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Pesquisa : D02.478.770 [Categoria DeCS]
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  1 / 3913 MEDLINE  
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Chiari, Egler
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[PMID]:29176759
[Au] Autor:Magalhães LMD; Viana A; de Jesus AC; Chiari E; Galvão L; Gomes JA; Gollob KJ; Dutra WO
[Ad] Endereço:Laboratório de Biologia das Interações Celulares, Departamento de Morfologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Belo Horizonte, Minas Gerais, Brazil.
[Ti] Título:Distinct Trypanosoma cruzi isolates induce activation and apoptosis of human neutrophils.
[So] Source:PLoS One;12(11):e0188083, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Neutrophils are critical players in the first line of defense against pathogens and in the activation of subsequent cellular responses. We aimed to determine the effects of the interaction of Trypanosoma cruzi with human neutrophils, using isolates of the two major discrete type units (DTUs) associated with Chagas' disease in Latin America (clone Col1.7G2 and Y strain, DTU I and II, respectively). Thus, we used CFSE-stained trypomastigotes to measure neutrophil-T. cruzi interaction, neutrophil activation, cytokine expression and death, after infection with Col1.7G2 and Y strain. Our results show that the frequency of CFSE+ neutrophils, indicative of interaction, and CFSE intensity on a cell-per-cell basis were similar when comparing Col1.7G2 and Y strains. Interaction with T. cruzi increased neutrophil activation, as measured by CD282, CD284, TNF and IL-12 expression, although at different levels between the two strains. No change in IL-10 expression was observed after interaction of neutrophils with either strain. We observed that exposure to Y and Col1.7G2 caused marked neutrophil death. This was specific to neutrophils, since interaction of either strain with monocytes did not cause death. Our further analysis showed that neutrophil death was a result of apoptosis, which was associated with an upregulation of TNF-receptor, TNF and FasLigand, but not of Fas. Induction of TNF-associated neutrophil apoptosis by the different T. cruzi isolates may act as an effective common mechanism to decrease the host's immune response and favor parasite survival.
[Mh] Termos MeSH primário: Apoptose
Ativação de Neutrófilo
Neutrófilos/citologia
Trypanosoma cruzi/isolamento & purificação
[Mh] Termos MeSH secundário: Adulto
Antígenos CD/metabolismo
Sobrevivência Celular
Proteína Ligante Fas/metabolismo
Fluoresceínas/metabolismo
Seres Humanos
Interleucina-10/metabolismo
Interleucina-12/metabolismo
Neutrófilos/metabolismo
Receptores do Fator de Necrose Tumoral/metabolismo
Succinimidas/metabolismo
Receptor 2 Toll-Like/metabolismo
Receptor 4 Toll-Like/metabolismo
Fator de Necrose Tumoral alfa/metabolismo
Adulto Jovem
Receptor fas/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (5-(6)-carboxyfluorescein diacetate succinimidyl ester); 0 (Antigens, CD); 0 (FAS protein, human); 0 (Fas Ligand Protein); 0 (Fluoresceins); 0 (Receptors, Tumor Necrosis Factor); 0 (Succinimides); 0 (Toll-Like Receptor 2); 0 (Toll-Like Receptor 4); 0 (Tumor Necrosis Factor-alpha); 0 (fas Receptor); 130068-27-8 (Interleukin-10); 187348-17-0 (Interleukin-12)
[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:171128
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0188083


  2 / 3913 MEDLINE  
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[PMID]:28456299
[Au] Autor:Li J; Kang T; Talab KMA; Zhu F; Li J
[Ad] Endereço:College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
[Ti] Título:Molecular and biochemical characterization of dimethachlone resistant isolates of Sclerotinia sclerotiorum.
[So] Source:Pestic Biochem Physiol;138:15-21, 2017 May.
[Is] ISSN:1095-9939
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Sclerotinia sclerotiorum is a necrotrophic fungal plant pathogen with a broad host range. The dicarboximide fungicide dimethachlone has been used to control this pathogen for more than a decade and resistance to dimethachlone has recently been reported in China. Compared with sensitive isolates, the three dimethachlone resistant isolates with resistance ratios of 78.3, 85.5, and 94.8 exhibited significantly (P<0.05) higher cell membrane permeability and peroxidase and polyphenol oxidase activities. Dimethachlone at 0.25µg/mL significantly increased cell membrane permeability and enhanced activity of the two enzymes in both resistant and sensitive isolates. There were no significant differences in glycerol or oxalate content between the resistant and sensitive isolates. Dimethachlone treatment increased glycerol content in the resistant isolates and reduced in the sensitive isolates (P<0.01). Sequencing of three genes involved in two-component signal pathway and of three genes in mitogen-activated protein (MAP) kinase cascade demonstrated that the dimethachlone resistant isolates HLJ4 and HLJ6 harbored point mutations of I232T and G1087D, respectively, in the deduce amino acid sequence of the histidine kinase (HK) gene Sshk. HLJ4 had a point mutation of P96L in the deduced amino acid sequence of the MAP kinase-kinase gene SsPbs. The expression levels of the Sshk gene were higher in HLJ4 and HLJ6 than in HLJ3 and the sensitive isolate HLJMG2, and transcription of the Sshk gene was up-regulated by dimethachlone for the three resistant isolates.
[Mh] Termos MeSH primário: Ascomicetos/efeitos dos fármacos
Clorobenzenos/farmacologia
Farmacorresistência Fúngica
Fungicidas Industriais/farmacologia
Succinimidas/farmacologia
[Mh] Termos MeSH secundário: Sequência de Bases
Catecol Oxidase/genética
Catecol Oxidase/metabolismo
Clonagem Molecular
Proteínas Fúngicas/genética
Proteínas Fúngicas/metabolismo
Regulação Enzimológica da Expressão Gênica
Regulação Fúngica da Expressão Gênica
Mutação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Chlorobenzenes); 0 (Fungal Proteins); 0 (Fungicides, Industrial); 0 (Succinimides); 0 (dimethachlon); EC 1.10.3.1 (Catechol Oxidase)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171128
[Lr] Data última revisão:
171128
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170501
[St] Status:MEDLINE


  3 / 3913 MEDLINE  
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[PMID]:28794626
[Au] Autor:Luo X; Peng X; Hou J; Wu S; Shen J; Wang L
[Ad] Endereço:Department of Gastroenterology.
[Ti] Título:Folic acid-functionalized polyethylenimine superparamagnetic iron oxide nanoparticles as theranostic agents for magnetic resonance imaging and PD-L1 siRNA delivery for gastric cancer.
[So] Source:Int J Nanomedicine;12:5331-5343, 2017.
[Is] ISSN:1178-2013
[Cp] País de publicação:New Zealand
[La] Idioma:eng
[Ab] Resumo:Programmed death ligand-1 (PD-L1), which is highly expressed in gastric cancers, interacts with programmed death-1 (PD-1) on T cells and is involved in T-cell immune resistance. To increase the therapeutic safety and accuracy of PD-1/PD-L1 blockade, RNA interference through targeted gene delivery was performed in our study. We developed folic acid (FA)- and disulfide (SS)-polyethylene glycol (PEG)-conjugated polyethylenimine (PEI) complexed with superparamagnetic iron oxide Fe O nanoparticles (SPIONs) as a siRNA-delivery system for PD-L1 knockdown. The characterization, binding ability, cytotoxicity, transfection efficiency, and cellular internalization of the polyplex were determined. At nitrogen:phosphate (N:P) ratios of 10 or above, the FA-PEG-SS-PEI-SPIONs bound to PD-L1 siRNA to form a polyplex with a diameter of approximately 120 nm. Cell-viability assays showed that the polyplex had minimal cytotoxicity at low N:P ratios. The FA-conjugated polyplex showed higher transfection efficiency and cellular internalization in the folate receptor-overexpressing gastric cancer cell line SGC-7901 than a non-FA-conjugated polyplex. Subsequently, we adopted the targeted FA-PEG-SS-PEI-SPION/siRNA polyplexes at an N:P ratio of 10 for function studies. Cellular magnetic resonance imaging (MRI) showed that the polyplex could also act as a -weighted contrast agent for cancer MRI. Furthermore, one of four PD-L1 siRNAs exhibited effective PD-L1 knockdown in PD-L1-overexpressing SGC-7901. To determine the effects of the functionalized polyplex on T-cell function, we established a coculture model of activated T cells and SGC-7901 cells and demonstrated changes in secreted cytokines. Our findings highlight the potential of this class of multifunctional theranostic nanoparticles for effective targeted PD-L1-knockdown therapy and MRI diagnosis in gastric cancers.
[Mh] Termos MeSH primário: Antígeno B7-H1/genética
Imagem por Ressonância Magnética/métodos
Nanopartículas de Magnetita/química
RNA Interferente Pequeno/administração & dosagem
Nanomedicina Teranóstica/métodos
[Mh] Termos MeSH secundário: Sobrevivência Celular/efeitos dos fármacos
Técnicas de Cocultura
Meios de Contraste/química
Compostos Férricos/química
Ácido Fólico/química
Técnicas de Transferência de Genes
Seres Humanos
Nanopartículas de Magnetita/administração & dosagem
Polietilenoglicóis/química
Polietilenoimina/química
RNA Interferente Pequeno/genética
Neoplasias Gástricas/diagnóstico por imagem
Neoplasias Gástricas/tratamento farmacológico
Neoplasias Gástricas/terapia
Succinimidas/química
Transfecção
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (B7-H1 Antigen); 0 (CD274 protein, human); 0 (Contrast Media); 0 (Ferric Compounds); 0 (Magnetite Nanoparticles); 0 (RNA, Small Interfering); 0 (Succinimides); 1K09F3G675 (ferric oxide); 30IQX730WE (Polyethylene Glycols); 85419-94-9 (polyethylene glycol bis(succinimidyl succinate)); 9002-98-6 (Polyethyleneimine); 935E97BOY8 (Folic Acid)
[Em] Mês de entrada:1711
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170811
[St] Status:MEDLINE
[do] DOI:10.2147/IJN.S137245


  4 / 3913 MEDLINE  
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[PMID]:28757182
[Au] Autor:Jacobsen MT; Fairhead M; Fogelstrand P; Howarth M
[Ad] Endereço:Department of Biochemistry, University of Oxford, South Parks Road, Oxford OX1 3QU, UK.
[Ti] Título:Amine Landscaping to Maximize Protein-Dye Fluorescence and Ultrastable Protein-Ligand Interaction.
[So] Source:Cell Chem Biol;24(8):1040-1047.e4, 2017 Aug 17.
[Is] ISSN:2451-9456
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Chemical modification of proteins provides great opportunities to control and visualize living systems. The most common way to modify proteins is reaction of their abundant amines with N-hydroxysuccinimide (NHS) esters. Here we explore the impact of amine number and positioning on protein-conjugate behavior using streptavidin-biotin, a central research tool. Dye-NHS modification of streptavidin severely damaged ligand binding, necessitating development of a new streptavidin-retaining ultrastable binding after labeling. Exploring the ideal level of dye modification, we engineered a panel bearing 1-6 amines per subunit: "amine landscaping." Surprisingly, brightness increased as amine number decreased, revealing extensive quenching following conventional labeling. We ultimately selected Flavidin (fluorophore-friendly streptavidin), combining ultrastable ligand binding with increased brightness after conjugation. Flavidin enhanced fluorescent imaging, allowing more sensitive and specific cell labeling in tissues. Flavidin should have wide application in molecular detection, providing a general insight into how to optimize simultaneously the behavior of the biomolecule and the chemical probe.
[Mh] Termos MeSH primário: Aminas/química
Biotina/química
Corantes Fluorescentes/química
Estreptavidina/química
[Mh] Termos MeSH secundário: Anticorpos/química
Anticorpos/imunologia
Biotina/metabolismo
Complexo CD3/imunologia
Citometria de Fluxo
Células HeLa
Seres Humanos
Ligantes
Microscopia Confocal
Mutagênese Sítio-Dirigida
Ligação Proteica
Estabilidade Proteica
Espectrometria de Fluorescência
Estreptavidina/genética
Estreptavidina/metabolismo
Succinimidas/química
Temperatura Ambiente
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Amines); 0 (Antibodies); 0 (CD3 Complex); 0 (Fluorescent Dyes); 0 (Ligands); 0 (Succinimides); 6SO6U10H04 (Biotin); 9013-20-1 (Streptavidin); MJE3791M4T (N-hydroxysuccinimide)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170801
[St] Status:MEDLINE


  5 / 3913 MEDLINE  
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[PMID]:28622387
[Au] Autor:Mazzocco P; Bernard S; Pujo-Menjouet L
[Ad] Endereço:Université de Lyon, Université Claude Bernard Lyon 1, CNRS UMR 5558, Laboratoire de Biométrie et Biologie Evolutive, Villeurbanne, France.
[Ti] Título:Estimates and impact of lymphocyte division parameters from CFSE data using mathematical modelling.
[So] Source:PLoS One;12(6):e0179768, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Carboxyfluorescein diacetate succinimidyl ester (CFSE) labelling has been widely used to track and study cell proliferation. Here we use mathematical modelling to describe the kinetics of immune cell proliferation after an in vitro polyclonal stimulation tracked with CFSE. This approach allows us to estimate a set of key parameters, including ones related to cell death and proliferation. We develop a three-phase model that distinguishes a latency phase, accounting for non-divided cell behaviour, a resting phase and the active phase of the division process. Parameter estimates are derived from model results, and numerical simulations are then compared to the dynamics of in vitro experiments, with different biological assumptions tested. Our model allows us to compare the dynamics of CD4+ and CD8+ cells, and to highlight their kinetic differences. Finally we perform a sensitivity analysis to quantify the impact of each parameter on proliferation kinetics. Interestingly, we find that parameter sensitivity varies with time and with cell generation. Our approach can help biologists to understand cell proliferation mechanisms and to identify potential pathological division processes.
[Mh] Termos MeSH primário: Linfócitos T CD4-Positivos/metabolismo
Linfócitos T CD8-Positivos/metabolismo
Divisão Celular/fisiologia
Fluoresceínas/química
Corantes Fluorescentes/química
Modelos Biológicos
Succinimidas/química
[Mh] Termos MeSH secundário: Linfócitos T CD4-Positivos/citologia
Linfócitos T CD8-Positivos/citologia
Seres Humanos
Coloração e Rotulagem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (5-(6)-carboxyfluorescein diacetate succinimidyl ester); 0 (Fluoresceins); 0 (Fluorescent Dyes); 0 (Succinimides)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170914
[Lr] Data última revisão:
170914
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170617
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0179768


  6 / 3913 MEDLINE  
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[PMID]:28613834
[Au] Autor:Warminski M; Sikorski PJ; Warminska Z; Lukaszewicz M; Kropiwnicka A; Zuberek J; Darzynkiewicz E; Kowalska J; Jemielity J
[Ad] Endereço:Division of Biophysics, Institute of Experimental Physics, Faculty of Physics, University of Warsaw , 02-093, Warsaw, Poland.
[Ti] Título:Amino-Functionalized 5' Cap Analogs as Tools for Site-Specific Sequence-Independent Labeling of mRNA.
[So] Source:Bioconjug Chem;28(7):1978-1992, 2017 Jul 19.
[Is] ISSN:1520-4812
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:mRNA is a template for protein biosynthesis, and consequently mRNA transport, translation, and turnover are key elements in the overall regulation of gene expression. Along with growing interest in the mechanisms regulating mRNA decay and localization, there is an increasing need for tools enabling convenient fluorescent labeling or affinity tagging of mRNA. We report new mRNA 5' cap analog-based tools that enable site-specific labeling of RNA within the cap using N-hydroxysuccinimide (NHS) chemistry. We explored two complementary methods: a co-transcriptional labeling method, in which the label is first attached to a cap analog and then incorporated into RNA by in vitro transcription, and a post-transcriptional labeling method, in which an amino-functionalized cap analog is incorporated into RNA followed by chemical labeling of the resulting transcript. After testing the biochemical properties of RNAs carrying the novel modified cap structures, we demonstrated the utility of fluorescently labeled RNAs in decapping assays, RNA decay assays, and RNA visualization in cells. Finally, we also demonstrated that mRNAs labeled by the reported method are translationally active. We envisage that the novel analogs will provide an alternative to radiolabeling of mRNA caps for in vitro studies and open possibilities for new applications related to the study of mRNA fates in vivo.
[Mh] Termos MeSH primário: Capuzes de RNA/química
RNA Mensageiro/química
Coloração e Rotulagem/métodos
Succinimidas/química
[Mh] Termos MeSH secundário: Animais
Sistema Livre de Células
Células HeLa
Seres Humanos
Biossíntese de Proteínas
Processamento Pós-Transcricional do RNA
Coelhos
Transcrição Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA Caps); 0 (RNA, Messenger); 0 (Succinimides); MJE3791M4T (N-hydroxysuccinimide)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170817
[Lr] Data última revisão:
170817
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170615
[St] Status:MEDLINE
[do] DOI:10.1021/acs.bioconjchem.7b00291


  7 / 3913 MEDLINE  
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[PMID]:28533682
[Au] Autor:Wang G; Gao X; Gu G; Shao Z; Li M; Wang P; Yang J; Cai X; Li Y
[Ad] Endereço:Department of Urology, Huashan Hospital, Fudan University.
[Ti] Título:Polyethylene glycol-poly(ε-benzyloxycarbonyl-l-lysine)-conjugated VEGF siRNA for antiangiogenic gene therapy in hepatocellular carcinoma.
[So] Source:Int J Nanomedicine;12:3591-3603, 2017.
[Is] ISSN:1178-2013
[Cp] País de publicação:New Zealand
[La] Idioma:eng
[Ab] Resumo:A polyethylene glycol-poly(ε-benzyloxycarbonyl-l-lysine) (PEG-SS-PLL) block copolymer based on a disulfide-linked, novel biodegradable catiomer bearing a PEG-sheddable shell was developed to avoid "PEG dilemma" in nanoparticle intracellular tracking of PEG-PLL where PEG was nondegradable. However, PEG-SS-PLL catiomers have not been used to deliver small interfering VEGF RNA (siVEGF) in antiangiogenesis gene therapy. In this study, we aimed to investigate whether this novel biodegradable catiomer can deliver siVEGF into cancer cells and at the same time have an antitumor effect in a xenograft mouse model. It was found that PEG-SS-PLL efficiently delivered siVEGF with negligible cytotoxicity, and significantly decreased the expression of VEGF at both the messenger-RNA and protein levels both in vitro and in vivo, and thus tumor growth was inhibited. Our findings demonstrated that PEG-SS-PLL/siVEGF could potentially be applied to antiangiogenesis gene therapy for hepatocellular carcinoma.
[Mh] Termos MeSH primário: Inibidores da Angiogênese/administração & dosagem
Sistemas de Liberação de Medicamentos/métodos
Terapia Genética/métodos
Polietilenoglicóis/química
Polilisina/análogos & derivados
RNA Interferente Pequeno/administração & dosagem
Fator A de Crescimento do Endotélio Vascular/genética
[Mh] Termos MeSH secundário: Inibidores da Angiogênese/genética
Inibidores da Angiogênese/farmacologia
Animais
Carcinoma Hepatocelular/genética
Carcinoma Hepatocelular/terapia
Feminino
Células Hep G2
Seres Humanos
Neoplasias Hepáticas/genética
Neoplasias Hepáticas/terapia
Lisina/química
Camundongos Endogâmicos BALB C
Nanopartículas/administração & dosagem
Nanopartículas/química
Polilisina/química
Polímeros/química
Succinimidas/química
Fator A de Crescimento do Endotélio Vascular/metabolismo
Ensaios Antitumorais Modelo de Xenoenxerto
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Angiogenesis Inhibitors); 0 (Polymers); 0 (RNA, Small Interfering); 0 (Succinimides); 0 (Vascular Endothelial Growth Factor A); 0 (polyethylene glycol-poly(epsilon-benzyloxycarbonyllysine)); 25104-18-1 (Polylysine); 30IQX730WE (Polyethylene Glycols); 85419-94-9 (polyethylene glycol bis(succinimidyl succinate)); K3Z4F929H6 (Lysine)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170911
[Lr] Data última revisão:
170911
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170524
[St] Status:MEDLINE
[do] DOI:10.2147/IJN.S131078


  8 / 3913 MEDLINE  
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[PMID]:28445029
[Au] Autor:Ward CC; Kleinman JI; Nomura DK
[Ad] Endereço:Departments of Chemistry, Molecular and Cell Biology, and Nutritional Sciences and Toxicology, 127 Morgan Hall, University of California, Berkeley , Berkeley, California 94720, United States.
[Ti] Título:NHS-Esters As Versatile Reactivity-Based Probes for Mapping Proteome-Wide Ligandable Hotspots.
[So] Source:ACS Chem Biol;12(6):1478-1483, 2017 Jun 16.
[Is] ISSN:1554-8937
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Most of the proteome is considered undruggable, oftentimes hindering translational efforts for drug discovery. Identifying previously unknown druggable hotspots in proteins would enable strategies for pharmacologically interrogating these sites with small molecules. Activity-based protein profiling (ABPP) has arisen as a powerful chemoproteomic strategy that uses reactivity-based chemical probes to map reactive, functional, and ligandable hotspots in complex proteomes, which has enabled inhibitor discovery against various therapeutic protein targets. Here, we report an alkyne-functionalized N-hydroxysuccinimide-ester (NHS-ester) as a versatile reactivity-based probe for mapping the reactivity of a wide range of nucleophilic ligandable hotspots, including lysines, serines, threonines, and tyrosines, encompassing active sites, allosteric sites, post-translational modification sites, protein interaction sites, and previously uncharacterized potential binding sites. Surprisingly, we also show that fragment-based NHS-ester ligands can be made to confer selectivity for specific lysine hotspots on specific targets including Dpyd, Aldh2, and Gstt1. We thus put forth NHS-esters as promising reactivity-based probes and chemical scaffolds for covalent ligand discovery.
[Mh] Termos MeSH primário: Sítios de Ligação
Ésteres/química
Ligantes
Sondas Moleculares/química
Proteoma/metabolismo
[Mh] Termos MeSH secundário: Aldeído-Desidrogenase Mitocondrial/metabolismo
Alquinos/química
Glutationa Transferase/metabolismo
Seres Humanos
Lisina/metabolismo
Succinimidas
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Alkynes); 0 (Esters); 0 (Ligands); 0 (Molecular Probes); 0 (Proteome); 0 (Succinimides); EC 1.2.1.3 (ALDH2 protein, human); EC 1.2.1.3 (Aldehyde Dehydrogenase, Mitochondrial); EC 2.5.1.- (glutathione S-transferase T1); EC 2.5.1.18 (Glutathione Transferase); K3Z4F929H6 (Lysine); MJE3791M4T (N-hydroxysuccinimide)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170919
[Lr] Data última revisão:
170919
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170427
[St] Status:MEDLINE
[do] DOI:10.1021/acschembio.7b00125


  9 / 3913 MEDLINE  
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[PMID]:28443656
[Au] Autor:Dovgan I; Ursuegui S; Erb S; Michel C; Kolodych S; Cianférani S; Wagner A
[Ad] Endereço:Laboratory of Functional ChemoSystems (UMR 7199), LabEx Medalis, University of Strasbourg , 67087 Strasbourg, France.
[Ti] Título:Acyl Fluorides: Fast, Efficient, and Versatile Lysine-Based Protein Conjugation via Plug-and-Play Strategy.
[So] Source:Bioconjug Chem;28(5):1452-1457, 2017 May 17.
[Is] ISSN:1520-4812
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:We report a plug-and-play strategy for the preparation of functionally enhanced antibodies with a defined average degree of conjugation (DoC). The first stage (plug) allows the controllable and efficient installation of azide groups on lysine residues of a native antibody using 4-azidobenzoyl fluoride. The second step (play) allows for versatile antibody functionalization with a single payload or combination of payloads, such as a toxin, a fluorophore, or an oligonucleotide, via copper-free strain-promoted azide-alkyne cycloaddition (SPAAC). It is notable that in comparison to a classical N-hydroxysuccinimide ester (NHS) strategy, benzoyl fluorides show faster and more efficient acylation of lysine residues in a PBS buffer. This translates into better control of the DoC and enables the efficient and fast functionalization of delicate biomolecules at low temperature.
[Mh] Termos MeSH primário: Anticorpos Monoclonais/química
Compostos de Benzil/química
Fluoretos/química
Imunoconjugados/química
Lisina/química
Receptor ErbB-2/imunologia
[Mh] Termos MeSH secundário: Acilação
Alquinos/química
Anticorpos Monoclonais/imunologia
Azidas/química
Química Click
Reação de Cicloadição
Corantes Fluorescentes/química
Seres Humanos
Imunoconjugados/imunologia
Estrutura Molecular
Oligonucleotídeos/química
Succinimidas/química
Toxinas Biológicas/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Alkynes); 0 (Antibodies, Monoclonal); 0 (Azides); 0 (Benzyl Compounds); 0 (Fluorescent Dyes); 0 (Immunoconjugates); 0 (Oligonucleotides); 0 (Succinimides); 0 (Toxins, Biological); EC 2.7.10.1 (ERBB2 protein, human); EC 2.7.10.1 (Receptor, ErbB-2); K3Z4F929H6 (Lysine); MJE3791M4T (N-hydroxysuccinimide); Q80VPU408O (Fluorides)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170817
[Lr] Data última revisão:
170817
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170427
[St] Status:MEDLINE
[do] DOI:10.1021/acs.bioconjchem.7b00141


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[PMID]:28420169
[Au] Autor:Khosravi F; Loeian SM; Panchapakesan B
[Ad] Endereço:Small Systems Laboratory, Department of Mechanical Engineering, Worcester Polytechnic Institute, Worcester, MA 01532, USA. fkhosravi@wpi.edu.
[Ti] Título:Ultrasensitive Label-Free Sensing of IL-6 Based on PASE Functionalized Carbon Nanotube Micro-Arrays with RNA-Aptamers as Molecular Recognition Elements.
[So] Source:Biosensors (Basel);7(2), 2017 Apr 17.
[Is] ISSN:2079-6374
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:This study demonstrates the rapid and label-free detection of Interleukin-6 (IL-6) using carbon nanotube micro-arrays with aptamer as the molecular recognition element. Single wall carbon nanotubes micro-arrays biosensors were manufactured using photo-lithography, metal deposition, and etching techniques. Nanotube biosensors were functionalized with 1-Pyrenebutanoic Acid Succinimidyl Ester (PASE) conjugated IL-6 aptamers. Real time response of the sensor conductance was monitored with increasing concentration of IL-6 (1 pg/mL to 10 ng/mL), exposure to the sensing surface in buffer solution, and clinically relevant spiked blood samples. Non-specific Bovine Serum Albumin (BSA), PBS samples, and anti-IgG functionalized devices gave similar signatures in the real time conductance versus time experiments with no significant change in sensor signal. Exposure of the aptamer functionalized nanotube surface to IL-6 decreased the conductance with increasing concentration of IL-6. Experiments based on field effect transistor arrays suggested shift in drain current versus gate voltage for 1 pg and 1 ng of IL-6 exposure. Non-specific BSA did not produce any appreciable shift in the I versus V suggesting specific interactions of IL-6 on PASE conjugated aptamer surface gave rise to the change in electrical signal. Both Z axis and phase image in an Atomic Force Microscope (AFM) suggested unambiguous molecular interaction of the IL-6 on the nanotube-aptamer surface at 1 pg/mL concentration. The concentration of 1 pg falls below the diagnostic gray zone for cancer (2.3 pg-4 ng/mL), which is an indicator of early stage cancer. Thus, nanotube micro-arrays could potentially be developed for creating multiplexed assays involving cancer biomarker proteins and possibly circulating tumor cells all in a single assay using PASE functionalization protocol.
[Mh] Termos MeSH primário: Aptâmeros de Nucleotídeos/química
Interleucina-6/sangue
Nanotubos de Carbono/química
Análise Serial de Proteínas/métodos
[Mh] Termos MeSH secundário: Animais
Biomarcadores Tumorais/sangue
Bovinos
Feminino
Seres Humanos
Masculino
Neoplasias/sangue
Pirenos/química
Sensibilidade e Especificidade
Soroalbumina Bovina/química
Succinimidas/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Aptamers, Nucleotide); 0 (Biomarkers, Tumor); 0 (IL6 protein, human); 0 (Interleukin-6); 0 (Nanotubes, Carbon); 0 (Pyrenes); 0 (Succinimides); 27432CM55Q (Serum Albumin, Bovine); 3443-45-6 (1-pyrenebutyrate)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170420
[St] Status:MEDLINE



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