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Pesquisa : G02.111.873.500 [Categoria DeCS]
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[PMID]:28463753
[Au] Autor:Walker A; Bergmann M; Camdereli J; Kaiser R; Lübke N; Timm J
[Ad] Endereço:Institute of Virology, Heinrich-Heine-University, University Hospital, Düsseldorf, Germany.
[Ti] Título:A genotype independent, full-genome reverse-transcription protocol for HCV genotyping and resistance testing.
[So] Source:J Clin Virol;91:42-48, 2017 06.
[Is] ISSN:1873-5967
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: HCV treatment options and cure rates have tremendously increased in the last decade. Although a pan-genotype HCV treatment has recently been approved, most DAA therapies are still genotype specific. Resistance-associated variants (RAVs) can limit the efficacy of DAA therapy and are associated with increased risk for therapy failure. With the approval of DAA regimens that recommend resistance testing prior to therapy, correct assessment of the genotype and testing for viruses with RAVs is clinically relevant. However, genotyping and resistance testing is generally done in costly and laborious separate reactions. OBJECTIVE: The aim of the study was to establish a genotype-independent full-genome reverse transcription protocol to generate a template for both genotyping and resistance testing and to implement it into our routine diagnostic setup. STUDY DESIGN: The complete HCV genome was reverse transcribed with a pan-genotype primer binding at the 3'end of the viral RNA. This cDNA served as template for transcription of the genotyping amplicon in the core region as well as for the resistance testing of NS3, NS5A, and NS5B. RESULTS: With the established RT-protocol the HCV core region was successfully amplified and genotyped from 124 out of 125 (99.2%) HCV-positive samples. The amplification efficiency of RAV containing regions in NS3, NS5A, NS5B was 96.2%, 96.6% and 94.4%, respectively. CONCLUSIONS: We developed a method for HCV full-genome cDNA synthesis and implemented it into a routine diagnostic setup. This cDNA can be used as template for genotyping amplicons covering the core or NS5B region as well as for resistance testing amplicons in NS3, NS5A and NS5B.
[Mh] Termos MeSH primário: Farmacorresistência Viral/genética
Genoma Viral/genética
Técnicas de Genotipagem
Hepacivirus/efeitos dos fármacos
Hepacivirus/genética
Transcrição Reversa
[Mh] Termos MeSH secundário: Antivirais/farmacologia
Antivirais/uso terapêutico
DNA Complementar
Genoma Viral/efeitos dos fármacos
Genótipo
Hepacivirus/isolamento & purificação
Hepatite C Crônica/sangue
Hepatite C Crônica/diagnóstico
Hepatite C Crônica/tratamento farmacológico
Seres Humanos
Reação em Cadeia da Polimerase
RNA Viral/sangue
RNA Viral/genética
Sensibilidade e Especificidade
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Antiviral Agents); 0 (DNA, Complementary); 0 (RNA, Viral)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180311
[Lr] Data última revisão:
180311
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170503
[St] Status:MEDLINE


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[PMID]:29307564
[Au] Autor:Niikura M; Inoue SI; Fukutomi T; Yamagishi J; Asahi H; Kobayashi F
[Ad] Endereço:Department of Infectious Diseases, Kyorin University School of Medicine, Tokyo 181-8611, Japan.
[Ti] Título:Comparative genomics and proteomic analyses between lethal and nonlethal strains of Plasmodium berghei.
[So] Source:Exp Parasitol;185:1-9, 2018 Feb.
[Is] ISSN:1090-2449
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Plasmodium berghei (Pb) XAT, a rodent malaria parasite, is an irradiation-attenuated variant derived from the lethal strain Pb NK65. Differences in genome sequence, protein structure and function between Pb XAT and Pb NK65 are currently unknown. In this study, to investigate genetic alterations in Pb XAT, we performed comparative genomics and proteomics analyses of nonlethal and lethal strains of Pb. We found mutations, such as a deletion mutation in rhoptry-associated protein (rap) 1, and deletion of rap2/3 and skeleton-binding protein 1 (sbp1), in Pb XAT. RAP1 is required for targeting of RAP2 to the rhoptries. However, the contribution of RAP2/3 to the lethality of Plasmodium is unclear. Therefore, we generated RAP1- and RAP2/3-deficient mutants of Pb ANKA, a reference strain of P. berghei. Furthermore, we investigated the effect of RAP1 and RAP2/3 deficiency on the outcome of infection. The parasitemia in mice infected with RAP1-deficient parasites was increased compared to that in control parasite-infected mice during the early phase of infection. However, mice infected with RAP1-deficient parasites survived longer than did control parasite-infected mice. Moreover, mice infected with RAP2/3-deficient parasites showed low levels of parasitemia and ultimately recovered from the infection The aim of this study was to investigate the effect of RAP2/3 expression on the outcome of infection with Pb XAT using a RAP2/3-expressing Pb XAT. Results showed that complementation of RAP2/3 expression in Pb XAT partially restored virulence. Our findings suggest that RAP1 and RAP2/3 contribute to virulence and a decrease in their expression explains the loss of virulence of the Pb XAT strain.
[Mh] Termos MeSH primário: Genômica
Malária/parasitologia
Plasmodium berghei/patogenicidade
Proteômica
[Mh] Termos MeSH secundário: Animais
Cromatografia Líquida
DNA de Protozoário/química
DNA de Protozoário/genética
Eritrócitos/parasitologia
Feminino
Malária/mortalidade
Camundongos
Camundongos Endogâmicos C57BL
Parasitemia/parasitologia
Plasmodium berghei/genética
Plasmodium berghei/metabolismo
Reação em Cadeia da Polimerase/métodos
Proteínas de Protozoários/genética
Transcrição Reversa
Deleção de Sequência
Organismos Livres de Patógenos Específicos
Espectrometria de Massas em Tandem
Virulência
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Protozoan); 0 (Protozoan Proteins); 0 (rhoptry associated protein, Plasmodium); 145184-78-7 (rhoptry-associated antigen-2, Plasmodium)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180214
[Lr] Data última revisão:
180214
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180109
[St] Status:MEDLINE


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[PMID]:29253877
[Au] Autor:Geldreich A; Haas G; Kubina J; Bouton C; Tanguy M; Erhardt M; Keller M; Ryabova L; Dimitrova M
[Ad] Endereço:Institut de Biologie Moléculaire des Plantes, CNRS UPR2357, Université de Strasbourg, Strasbourg, France.
[Ti] Título:Formation of large viroplasms and virulence of Cauliflower mosaic virus in turnip plants depend on the N-terminal EKI sequence of viral protein TAV.
[So] Source:PLoS One;12(12):e0189062, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Cauliflower mosaic virus (CaMV) TAV protein (TransActivator/Viroplasmin) plays a pivotal role during the infection cycle since it activates translation reinitiation of viral polycistronic RNAs and suppresses RNA silencing. It is also the major component of cytoplasmic electron-dense inclusion bodies (EDIBs) called viroplasms that are particularly evident in cells infected by the virulent CaMV Cabb B-JI isolate. These EDIBs are considered as virion factories, vehicles for CaMV intracellular movement and reservoirs for CaMV transmission by aphids. In this study, focused on different TAV mutants in vivo, we demonstrate that three physically separated domains collectively participate to the formation of large EDIBs: the N-terminal EKI motif, a sequence of the MAV domain involved in translation reinitiation and a C-terminal region encompassing the zinc finger. Surprisingly, EKI mutant TAVm3, corresponding to a substitution of the EKI motif at amino acids 11-13 by three alanines (AAA), which completely abolished the formation of large viroplasms, was not lethal for CaMV but highly reduced its virulence without affecting the rate of systemic infection. Expression of TAVm3 in a viral context led to formation of small irregularly shaped inclusion bodies, mild symptoms and low levels of viral DNA and particles accumulation, despite the production of significant amounts of mature capsid proteins. Unexpectedly, for CaMV-TAVm3 the formation of viral P2-containing electron-light inclusion body (ELIB), which is essential for CaMV aphid transmission, was also altered, thus suggesting an indirect role of the EKI tripeptide in CaMV plant-to-plant propagation. This important functional contribution of the EKI motif in CaMV biology can explain the strict conservation of this motif in the TAV sequences of all CaMV isolates.
[Mh] Termos MeSH primário: Brassica napus/virologia
Caulimovirus/metabolismo
Caulimovirus/patogenicidade
Transativadores/química
Transativadores/metabolismo
[Mh] Termos MeSH secundário: Motivos de Aminoácidos
Sequência de Aminoácidos
Caulimovirus/ultraestrutura
Corpos de Inclusão Viral/metabolismo
Corpos de Inclusão Viral/ultraestrutura
Proteínas Mutantes/metabolismo
Fenótipo
Domínios Proteicos
Protoplastos/metabolismo
Transcrição Reversa/genética
Relação Estrutura-Atividade
Virulência
Replicação Viral
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Mutant Proteins); 0 (Trans-Activators); 0 (gene VI protein, Cauliflower mosaic virus)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180108
[Lr] Data última revisão:
180108
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171219
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0189062


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[PMID]:29223150
[Au] Autor:Filippova JA; Semenov DV; Juravlev ES; Komissarov AB; Richter VA; Stepanov GA
[Ad] Endereço:Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Novosibirsk, 630090, Russia. stepanovga@niboch.nsc.ru.
[Ti] Título:Modern Approaches for Identification of Modified Nucleotides in RNA.
[So] Source:Biochemistry (Mosc);82(11):1217-1233, 2017 Nov.
[Is] ISSN:1608-3040
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:This review considers approaches for detection of modified monomers in the RNA structure of living organisms. Recently, some data on dynamic alterations in the pool of modifications of the key RNA species that depend on external factors affecting the cells and physiological conditions of the whole organism have been accumulated. The recent studies have presented experimental data on relationship between the mechanisms of formation of modified/minor nucleotides of RNA in mammalian cells and the development of various pathologies. The development of novel methods for detection of chemical modifications of RNA nucleotides in the cells of living organisms and accumulation of knowledge on the contribution of modified monomers to metabolism and functioning of individual RNA species establish the basis for creation of novel diagnostic and therapeutic approaches. This review includes a short description of routine methods for determination of modified nucleotides in RNA and considers in detail modern approaches that enable not only detection but also quantitative assessment of the modification level of various nucleotides in individual RNA species.
[Mh] Termos MeSH primário: Nucleotídeos/química
Processamento Pós-Transcricional do RNA
RNA/genética
[Mh] Termos MeSH secundário: Animais
Técnicas de Química Analítica/instrumentação
Técnicas de Química Analítica/métodos
Cromatografia Líquida de Alta Pressão
Técnicas Genéticas
Seres Humanos
Espectrometria de Massas
Métodos
Nucleotídeos/análise
Transcrição Reversa
Ribonucleases/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (Nucleotides); 63231-63-0 (RNA); EC 3.1.- (Ribonucleases)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180103
[Lr] Data última revisão:
180103
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171211
[St] Status:MEDLINE
[do] DOI:10.1134/S0006297917110013


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[PMID]:29016661
[Au] Autor:Yan YW; Zou B; Zhu T; Hozzein WN; Quan ZX
[Ad] Endereço:Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering, School of Life Sciences, Fudan University, Shanghai, People's Republic of China.
[Ti] Título:Modified RNA-seq method for microbial community and diversity analysis using rRNA in different types of environmental samples.
[So] Source:PLoS One;12(10):e0186161, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:RNA-seq-based SSU (small subunit) rRNA (ribosomal RNA) analysis has provided a better understanding of potentially active microbial community within environments. However, for RNA-seq library construction, high quantities of purified RNA are typically required. We propose a modified RNA-seq method for SSU rRNA-based microbial community analysis that depends on the direct ligation of a 5' adaptor to RNA before reverse-transcription. The method requires only a low-input quantity of RNA (10-100 ng) and does not require a DNA removal step. The method was initially tested on three mock communities synthesized with enriched SSU rRNA of archaeal, bacterial and fungal isolates at different ratios, and was subsequently used for environmental samples of high or low biomass. For high-biomass salt-marsh sediments, enriched SSU rRNA and total nucleic acid-derived RNA-seq datasets revealed highly consistent community compositions for all of the SSU rRNA sequences, and as much as 46.4%-59.5% of 16S rRNA sequences were suitable for OTU (operational taxonomic unit)-based community and diversity analyses with complete coverage of V1-V2 regions. OTU-based community structures for the two datasets were also highly consistent with those determined by all of the 16S rRNA reads. For low-biomass samples, total nucleic acid-derived RNA-seq datasets were analyzed, and highly active bacterial taxa were also identified by the OTU-based method, notably including members of the previously underestimated genus Nitrospira and phylum Acidobacteria in tap water, members of the phylum Actinobacteria on a shower curtain, and members of the phylum Cyanobacteria on leaf surfaces. More than half of the bacterial 16S rRNA sequences covered the complete region of primer 8F, and non-coverage rates as high as 38.7% were obtained for phylum-unclassified sequences, providing many opportunities to identify novel bacterial taxa. This modified RNA-seq method will provide a better snapshot of diverse microbial communities, most notably by OTU-based analysis, even communities with low-biomass samples.
[Mh] Termos MeSH primário: Archaea/genética
Bactérias/genética
Fungos/genética
Metagenoma
Filogenia
RNA Ribossômico 16S/genética
Análise de Sequência de DNA/métodos
[Mh] Termos MeSH secundário: Archaea/classificação
Bactérias/classificação
Conjuntos de Dados como Assunto
Água Potável/microbiologia
Fungos/classificação
Biblioteca Gênica
Sequenciamento de Nucleotídeos em Larga Escala
Consórcios Microbianos/genética
Folhas de Planta/microbiologia
Transcrição Reversa
Zonas Úmidas
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Drinking Water); 0 (RNA, Ribosomal, 16S)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171031
[Lr] Data última revisão:
171031
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171011
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0186161


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[PMID]:28977401
[Au] Autor:Weinberg Z; Lünse CE; Corbino KA; Ames TD; Nelson JW; Roth A; Perkins KR; Sherlock ME; Breaker RR
[Ad] Endereço:HHMI, Yale University, Box 208103, New Haven, CT 06520-8103, USA.
[Ti] Título:Detection of 224 candidate structured RNAs by comparative analysis of specific subsets of intergenic regions.
[So] Source:Nucleic Acids Res;45(18):10811-10823, 2017 Oct 13.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The discovery of structured non-coding RNAs (ncRNAs) in bacteria can reveal new facets of biology and biochemistry. Comparative genomics analyses executed by powerful computer algorithms have successfully been used to uncover many novel bacterial ncRNA classes in recent years. However, this general search strategy favors the discovery of more common ncRNA classes, whereas progressively rarer classes are correspondingly more difficult to identify. In the current study, we confront this problem by devising several methods to select subsets of intergenic regions that can concentrate these rare RNA classes, thereby increasing the probability that comparative sequence analysis approaches will reveal their existence. By implementing these methods, we discovered 224 novel ncRNA classes, which include ROOL RNA, an RNA class averaging 581 nt and present in multiple phyla, several highly conserved and widespread ncRNA classes with properties that suggest sophisticated biochemical functions and a multitude of putative cis-regulatory RNA classes involved in a variety of biological processes. We expect that further research on these newly found RNA classes will reveal additional aspects of novel biology, and allow for greater insights into the biochemistry performed by ncRNAs.
[Mh] Termos MeSH primário: RNA Bacteriano/química
RNA não Traduzido/química
Sequências Reguladoras de Ácido Ribonucleico
[Mh] Termos MeSH secundário: Integrons
Motivos de Nucleotídeos
Plasmídeos/genética
Transcrição Reversa
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Bacterial); 0 (RNA, Untranslated); 0 (Regulatory Sequences, Ribonucleic Acid)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171107
[Lr] Data última revisão:
171107
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171005
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx699


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[PMID]:28945787
[Au] Autor:Calvert AE; Biggerstaff BJ; Tanner NA; Lauterbach M; Lanciotti RS
[Ad] Endereço:Arboviral Diseases Branch, Division of Vector-Borne Diseases, U.S. Centers for Disease Control and Prevention, Fort Collins, CO, United States of America.
[Ti] Título:Rapid colorimetric detection of Zika virus from serum and urine specimens by reverse transcription loop-mediated isothermal amplification (RT-LAMP).
[So] Source:PLoS One;12(9):e0185340, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Zika virus (ZIKV) has emerged as a major global public health concern in the last two years due to its link as a causative agent of human birth defects. Its rapid expansion into the Western Hemisphere as well as the ability to be transmitted from mother to fetus, through sexual transmission and possibly through blood transfusions has increased the need for a rapid and expansive public health response to this unprecedented epidemic. A non-invasive and rapid ZIKV diagnostic screening assay that can be performed in a clinical setting throughout pregnancy is vital for prenatal care of women living in areas of the world where exposure to the virus is possible. To meet this need we have developed a sensitive and specific reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) assay to detect ZIKV RNA in urine and serum with a simple visual detection. RT-LAMP results were shown to have a limit of detection 10-fold higher than qRT-PCR. As little as 1.2 RNA copies/µl was detected by RT-LAMP from a panel of 178 diagnostic specimens. The assay was shown to be highly specific for ZIKV RNA when tested with diagnostic specimens positive for dengue virus (DENV) and chikungunya virus (CHIKV). The assay described here illustrates the potential for a fast, reliable, sensitive and specific assay for the detection of ZIKV from urine or serum that can be performed in a clinical or field setting with minimal equipment and technological expertise.
[Mh] Termos MeSH primário: Colorimetria/métodos
Zika virus/isolamento & purificação
[Mh] Termos MeSH secundário: Primers do DNA/genética
Feminino
Seres Humanos
Técnicas de Amplificação de Ácido Nucleico/métodos
Testes Imediatos
Valor Preditivo dos Testes
Gravidez
Complicações Infecciosas na Gravidez/diagnóstico
RNA Viral/sangue
RNA Viral/genética
RNA Viral/urina
Reação em Cadeia da Polimerase em Tempo Real/métodos
Transcrição Reversa
Zika virus/genética
Infecção pelo Zika virus/complicações
Infecção pelo Zika virus/diagnóstico
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Primers); 0 (RNA, Viral)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171016
[Lr] Data última revisão:
171016
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170926
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0185340


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[PMID]:28845978
[Au] Autor:Alenko A; Fleming AM; Burrows CJ
[Ad] Endereço:Department of Chemistry, University of Utah , 315 South 1400 East, Salt Lake City, Utah 84112-0850, United States.
[Ti] Título:Reverse Transcription Past Products of Guanine Oxidation in RNA Leads to Insertion of A and C opposite 8-Oxo-7,8-dihydroguanine and A and G opposite 5-Guanidinohydantoin and Spiroiminodihydantoin Diastereomers.
[So] Source:Biochemistry;56(38):5053-5064, 2017 Sep 26.
[Is] ISSN:1520-4995
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Reactive oxygen species, both endogenous and exogenous, can damage nucleobases of RNA and DNA. Among the nucleobases, guanine has the lowest redox potential, making it a major target of oxidation. Although RNA is more prone to oxidation than DNA is, oxidation of guanine in RNA has been studied to a significantly lesser extent. One of the reasons for this is that many tools that were previously developed to study oxidation of DNA cannot be used on RNA. In the study presented here, the lack of a method for seeking sites of modification in RNA where oxidation occurs is addressed. For this purpose, reverse transcription of RNA containing major products of guanine oxidation was used. Extension of a DNA primer annealed to an RNA template containing 8-oxo-7,8-dihydroguanine (OG), 5-guanidinohydantoin (Gh), or the R and S diastereomers of spiroiminodihydantoin (Sp) was studied under standing start conditions. SuperScript III reverse transcriptase is capable of bypassing these lesions in RNA inserting predominantly A opposite OG, predominantly G opposite Gh, and almost an equal mixture of A and G opposite the Sp diastereomers. These data should allow RNA sequencing of guanine oxidation products by following characteristic mutation signatures formed by the reverse transcriptase during primer elongation past G oxidation sites in the template RNA strand.
[Mh] Termos MeSH primário: Guanidinas/química
Guanina/análogos & derivados
Guanosina/análogos & derivados
Hidantoínas/química
RNA/química
Transcrição Reversa
Compostos de Espiro/química
[Mh] Termos MeSH secundário: Adenina/química
Guanina/química
Guanosina/química
Guanosina/genética
Cinética
Oxirredução
DNA Polimerase Dirigida por RNA/química
DNA Polimerase Dirigida por RNA/metabolismo
Estereoisomerismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Guanidines); 0 (Hydantoins); 0 (Spiro Compounds); 0 (guanidinohydantoin); 0 (spiroiminodihydantoin); 12133JR80S (Guanosine); 5614-64-2 (8-hydroxyguanine); 5Z93L87A1R (Guanine); 63231-63-0 (RNA); EC 2.7.7.49 (RNA-Directed DNA Polymerase); JAC85A2161 (Adenine)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171010
[Lr] Data última revisão:
171010
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170829
[St] Status:MEDLINE
[do] DOI:10.1021/acs.biochem.7b00730


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[PMID]:28835460
[Au] Autor:Achleitner M; Kleefisch M; Hennig A; Peschke K; Polikarpova A; Oertel R; Gabriel B; Schulze L; Lindeman D; Gerbaulet A; Fiebig U; Lee-Kirsch MA; Roers A; Behrendt R
[Ad] Endereço:Institute for Immunology, Medical Faculty Carl Gustav Carus, Technical University of Dresden, 01307 Dresden, Germany.
[Ti] Título:Lack of Trex1 Causes Systemic Autoimmunity despite the Presence of Antiretroviral Drugs.
[So] Source:J Immunol;199(7):2261-2269, 2017 Oct 01.
[Is] ISSN:1550-6606
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Biallelic mutations of three prime repair exonuclease 1 (TREX1) cause the lupus-like disease Aicardi-Goutières syndrome in which accumulation of a yet unknown endogenous DNA substrate of TREX1 triggers a cyclic GMP-AMP synthase-dependent type I IFN response and systemic autoimmunity. Products of reverse transcription originating from endogenous retroelements have been suggested to be a major substrate for TREX1, and reverse transcriptase inhibitors (RTIs) were proposed as a therapeutic option in autoimmunity ensuing from defects of TREX1. In this study, we treated mice with RTIs. The serum RTI levels reached were sufficient to block retrotransposition of endogenous retroelements. However, the treatment did not reduce the spontaneous type I IFN response and did not ameliorate lethal inflammation. Furthermore, long interspersed nuclear elements 1 retrotransposition was not enhanced in the absence of Trex1. Our data do not support the concept of retroelement-derived cDNA as key triggers of systemic autoimmunity in Trex1-deficient humans and mice and motivate the continuing search for the pathogenic IFN-inducing Trex1 substrate.
[Mh] Termos MeSH primário: Autoimunidade
Exodesoxirribonucleases/metabolismo
Fosfoproteínas/metabolismo
Inibidores da Transcriptase Reversa/sangue
[Mh] Termos MeSH secundário: Animais
Doenças Autoimunes do Sistema Nervoso/imunologia
DNA Complementar
Exodesoxirribonucleases/deficiência
Exodesoxirribonucleases/genética
Células HeLa
Seres Humanos
Inflamação
Interferon Tipo I/biossíntese
Interferon Tipo I/imunologia
Camundongos
Mutação
Malformações do Sistema Nervoso/imunologia
Fosfoproteínas/deficiência
Fosfoproteínas/genética
Retroelementos
Inibidores da Transcriptase Reversa/efeitos adversos
Inibidores da Transcriptase Reversa/uso terapêutico
Transcrição Reversa
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Complementary); 0 (Interferon Type I); 0 (Phosphoproteins); 0 (Retroelements); 0 (Reverse Transcriptase Inhibitors); EC 3.1.- (Exodeoxyribonucleases); EC 3.1.16.- (three prime repair exonuclease 1)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171024
[Lr] Data última revisão:
171024
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:170825
[St] Status:MEDLINE
[do] DOI:10.4049/jimmunol.1700714


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[PMID]:28703578
[Au] Autor:Wulff TF; Argüello RJ; Molina Jordàn M; Roura Frigolé H; Hauquier G; Filonava L; Camacho N; Gatti E; Pierre P; Ribas de Pouplana L; Torres AG
[Ad] Endereço:Institute for Research in Biomedicine (IRB Barcelona), The Barcelona Institute of Science and Technology , Parc Científic de Barcelona, C/Baldiri Reixac 10, 08028 Barcelona, Catalonia, Spain.
[Ti] Título:Detection of a Subset of Posttranscriptional Transfer RNA Modifications in Vivo with a Restriction Fragment Length Polymorphism-Based Method.
[So] Source:Biochemistry;56(31):4029-4038, 2017 Aug 08.
[Is] ISSN:1520-4995
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Transfer RNAs (tRNAs) are among the most heavily modified RNA species. Posttranscriptional tRNA modifications (ptRMs) play fundamental roles in modulating tRNA structure and function and are being increasingly linked to human physiology and disease. Detection of ptRMs is often challenging, expensive, and laborious. Restriction fragment length polymorphism (RFLP) analyses study the patterns of DNA cleavage after restriction enzyme treatment and have been used for the qualitative detection of modified bases on mRNAs. It is known that some ptRMs induce specific and reproducible base "mutations" when tRNAs are reverse transcribed. For example, inosine, which derives from the deamination of adenosine, is detected as a guanosine when an inosine-containing tRNA is reverse transcribed, amplified via polymerase chain reaction (PCR), and sequenced. ptRM-dependent base changes on reverse transcription PCR amplicons generated as a consequence of the reverse transcription reaction might create or abolish endonuclease restriction sites. The suitability of RFLP for the detection and/or quantification of ptRMs has not been studied thus far. Here we show that different ptRMs can be detected at specific sites of different tRNA types by RFLP. For the examples studied, we show that this approach can reliably estimate the modification status of the sample, a feature that can be useful in the study of the regulatory role of tRNA modifications in gene expression.
[Mh] Termos MeSH primário: Adenosina Desaminase/metabolismo
Modelos Biológicos
Polimorfismo de Fragmento de Restrição
Processamento Pós-Transcricional do RNA
RNA de Transferência de Alanina/metabolismo
RNA de Transferência de Treonina/metabolismo
[Mh] Termos MeSH secundário: Adenosina/metabolismo
Adenosina Desaminase/química
Adenosina Desaminase/genética
Análise do Polimorfismo de Comprimento de Fragmentos Amplificados
Pareamento de Bases
Biologia Computacional
Desaminação
Sistemas Especialistas
Células HeLa
Seres Humanos
Concentração de Íons de Hidrogênio
Inosina/metabolismo
Interferência de RNA
RNA Interferente Pequeno/metabolismo
RNA de Transferência de Alanina/antagonistas & inibidores
RNA de Transferência de Treonina/antagonistas & inibidores
RNA de Transferência de Valina/antagonistas & inibidores
RNA de Transferência de Valina/metabolismo
Transcrição Reversa
Especificidade por Substrato
[Pt] Tipo de publicação:JOURNAL ARTICLE; VALIDATION STUDIES
[Nm] Nome de substância:
0 (RNA, Small Interfering); 0 (RNA, Transfer, Ala); 0 (RNA, Transfer, Thr); 0 (RNA, Transfer, Val); 5A614L51CT (Inosine); EC 3.5.4.4 (ADAT2 protein, human); EC 3.5.4.4 (Adenosine Deaminase); K72T3FS567 (Adenosine)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170818
[Lr] Data última revisão:
170818
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170714
[St] Status:MEDLINE
[do] DOI:10.1021/acs.biochem.7b00324



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