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  1 / 18867 MEDLINE  
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[PMID]:28450830
[Au] Autor:Elliott KL; Kersigo J; Pan N; Jahan I; Fritzsch B
[Ad] Endereço:Department of Biology, University of IowaIowa City, IA, USA.
[Ti] Título:Spiral Ganglion Neuron Projection Development to the Hindbrain in Mice Lacking Peripheral and/or Central Target Differentiation.
[So] Source:Front Neural Circuits;11:25, 2017.
[Is] ISSN:1662-5110
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:We investigate the importance of the degree of peripheral or central target differentiation for mouse auditory afferent navigation to the organ of Corti and auditory nuclei in three different mouse models: first, a mouse in which the differentiation of hair cells, but not central auditory nuclei neurons is compromised ( ); second, a mouse in which hair cell defects are combined with a delayed defect in central auditory nuclei neurons ( ), and third, a mouse in which both hair cells and central auditory nuclei are absent ( ). Our results show that neither differentiated peripheral nor the central target cells of inner ear afferents are needed (hair cells, cochlear nucleus neurons) for segregation of vestibular and cochlear afferents within the hindbrain and some degree of base to apex segregation of cochlear afferents. These data suggest that inner ear spiral ganglion neuron processes may predominantly rely on temporally and spatially distinct molecular cues in the region of the targets rather than interaction with differentiated target cells for a crude topological organization. These developmental data imply that auditory neuron navigation properties may have evolved before auditory nuclei.
[Mh] Termos MeSH primário: Fatores de Transcrição Hélice-Alça-Hélice Básicos/deficiência
Diferenciação Celular/genética
Células Ciliadas Auditivas/fisiologia
Malformações do Sistema Nervoso/patologia
Fator de Transcrição PAX2/deficiência
Rombencéfalo/patologia
Gânglio Espiral da Cóclea
[Mh] Termos MeSH secundário: Animais
Animais Recém-Nascidos
Vias Auditivas/embriologia
Fatores de Transcrição Hélice-Alça-Hélice Básicos/genética
Núcleo Coclear/citologia
Núcleo Coclear/embriologia
Núcleo Coclear/crescimento & desenvolvimento
Embrião de Mamíferos
Camundongos
Camundongos Knockout
Malformações do Sistema Nervoso/genética
Fator de Transcrição PAX2/genética
Gânglio Espiral da Cóclea/embriologia
Gânglio Espiral da Cóclea/crescimento & desenvolvimento
Gânglio Espiral da Cóclea/patologia
beta-Galactosidase/genética
beta-Galactosidase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Atoh1 protein, mouse); 0 (Basic Helix-Loop-Helix Transcription Factors); 0 (PAX2 Transcription Factor); 0 (Pax2 protein, mouse); EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180307
[Lr] Data última revisão:
180307
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170429
[St] Status:MEDLINE
[do] DOI:10.3389/fncir.2017.00025


  2 / 18867 MEDLINE  
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[PMID]:28470615
[Au] Autor:Phan T; Huynh P; Truong T; Nguyen H
[Ad] Endereço:VNUHCM-University of Science, 227 Nguyen Van Cu, District 5, Hochiminh City, Vietnam.
[Ti] Título:A Generic Protocol for Intracellular Expression of Recombinant Proteins in Bacillus subtilis.
[So] Source:Methods Mol Biol;1586:325-334, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Bacillus subtilis (B. subtilis) is a potential and attractive host for the production of recombinant proteins. Different expression systems for B. subtilis have been developed recently, and various target proteins have been recombinantly synthesized and purified using this host. In this chapter, we introduce a generic protocol to express a recombinant protein in B. subtilis. It includes protocols for (1) using our typical expression vector (plasmid pHT254) to introduce a target gene, (2) transformation of the target vector into B. subtilis, and (3) evaluation of the actual expression of a recombinant protein.
[Mh] Termos MeSH primário: Bacillus subtilis/genética
Proteínas de Bactérias/genética
Clonagem Molecular/métodos
Vetores Genéticos/genética
beta-Galactosidase/genética
[Mh] Termos MeSH secundário: Bacillus subtilis/crescimento & desenvolvimento
Expressão Gênica
Genes Bacterianos
Plasmídeos/genética
Regiões Promotoras Genéticas
Proteínas Recombinantes/genética
Transformação Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (Recombinant Proteins); EC 3.2.1.23 (beta-Galactosidase)
[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_21


  3 / 18867 MEDLINE  
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[PMID]:29235592
[Au] Autor:Li Y; Ogorzalek TL; Wei S; Zhang X; Yang P; Jasensky J; Brooks CL; Marsh ENG; Chen Z
[Ad] Endereço:Department of Chemistry, University of Michigan, Ann Arbor, Michigan 48109, USA. zhanc@umich.edu.
[Ti] Título:Effect of immobilization site on the orientation and activity of surface-tethered enzymes.
[So] Source:Phys Chem Chem Phys;20(2):1021-1029, 2018 Jan 03.
[Is] ISSN:1463-9084
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Tethering peptides and proteins to abiotic surfaces has the potential to create biomolecule-functionalized surfaces with useful properties. Commonly used methods of immobilization lack control over the orientation in which biological molecules are covalently or physically bound to the surface, leading to sub-optimal materials. Here we use an engineered beta-galactosidase that can be chemically immobilized on a surface with a well-defined orientation through unique surface-accessible cysteine residues. A combined study using sum frequency generation (SFG) vibrational spectroscopy and coarse grained molecular dynamics (MD) simulations was performed to determine the effects of enzyme immobilization site and abiotic surface chemistry on enzyme surface orientation, surface coverage, and catalytic activity. Two beta-galactosidase variants that were immobilized through cysteine introduced at positions 227 and 308 were studied. In both cases, when the abiotic surface was made more hydrophilic, the enzyme surface coverage decreased, but the activity increased. MD simulations indicated that this is due to the weakened interactions between the immobilized enzyme and the more hydrophilic surface. These studies provide improved understanding of how enzyme-surface interactions can be optimized to maximize the catalytic activity of surface tethered enzymes.
[Mh] Termos MeSH primário: Enzimas Imobilizadas/química
Simulação de Dinâmica Molecular
beta-Galactosidase/química
[Mh] Termos MeSH secundário: Cisteína
Peptídeos/química
Análise Espectral
Propriedades de Superfície
Vibração
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Enzymes, Immobilized); 0 (Peptides); EC 3.2.1.23 (beta-Galactosidase); K848JZ4886 (Cysteine)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180209
[Lr] Data última revisão:
180209
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171214
[St] Status:MEDLINE
[do] DOI:10.1039/c7cp06063g


  4 / 18867 MEDLINE  
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[PMID]:29323842
[Au] Autor:Kuznetsova TV; Smimova MS; Leonovich OA; Gordeichuk IV; Biriukova IK; Zylkova MV; Tyn'o YY; Belyakova AV; Shevelev AB
[Ti] Título:A new method of producing NS5A antigen of hepatitis C virus.
[So] Source:Vopr Virusol;62(1):17-25, 2017.
[Is] ISSN:0507-4088
[Cp] País de publicação:Russia (Federation)
[La] Idioma:eng
[Ab] Resumo:A task of creating a universal platform for engineering affordable recombinant producers of viral proteins conserving immunogenicity has not been solved yet. High toxicity of the viral proteins for the host cells, low yield and abnormal folding of the products often present severe obstacles to obtaining producers of the viral proteins. In this work, we report a new method of engineering and screening of deletion libraries from the viral antigen genes. This method allows selection of artificial derivatives of these genes adapted for expression in microbial producer cells. The method involves PCR amplification of the gene fragments using a system of randomized and adapter primers, which allows the spontaneous formation of duplexes from the random primers in the absence of the template DNA to be prevented. For selecting variants capable of in vivo expression, the obtained PCR products are cloned to a special vector of a direct phenotypical selection pQL30. It contains E. coli ß-galactosidase gene with an inserted polylinker producing a frame-shift mutation. Using this screening method, an artificial variant of hepatitis C (HCV) NS5a gene with optimal biotechnological properties was established. 27 clinical specimens of 1670 bp long HCV1b NS5a fragments were used as a source gene. A PCR bank of the deletion derivatives was produced. 40 LacZ-positive clones based on pQL30 vector with a 50-700 bp long insertion were selected. The LacZ activity of the cell lysates and the immunogenicity of the products were tested. As a result, a single clone encoding a soluble protein with Mr = 114 kDa was selected. Its yield reached 0.3% of the total cell protein. It was highly reactive with sera of HCV 1b infected patients but not with sera of the healthy donors.
[Mh] Termos MeSH primário: Antígenos Virais/genética
Escherichia coli/genética
Hepacivirus/genética
Hepatite C/diagnóstico
Engenharia de Proteínas/métodos
Proteínas Recombinantes/genética
Proteínas não Estruturais Virais/genética
[Mh] Termos MeSH secundário: Antígenos Virais/biossíntese
Antígenos Virais/imunologia
Antígenos Virais/isolamento & purificação
Proteínas de Bactérias/genética
Proteínas de Bactérias/metabolismo
Primers do DNA/síntese química
Primers do DNA/genética
Escherichia coli/metabolismo
Mutação da Fase de Leitura
Biblioteca Gênica
Vetores Genéticos/química
Vetores Genéticos/metabolismo
Hepacivirus/imunologia
Hepatite C/imunologia
Hepatite C/virologia
Seres Humanos
Soros Imunes/química
Óperon Lac
Proteínas Recombinantes/biossíntese
Proteínas Recombinantes/imunologia
Proteínas Recombinantes/isolamento & purificação
Proteínas não Estruturais Virais/biossíntese
Proteínas não Estruturais Virais/imunologia
Proteínas não Estruturais Virais/isolamento & purificação
beta-Galactosidase/genética
beta-Galactosidase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antigens, Viral); 0 (Bacterial Proteins); 0 (DNA Primers); 0 (Immune Sera); 0 (NS-5 protein, hepatitis C virus); 0 (Recombinant Proteins); 0 (Viral Nonstructural Proteins); EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180208
[Lr] Data última revisão:
180208
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:180112
[St] Status:MEDLINE


  5 / 18867 MEDLINE  
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[PMID]:29176033
[Au] Autor:Kandhaya-Pillai R; Miro-Mur F; Alijotas-Reig J; Tchkonia T; Kirkland JL; Schwartz S
[Ad] Endereço:Aging Basic Research Group, Molecular Biology and Biochemistry Research Center for Nanomedicine, CIBBIM-Nanomedicine, Vall d'Hebron University Hospital, Research Institute (VHIR), Barcelona, Spain.
[Ti] Título:TNFα-senescence initiates a STAT-dependent positive feedback loop, leading to a sustained interferon signature, DNA damage, and cytokine secretion.
[So] Source:Aging (Albany NY);9(11):2411-2435, 2017 Nov 22.
[Is] ISSN:1945-4589
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Cellular senescence is a cell fate program that entails essentially irreversible proliferative arrest in response to damage signals. Tumor necrosis factor-alpha (TNFα), an important pro-inflammatory cytokine secreted by some types of senescent cells, can induce senescence in mouse and human cells. However, downstream signaling pathways linking TNFα-related inflammation to senescence are not fully characterized. Using human umbilical vein endothelial cells (HUVECs) as a model, we show that TNFα induces permanent growth arrest and increases p21CIP1, p16INK4A, and SA-ß-gal, accompanied by persistent DNA damage and ROS production. By gene expression profiling, we identified the crucial involvement of inflammatory and JAK/STAT pathways in TNFα-mediated senescence. We found that TNFα activates a STAT-dependent autocrine loop that sustains cytokine secretion and an interferon signature to lock cells into senescence. Furthermore, we show STAT1/3 activation is necessary for cytokine and ROS production during TNFα-induced senescence. However, inhibition of STAT1/3 did not rescue cells from proliferative arrest, but rather suppressed cell cycle regulatory genes and altered TNFα-induced senescence. Our findings suggest a positive feedback mechanism via the STAT pathway that sustains cytokine production and reveal a reciprocal regulatory role of JAK/STAT in TNFα-mediated senescence.
[Mh] Termos MeSH primário: Senescência Celular/efeitos dos fármacos
Citocinas/metabolismo
Dano ao DNA
Células Endoteliais da Veia Umbilical Humana/efeitos dos fármacos
Fatores Reguladores de Interferon/metabolismo
Fator de Transcrição STAT1/metabolismo
Fator de Transcrição STAT3/metabolismo
Fator de Necrose Tumoral alfa/farmacologia
[Mh] Termos MeSH secundário: Pontos de Checagem do Ciclo Celular/efeitos dos fármacos
Proliferação Celular/efeitos dos fármacos
Células Cultivadas
Inibidor p16 de Quinase Dependente de Ciclina/metabolismo
Inibidor de Quinase Dependente de Ciclina p21/metabolismo
Citocinas/genética
Citocinas/secreção
Retroalimentação Fisiológica
Regulação da Expressão Gênica
Células Endoteliais da Veia Umbilical Humana/metabolismo
Células Endoteliais da Veia Umbilical Humana/patologia
Células Endoteliais da Veia Umbilical Humana/secreção
Seres Humanos
Fatores Reguladores de Interferon/genética
Janus Quinases/metabolismo
Espécies Reativas de Oxigênio/metabolismo
Transdução de Sinais/efeitos dos fármacos
Fatores de Tempo
beta-Galactosidase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (CDKN1A protein, human); 0 (Cyclin-Dependent Kinase Inhibitor p16); 0 (Cyclin-Dependent Kinase Inhibitor p21); 0 (Cytokines); 0 (Interferon Regulatory Factors); 0 (P16 protein, human); 0 (Reactive Oxygen Species); 0 (STAT1 Transcription Factor); 0 (STAT1 protein, human); 0 (STAT3 Transcription Factor); 0 (STAT3 protein, human); 0 (Tumor Necrosis Factor-alpha); EC 2.7.10.2 (Janus Kinases); EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180205
[Lr] Data última revisão:
180205
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171128
[St] Status:MEDLINE
[do] DOI:10.18632/aging.101328


  6 / 18867 MEDLINE  
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[PMID]:28743271
[Au] Autor:Tran DTM; Phan TTP; Huynh TK; Dang NTK; Huynh PTK; Nguyen TM; Truong TTT; Tran TL; Schumann W; Nguyen HD
[Ad] Endereço:Center for Bioscience and Biotechnology, VNUHCMC-University of Science, 227 Nguyen Van Cu Dist. 5, Hochiminh, Vietnam.
[Ti] Título:Development of inducer-free expression plasmids based on IPTG-inducible promoters for Bacillus subtilis.
[So] Source:Microb Cell Fact;16(1):130, 2017 Jul 25.
[Is] ISSN:1475-2859
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Besides Escherichia coli, Bacillus subtilis is an important bacterial species for the production of recombinant proteins. Recombinant genes are inserted into shuttle expression vectors which replicate in both E. coli and in B. subtilis. The ligation products are first transformed into E. coli cells, analyzed for correct insertions, and the correct recombinant plasmids are then transformed into B. subtilis. A major problem using E. coli cells can be the strong basal level of expression of the recombinant protein which may interfere with the stability of the cells. To minimize this problem, we developed strong expression vectors being repressed in E. coli and inducer-free in B. subtilis. RESULTS: In general, induction of IPTG-inducible expression vectors is determined by the regulatory lacI gene encoding the LacI repressor in combination with the lacO operator on the promoter. To investigate the inducer-free properties of the vectors, we constructed inducer-free expression plasmids by removing the lacI gene and characterized their properties. First, we examined the ability to repress a reporter gene in E. coli, which is a prominent property facilitating the construction of the expression vectors carrying a target gene. The ß-galactosidase (bgaB gene) basal levels expressed from Pgrac01-bgaB could be repressed at least twice in the E. coli cloning strain. Second, the inducer-free production of BgaB from four different plasmids with the Pgrac01 promoter in B. subtilis was investigated. As expected, BgaB expression levels of inducer-free constructs are at least 37 times higher than that of the inducible constructs in the absence of IPTG, and comparable to those in the presence of the inducer. Third, using efficient IPTG-inducible expression vectors containing the strong promoter Pgrac100, we could convert them into inducer-free expression plasmids. The BgaB production levels from the inducer-free plasmid in the absence of the inducer were at least 4.5 times higher than that of the inducible vector using the same promoter. Finally, we used gfp as a reporter gene in combination with the two promoters Pgrac01 and Pgrac100 to test the new vector types. The GFP expression levels could be repressed at least 1.5 times for the Pgrac01-gfp+ inducer-free construct in E. coli. The inducer-free constructs Pgrac01-gfp+ and Pgrac100-gfp+ allowed GFP expression at high levels from 23 × 10 to 32 × 10 RFU units and 9-13% of total intracellular proteins. We could reconfirm the two major advantages of the new inducer-free expression plasmids: (1) Strong repression of the target gene expression in the E. coli cloning strain, and (2) production of the target protein at high levels in B. subtilis in the absence of the inducer. CONCLUSIONS: We propose a general strategy to generate inducer-free expression vector by using IPTG-inducible vectors, and more specifically we developed inducer-free expression plasmids using IPTG-inducible promoters in the absence of the LacI repressor. These plasmids could be an excellent choice for high-level production of recombinant proteins in B. subtilis without the addition of inducer and at the same time maintaining a low basal level of the recombinant proteins in E. coli. The repression of the recombinant gene expression would facilitate cloning of genes that potentially inhibit the growth of E. coli cloning strains. The inducer-free expression plasmids will be extended versions of the current available IPTG-inducible expression vectors for B. subtilis, in which all these vectors use the same cognate promoters. These inducer-free and previously developed IPTG-inducible expression plasmids will be a useful cassette to study gene expression at a small scale up to a larger scale up for the production of recombinant proteins.
[Mh] Termos MeSH primário: Bacillus subtilis/metabolismo
Expressão Gênica/efeitos dos fármacos
Isopropiltiogalactosídeo/farmacologia
Plasmídeos/metabolismo
[Mh] Termos MeSH secundário: Escherichia coli/metabolismo
Genes Reporter
Proteínas de Fluorescência Verde/genética
Proteínas de Fluorescência Verde/metabolismo
Plasmídeos/genética
Regiões Promotoras Genéticas
beta-Galactosidase/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
147336-22-9 (Green Fluorescent Proteins); 367-93-1 (Isopropyl Thiogalactoside); EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180205
[Lr] Data última revisão:
180205
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170727
[St] Status:MEDLINE
[do] DOI:10.1186/s12934-017-0747-0


  7 / 18867 MEDLINE  
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[PMID]:28470628
[Au] Autor:Zhou Y; Liu K; Zhang J; Chu J; He B
[Ad] Endereço:College of Biotechnology and Pharmaceutical Engineering, Nanjing Tech University, Nanjing, China.
[Ti] Título:Mg -induced stabilization of ß-galactosidase from Bacillus megaterium and its application in the galactosylation of natural products.
[So] Source:Biotechnol Lett;39(8):1175-1181, 2017 Aug.
[Is] ISSN:1573-6776
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:OBJECTIVE: To improve the stability of ß-galactosidase from Bacillus megaterium YZ08 (BMG) in aqueous hydrophilic solvents and promote its application in the galactosylation of natural products. RESULTS: The addition of 5 mM Mg significantly enhanced the stability of BMG in aqueous hydrophilic solvents, and the half-lives of BMG in these solutions reached 56 min to 208 h, while they were only 7 min to 5.9 h without addition of Mg . Studies on the kinetic parameters in buffer solution and 30% dimethyl sulfoxide (DMSO) indicated that the affinity of BMG to 2-nitrophenyl-ß-D-galactopyranoside and its catalytic efficiency (κ /K ) increased with the addition of Mg . Furthermore, the addition of Mg facilitated galactosylation reactions in 30% DMSO and increased product conversions by 24-41% due to the reversal of the thermodynamic equilibrium of hydrolysis. CONCLUSION: A convenient approach was established to improve the stability of BMG in aqueous hydrophilic solvents.
[Mh] Termos MeSH primário: Proteínas de Bactérias/química
Produtos Biológicos/metabolismo
Magnésio/química
beta-Galactosidase/química
[Mh] Termos MeSH secundário: Bacillus megaterium
Proteínas de Bactérias/metabolismo
Produtos Biológicos/química
Dimetil Sulfóxido/química
Estabilidade Enzimática
Temperatura Alta
Fenômenos de Química Orgânica
beta-Galactosidase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (Biological Products); EC 3.2.1.23 (beta-Galactosidase); I38ZP9992A (Magnesium); YOW8V9698H (Dimethyl Sulfoxide)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180202
[Lr] Data última revisão:
180202
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE
[do] DOI:10.1007/s10529-017-2344-z


  8 / 18867 MEDLINE  
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[PMID]:27778280
[Au] Autor:Chandrasekar B; Hong TN; van der Hoorn RA
[Ad] Endereço:The Plant Chemetics Laboratory, Max Planck Institute for Plant Breeding Research, Carl-von-Linne Weg 10, 50829, Cologne, Germany.
[Ti] Título:Inhibitor Discovery by Convolution ABPP.
[So] Source:Methods Mol Biol;1491:47-56, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Activity-based protein profiling (ABPP) has emerged as a powerful proteomic approach to study the active proteins in their native environment by using chemical probes that label active site residues in proteins. Traditionally, ABPP is classified as either comparative or competitive ABPP. In this protocol, we describe a simple method called convolution ABPP, which takes benefit from both the competitive and comparative ABPP. Convolution ABPP allows one to detect if a reduced signal observed during comparative ABPP could be due to the presence of inhibitors. In convolution ABPP, the proteomes are analyzed by comparing labeling intensities in two mixed proteomes that were labeled either before or after mixing. A reduction of labeling in the mix-and-label sample when compared to the label-and-mix sample indicates the presence of an inhibitor excess in one of the proteomes. This method is broadly applicable to detect inhibitors in proteomes against any proteome containing protein activities of interest. As a proof of concept, we applied convolution ABPP to analyze secreted proteomes from Pseudomonas syringae-infected Nicotiana benthamiana leaves to display the presence of a beta-galactosidase inhibitor.
[Mh] Termos MeSH primário: Inibidores Enzimáticos/química
Proteômica/métodos
[Mh] Termos MeSH secundário: Inibidores Enzimáticos/farmacologia
Sondas Moleculares/química
Proteínas/química
Pseudomonas syringae/isolamento & purificação
Tabaco/química
beta-Galactosidase/antagonistas & inibidores
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Enzyme Inhibitors); 0 (Molecular Probes); 0 (Proteins); EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1801
[Cu] Atualização por classe:180116
[Lr] Data última revisão:
180116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161026
[St] Status:MEDLINE


  9 / 18867 MEDLINE  
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[PMID]:29198188
[Au] Autor:Pimentel G; Burton KJ; Rosikiewicz M; Freiburghaus C; von Ah U; Münger LH; Pralong FP; Vionnet N; Greub G; Badertscher R; Vergères G
[Ad] Endereço:1Agroscope,Schwarzenburgstrasse 161,3003 Bern,Switzerland.
[Ti] Título:Blood lactose after dairy product intake in healthy men.
[So] Source:Br J Nutr;118(12):1070-1077, 2017 Dec.
[Is] ISSN:1475-2662
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The absence of a dedicated transport for disaccharides in the intestine implicates that the metabolic use of dietary lactose relies on its prior hydrolysis at the intestinal brush border. Consequently, lactose in blood or urine has mostly been associated with specific cases in which the gastrointestinal barrier is damaged. On the other hand, lactose appears in the blood of lactating women and has been detected in the blood and urine of healthy men, indicating that the presence of lactose in the circulation of healthy subjects is not incompatible with normal physiology. In this cross-over study we have characterised the postprandial kinetics of lactose, and its major constituent, galactose, in the serum of fourteen healthy men who consumed a unique dose of 800 g milk or yogurt. Genetic testing for lactase persistence and microbiota profiling of the subjects were also performed. Data revealed that lactose does appear in serum after dairy intake, although with delayed kinetics compared with galactose. Median serum concentrations of approximately 0·02 mmol/l lactose and approximately 0·2 mmol/l galactose were observed after the ingestion of milk and yogurt respectively. The serum concentrations of lactose were inversely correlated with the concentrations of galactose, and the variability observed between the subjects' responses could not be explained by the presence of the lactase persistence allele. Finally, lactose levels have been associated with the abundance of the Veillonella genus in faecal microbiota. The measurement of systemic lactose following dietary intake could provide information about lactose metabolism and nutrient transport processes under normal or pathological conditions.
[Mh] Termos MeSH primário: Dieta
Lactose/sangue
Leite
Iogurte
[Mh] Termos MeSH secundário: Adolescente
Adulto
Alelos
Animais
Estudos Cross-Over
Método Duplo-Cego
Fezes/microbiologia
Galactose/sangue
Microbioma Gastrointestinal
Seres Humanos
Intestinos/metabolismo
Intestinos/microbiologia
Masculino
Período Pós-Prandial
Veillonella/isolamento & purificação
Adulto Jovem
beta-Galactosidase/genética
beta-Galactosidase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RANDOMIZED CONTROLLED TRIAL
[Nm] Nome de substância:
EC 3.2.1.23 (beta-Galactosidase); J2B2A4N98G (Lactose); X2RN3Q8DNE (Galactose)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:171227
[Lr] Data última revisão:
171227
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171205
[St] Status:MEDLINE
[do] DOI:10.1017/S0007114517003245


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[PMID]:27771213
[Au] Autor:Søndergaard RH; Follin B; Lund LD; Juhl M; Ekblond A; Kastrup J; Haack-Sørensen M
[Ad] Endereço:Cardiology Stem Cell Centre, The Heart Centre, Rigshospitalet, Copenhagen University Hospital, Copenhagen, Denmark.
[Ti] Título:Senescence and quiescence in adipose-derived stromal cells: Effects of human platelet lysate, fetal bovine serum and hypoxia.
[So] Source:Cytotherapy;19(1):95-106, 2017 01.
[Is] ISSN:1477-2566
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND AIMS: Adipose-derived stromal cells (ASCs) are attractive sources for cell-based therapies. The hypoxic niche of ASCs in vivo implies that cells will benefit from hypoxia during in vitro expansion. Human platelet lysate (hPL) enhances ASC proliferation rates, compared with fetal bovine serum (FBS) at normoxia. However, the low proliferation rates of FBS-expanded ASCs could be signs of senescence or quiescence. We aimed to determine the effects of hypoxia and hPL on the expansion of ASCs and whether FBS-expanded ASCs are senescent or quiescent. METHODS: ASCs expanded in FBS or hPL at normoxia or hypoxia until passage 7 (P7), or in FBS until P5 followed by culture in hPL until P7, were evaluated by proliferation rates, cell cycle analyses, gene expression and ß-galactosidase activity. RESULTS: hPL at normoxia and hypoxia enhanced proliferation rates and expression of cyclins, and decreased G0/G1 fractions and expression of p21 and p27, compared with FBS. The shift from FBS to hPL enhanced cyclin levels, decreased p21 and p27 levels and tended to decrease G0/G1 fractions. CONCLUSION: Hypoxia does not add to the effect of hPL during ASC expansion with regard to proliferation, cell cycle regulation and expression of cyclins, p21 and p27. hPL rejuvenates FBS-expanded ASCs with regard to cell cycle regulation and expression of cyclins, p21 and p27. This indicates a reversible arrest. Therefore, we conclude that ASCs expanded until P7 are not senescent regardless of culture conditions.
[Mh] Termos MeSH primário: Tecido Adiposo/citologia
Plaquetas/química
Técnicas de Cultura de Células/métodos
Células Estromais/citologia
[Mh] Termos MeSH secundário: Adulto
Animais
Bovinos
Ciclo Celular
Hipóxia Celular
Proliferação Celular
Células Cultivadas
Senescência Celular
Ciclinas/genética
Ciclinas/metabolismo
Feminino
Regulação da Expressão Gênica
Seres Humanos
Imunofenotipagem
Masculino
Soro
Células Estromais/fisiologia
beta-Galactosidase/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Cyclins); EC 3.2.1.23 (beta-Galactosidase)
[Em] Mês de entrada:1712
[Cu] Atualização por classe:171207
[Lr] Data última revisão:
171207
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161025
[St] Status:MEDLINE



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