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Id: lil-734665
Autor: Melo, A; Artigas, C. G; Fritz, C; Díaz, P; Muñoz, S; Brebi, P; Roa, J. C.
Título: Hipermetilación del gen supresor de tumores p53 en pacientes pediátricos con leucemia linfoblástica aguda / Hypermethylation of tumor suppressor gene p53 in Chilean pediatrics patients under 15 years with acute lymphoblastic leukemia
Fonte: Int. j. morphol;32(4):1243-1247, Dec. 2014. ilus.
Idioma: es.
Projeto: Universidad de La Frontera. Dirección de Investigación y Desarrollo; . CORFO-CEGIN; . Scientific and Technological Bioresource Nucleus.
Resumo: La leucemia linfoblástica aguda (LLA) es la neoplasia maligna hematooncólogica más frecuente en pacientes pediátricos contando hasta 75% de las leucemias y 32-35% del total de cánceres infantiles. Aunque la LLA es considerada una enfermedad con base genética, es cada vez más evidente que alteraciones epigenéticas desempeñan un rol central en su patogénia y progresión. La hipermetilación de regiones promotoras de genes es asociada con la pérdida de función génica. El gen supresor de tumores p53 (GST), es uno de los principales genes en el ciclo celular y apoptosis. El objetivo de este trabajo fue determinar el estado de metilación en la región del promotor-exón 1 del GST p53 y la asociación con la supervivencia en menores de 15 años con LLA. Se analizaron 40 pacientes provenientes de la Región de la Araucanía-Chile. La hipermetilación del p53 se determinó combinando enzimas de restricción sensibles a metilación (HpaII y EcoR II) y reacción en cadena de la polimerasa. Los resultados indicaron que 15/40 casos (37,5%) presentaron hipermetilación. Se encontró una diferencia estadística en la supervivencia según estado de metilación de p53 en el grupo de niñas (p=0,02). Considerando el total de pacientes, una tendencia a mejor supervivencia cuando los recuentos de leucocitos fueron <30.000/mm3 (p=0,08). Se encontró frecuentemente hipermetilado el gen p53 en la región del promotor-exon1. Esto indicaría que la hipermetilación del GST p53 puede ser un evento importante en la patogénesis de la LLA.

Acute lymphoblastic leukemia (ALL) is the most common hematology oncology malignancy in pediatric patients counting up to 75% of leukemias and 32­35% of all childhood cancers. Although ALL is considered a disease with a genetic basis, it is increasingly clear that epigenetic alterations play a central role in the pathogenesis and work was to determine the methylation status in promoter-exon1 of the TSG-p53 and association with survival in children under 15 years with ALL. In our study 40 patients from the Araucanía Region, Chile were analyzed. Hypermethylation of p53 was determined by combining restriction enzymes sensitive to methylation (HpaII and EcoR II) and polymerase chain reaction. Results indicated that 15/40 cases (37.5%) showed hypermethylation. Statistical difference was found in survival according to p53 methylation status in the girls group (p=0.02). Considering all patients, there was a trend to improved survival when leukocyte counts were <30.000/ul (p=0.08). We found the p53 gene frequently hypermethylated in the promoter-exon1 region. This would indicate that TSG p53 hypermethylation may be an important event in the pathogenesis of ALL.
Descritores: Genes p53
Metilação de DNA
Leucemia-Linfoma Linfoblástico de Células Precursoras/genética
-Medula Óssea
Enzimas de Restrição do DNA
Análise de Sobrevida
Regiões Promotoras Genéticas
Epigênese Genética
Distribuição por Idade e Sexo
Reação em Cadeia da Polimerase Multiplex
Contagem de Leucócitos
Limites: Humanos
Masculino
Feminino
Lactente
Pré-Escolar
Criança
Adolescente
Responsável: CL1.1 - Biblioteca Central


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Id: lil-749718
Autor: Jung, Lenice Roteia Cardoso; Bomfim, Maria Rosa Quaresma; Kroon, Erna Geessien; Nunes, Álvaro Cantini.
Título: Identification of Leptospira serovars by RFLP of the RNA polymerase beta subunit gene (rpoB)
Fonte: Braz. j. microbiol;46(2):465-476, Apr-Jun/2015. tab, graf.
Idioma: en.
Resumo: Leptospires are usually classified by methods based on DNA-DNA hybridization and the conventional cross-agglutination absorption test, which uses polyclonal antibodies against lipopolysaccharides. In this study, the amplification of the rpoB gene, which encodes the beta-subunit of RNA polymerase, was used as an alternative tool to identify Leptospira. DNA extracts from sixty-eight serovars were obtained, and the hypervariable region located between 1990 and 2500-bp in the rpoB gene was amplified by polymerase chain reaction (PCR). The 600-bp amplicons of the rpoB gene were digested with the restriction endonucleases TaqI, Tru1I, Sau3AI and MslI, and the restriction fragments were separated by 6% polyacrylamide gel electrophoresis. Thirty-five fragment patters were obtained from the combined data of restriction fragment length polymorphism (PCR-RFLP) analysis and used to infer the phylogenetic relationships among the Leptospira species and serovars. The species assignments obtained were in full agreement with the established taxonomic classifications. Twenty-two serovars were effectively identified based on differences in their molecular profiles. However, the other 46 serovars remained clustered in groups that included more than one serovar of different species. This study demonstrates the value of RFLP analysis of PCR-amplified rpoB as an initial method for identifying Leptospira species and serovars.
Descritores: RNA Polimerases Dirigidas por DNA/genética
Leptospira/classificação
Leptospira/genética
Técnicas de Diagnóstico Molecular/métodos
Reação em Cadeia da Polimerase
Polimorfismo de Fragmento de Restrição
-Análise por Conglomerados
Enzimas de Restrição do DNA/metabolismo
Eletroforese em Gel de Poliacrilamida
Genótipo
Leptospira/isolamento & purificação
Dados de Sequência Molecular
Filogenia
Análise de Sequência de DNA
Sorogrupo
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Id: lil-607558
Autor: García-Agudo, Lidia; Jesús, Iría; Rodríguez-Iglesias, Manuel; García-Martos, Pedro.
Título: Evaluation of INNO-LiPA mycobacteria v2 assay for identification of rapidly growing mycobacteria
Fonte: Braz. j. microbiol;42(3):1220-1226, July-Sept. 2011. tab.
Idioma: en.
Resumo: A total of 54 rapidly growing mycobacteria (RGM) isolated from patients attended in the two hospitals of Cádiz Bay (Spain) were selected during a seven-year-period (2000-2006) in order to evaluate the INNO-LiPA Mycobacteria v2 assay for mycobacterial identification, based on the reverse hybridization principle. The strains were cultured in Lõwenstein-Jensen and Middlebrook 7H9 media and identified to the species level by sequencing of the 16S rRNA, PCR-restriction enzyme analysis of the hsp65 gene, conventional tests and INNO-LiPA Mycobacteria v2 assay. By the molecular methods we identified a total of 12 different species: 23 Mycobacterium fortuitum, 11 M. chelonae, 10 M. abscessus, 2 M. senegalense, 1 M. alvei, 1 M. brumae, 1 M. mageritense, 1 M. mucogenicum, 1 M. neoaurum, 1 M. peregrinum, 1 M. septicum and 1 M. smegmatis. Fifty two strains (96.3 percent) were correctly identified by conventional techniques and 47 strains (87.0 percent) by INNO-LiPA Mycobacteria v2 assay. We find INNO-LiPA Mycobacteria v2 assay simple to perform but it provides few advantages in comparison with conventional methods and sometimes needs complementary tests to identify Mycobacterium fortuitum complex, M. chelonae complex and specific species due to the great heterogeneity in the RGM group.
Descritores: Sequência de Bases
Enzimas de Restrição do DNA
Ativação Enzimática
Hibridização Genética
Técnicas In Vitro
Micobactérias não Tuberculosas/genética
Micobactérias não Tuberculosas/isolamento & purificação
Reação em Cadeia da Polimerase
-Marcadores Genéticos
Genética Microbiana
Métodos
Pacientes
Técnicas
Limites: Humanos
Tipo de Publ: Estudo Comparativo
Estudo de Avaliação
Responsável: BR32.1 - Serviço de Biblioteca e Informação Biomédica


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Id: lil-597204
Autor: Senna, Simone Gonçalves; Marsico, Ana Grazia; Vieira, Gisele Betzler de Oliveira; Sobral, Luciana Fonseca; Suffys, Philip Noel; Fonseca, Leila de Souza.
Título: Identificação de micobactérias não tuberculosas isoladas de sítios estéreis em pacientes em um hospital universitário na cidade do Rio de Janeiro / Identification of nontuberculous mycobacteria isolated from clinical sterile sites in patients at a university hospital in the city of Rio de Janeiro, Brazil
Fonte: J. bras. pneumol;37(4):521-526, jul.-ago. 2011. ilus, tab.
Idioma: pt.
Resumo: OBJETIVO: Identificar micobactérias não tuberculosas (MNT) isoladas de sítios estéreis em pacientes internados no Hospital Universitário Clementino Fraga Filho, Rio de Janeiro (RJ) entre 2001 e 2006. MÉTODOS: Durante o período do estudo, 34 isolados de MNT de sítios estéreis de 14 pacientes, a maioria HIV positivos, foram submetidos a identificação fenotípica e hsp65 PCR-restriction enzyme analysis (PRA, análise por enzimas de restrição por PCR do gene hsp65). RESULTADOS: A maioria dos isolados foi identificada como Mycobacterium avium, seguida por M. monacense, M. kansasii e M. abscessus em menores proporções. CONCLUSÕES: A combinação de PRA, um método relativamente simples e de baixo custo, com algumas características fenotípicas pode fornecer a identificação correta de MNT na rotina de laboratórios clínicos.

OBJECTIVE: To identify nontuberculous mycobacteria (NTM) isolated from sterile sites in patients hospitalized between 2001 and 2006 at the Clementino Fraga Filho University Hospital, located in the city of Rio de Janeiro, Brazil. METHODS: During the study period, 34 NTM isolates from sterile sites of 14 patients, most of whom were HIV-positive, were submitted to phenotypic identification and hsp65 PCR-restriction enzyme analysis (PRA). RESULTS: Most isolates were identified as Mycobacterium avium, followed by M. monacense, M. kansasii, and M. abscessus. CONCLUSIONS: The combination of PRA, a relatively simple and inexpensive method, with the evaluation of a few phenotypic characteristics can allow NTM to be accurately identified in the routine of clinical laboratories.
Descritores: Proteínas de Bactérias/análise
/análise
CHAPERONIN ACCESSORY NERVE/análise
Genes Bacterianos/genética
Infecções por Mycobacterium não Tuberculosas/microbiologia
Micobactérias não Tuberculosas/genética
Reação em Cadeia da Polimerase/métodos
Mapeamento por Restrição/métodos
-Técnicas Bacteriológicas
Brasil
Enzimas de Restrição do DNA
DNA Bacteriano/análise
Hospitais Universitários
Pacientes Internados
Complexo Mycobacterium avium/isolamento & purificação
Infecção por Mycobacterium avium-intracellulare/microbiologia
Micobactérias não Tuberculosas/isolamento & purificação
Limites: Adolescente
Adulto
Humanos
Pessoa de Meia-Idade
Adulto Jovem
Responsável: BR1.1 - BIREME


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Miagostovich, Marize P
Leite, José Paulo G
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Id: lil-570683
Autor: Rose, Tatiana L; Miagostovich, Marize P; Leite, José Paulo G.
Título: Rotavirus A genotype G1P[8]: a novel method to distinguish wild-type strains from the Rotarix® vaccine strain
Fonte: Mem. Inst. Oswaldo Cruz;105(8):1068-1072, Dec. 2010. ilus.
Idioma: en.
Projeto: FAPERJ. FCC; . CNPq.
Resumo: Rotaviruses are important enteric pathogens for humans and animals. Group A rotaviruses (RV-A) are the most common agents of severe gastroenteritis in infants and young children and vaccination is the most effective method to reduce RV-A-associated diseases. G1P[8], the most prevalent RV-A genotype worldwide, is included in the RV-A vaccine Rotarix®. The discrimination between wild-type G1P[8] and vaccine G1P[8] strains is an important topic in the study of RV-A epidemiology to manage outbreaks and to define control measures for vaccinated children. In this study, we developed a novel method to segregate the wild-type and vaccine strains using restriction endonucleases. The dsRNA from the Rotarix® vaccine was sequenced and the NSP3 gene was selected as the target gene. The vaccine strain has a restriction pattern that is different than that of wild-type RV-A G1P[8] isolates after digestion with the restriction endonuclease BspHI. This pattern could be used as a marker for the differentiation of wild-type G1P[8] strains from the vaccine strain.
Descritores: Fezes
Vacinas contra Rotavirus
Rotavirus
-Enzimas de Restrição do DNA
Genótipo
Polimorfismo de Fragmento de Restrição
Infecções por Rotavirus
Rotavirus
Vacinas Atenuadas
Limites: Humanos
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Id: lil-556001
Autor: Nes, Fernanda De; Riboldi, Gustavo Pelicioli; Frazzon, Ana Paula Guedes; d'Azevedo, Pedro Alves; Frazzon, Jeverson.
Título: Antimicrobial resistance and investigation of the molecular epidemiology of Listeria monocytogenes in dairy products / A resistência antimicrobiana e investigação de epidemiologia molecular de Listeria monocytogenes em produtos lácteos
Fonte: Rev. Soc. Bras. Med. Trop;43(4):382-385, jul.-ago. 2010. ilus.
Idioma: en.
Resumo: INTRODUCTION: Listeria monocytogenes is a ubiquitous microorganism in nature and is responsible for listeriosis, an infectious disease caused by consumption of contaminated food. METHODS: Molecular characterization was performed on 19 strains of Listeria monocytogenes (serovars 1/2a, 1/2b, 4b and 4c), isolated from dairy products in Rio Grande do Sul, Brazil. The molecular techniques applied were random amplification of polymorphic DNA and restriction enzyme analysis. In addition to the molecular analysis, the antimicrobial resistance profile was determined. RESULTS: The strains studied showed a low degree of diversity. In relation to the antimicrobial resistance profile of those microorganisms from the samples analyzed, all of them were susceptible to the antimicrobials tested. CONCLUSIONS: The molecular techniques that were used presented good discriminatory power for the strains studied. Furthermore, all of the samples that were analyzed were susceptible to the antimicrobials tested.

INTRODUÇÃO: Listeria monocytogenes é um microrganismo que se encontra disseminado na natureza, sendo responsável por causar listeriose, uma doença infecciosa causada pelo consumo de alimentos contaminados. MÉTODOS: A análise molecular de 19 linhagens de Listeria monocytogenes, sorovares 1/2a, 1/2b, 4b, 4c, isoladas de produtos lácteos do Rio Grande do Sul, Brasil. As técnicas moleculares aplicadas foram: Amplificação Randômica do DNA Polimórfico e Análise por Enzimas de Restrição. Além da análise molecular foi realizado o perfil de resistência antimicrobiana. RESULTADOS: As linhagens estudadas mostraram baixo grau de diversidade, em relação ao perfil de resistência antimicrobiana desses microrganismos das amostras analisadas todas foram susceptíveis aos antimicrobianos testados. CONCLUSÕES: As técnicas moleculares estudadas apresentaram um bom poder de discriminação para as linhagens estudadas. Além disso, todas as amostras analisadas foram susceptíveis aos antimicrobianos analisados.
Descritores: Laticínios/microbiologia
Microbiologia de Alimentos
Listeria monocytogenes/genética
-Antibacterianos/farmacologia
Brasil
Enzimas de Restrição do DNA
Listeria monocytogenes/efeitos dos fármacos
Testes de Sensibilidade Microbiana
Epidemiologia Molecular
Técnica de Amplificação ao Acaso de DNA Polimórfico
Responsável: BR1.1 - BIREME


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Richtzenhain, Leonardo José
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Id: lil-549421
Autor: Souza, Sibele Pinheiro de; Asano, Karen Miyuki; Sandri, Thaisa Lucas; Barros, Iracema Nunes de; Richtzenhain, Leonardo José; Brandão, Paulo Eduardo.
Título: Differentiation of Bovine coronavirus (BCoV) genotypes by a restriction enzyme assay
Fonte: Braz. j. microbiol;41(3):810-812, Oct. 2010. ilus.
Idioma: en.
Projeto: Fundação de Amparo à Pesquisa do Estado de São Paulo.
Resumo: This article reports the use of the GsuI restriction enzyme to differentiate genotypes of Bovine Coronavirus (BCoV), based on an 18-nucleotide deletion of S1-coding region found in one of the two genotypes. It was concluded that this assay can be used as a rapid tool for BCoV genotypes differentiation.
Descritores: Coronavirus Bovino/isolamento & purificação
Coronavirus Bovino/patogenicidade
Enzimas de Restrição do DNA
Ativadores de Enzimas
-Genótipo
Métodos
Técnicas
Virulência
Limites: Animais
Bovinos
Tipo de Publ: Relatório Técnico
Responsável: BR32.1 - Serviço de Biblioteca e Informação Biomédica


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Id: lil-534166
Autor: Scheidegger, EMD; Fracalanzza, SAP; Teixeira, LM; Cardarelli-Leite, P.
Título: RFLP analysis of a PCR-amplified fragment of the 16S rRNA gene as a tool to identify Enterococcus strains
Fonte: Mem. Inst. Oswaldo Cruz;104(7):1003-1008, Nov. 2009. tab, ilus.
Idioma: en.
Resumo: Restriction fragment length polymorphism (RFLP) analysis of a PCR-amplified fragment of the 16S rRNA gene was performed on reference strains belonging to 21 different enterococcal species and on 75 Enterococcus isolates recovered from poultry meat, pasteurised milk and fresh cheese. PCR amplification generated a 275 bp fragment, which was digested with three restriction endonucleases (DdeI, HaeIII, HinfI). The strains were divided into five groups (groups A-E) on the basis of their restriction patterns. Five biochemical tests (arabinose, arginine, manitol, methyl-â-D-glucopyranoside and raffinose) were then performed in addition to RFLP analysis to narrow the identification of enterococcal strains to the species level. PCR-RFLP, in conjunction with the selected biochemical tests, allowed the precise identification of the 21 species of Enterococcus included in the present study. This proposed method is relatively simple and rapid and can be useful as an adjunct tool for accurate identification of Enterococcus.
Descritores: DNA Bacteriano/genética
Enterococcus/genética
Polimorfismo de Fragmento de Restrição/genética
/genética
RNA, RIBOSOMAL, ABNORMALITIES, MULTIPLES/genética
-Sequência de Bases
Enzimas de Restrição do DNA
Enterococcus/classificação
Enterococcus/isolamento & purificação
Microbiologia de Alimentos
Reação em Cadeia da Polimerase
Limites: Animais
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


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Id: lil-531897
Autor: Tosto, Daniela; Hopp, Esteban.
Título: Characterization of the nuclear ribosomal DNA unit in Oxalis tuberosa (Oxalidacea) and related species
Fonte: Electron. j. biotechnol;11(3):11-22, July 2008. ilus, graf.
Idioma: en.
Projeto: Instituto Nacional de Tecnología Agropecuaria. Filogeografía e implementación de un sistema de monitoreo molecular de la identidad genética de cultivos andinos.
Resumo: Oxalis tuberosa is an octoploid Andean tuber crop called oca that belongs to the worldwide distributed genus Oxalis. The genus is very heterogeneous and its systematics is still problematic. It has been proposed that O. tuberosa evolved by polyploidization of a still not defined ancestor that belongs to an alliance of species sharing the same basic chromosome number (x = 8). Nuclear rDNA units of O. tuberosa and a selected group of four related diploid species were characterised by RFLP using different restriction endonucleases and southern hybridization probes to produce a restriction map for EcoRI and BamHI. The major rDNA unit length in O. tuberosa was estimated at 10.7 kbp. As expected, restriction site variation was observed mainly in the intergenic spacer (IGS), but was also detected in coding regions. Restriction site mapping organization of the transcribed rDNA unit of O. tuberosa is very similar to O. oblongiformis. Nucleotide sequencing of a region of O. peduncularis IGS generated a complex organization pattern of repeats and subrepeats. Diploid species O. peduncularis, O. tabaconasensis and O. aff. villosula exhibited a ladder pattern that is a consequence of a 170 bp subrepeat unit indicating that these species share organization similarity and sequence homology. The variation pattern provided information to compare among diploid species, although it did not help to clarify taxonomic relationships between O. tuberosa and the putative diploid ancestors analysed in this study. Nonetheless, the RFLP pattern exhibited by O. tuberosa for the IGS region was quite unique and will be a useful tool to prospect in other related species.
Descritores: DNA
Plantas/genética
Tabaco/genética
-Sequência de Bases
Enzimas de Restrição do DNA
Reação em Cadeia da Polimerase
Responsável: CL1.1 - Biblioteca Central


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Id: lil-522319
Autor: Shahinnia, Fahimeh; Sayed-Tabatabaei, Badraldin Ebrahim.
Título: Conversion of barley SNPs into PCR-based markers using dCAPS method
Fonte: Genet. mol. biol;32(3):564-567, 2009. ilus, tab.
Idioma: en.
Resumo: Molecular genetic research relies heavily on the ability to detect polymorphisms in DNA. Single nucleotide polymorphisms (SNPs) are the most frequent form of DNA variation in the genome. In combination with a PCR assay, the corresponding SNP can be analyzed as a derived cleaved amplified polymorphic sequence (dCAPS) marker. The dCAPS method exploits the well-known specificity of a restriction endonuclease for its recognition site and can be used to virtually detect any SNP. Here, we describe the use of the dCAPS method for detecting single-nucleotide changes by means of a barley EST, CK569932, PCR-based marker.
Descritores: Hordeum/genética
Polimorfismo de Nucleotídeo Único
-Enzimas de Restrição do DNA
Genoma
Reação em Cadeia da Polimerase
Responsável: BR26.1 - Biblioteca Central



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