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Pesquisa : A11.284.835.168 [Categoria DeCS]
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  1 / 18733 MEDLINE  
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[PMID]:28470613
[Au] Autor:Rues RB; Gräwe A; Henrich E; Bernhard F
[Ad] Endereço:Centre for Biomolecular Magnetic Resonance, Institute for Biophysical Chemistry, Goethe-University of Frankfurt/Main, Max-von-Laue-Str. 9, 60438, Frankfurt/Main, Germany.
[Ti] Título:Membrane Protein Production in E. coli Lysates in Presence of Preassembled Nanodiscs.
[So] Source:Methods Mol Biol;1586:291-312, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Cell-free expression allows to synthesize membrane proteins in completely new formats that can relatively easily be customized for particular applications. Amphiphilic superstructures such as micelles, lipomicelles, or nanodiscs can be provided as nano-devices for the solubilization of membrane proteins. Defined empty bilayers in the form of nanodiscs offer native like environments for membrane proteins, supporting functional folding, proper oligomeric assembly as well as stability. Even very difficult and detergent-sensitive membrane proteins can be addressed by the combination of nanodisc technology with efficient cell-free expression systems as the direct co-translational insertion of nascent membrane proteins into supplied preassembled nanodiscs is possible. This chapter provides updated protocols for the synthesis of membrane proteins in presence of preassembled nanodiscs suitable for emerging applications such as screening of lipid effects on membrane protein function and the modulation of oligomeric complex formation.
[Mh] Termos MeSH primário: Sistema Livre de Células/metabolismo
Escherichia coli/genética
Bicamadas Lipídicas/química
Proteínas de Membrana/genética
Nanoestruturas/química
Biologia Sintética/métodos
[Mh] Termos MeSH secundário: Detergentes/química
Expressão Gênica
Lipídeos/química
Proteínas de Membrana/química
Proteínas de Membrana/isolamento & purificação
Dobramento de Proteína
Solubilidade
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Detergents); 0 (Lipid Bilayers); 0 (Lipids); 0 (Membrane Proteins)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180220
[Lr] Data última revisão:
180220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE
[do] DOI:10.1007/978-1-4939-6887-9_19


  2 / 18733 MEDLINE  
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[PMID]:27770630
[Au] Autor:Jang J; Park HJ; Kim SW; Kim H; Park JY; Na SJ; Kim HJ; Park MN; Choi SH; Park SH; Kim SW; Kwon SM; Kim PJ; Cho DW
[Ad] Endereço:Department of Mechanical Engineering, Pohang University of Science and Technology (POSTECH), 77 Cheongam-ro, Nam-gu, Pohang, Kyungbuk, 37673, South Korea.
[Ti] Título:3D printed complex tissue construct using stem cell-laden decellularized extracellular matrix bioinks for cardiac repair.
[So] Source:Biomaterials;112:264-274, 2017 01.
[Is] ISSN:1878-5905
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Stem cell therapy is a promising therapeutic method for the treatment of ischemic heart diseases; however, some challenges prohibit the efficacy after cell delivery due to hostile microenvironment of the injured myocardium. 3D printed pre-vascularized stem cell patch can enhance the therapeutic efficacy for cardiac repair through promotion of rapid vascularization after patch transplantation. In this study, stem cell-laden decellularized extracellular matrix bioinks are used in 3D printing of pre-vascularized and functional multi-material structures. The printed structure composed of spatial patterning of dual stem cells improves cell-to-cell interactions and differentiation capability and promotes functionality for tissue regeneration. The developed stem cell patch promoted strong vascularization and tissue matrix formation in vivo. The patterned patch exhibited enhanced cardiac functions, reduced cardiac hypertrophy and fibrosis, increased migration from patch to the infarct area, neo-muscle and capillary formation along with improvements in cardiac functions. Therefore, pre-vascularized stem cell patch provides cardiac niche-like microenvironment, resulting in beneficial effects on cardiac repair.
[Mh] Termos MeSH primário: Matriz Extracelular/química
Regeneração Tecidual Guiada/instrumentação
Infarto do Miocárdio/terapia
Impressão Tridimensional
Transplante de Células-Tronco/instrumentação
Tecidos Suporte
[Mh] Termos MeSH secundário: Animais
Sistema Livre de Células/química
Células Cultivadas
Regeneração Tecidual Guiada/métodos
Seres Humanos
Tinta
Camundongos
Camundongos Endogâmicos BALB C
Camundongos Nus
Infarto do Miocárdio/patologia
Infarto do Miocárdio/fisiopatologia
Ratos
Ratos Endogâmicos F344
Transplante de Células-Tronco/métodos
Suínos
Engenharia Tecidual/instrumentação
Engenharia Tecidual/métodos
Resultado do Tratamento
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Em] Mês de entrada:1712
[Cu] Atualização por classe:180106
[Lr] Data última revisão:
180106
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161023
[St] Status:MEDLINE


  3 / 18733 MEDLINE  
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[PMID]:29223161
[Au] Autor:Wang J; Liu Y; Zhang J; Han Z; Wang W; Liu Y; Wei D; Huang W
[Ad] Endereço:Food Safety Research Center, Hebei North University, Zhangjiakou, 075000, China. xuanyuanjian0228@126.com.
[Ti] Título:Cell-Free Expression, Purification, and Characterization of the Functional ß -Adrenergic Receptor for Multianalyte Detection of ß-Agonists.
[So] Source:Biochemistry (Mosc);82(11):1346-1353, 2017 Nov.
[Is] ISSN:1608-3040
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Large-scale expression of ß -adrenergic receptor (ß -AR) in functional form is necessary for establishment of receptor assays for detecting illegally abused ß-adrenergic agonists (ß-agonists). Cell-based heterologous expression systems have manycritical difficulties in synthesizing this membrane protein, such as low protein yields and aberrant folding. To overcome these challenges, the main objective of the present work was to synthesize large amounts of functional ß -AR in a cell-free system based on Escherichia coli extracts. A codon-optimized porcine ß -AR gene (codon adaptation index: 0.96) suitable for high expression in E. coli was synthesized and transcribed to the cell-free system, which contributed to increase the expression up to 1.1 mg/ml. After purification using Ni-affinity chromatography, the bioactivity of the purified receptor was measured by novel enzyme-linked receptor assays. It was determined that the relative affinities of the purified ß -AR for ß-agonists in descending order were as follows: clenbuterol > salbutamol > ractopamine. Moreover, their IC values were 45.99, 60.38, and 78.02 µg/liter, respectively. Although activity of the cell-free system was slightly lower than activity of systems based on insect and mammalian cells, this system should allow production of ß -AR in bulk amounts sufficient for the development of multianalyte screening methods for detecting ß-agonist residues.
[Mh] Termos MeSH primário: Agonistas de Receptores Adrenérgicos beta 2/análise
Sistema Livre de Células/metabolismo
Receptores Adrenérgicos beta 2/biossíntese
[Mh] Termos MeSH secundário: Animais
Escherichia coli/metabolismo
Receptores Adrenérgicos beta 2/genética
Receptores Adrenérgicos beta 2/isolamento & purificação
Suínos/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Adrenergic beta-2 Receptor Agonists); 0 (Receptors, Adrenergic, beta-2)
[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/S0006297917110128


  4 / 18733 MEDLINE  
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[PMID]:29180822
[Au] Autor:Michelini F; Pitchiaya S; Vitelli V; Sharma S; Gioia U; Pessina F; Cabrini M; Wang Y; Capozzo I; Iannelli F; Matti V; Francia S; Shivashankar GV; Walter NG; d'Adda di Fagagna F
[Ad] Endereço:IFOM-The FIRC Institute of Molecular Oncology, Milan 20139, Italy.
[Ti] Título:Damage-induced lncRNAs control the DNA damage response through interaction with DDRNAs at individual double-strand breaks.
[So] Source:Nat Cell Biol;19(12):1400-1411, 2017 Dec.
[Is] ISSN:1476-4679
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The DNA damage response (DDR) preserves genomic integrity. Small non-coding RNAs termed DDRNAs are generated at DNA double-strand breaks (DSBs) and are critical for DDR activation. Here we show that active DDRNAs specifically localize to their damaged homologous genomic sites in a transcription-dependent manner. Following DNA damage, RNA polymerase II (RNAPII) binds to the MRE11-RAD50-NBS1 complex, is recruited to DSBs and synthesizes damage-induced long non-coding RNAs (dilncRNAs) from and towards DNA ends. DilncRNAs act both as DDRNA precursors and by recruiting DDRNAs through RNA-RNA pairing. Together, dilncRNAs and DDRNAs fuel DDR focus formation and associate with 53BP1. Accordingly, inhibition of RNAPII prevents DDRNA recruitment, DDR activation and DNA repair. Antisense oligonucleotides matching dilncRNAs and DDRNAs impair site-specific DDR focus formation and DNA repair. We propose that DDR signalling sites, in addition to sharing a common pool of proteins, individually host a unique set of site-specific RNAs necessary for DDR activation.
[Mh] Termos MeSH primário: Quebras de DNA de Cadeia Dupla
Dano ao DNA
Reparo do DNA
RNA Longo não Codificante/metabolismo
[Mh] Termos MeSH secundário: Transportadores de Cassetes de Ligação de ATP/metabolismo
Animais
Proteínas de Ciclo Celular/metabolismo
Linhagem Celular
Sistema Livre de Células
Dano ao DNA/genética
Dano ao DNA/fisiologia
Reparo do DNA/genética
Reparo do DNA/fisiologia
Proteína Homóloga a MRE11/metabolismo
Camundongos
Modelos Biológicos
Proteínas Nucleares/metabolismo
Oligonucleotídeos Antissenso/genética
RNA Polimerase II/metabolismo
RNA Longo não Codificante/genética
Transcrição Genética
Proteína 1 de Ligação à Proteína Supressora de Tumor p53/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (ATP-Binding Cassette Transporters); 0 (Cell Cycle Proteins); 0 (Mre11a protein, mouse); 0 (Nijmegen breakage syndrome 1 protein, mouse); 0 (Nuclear Proteins); 0 (Oligonucleotides, Antisense); 0 (RNA, Long Noncoding); 0 (Rad50 protein, mouse); 0 (Trp53bp1 protein, mouse); 0 (Tumor Suppressor p53-Binding Protein 1); EC 2.7.7.- (RNA Polymerase II); EC 3.1.- (MRE11 Homologue Protein)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:171219
[Lr] Data última revisão:
171219
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171129
[St] Status:MEDLINE
[do] DOI:10.1038/ncb3643


  5 / 18733 MEDLINE  
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[PMID]:28918745
[Au] Autor:Sogorin EA; Selikhanov GK; Agalarov SC
[Ad] Endereço:Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, 142290, Russia. sultan@vega.protres.ru.
[Ti] Título:Coupling of Translation Initiation and Termination Does Not Depend on the Mode of Initiation.
[So] Source:Biochemistry (Mosc);82(7):816-820, 2017 Jul.
[Is] ISSN:1608-3040
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Recently we described a novel phenomenon observed during eukaryotic translation in a cell-free system: the coupling of initiation and termination on different mRNA molecules. Here we show that the phenomenon does not depend on a special mode of initiation. The mRNAs with certain leader sequences known to require different determinants for successful initiation were examined. Even in a case of using the intergenic internal ribosome entry site (IRES) of cricket paralysis virus RNA as the leader sequence, while no initiation factors are required, the effect of coupling is well expressed, including trials in the presence of hippuristanol as an inhibitor of eIF4A. Thus, the effect persists in the absence of scanning and does not depend on initiator tRNA and eIF2. The results suggest that the initiation factors are not involved in the coupling mechanism.
[Mh] Termos MeSH primário: RNA Mensageiro/metabolismo
[Mh] Termos MeSH secundário: Regiões 5' não Traduzidas
Sistema Livre de Células/metabolismo
Fator de Iniciação 2 em Eucariotos/antagonistas & inibidores
Fator de Iniciação 2 em Eucariotos/metabolismo
Fator de Iniciação 4A em Eucariotos/antagonistas & inibidores
Fator de Iniciação 4A em Eucariotos/metabolismo
Genes Reporter
Sítios Internos de Entrada Ribossomal/genética
Iniciação Traducional da Cadeia Peptídica
Terminação Traducional da Cadeia Peptídica
Plasmídeos/metabolismo
Esteróis/química
Esteróis/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (5' Untranslated Regions); 0 (Eukaryotic Initiation Factor-2); 0 (Internal Ribosome Entry Sites); 0 (RNA, Messenger); 0 (Sterols); 0 (hippuristanol); EC 2.7.7.- (Eukaryotic Initiation Factor-4A)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170926
[Lr] Data última revisão:
170926
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170919
[St] Status:MEDLINE
[do] DOI:10.1134/S0006297917070069


  6 / 18733 MEDLINE  
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[PMID]:28911115
[Au] Autor:Gross L; Vicens Q; Einhorn E; Noireterre A; Schaeffer L; Kuhn L; Imler JL; Eriani G; Meignin C; Martin F
[Ad] Endereço:Université de Strasbourg, CNRS, Architecture et Réactivité de l'ARN, UPR 9002, F-67000 Strasbourg, France.
[Ti] Título:The IRES5'UTR of the dicistrovirus cricket paralysis virus is a type III IRES containing an essential pseudoknot structure.
[So] Source:Nucleic Acids Res;45(15):8993-9004, 2017 Sep 06.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Cricket paralysis virus (CrPV) is a dicistrovirus. Its positive-sense single-stranded RNA genome contains two internal ribosomal entry sites (IRESs). The 5' untranslated region (5'UTR) IRES5'UTR mediates translation of non-structural proteins encoded by ORF1 whereas the well-known intergenic region (IGR) IRESIGR is required for translation of structural proteins from open reading frame 2 in the late phase of infection. Concerted action of both IRES is essential for host translation shut-off and viral translation. IRESIGR has been extensively studied, in contrast the IRES5'UTR remains largely unexplored. Here, we define the minimal IRES element required for efficient translation initiation in drosophila S2 cell-free extracts. We show that IRES5'UTR promotes direct recruitment of the ribosome on the cognate viral AUG start codon without any scanning step, using a Hepatitis-C virus-related translation initiation mechanism. Mass spectrometry analysis revealed that IRES5'UTR recruits eukaryotic initiation factor 3, confirming that it belongs to type III class of IRES elements. Using Selective 2'-hydroxyl acylation analyzed by primer extension and DMS probing, we established a secondary structure model of 5'UTR and of the minimal IRES5'UTR. The IRES5'UTR contains a pseudoknot structure that is essential for proper folding and ribosome recruitment. Overall, our results pave the way for studies addressing the synergy and interplay between the two IRES from CrPV.
[Mh] Termos MeSH primário: Regiões 5´ não Traduzidas
Dicistroviridae/genética
Sítios Internos de Entrada Ribossomal
Biossíntese de Proteínas
RNA Viral/química
Proteínas Virais/química
[Mh] Termos MeSH secundário: Animais
Sequência de Bases
Linhagem Celular
Sistema Livre de Células/metabolismo
Dicistroviridae/crescimento & desenvolvimento
Dicistroviridae/metabolismo
Drosophila melanogaster/virologia
Gryllidae/virologia
Interações Hospedeiro-Patógeno
Conformação de Ácido Nucleico
Fases de Leitura Aberta
RNA Viral/genética
RNA Viral/metabolismo
Ribossomos/genética
Ribossomos/metabolismo
Proteínas Virais/genética
Proteínas Virais/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (5' Untranslated Regions); 0 (Internal Ribosome Entry Sites); 0 (RNA, Viral); 0 (Viral Proteins)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171012
[Lr] Data última revisão:
171012
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170916
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkx622


  7 / 18733 MEDLINE  
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[PMID]:28910365
[Au] Autor:Tug S; Tross AK; Hegen P; Neuberger EWI; Helmig S; Schöllhorn W; Simon P
[Ad] Endereço:Department of Sports Medicine, Disease Prevention and Rehabilitation, Johannes Gutenberg-University Mainz, Mainz, Germany.
[Ti] Título:Acute effects of strength exercises and effects of regular strength training on cell free DNA concentrations in blood plasma.
[So] Source:PLoS One;12(9):e0184668, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Creatine kinase (CK) is a marker for muscle cell damage with limited potential as marker for training load in strength training. Recent exercise studies identified cell free DNA (cfDNA) as a marker for aseptic inflammation and cell damage. Here we overserved in a pilot study the acute effects during strength exercise and chronic effects of regular strength training on cfDNA concentrations over a period of four weeks in three training groups applying conservation training (CT) at 60% of the 1 repetition maximum, high intensity-low repetition training (HT) at 90% of the 1 repetition maximum and differential training (DT) at 60% of the 1 repetition maximum. EDTA-plasma samples were collected before every training session, and on the first and last training day repeatedly after every set of exercises. CfDNA increased significantly by 1.62-fold (mean (±SD) before first exercise: 8.31 (2.84) ng/ml, after last exercise 13.48 (4.12) ng/ml) across all groups within a single training session (p<0.001). The increase was 1.77-fold higher (mean (±SD) before first exercise: 12.23 (6.29) ng/ml, after last exercise 17.73 (11.24) ng/ml) in HT compared to CT (mean (±SD) before first exercise: 6.79 (1.28) ng/ml, after last exercise 10.05 (2.89) ng/ml) (p = 0.01). DNA size analysis suggested predominant release of short, mononucleosomal DNA-fragments in the acute exercise setting, while we detected an increase of mostly longer, polynucleosomal cfDNA-fragments at rest before the training session only at day two with a subsequent return to baseline (p<0.001). In contrast, training procedures did not cause any alterations in CK. Our results suggest that during strength exercise short-fragmented cfDNA is released, reflecting a fast, aseptic inflammatory response, while elevation of longer fragments at baseline on day two seemed to reflect mild cellular damage due to a novel training regime. We critically discuss the implications of our findings for future evaluations of cfDNA as a marker for training load in strength training.
[Mh] Termos MeSH primário: DNA/sangue
Exercício/fisiologia
Resistência Física/genética
[Mh] Termos MeSH secundário: Adaptação Fisiológica
Adulto
Sistema Livre de Células
Creatina Quinase
Dano ao DNA
Seres Humanos
Masculino
Projetos Piloto
Treinamento de Resistência
Adulto Jovem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
9007-49-2 (DNA); EC 2.7.3.2 (Creatine Kinase)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171013
[Lr] Data última revisão:
171013
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170915
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0184668


  8 / 18733 MEDLINE  
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[PMID]:28887129
[Au] Autor:Hasinoff BB; Patel D
[Ad] Endereço:College of Pharmacy, Apotex Centre, University of Manitoba, Winnipeg, Manitoba R3E 0T5, Canada. Electronic address: B_Hasinoff@UManitoba.ca.
[Ti] Título:Disulfiram is a slow-binding partial noncompetitive inhibitor of 20S proteasome activity.
[So] Source:Arch Biochem Biophys;633:23-28, 2017 Nov 01.
[Is] ISSN:1096-0384
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The alcohol abuse drug disulfiram has also been shown to exhibit potent cell growth inhibitory and anticancer activity. While a number of cellular and animal studies have suggested that disulfiram exhibits its anticancer activity through interaction with the proteasome, direct evidence for inhibition of proteasome activity is lacking. In this study we show that disulfiram potently inhibits the chymotrypsin-like activity of purified human 20S proteasome at low micromolar pharmacological concentrations. The enzyme progress curves displayed characteristics of a slow-binding reaction, similar to that observed for the FDA-approved proteasomal-targeted anticancer drugs bortezomib and carfilzomib. The apparent second order rate constant for reaction with 20s proteasome that was derived from an analysis of the progress curves was about 250-fold smaller than for bortezomib and carfilzomib. The concentration dependence of the enzyme kinetics was consistent with partial noncompetitive inhibition, whereby the putative disulfiram-proteasome adduct retains, partial but decreased enzyme activity. Disulfiram, which is known to have a high affinity for protein thiols, likely reacted with a non-critical cysteine residue, and not at the proteasome substrate binding site.
[Mh] Termos MeSH primário: Inibidores de Acetaldeído Desidrogenases/farmacologia
Dissulfiram/farmacologia
Complexo de Endopeptidases do Proteassoma/efeitos dos fármacos
Inibidores de Proteassoma/farmacologia
[Mh] Termos MeSH secundário: Antineoplásicos/farmacologia
Bortezomib/farmacologia
Sistema Livre de Células/efeitos dos fármacos
Sistema Livre de Células/enzimologia
Seres Humanos
Cinética
Oligopeptídeos/farmacologia
Complexo de Endopeptidases do Proteassoma/metabolismo
Ligação Proteica
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Acetaldehyde Dehydrogenase Inhibitors); 0 (Antineoplastic Agents); 0 (Oligopeptides); 0 (Proteasome Inhibitors); 69G8BD63PP (Bortezomib); 72X6E3J5AR (carfilzomib); EC 3.4.25.1 (Proteasome Endopeptidase Complex); TR3MLJ1UAI (Disulfiram)
[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:170910
[St] Status:MEDLINE


  9 / 18733 MEDLINE  
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[PMID]:28857158
[Au] Autor:Henriksen SD; Madsen PH; Larsen AC; Johansen MB; Pedersen IS; Krarup H; Thorlacius-Ussing O
[Ad] Endereço:Department of Gastrointestinal Surgery, Aalborg University Hospital, Denmark.
[Ti] Título:Promoter hypermethylation in plasma-derived cell-free DNA as a prognostic marker for pancreatic adenocarcinoma staging.
[So] Source:Int J Cancer;141(12):2489-2497, 2017 Dec 15.
[Is] ISSN:1097-0215
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Correct staging of pancreatic cancer is paramount, as treatment is stage specific. However, minimally invasive tools to facilitate staging are lacking. DNA promoter hypermethylation is a hallmark of cancer. The aim of this study is to evaluate promoter hypermethylation in cell-free DNA as a prognostic marker for stage classification of pancreatic adenocarcinoma. Consecutive patients with pancreatic adenocarcinoma were prospectively included. Plasma samples were obtained before diagnostic work-up and treatment. Patients were staged according to the TNM classification. Methylation-specific PCR of 28 genes was performed. Prognostic prediction models for staging of pancreatic adenocarcinoma were developed by multivariable logistic regression analysis using stepwise backwards elimination. Ninety-five patients with pancreatic adenocarcinoma were included. The mean number of hypermethylated genes was identical for stage I, II and III disease (7.09 (95% CI; 5.51-8.66), 7.00 (95% CI; 5.93-8.07) and 6.77 (95% CI; 5.08-8.46)), respectively, and highly significantly different from stage IV disease (10.24 (95% CI; 8.88-11.60)). The prediction model (SEPT9v2, SST, ALX4, CDKN2B, HIC1, MLH1, NEUROG1, and BNC1) enabled the differentiation of stage IV from stage I-III disease (AUC of 0.87 (cut point 0.55; sensitivity 74%, specificity 87%)). Model (MLH1, SEPT9v2, BNC1, ALX4, CDKN2B, NEUROG1, WNT5A, and TFPI2) enabled the differentiation of stage I-II from stage III-IV disease (AUC of 0.82 (cut point 0.66; sensitivity 73%, specificity 80%)). Cell-free DNA promoter hypermethylation has the potential to be blood-based prognostic markers for pancreatic adenocarcinoma, as panels of hypermethylated genes enables the differentiation according to cancer stage. However, further validation is required.
[Mh] Termos MeSH primário: Metilação de DNA
DNA/genética
Neoplasias Pancreáticas/patologia
Regiões Promotoras Genéticas
[Mh] Termos MeSH secundário: Idoso
Sistema Livre de Células
Feminino
Redes Reguladoras de Genes
Seres Humanos
Modelos Logísticos
Masculino
Meia-Idade
Estadiamento de Neoplasias
Neoplasias Pancreáticas/genética
Prognóstico
Estudos Prospectivos
[Pt] Tipo de publicação:JOURNAL ARTICLE; OBSERVATIONAL STUDY
[Nm] Nome de substância:
9007-49-2 (DNA)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171027
[Lr] Data última revisão:
171027
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170901
[St] Status:MEDLINE
[do] DOI:10.1002/ijc.31024


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[PMID]:28829813
[Au] Autor:Li Y; Xu H; Su S; Ye J; Chen J; Jin X; Lin Q; Zhang D; Ye C; Chen C
[Ad] Endereço:Department of Pulmonary and Critical Care Medicine, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, China.
[Ti] Título:Clinical validation of a highly sensitive assay to detect EGFR mutations in plasma cell-free DNA from patients with advanced lung adenocarcinoma.
[So] Source:PLoS One;12(8):e0183331, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Circulating tumor DNA (ctDNA) is a promising biomarker for noninvasive epidermal growth factor receptor (EGFR) mutations detection in lung cancer patients, but the existing methods have limitations in sensitivity or in availability. In this study, we evaluated the performance of a novel assay called ADx-SuperARMS in detecting EGFR mutations in plasma cell-free DNA from patients with advanced lung adenocarcinoma. METHODS: A total of 109 patients with metastatic advanced adenocarcinoma were recruited who provided both blood samples and matched tumor tissue samples. EGFR mutation status in plasma samples were tested with ADx-SuperARMS EGFR assay and tumor tissue samples were tested with ADx-ARMS EGFR assay. The clinical sensitivity, specificity, positive prediction value (PPV), and negative prediction value (NPV) of ADx-SuperARMS EGFR assay were calculated by using EGFR mutation status in tumor tissue as standard reference. A receiver operating characteristic (ROC) analysis was implemented and an area under the curve (AUC) was calculated to evaluate sensitivity and specificity of exon 19 deletion (E19Del) and L858R mutation detection. The objective response rate (ORR) were calculated according to the EGFR mutation status determined by ADx-superARMS as well. RESULTS: 0.2% analytical sensitivity and 100% specificity of the ADx-SuperARMS EGFR assays for EGFR E19Del, L858R, and T790M mutants were confirmed by using a series of diluted cell line DNA. In the clinical study, EGFR mutations were detected in 45.9% (50/109) of the plasma samples and in 56.9% (62/109) of the matched tumor tissue samples. The sensitivity, specificity, PPV and NPV of the ADx-SuperARMS EGFR assay for plasma EGFR mutation detection were 82.0% (50/61), 100% (48/48), 100% (50/50), and 81.4% (48/59), respectively. In ROC analysis, ADx-SuperARMS achieved sensitivity and specificity of 88% and 99% in E19Dels as well as sensitivity and specificity of 89% and 100% in L858R, respectively. Among the 35 patients who were plasma EGFR mutation positive and treated with first generation of EGFR-tyrosine kinase inhibitors (TKIs), 23 (65.7%) achieved partial response, 11 (31.4%) sustained disease, and 1 (2.9%) progressive disease. The ORR and disease control rate (DCR) were 65.7% and 97.1%, respectively. CONCLUSIONS: ADx-SuperARMS EGFR assay is likely to be a highly sensitive and specific method to noninvasively detect plasma EGFR mutations of patients with advanced lung adenocarcinoma. The EGFR mutations detected by ADx-SuperARMS EGFR assay could predict the efficacy of the treatment with first generation of EGFR-TKIs. Hence, EGFR blood testing with ADx-SuperARMS could address the unmet clinical needs.
[Mh] Termos MeSH primário: Adenocarcinoma/genética
Sistema Livre de Células
Neoplasias Pulmonares/genética
Receptor do Fator de Crescimento Epidérmico/genética
[Mh] Termos MeSH secundário: Feminino
Seres Humanos
Masculino
Meia-Idade
Sensibilidade e Especificidade
[Pt] Tipo de publicação:JOURNAL ARTICLE; VALIDATION STUDIES
[Nm] Nome de substância:
EC 2.7.10.1 (Receptor, Epidermal Growth Factor)
[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:170823
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0183331



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