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[PMID]:28884816
[Au] Autor:Ferreira R; Gatto F; Nielsen J
[Ad] Endereço:Department of Biology and Biological Engineering, Systems and Synthetic Biology, Chalmers University of Technology, Gothenburg, Sweden.
[Ti] Título:Exploiting off-targeting in guide-RNAs for CRISPR systems for simultaneous editing of multiple genes.
[So] Source:FEBS Lett;591(20):3288-3295, 2017 Oct.
[Is] ISSN:1873-3468
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Bioinformatics tools to design guide-RNAs (gRNAs) in Clustered Regularly Interspaced Short Palindromic Repeats systems mostly focused on minimizing off-targeting to enhance efficacy of genome editing. However, there are circumstances in which off-targeting might be desirable to target multiple genes simultaneously with a single gRNA. We termed these gRNAs as promiscuous gRNAs. Here, we present a computational workflow to identify promiscuous gRNAs that putatively bind to the region of interest for a defined list of genes in a genome. We experimentally validated two promiscuous gRNA for gene deletion, one targeting FAA1 and FAA4 and one targeting PLB1 and PLB2, thus demonstrating that multiplexed genome editing through design of promiscuous gRNA can be performed in a time and cost-effective manner.
[Mh] Termos MeSH primário: Sistemas CRISPR-Cas
Biologia Computacional/métodos
Edição de Genes/métodos
Genoma Fúngico
RNA Guia/genética
Saccharomyces cerevisiae/genética
[Mh] Termos MeSH secundário: Acil Coenzima A/genética
Acil Coenzima A/metabolismo
Sequência de Bases
Repetições Palindrômicas Curtas Agrupadas e Regularmente Espaçadas
Coenzima A Ligases/genética
Coenzima A Ligases/metabolismo
Deleção de Genes
Expressão Gênica
Lisofosfolipase/genética
Lisofosfolipase/metabolismo
Proteínas de Membrana/genética
Proteínas de Membrana/metabolismo
RNA Guia/metabolismo
Saccharomyces cerevisiae/metabolismo
Proteínas de Saccharomyces cerevisiae/genética
Proteínas de Saccharomyces cerevisiae/metabolismo
[Pt] Tipo de publicação:LETTER
[Nm] Nome de substância:
0 (Acyl Coenzyme A); 0 (Membrane Proteins); 0 (RNA, Guide); 0 (Saccharomyces cerevisiae Proteins); EC 3.1.1.5 (Lysophospholipase); EC 3.1.1.5 (PLB1 protein, S cerevisiae); EC 3.1.1.5 (PLB2 protein, S cerevisiae); EC 6.2.1.- (Coenzyme A Ligases); EC 6.2.1.- (Faa1 protein, S cerevisiae); EC 6.2.1.3 (FAA4 protein, S cerevisiae)
[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:170909
[St] Status:MEDLINE
[do] DOI:10.1002/1873-3468.12835


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[PMID]:28645872
[Au] Autor:Lv D; Zhou D; Zhang Y; Zhang S; Zhu YM
[Ad] Endereço:Department of Epidemiology & Biostatistics, Zhejiang University School of Public Health, Hangzhou 310058, China; Research Center of Clinical Pharmacy, State Key Laboratory for Diagnosis and Treatment of Infectious Disease, First Affiliated Hospital, Zhejiang University, Hangzhou 310058, China.
[Ti] Título:Two obesity susceptibility loci in LYPLAL1 and ETV5 independently associated with childhood hypertension in Chinese population.
[So] Source:Gene;627:284-289, 2017 Sep 05.
[Is] ISSN:1879-0038
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:AIMS: Genome-wide association studies have identified novel obesity-associated susceptibility loci. Associations of these variants with childhood obesity have been studied in our previous research. The purpose of this study is to investigate if these loci are associated with hypertension being independent of obesity in Chinese children and adolescents. METHODS: Nineteen candidate SNPs were genotyped using Sequenom MassARRAY platform among Chinese children (N=2954, 514 hypertension and 2440 controls, aged 7-17years). Dietary behaviors were assessed through face to face investigations. RESULTS: Of the nineteen obese related SNPs, ten SNPs were found to be associated with systolic blood pressure (SBP) or diastolic blood pressure (DBP) in Chinese children. After adjusting for age, sex and WHtR, rs2605100 in LYPLAL1was found to be associated with high blood pressure (HBP) under dominant model (P=0.024) with the OR of 1.274 (95% CI =1.033-1.572, effect genotype=GG). The distribution of genotype of rs7647305 in ETV5 showed significant difference between HBP and non-HBP subjects under dominant model (P=0.011) with the OR of 0.654 (95% CI=0.471-0.909, effect genotype=CC). Using rs2605100 and rs7647305, the genetic risk score (GRS) analysis showed that, after adjusted for age, sex and WHtR, subjects carrying one or two risk alleles had the risks of hypertension with the ORs 1.797 (95% CI, 1.168-2.765), 2.149 (95% CI, 1.375-3.357) comparing with the subjects with non-risk-allele. CONCLUSIONS: Genetic variations of obesity-associated loci, LYPLAL1 rs2605100 and ETV5 rs7647305 independently associate with the risk of childhood hypertension in China.
[Mh] Termos MeSH primário: Proteínas de Ligação a DNA/genética
Hipertensão/genética
Lisofosfolipase/genética
Obesidade/genética
Polimorfismo de Nucleotídeo Único
Fatores de Transcrição/genética
[Mh] Termos MeSH secundário: Adolescente
Estudos de Casos e Controles
Criança
China
Feminino
Seres Humanos
Masculino
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA-Binding Proteins); 0 (ETV5 protein, human); 0 (Transcription Factors); EC 3.1.1.5 (Lysophospholipase); EC 3.1.2.- (LYPLAL1 protein, human)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170918
[Lr] Data última revisão:
170918
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170625
[St] Status:MEDLINE


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[PMID]:28594920
[Au] Autor:Dold L; Luda C; Schwarze-Zander C; Boesecke C; Hansel C; Nischalke HD; Lutz P; Mohr R; Wasmuth JC; Strassburg CP; Trebicka J; Rockstroh JK; Spengler U
[Ad] Endereço:Department of Internal Medicine I, Rheinische Friedrich-Wilhelms University Bonn, Bonn, Germany.
[Ti] Título:Genetic polymorphisms associated with fatty liver disease and fibrosis in HIV positive patients receiving combined antiretroviral therapy (cART).
[So] Source:PLoS One;12(6):e0178685, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Hepatic steatosis can occur with any antiretroviral therapy (cART). Although single nucleotide polymorphisms (SNPs) have been identified to predispose to alcoholic and non-alcoholic fatty liver disease, their role for treatment-associated steatosis in HIV-positive patients remains unclear. We determined the frequency of PNPLA3 (rs738409), CSPG3/NCAN (rs2228603), GCKR (rs780094), PPP1R3B (rs4240624), TM6SF (rs8542926), LYPLAL1 (rs12137855) and MBOAT7 (rs626283) by RT-PCR in 117 HIV-positive patients on cART and stratified participants based on their "controlled attenuation parameter" (CAP) into probable (CAP: 215-300 dB/m) and definite (CAP >300 dB/m) hepatic steatosis. We analyzed CAP values and routine metabolic parameters according to the allele frequencies. Sixty-five (55.6%) and 13 (11.1%) patients were allocated to probable and definite steatosis. CAP values (p = 0.012) and serum triglycerides (p = 0.043) were increased in carriers of the GCKR (rs780094) A allele. Cox logistic regression identified triglycerides (p = 0.006), bilirubin (p = 0.021) and BMI (p = 0.068), but not the genetic parameters as risk factors for the occurrence of hepatic steatosis. Taken together, according to the limited sample size, this exploratory study generates the hypothesis that genetic polymorphisms seem to exert minor effects on the risk for fatty liver disease in HIV-positive patients on cART. Nevertheless, SNPs may modify metabolic complications once metabolic abnormalities have developed. Hence, subsequent analysis of a larger cohort is needed.
[Mh] Termos MeSH primário: Fígado Gorduroso/genética
Cirrose Hepática/genética
[Mh] Termos MeSH secundário: Aciltransferases/genética
Proteínas Adaptadoras de Transdução de Sinal/genética
Adulto
Idoso
Idoso de 80 Anos ou mais
Antirretrovirais/uso terapêutico
Bilirrubina/sangue
Índice de Massa Corporal
Proteoglicanas de Sulfatos de Condroitina/genética
Feminino
Predisposição Genética para Doença/genética
Soropositividade para HIV/tratamento farmacológico
Soropositividade para HIV/genética
Seres Humanos
Lipase/genética
Lisofosfolipase/genética
Masculino
Proteínas de Membrana/genética
Meia-Idade
Polimorfismo de Nucleotídeo Único/genética
Modelos de Riscos Proporcionais
Proteína Fosfatase 1/genética
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Triglicerídeos/sangue
Adulto Jovem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Adaptor Proteins, Signal Transducing); 0 (Anti-Retroviral Agents); 0 (CSPG4 protein, human); 0 (Chondroitin Sulfate Proteoglycans); 0 (GCKR protein, human); 0 (Membrane Proteins); 0 (Triglycerides); EC 2.3.- (Acyltransferases); EC 2.3.- (MBOAT7 protein, human); EC 3.1.1.3 (Lipase); EC 3.1.1.3 (adiponutrin, human); EC 3.1.1.5 (Lysophospholipase); EC 3.1.2.- (LYPLAL1 protein, human); EC 3.1.3.16 (PPP1R3B protein, human); EC 3.1.3.16 (Protein Phosphatase 1); RFM9X3LJ49 (Bilirubin)
[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:170609
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0178685


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[PMID]:28322448
[Au] Autor:Lavin J; Min JY; Lidder AK; Huang JH; Kato A; Lam K; Meen E; Chmiel JS; Norton J; Suh L; Mahdavinia M; Hulse KE; Conley DB; Chandra RK; Shintani-Smith S; Kern RC; Schleimer RP; Tan BK
[Ad] Endereço:Department of Otolaryngology-Head and Neck SurgeryDivision of Pediatric Otolaryngology, Ann & Robert H. Lurie Children's Hospital of Chicago, Northwestern University Feinberg School of Medicine, Chicago, Illinois, U.S.A.
[Ti] Título:Superior turbinate eosinophilia correlates with olfactory deficit in chronic rhinosinusitis patients.
[So] Source:Laryngoscope;127(10):2210-2218, 2017 Oct.
[Is] ISSN:1531-4995
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:OBJECTIVE: To evaluate if molecular markers of eosinophilia in olfactory-enriched mucosa are associated with olfactory dysfunction. STUDY DESIGN: Cross-sectional study of tissue biopsies from 99 patients, and an additional 30 patients who underwent prospective olfactory testing prior to sinonasal procedures. METHODS: Tissue biopsies were processed for analysis of inflammatory markers using quantitative real time polymerase chain reaction (qRT-PCR). Ipsilateral olfactory performance was assessed using the Sniffin' Sticks (Burghart, Wedel, Germany) threshold component and the University of Pennsylvania Smell Identification Test (Sensonics, Haddon Heights, NJ). Age-adjusted data was correlated with inflammatory marker expression and clinical measures of obstruction from computed tomography and endoscopy. RESULTS: Gene expression of the eosinophil marker CLC (Charcot Leyden crystal protein) was elevated in superior turbinate (ST) tissue in chronic rhinosinusitis (CRS) with nasal polyps (CRSwNP) compared to ST and inferior turbinate tissue in CRS without nasal polyps (CRSsNP) and control patients (all P < 0.001, respectively). CLC in ST tissue was correlated with IL-5 and eotaxin-1 expression (all P < 0.001; P = 0.65, and 0.49, respectively). CLC expression was strongly correlated with eosinophilic cationic protein levels (P < 0.001; r = -0.76), and ST CLC expression was inversely related to olfactory threshold (P = 0.002, r = -0.57) and discrimination scores (P = 0.05, r = -0.42). In multiple linear regression of CLC gene expression, polyp status, and radiographic and endoscopic findings with olfactory threshold, CLC was the only significantly correlated variable (P < 0.05). CONCLUSION: Markers of eosinophils are elevated in the ST of patients with CRSwNP and correlate with olfactory loss. These findings support the hypothesis that olfactory dysfunction in CRS correlates local eosinophil influx into the olfactory cleft. LEVEL OF EVIDENCE: NA. Laryngoscope, 127:2210-2218, 2017.
[Mh] Termos MeSH primário: Eosinofilia/complicações
Transtornos do Olfato/etiologia
Rinite/complicações
Sinusite/complicações
[Mh] Termos MeSH secundário: Adulto
Idoso
Quimiocina CCL11/análise
Doença Crônica
Estudos Transversais
Proteína Catiônica de Eosinófilo/sangue
Eosinofilia/sangue
Eosinofilia/patologia
Feminino
Glicoproteínas/análise
Seres Humanos
Interleucina-5/análise
Lisofosfolipase/análise
Masculino
Meia-Idade
Pólipos Nasais/sangue
Pólipos Nasais/complicações
Pólipos Nasais/patologia
Estudos Prospectivos
Rinite/sangue
Rinite/patologia
Sinusite/sangue
Sinusite/patologia
Conchas Nasais/patologia
Adulto Jovem
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (CCL11 protein, human); 0 (Chemokine CCL11); 0 (Glycoproteins); 0 (IL5 protein, human); 0 (Interleukin-5); EC 3.1.1.5 (Lysophospholipase); EC 3.1.1.5 (lysolecithin acylhydrolase); EC 3.1.27.- (Eosinophil Cationic Protein)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170929
[Lr] Data última revisão:
170929
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170322
[St] Status:MEDLINE
[do] DOI:10.1002/lary.26555


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[PMID]:27940286
[Au] Autor:Su L; Ji D; Tao X; Yu L; Wu J; Xia Y
[Ad] Endereço:State Key Laboratory of Food Science and Technology, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China; School of Biotechnology and Key Laboratory of Industrial Biotechnology Ministry of Education, Jiangnan University, 1800 Lihu Avenue, Wuxi, 214122, China.
[Ti] Título:Recombinant expression, characterization, and application of a phospholipase B from Fusarium oxysporum.
[So] Source:J Biotechnol;242:92-100, 2017 Jan 20.
[Is] ISSN:1873-4863
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:In this study, a gene encoding a putative lipase from Fusarium oxysporum was optimized via codon optimization and expressed in Pichia pastoris KM71. The gene product was identified as a phospholipase B (PLB). The engineered P. pastoris was further cultured in a 3.6-L bioreactor. After optimization of the induction conditions, this system produced 6.6mgmL protein and 6503.8UmL PLB activity in the culture medium. Efficient expression of this PLB in P. pastoris should reduce the costs of production and application. The purified enzyme, with a specific activity of 1170Umg , was optimally active at pH 5.0 and 55°C. The results of a degumming experiment performed using the recombinant PLB showed that the phosphorus content of a test oil was decreased from 75.88ppm to 3.3ppm in 2h under optimal reaction conditions. This study provides a basis for the industrial use of F. oxysporum PLB in oil degumming applications.
[Mh] Termos MeSH primário: Fusarium/enzimologia
Lisofosfolipase/genética
Lisofosfolipase/metabolismo
[Mh] Termos MeSH secundário: Proteínas de Bactérias/genética
Proteínas de Bactérias/metabolismo
Técnicas de Cultura Celular por Lotes
Reatores Biológicos
Contagem de Células
Clonagem Molecular
Ativação Enzimática
Estabilidade Enzimática
Fusarium/genética
Lipase/genética
Lipase/metabolismo
Lisofosfolipase/biossíntese
Lisofosfolipase/química
Petróleo/metabolismo
Pichia/genética
Pichia/metabolismo
Engenharia de Proteínas
Proteínas Recombinantes/biossíntese
Proteínas Recombinantes/química
Proteínas Recombinantes/genética
Proteínas Recombinantes/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Bacterial Proteins); 0 (Petroleum); 0 (Recombinant Proteins); EC 3.1.1.3 (Lipase); EC 3.1.1.3 (thermostable lipase); EC 3.1.1.5 (Lysophospholipase)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170411
[Lr] Data última revisão:
170411
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161213
[St] Status:MEDLINE


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[PMID]:27926815
[Au] Autor:Zhu H; Liu P; Du J; Wang J; Jing Y; Zhang J; Gu W; Wang W; Meng Q
[Ad] Endereço:2​Laboratory of Animal Improvement and Reproduction, Institute of Animal Science, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, PR China 1​Jiangsu Key Laboratory for Biodiversity & Biotechnology and Jiangsu Key Laboratory for Aquatic Crustacean Diseases, College of Life Sciences,
[Ti] Título:Identification of lysophospholipase protein from Spiroplasma eriocheiris and verification of its function.
[So] Source:Microbiology;163(2):175-184, 2017 Feb.
[Is] ISSN:1465-2080
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Spiroplasma eriocheiris is known to cause tremor disease in the Chinese mitten crab Eriocheir sinensis; however, the molecular characterization of this pathogen is still unclear. S. eriocheiris has the ability to invade and survive within mouse 3T6 cells. The invasion process may require causing damage to the host cell membrane by chemical, physical or enzymatic means. In this study, we systematically characterized a novel lysophospholipase (lysoPL) of S. eriocheiris TDA-040725-5T. The gene that encodes lysoPL in S. eriocheiris (SE-LysoPL) was cloned, sequenced and expressed in Escherichia coli BL21 (DE3). Enzymatic assays revealed that the purified recombinant SE-LysoPL hydrolysed long-chain acyl esterases at pH 7 and 30 °C. SE-LysoPL was detected in the membrane and cytoplasmic protein fractions using the SE-LysoPL antibody in Western blot. The virulence ability of S. eriocheiris was effectively reduced at the early stage of infection (m.o.i.=100) by the SE-LysoPL antibody neutralization test. To the best of our knowledge, this is the first study to identify and characterize a gene from S. eriocheiris encoding a protein exhibiting lysoPL and esterase activities. Our findings indicate that SE-LysoPL plays important roles in the pathogenicity of S. eriocheiris.
[Mh] Termos MeSH primário: Anticorpos Neutralizantes/imunologia
Braquiúros/microbiologia
Lisofosfolipase/genética
Lisofosfolipase/imunologia
Spiroplasma/patogenicidade
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Animais
Linhagem Celular
Clonagem Molecular
Escherichia coli/genética
Escherichia coli/metabolismo
Camundongos
Alinhamento de Sequência
Spiroplasma/enzimologia
Spiroplasma/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antibodies, Neutralizing); EC 3.1.1.5 (Lysophospholipase)
[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:161208
[St] Status:MEDLINE
[do] DOI:10.1099/mic.0.000407


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[PMID]:27876784
[Au] Autor:He Y; Li L; Hu F; Chen W; Lei H; Chen X; Cai W; Tang X
[Ad] Endereço:Department of Infectious Diseases, Guangzhou Eighth People's Hospital, Guangzhou Medical University, Guangzhou 510060, Guangdong Province, China.
[Ti] Título:Expression and characterization of a Talaromyces marneffei active phospholipase B expressed in a Pichia pastoris expression system.
[So] Source:Emerg Microbes Infect;5(11):e120, 2016 Nov 23.
[Is] ISSN:2222-1751
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Phospholipase B is a virulence factor for several clinically important pathogenic fungi, including Candida albicans, Cryptococcus neoformans and Aspergillus fumigatus, but its role in the thermally dimorphic fungus Talaromyces marneffei remains unclear. Here, we provide the first report of the expression of a novel phospholipase gene, designated TmPlb1, from T. marneffei in the eukaryotic expression system of Pichia pastoris GS115. Sensitive real-time quantitative reverse-transcription PCR (qRT-PCR) demonstrated that the expression of TmPlb1 increased 1.85-fold in the yeast phase compared with the mycelial phase. TmPlb1 contains an open reading frame (ORF) of 732 bp that encodes a protein of 243 amino acids. The conserved serine, aspartate and histidine catalytic triad and the G-X-S-X-G domain of TmPLB1 provide the structural basis for its molecular activity. The ORF of TmPlb1 was successfully cloned into a pPIC9K vector containing an α-mating factor secretion signal that allowed the secretory expression of TmPLB1 in P. pastoris. The heterologous protein expression began 12 h after methanol induction and peaked at 96 h. Through analysis with SDS-polyacrylamide gel electrophoresis (SDS-PAGE), western blotting and mass spectrometry, we confirmed that TmPLB1 was successfully expressed. Through Ni-affinity chromatography, TmPLB1 was highly purified, and its concentration reached 240.4 mg/L of culture medium. With specific substrates, the phospholipase A1 and phospholipase A2 activities of TmPLB1 were calculated to be 5.96 and 1.59 U/mg, respectively. The high purity and activity of the TmPLB1 obtained here lay a solid foundation for further investigation.
[Mh] Termos MeSH primário: Lisofosfolipase/genética
Lisofosfolipase/metabolismo
Talaromyces/enzimologia
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Western Blotting
Domínio Catalítico
Cromatografia de Afinidade
Sequência Conservada
Eletroforese em Gel de Poliacrilamida
Perfilação da Expressão Gênica
Lisofosfolipase/isolamento & purificação
Espectrometria de Massas
Dados de Sequência Molecular
Fases de Leitura Aberta
Filogenia
Pichia/genética
Pichia/metabolismo
Reação em Cadeia da Polimerase em Tempo Real
Proteínas Recombinantes/genética
Proteínas Recombinantes/isolamento & purificação
Proteínas Recombinantes/metabolismo
Alinhamento de Sequência
Talaromyces/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Recombinant Proteins); EC 3.1.1.5 (Lysophospholipase)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:170220
[Lr] Data última revisão:
170220
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161124
[St] Status:MEDLINE
[do] DOI:10.1038/emi.2016.119


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[PMID]:27793292
[Au] Autor:Chadda R; Robertson JL
[Ad] Endereço:The University of Iowa, Iowa City, IA, United States.
[Ti] Título:Measuring Membrane Protein Dimerization Equilibrium in Lipid Bilayers by Single-Molecule Fluorescence Microscopy.
[So] Source:Methods Enzymol;581:53-82, 2016.
[Is] ISSN:1557-7988
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Dimerization of membrane protein interfaces occurs during membrane protein folding and cell receptor signaling. Here, we summarize a method that allows for measurement of equilibrium dimerization reactions of membrane proteins in lipid bilayers, by measuring the Poisson distribution of subunit capture into liposomes by single-molecule photobleaching analysis. This strategy is grounded in the fact that given a comparable labeling efficiency, monomeric or dimeric forms of a membrane protein will give rise to distinctly different photobleaching probability distributions. These methods have been used to verify the dimer stoichiometry of the Fluc F ion channel and the dimerization equilibrium constant of the ClC-ec1 Cl /H antiporter in lipid bilayers. This approach can be applied to any membrane protein system provided it can be purified, fluorescently labeled in a quantitative manner, and verified to be correctly folded by functional assays, even if the structure is not yet known.
[Mh] Termos MeSH primário: Glicoproteínas/isolamento & purificação
Bicamadas Lipídicas/química
Lisofosfolipase/isolamento & purificação
Microscopia de Fluorescência/métodos
Imagem Individual de Molécula/métodos
[Mh] Termos MeSH secundário: Dimerização
Glicoproteínas/química
Seres Humanos
Canais Iônicos/química
Lipossomos/química
Lisofosfolipase/química
Fotodegradação
Multimerização Proteica
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Glycoproteins); 0 (Ion Channels); 0 (Lipid Bilayers); 0 (Liposomes); EC 3.1.1.5 (Lysophospholipase); EC 3.1.1.5 (lysolecithin acylhydrolase)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170825
[Lr] Data última revisão:
170825
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161030
[St] Status:MEDLINE


  9 / 689 MEDLINE  
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[PMID]:27780639
[Au] Autor:Shah P; Cheasty A; Foxton C; Raynham T; Farooq M; Gutierrez IF; Lejeune A; Pritchard M; Turnbull A; Pang L; Owen P; Boyd S; Stowell A; Jordan A; Hamilton NM; Hitchin JR; Stockley M; MacDonald E; Quesada MJ; Trivier E; Skeete J; Ovaa H; Moolenaar WH; Ryder H
[Ad] Endereço:Cancer Research Technology, Discovery Laboratories, Babraham Research Campus, Cambridge CB22 3AT, UK.
[Ti] Título:Discovery of potent inhibitors of the lysophospholipase autotaxin.
[So] Source:Bioorg Med Chem Lett;26(22):5403-5410, 2016 11 15.
[Is] ISSN:1464-3405
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The autotaxin-lysophosphatidic acid (ATX-LPA) axis has been implicated in several disease conditions including inflammation, fibrosis and cancer. This makes ATX an attractive drug target and its inhibition may lead to useful therapeutic agents. Through a high throughput screen (HTS) we identified a series of small molecule inhibitors of ATX which have subsequently been optimized for potency, selectivity and developability properties. This has delivered drug-like compounds such as 9v (CRT0273750) which modulate LPA levels in plasma and are suitable for in vivo studies. X-ray crystallography has revealed that these compounds have an unexpected binding mode in that they do not interact with the active site zinc ions but instead occupy the hydrophobic LPC pocket extending from the active site of ATX together with occupying the LPA 'exit' channel.
[Mh] Termos MeSH primário: Inibidores Enzimáticos/química
Inibidores Enzimáticos/farmacologia
Lisofosfolipase/antagonistas & inibidores
Lisofosfolipídeos/metabolismo
Diester Fosfórico Hidrolases/metabolismo
[Mh] Termos MeSH secundário: Animais
Antineoplásicos/química
Antineoplásicos/farmacocinética
Antineoplásicos/farmacologia
Cristalografia por Raios X
Inibidores Enzimáticos/farmacocinética
Seres Humanos
Lisofosfolipase/metabolismo
Camundongos
Simulação de Acoplamento Molecular
Terapia de Alvo Molecular
Neoplasias/tratamento farmacológico
Neoplasias/enzimologia
Piridinas/química
Piridinas/farmacocinética
Piridinas/farmacologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antineoplastic Agents); 0 (Enzyme Inhibitors); 0 (Lysophospholipids); 0 (Pyridines); EC 3.1.1.5 (Lysophospholipase); EC 3.1.4.- (Phosphoric Diester Hydrolases); EC 3.1.4.39 (alkylglycerophosphoethanolamine phosphodiesterase); PG6M3969SG (lysophosphatidic acid)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:171124
[Lr] Data última revisão:
171124
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161027
[St] Status:MEDLINE


  10 / 689 MEDLINE  
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[PMID]:27764108
[Au] Autor:Aminnejad M; Cogliati M; Duan S; Arabatzis M; Tintelnot K; Castañeda E; Lazéra M; Velegraki A; Ellis D; Sorrell TC; Meyer W
[Ad] Endereço:Molecular Mycology Research Laboratory, Centre for Infectious Diseases and Microbiology, Sydney Medical School - Westmead Hospital, Marie Bashir Institute for Infectious Diseases and Biosecurity, The University of Sydney, Westmead Institute for Medical Research, Sydney, Australia.
[Ti] Título:Identification and Characterization of VNI/VNII and Novel VNII/VNIV Hybrids and Impact of Hybridization on Virulence and Antifungal Susceptibility Within the C. neoformans/C. gattii Species Complex.
[So] Source:PLoS One;11(10):e0163955, 2016.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Cryptococcus neoformans and C. gattii are pathogenic basidiomycetous yeasts and the commonest cause of fungal infection of the central nervous system. Cryptococci are typically haploid but several inter-species, inter-varietal and intra-varietal hybrids have been reported. It has a bipolar mating system with sexual reproduction occurring normally between two individuals with opposite mating types, α and a. This study set out to characterize hybrid isolates within the C. neoformans/C. gattii species complex: seven unisexual mating intra-varietal VNI/VNII (αAAα) and six novel inter-varietal VNII/VNIV (aADα). The URA5-RFLP pattern for VNII/VNIV (aADα) differs from the VNIII (αADa) hybrids. Analysis of the allelic patterns of selected genes for AD hybrids showed 79% or more heterozygosis for the studied loci except for CBS132 (VNIII), which showed 50% of heterozygosity. MALDI-TOF MS was applied to hybrids belonging to different sero/mating type allelic patterns. All hybrid isolates were identified as belonging to the same hybrid group with identification scores ranging between 2.101 to 2.634. All hybrids were virulent when tested in the Galleria mellonella (wax moth) model, except for VNII/VNIV (aADα) hybrids. VNI/VGII hybrids were the most virulent hybrids. Hybrids recovered from larvae manifested a significant increase in capsule and total cell size and produced a low proportion (5-10%) of giant cells compared with the haploid control strains. All strains expressed the major virulence factors-capsule, melanin and phospholipase B-and grew well at 37°C. The minimal inhibitory concentration of nine drugs was measured by micro-broth dilution and compared with published data on haploid strains. MICs were similar amongst hybrids and haploid parental strains. This is the first study reporting natural same sex αAAα intra-varietal VNI/VNII hybrids and aADα inter-varietal VNII/VNIV hybrids.
[Mh] Termos MeSH primário: Antifúngicos/farmacologia
Cryptococcus gattii/fisiologia
Cryptococcus neoformans/fisiologia
Hibridização Genética/genética
Virulência/efeitos dos fármacos
[Mh] Termos MeSH secundário: Cryptococcus gattii/genética
Cryptococcus neoformans/genética
Proteínas Fúngicas/genética
Proteínas Fúngicas/metabolismo
Genes Fúngicos Tipo Acasalamento/genética
Haploidia
Lisofosfolipase/metabolismo
Melaninas/metabolismo
Testes de Sensibilidade Microbiana
Reação em Cadeia da Polimerase Multiplex
Técnicas de Tipagem Micológica
Sorogrupo
Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz
Virulência/genética
Fatores de Virulência/análise
Fatores de Virulência/genética
Fatores de Virulência/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Antifungal Agents); 0 (CAP59 protein, Cryptococcus neoformans); 0 (Fungal Proteins); 0 (Melanins); 0 (Virulence Factors); EC 3.1.1.5 (Lysophospholipase)
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170606
[Lr] Data última revisão:
170606
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161021
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0163955



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