in group A(P<0.05).The value of PETCO2 increased, and that of pH decreased