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Pesquisa : E01.370.225.500.620.670 [Categoria DeCS]
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[PMID]:28456689
[Au] Autor:Mandic A; Strebinger D; Regali C; Phillips NE; Suter DM
[Ad] Endereço:The Institute of Bioengineering (IBI), School of Life Sciences, Swiss Federal Institute of Technology Lausanne (EPFL), 1015 Lausanne, Switzerland.
[Ti] Título:A novel method for quantitative measurements of gene expression in single living cells.
[So] Source:Methods;120:65-75, 2017 May 01.
[Is] ISSN:1095-9130
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Gene expression is at the heart of virtually any biological process, and its deregulation is at the source of numerous pathological conditions. While impressive progress has been made in genome-wide measurements of mRNA and protein expression levels, it is still challenging to obtain highly quantitative measurements in single living cells. Here we describe a novel approach based on internal tagging of endogenous proteins with a reporter allowing luminescence and fluorescence time-lapse microscopy. Using luminescence microscopy, fluctuations of protein expression levels can be monitored in single living cells with high sensitivity and temporal resolution over extended time periods. The integrated protein decay reporter allows measuring protein degradation rates in the absence of protein synthesis inhibitors, and in combination with absolute protein levels allows determining absolute amounts of proteins synthesized over the cell cycle. Finally, the internal tag can be excised by inducible expression of Cre recombinase, which enables to estimate endogenous mRNA half-lives. Our method thus opens new avenues in quantitative analysis of gene expression in single living cells.
[Mh] Termos MeSH primário: Imagem Molecular/métodos
Proteínas/genética
Análise de Célula Única/métodos
Coloração e Rotulagem/métodos
Transcrição Genética
[Mh] Termos MeSH secundário: Animais
Linhagem Celular
Genes Reporter/genética
Vetores Genéticos/genética
Meia-Vida
Integrases/genética
Lentivirus/genética
Luminescência
Camundongos
Microscopia de Fluorescência/métodos
Imagem Molecular/instrumentação
Proteínas/química
Proteínas/metabolismo
Proteólise
RNA Mensageiro/genética
RNA Mensageiro/metabolismo
Análise de Célula Única/instrumentação
Coloração e Rotulagem/instrumentação
Imagem com Lapso de Tempo/instrumentação
Imagem com Lapso de Tempo/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Proteins); 0 (RNA, Messenger); EC 2.7.7.- (Cre recombinase); EC 2.7.7.- (Integrases)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170501
[St] Status:MEDLINE


  2 / 62903 MEDLINE  
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[PMID]:28454775
[Au] Autor:Eggert F; Kulikov K; Domnick C; Leifels P; Kath-Schorr S
[Ad] Endereço:LIMES Institute, Chemical Biology & Medicinal Chemistry Unit, University of Bonn, Gerhard-Domagk-Straße 1, 53121 Bonn, Germany.
[Ti] Título:Iluminated by foreign letters - Strategies for site-specific cyclopropene modification of large functional RNAs via in vitro transcription.
[So] Source:Methods;120:17-27, 2017 May 01.
[Is] ISSN:1095-9130
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The synthesis of sequence-specifically modified long RNA molecules, which cannot entirely be prepared via solid phase synthesis methods is experimentally challenging. We are using a new approach based on an expanded genetic alphabet preparing site-specifically modified RNA molecules via standard in vitro transcription. In this report, the site-specific labeling of functional RNAs, in particular ribozymes and a long non-coding RNA with cyclopropene moieties, is presented. We provide detailed instructions for RNA labeling via in vitro transcription and include required analytical methods to verify production and identity of the transcript. We further present post-transcriptional inverse electron demand Diels-Alder cycloaddition reactions on the cyclopropene-modified sequences and discuss applications of the genetic alphabet expansion transcription for in vitro preparation of labeled functional RNAs with complex foldings. In detail, the glmS and CPEB3 ribozymes were site-specifically decorated with methyl cyclopropene moieties using the unnatural TPT3 triphosphate and were proven to be still functional. In addition, the structurally complex A region of the Xist lncRNA (401nt) was site-specifically modified with methyl cyclopropene and detected by fluorescence after cycloaddition reaction with a tetrazine-BODIPY conjugate.
[Mh] Termos MeSH primário: Reação de Cicloadição/métodos
Ciclopropanos/química
RNA Catalítico/química
RNA Longo não Codificante/química
Coloração e Rotulagem/métodos
[Mh] Termos MeSH secundário: Elétrons
Corantes Fluorescentes/química
Técnicas In Vitro/métodos
Nucleotídeos/química
Processamento Pós-Transcricional do RNA
Transcrição Genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cyclopropanes); 0 (Fluorescent Dyes); 0 (Nucleotides); 0 (RNA, Catalytic); 0 (RNA, Long Noncoding); J6UJO23JGU (1-methylcyclopropene)
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180309
[Lr] Data última revisão:
180309
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170430
[St] Status:MEDLINE


  3 / 62903 MEDLINE  
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[PMID]:29203745
[Au] Autor:Hryn VH; Sherstyuk OO; Svintsytska NL; Piliuhin АV; Ustenko RL
[Ad] Endereço:Higher State Educational Establishment Of Ukraine "Ukrainian Medical Stomatological Academy", Poltava, Ukraine.
[Ti] Título:The use of morphological study technique for investigation of labial and palatine glands.
[So] Source:Wiad Lek;70(5):934-938, 2017.
[Is] ISSN:0043-5147
[Cp] País de publicação:Poland
[La] Idioma:eng
[Ab] Resumo:INTRODUCTION: Due to the deterioration of environmental conditions that promotes the onset of inflammatory and autoimmune diseases, and the progress in the diagnosis, the frequency of registration of intercurrent pathology of salivary glands has markedly risen in recent years, demonstrating the increased scientific interest in the research of the common and distinctive features of their structure. THE AIM: The paper was aimed at the development of the method of morphological study of human minor salivary (labial and palatine) glands by the use of plastic wax reconstruction to obtain the plastic model of the acini and ducts of human minor salivary glands. MATERIALS AND METHODS: Specimens of the glandular area of the hard palate mucosa and labial mucosa in its middle third have been studied. To gain the objective of the investigation the technique for morphological study of the human minor salivary (labial and palatine) glands is to be developed, encompassing the analysis of the spatial organization of the glandular epithelium of the labial and palatine glands together with blood microcirculatory flow by fixing the obtained specimens of the minor salivary glands in 4% glutaraldehyde solution and osmium tetroxide with subsequent embedding into the Epon-812, staining the serial semi-thin sections with phosphate buffered 0,1% toluidine blue solution, photomacrography of the distinguished boundaries of the investigated structures and obtaining of photoreconstructions. RESULTS AND CONCLUSIONS: Thus, the use of suggested technique enables to obtain the megascopic reconstruction of the acini and ducts of the labial and palatine glands, which can be studied from different sides, getting the full visualization of the shape and size, as well as to explore the glands' inner configuration, the geometry of the lumen of the epithelial excretory ducts, to determine changes in the thickness of the wall, to get a visual representation of the microtopographic interactions between the different parts of blood microcirculatory flow and excretory ducts of the minor salivary glands.
[Mh] Termos MeSH primário: Células Epiteliais/citologia
Microcirculação/fisiologia
Glândulas Salivares Menores/citologia
[Mh] Termos MeSH secundário: Seres Humanos
Imuno-Histoquímica
Microscopia Eletrônica de Varredura
Coloração e Rotulagem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1803
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171206
[St] Status:MEDLINE


  4 / 62903 MEDLINE  
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[PMID]:27770402
[Au] Autor:Soufi M; Kamali-Asl A; Geramifar P; Rahmim A
[Ad] Endereço:Department of Radiation Medicine Engineering, Shahid Beheshti University, Tehran, Iran.
[Ti] Título:A Novel Framework for Automated Segmentation and Labeling of Homogeneous Versus Heterogeneous Lung Tumors in [ F]FDG-PET Imaging.
[So] Source:Mol Imaging Biol;19(3):456-468, 2017 06.
[Is] ISSN:1860-2002
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:PURPOSE: Determination of intra-tumor high-uptake area using 2-deoxy-2-[ F]fluoro-D-glucose ([ F]FDG) positron emission tomography (PET) imaging is an important consideration for dose painting in radiation treatment applications. The aim of our study was to develop a framework towards automated segmentation and labeling of homogeneous vs. heterogeneous tumors in clinical lung [ F]FDG-PET with the capability of intra-tumor high-uptake region delineation. PROCEDURES: We utilized and extended a fuzzy random walk PET tumor segmentation algorithm to delineate intra-tumor high-uptake areas. Tumor textural feature (TF) analysis was used to find a relationship between tumor type and TF values. Segmentation accuracy was evaluated quantitatively utilizing 70 clinical [ F]FDG-PET lung images of patients with a total of 150 solid tumors. For volumetric analysis, the Dice similarity coefficient (DSC) and Hausdorff distance (HD) measures were extracted with respect to gold-standard manual segmentation. A multi-linear regression model was also proposed for automated tumor labeling based on TFs, including cross-validation analysis. RESULTS: Two-tailed t test analysis of TFs between homogeneous and heterogeneous tumors revealed significant statistical difference for size-zone variability (SZV), intensity variability (IV), zone percentage (ZP), proposed parameters II and III, entropy and tumor volume (p < 0.001), dissimilarity, high intensity emphasis (HIE), and SUV (p < 0.01). Lower statistical differences were observed for proposed parameter I (p = 0.02), and no significant differences were observed for SUV and SUV . Furthermore, the Spearman rank analysis between visual tumor labeling and TF analysis depicted a significant correlation for SZV, IV, entropy, parameters II and III, and tumor volume (0.68 ≤ ρ ≤ 0.84) and moderate correlation for ZP, HIE, homogeneity, dissimilarity, parameter I, and SUV (0.22 ≤ ρ ≤ 0.52), while no correlations were observed for SUV and SUV (ρ < 0.08). The multi-linear regression model for automated tumor labeling process resulted in R and RMSE values of 0.93 and 0.14, respectively (p < 0.001), and generated tumor labeling sensitivity and specificity of 0.93 and 0.89. With respect to baseline random walk segmentation, the results showed significant (p < 0.001) mean DSC, HD, and SUV error improvements of 21.4 ± 11.5 %, 1.4 ± 0.8 mm, and 16.8 ± 8.1 % in homogeneous tumors and 7.4 ± 4.4 %, 1.5 ± 0.6 mm, and 7.9 ± 2.7 % in heterogeneous lesions. In addition, significant (p < 0.001) mean DSC, HD, and SUV error improvements were observed for tumor sub-volume delineations, namely 5 ± 2 %, 1.5 ± 0.6 mm, and 7 ± 3 % for the proposed Fuzzy RW method compared to RW segmentation. CONCLUSION: We proposed and demonstrated an automatic framework for significantly improved segmentation and labeling of homogeneous vs. heterogeneous tumors in lung [ F]FDG-PET images.
[Mh] Termos MeSH primário: Algoritmos
Fluordesoxiglucose F18/química
Processamento de Imagem Assistida por Computador
Neoplasias Pulmonares/diagnóstico por imagem
Tomografia por Emissão de Pósitrons
[Mh] Termos MeSH secundário: Automação
Lógica Fuzzy
Seres Humanos
Modelos Lineares
Neoplasias Pulmonares/patologia
Coloração e Rotulagem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0Z5B2CJX4D (Fluorodeoxyglucose F18)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180308
[Lr] Data última revisão:
180308
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161023
[St] Status:MEDLINE
[do] DOI:10.1007/s11307-016-1015-0


  5 / 62903 MEDLINE  
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[PMID]:28470515
[Au] Autor:Chan LL; McCulley KJ; Kessel SL
[Ad] Endereço:Department of Technology R&D, Nexcelom Bioscience LLC, 360 Merrimack Street, Building 9, Lawrence, MA, 01843, USA. lchan@nexcelom.com.
[Ti] Título:Assessment of Cell Viability with Single-, Dual-, and Multi-Staining Methods Using Image Cytometry.
[So] Source:Methods Mol Biol;1601:27-41, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The ability to accurately measure cell viability is important for any cell-based assay. Traditionally, viability measurements have been performed using the trypan blue exclusion method on a hemacytometer, which allows researchers to visually distinguish viable from nonviable cells. While the trypan blue method can work for cell lines or primary cells that have been rigorously purified, in more complex samples such as PBMCs, bone marrow, whole blood, or any sample with low viability, this method can lead to errors. In recent years, advances in optics and fluorescent dyes have led to the development of automated benchtop image-based cell counters for rapid cell concentration and viability measurement. In this work, we demonstrate the use of image-based cytometry for cell viability detection using single-, dual-, or multi-stain techniques. Single-staining methods using nucleic acid stains such as EB, PI, 7-AAD, DAPI, SYTOX Green, and SYTOX Red, and enzymatic stains such as CFDA and Calcein AM, were performed. Dual-staining methods using AO/PI, CFDA/PI, Calcein AM/PI, Hoechst/PI, Hoechst/DRAQ7, and DRAQ5/DAPI that enumerate viable and nonviable cells were also performed. Finally, Hoechst/Calcein AM/PI was used for a multi-staining method. Fluorescent viability staining allows exclusion of cellular debris and nonnucleated cells from analysis, which can eliminate the need to perform purification steps during sample preparation and improve efficiency. Image cytometers increase speed and throughput, capture images for visual confirmation of results, and can greatly simplify cell count and viability measurements.
[Mh] Termos MeSH primário: Sobrevivência Celular
Citometria por Imagem/métodos
Coloração e Rotulagem/métodos
[Mh] Termos MeSH secundário: Contagem de Células
Corantes Fluorescentes/química
Células HeLa
Seres Humanos
Células Jurkat
Células MCF-7
Ácidos Nucleicos/química
Fatores de Tempo
Azul Tripano/química
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Fluorescent Dyes); 0 (Nucleic Acids); I2ZWO3LS3M (Trypan Blue)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180219
[Lr] Data última revisão:
180219
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170505
[St] Status:MEDLINE
[do] DOI:10.1007/978-1-4939-6960-9_3


  6 / 62903 MEDLINE  
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[PMID]:29300923
[Au] Autor:Zempo B; Karigo T; Kanda S; Akazome Y; Oka Y
[Ad] Endereço:Department of Biological Sciences, Graduate School of Science, The University of Tokyo, Tokyo, Japan.
[Ti] Título:Morphological Analysis of the Axonal Projections of EGFP-Labeled Esr1-Expressing Neurons in Transgenic Female Medaka.
[So] Source:Endocrinology;159(2):1228-1241, 2018 02 01.
[Is] ISSN:1945-7170
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Some hypothalamic neurons expressing estrogen receptor α (Esr1) are thought to transmit a gonadal estrogen feedback signal to gonadotropin-releasing hormone 1 (GnRH1) neurons, which is the final common pathway for feedback regulation of reproductive functions. Moreover, estrogen-sensitive neurons are suggested to control sexual behaviors in coordination with reproduction. In mammals, hypothalamic estrogen-sensitive neurons release the peptide kisspeptin and regulate GnRH1 neurons. However, a growing body of evidence in nonmammalian species casts doubt on the regulation of GnRH1 neurons by kisspeptin neurons. As a step toward understanding how estrogen regulates neuronal circuits for reproduction and sex behavior in vertebrates in general, we generated a transgenic (Tg) medaka that expresses enhanced green fluorescent protein (EGFP) specifically in esr1-expressing neurons (esr1 neurons) and analyzed their axonal projections. We found that esr1 neurons in the preoptic area (POA) project to the gnrh1 neurons. We also demonstrated by transcriptome and histological analyses that these esr1 neurons are glutamatergic or γ-aminobutyric acidergic (GABAergic) but not kisspeptinergic. We therefore suggest that glutamatergic and GABAergic esr1 neurons in the POA regulate gnrh1 neurons. This hypothesis is consistent with previous studies in mice that found that glutamatergic and GABAergic transmission is critical for estrogen-dependent changes in GnRH1 neuron firing. Thus, we propose that this neuronal circuit may provide an evolutionarily conserved mechanism for regulation of reproduction. In addition, we showed that telencephalic esr1 neurons project to medulla, which may control sexual behavior. Moreover, we found that some POA-esr1 neurons coexpress progesterone receptors. These neurons may form the neuronal circuits that regulate reproduction and sex behavior in response to the serum estrogen/progesterone.
[Mh] Termos MeSH primário: Axônios/fisiologia
Receptor alfa de Estrogênio/genética
Proteínas de Fluorescência Verde/genética
Neurônios/metabolismo
Oryzias
[Mh] Termos MeSH secundário: Animais
Animais Geneticamente Modificados
Receptor alfa de Estrogênio/metabolismo
Feminino
Proteínas de Fluorescência Verde/metabolismo
Rede Nervosa/metabolismo
Oryzias/genética
Oryzias/metabolismo
Área Pré-Óptica/metabolismo
Progesterona/metabolismo
Receptores de Progesterona/genética
Receptores de Progesterona/metabolismo
Coloração e Rotulagem
Telencéfalo/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Nm] Nome de substância:
0 (Estrogen Receptor alpha); 0 (Receptors, Progesterone); 0 (enhanced green fluorescent protein); 147336-22-9 (Green Fluorescent Proteins); 4G7DS2Q64Y (Progesterone)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180215
[Lr] Data última revisão:
180215
[Sb] Subgrupo de revista:AIM; IM
[Da] Data de entrada para processamento:180105
[St] Status:MEDLINE
[do] DOI:10.1210/en.2017-00873


  7 / 62903 MEDLINE  
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[PMID]:25815882
[Au] Autor:Chen H; Ma S; Wu X; Yang W; Zhao T
[Ti] Título:Diagnose human colonic tissues by terahertz near-field imaging.
[So] Source:J Biomed Opt;20(3):036017, 2015 Mar.
[Is] ISSN:1560-2281
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Based on a terahertz (THz) pipe-based near-field imaging system, we demonstrate the capability of THz imaging to diagnose freshly surgically excised human colonic tissues. Through THz near-field scanning the absorbance of the colonic tissues, the acquired images can clearly distinguish cancerous tissues from healthy tissues fast and automatically without pathological hematoxylin and eosin stain diagnosis. A statistical study on 58 specimens (20 healthy tissues and 38 tissues with tumor) from 31 patients (mean age: 59 years; range: 46 to 79 years) shows that the corresponding diagnostic sensitivity and specificity on colonic tissues are both 100%. Due to its capability to perform quantitative analysis, our study indicates the potential of the THz pipe-based near-field imaging for future automation on human tumor pathological examinations.
[Mh] Termos MeSH primário: Colo/diagnóstico por imagem
Neoplasias do Colo/diagnóstico por imagem
Imagem Terahertz/métodos
[Mh] Termos MeSH secundário: Idoso
Diagnóstico Diferencial
Seres Humanos
Meia-Idade
Sensibilidade e Especificidade
Manejo de Espécimes/métodos
Coloração e Rotulagem
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180214
[Lr] Data última revisão:
180214
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:150328
[St] Status:MEDLINE
[do] DOI:10.1117/1.JBO.20.3.036017


  8 / 62903 MEDLINE  
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[PMID]:28456964
[Au] Autor:Li Y; O'Neill C
[Ad] Endereço:Human Reproduction Unit, Northern Clinical School, Sydney Medical School, University of Sydney, Sydney, NSW, 2065, Australia.
[Ti] Título:Immunological Staining of Global Changes in DNA Methylation in the Early Mammalian Embryo.
[So] Source:Methods Mol Biol;1605:161-169, 2017.
[Is] ISSN:1940-6029
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The structural complexity of chromatin can make antibody access to some nuclear antigens energetically unfavorable. This can limit the usefulness and reliability of immunostaining unless validated methods of epitope retrieval are applied. We found that denaturation of chromatin by sequential use of acidification and tryptic digestion of fixed cells is required to reliably detect DNA methylation in the embryo. Using this method to unmask the epitope revealed an unexpected pattern of reprogramming of global patterns of DNA methylation in the preimplantation embryo. This paper provides a detailed description of the procedures required for immunological detection of 5-methylcytosine in the early embryo.
[Mh] Termos MeSH primário: Blastocisto/química
Metilação de DNA
[Mh] Termos MeSH secundário: 5-Metilcitosina
Animais
Cromatina/química
Epigênese Genética
Feminino
Camundongos
Coloração e Rotulagem
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Chromatin); 6R795CQT4H (5-Methylcytosine)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180212
[Lr] Data última revisão:
180212
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170501
[St] Status:MEDLINE
[do] DOI:10.1007/978-1-4939-6988-3_11


  9 / 62903 MEDLINE  
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[PMID]:28625736
[Au] Autor:Ducharme J; Auclair K
[Ad] Endereço:Department of Chemistry, McGill University, 801 Sherbrooke Street West, Montreal, Quebec H3A 0B8, Canada.
[Ti] Título:Use of bioconjugation with cytochrome P450 enzymes.
[So] Source:Biochim Biophys Acta;1866(1):32-51, 2018 01.
[Is] ISSN:0006-3002
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Bioconjugation, defined as chemical modification of biomolecules, is widely employed in biological and biophysical studies. It can expand functional diversity and enable applications ranging from biocatalysis, biosensing and even therapy. This review summarizes how chemical modifications of cytochrome P450 enzymes (P450s or CYPs) have contributed to improving our understanding of these enzymes. Genetic modifications of P450s have also proven very useful but are not covered in this review. Bioconjugation has served to gain structural information and investigate the mechanism of P450s via photoaffinity labeling, mechanism-based inhibition (MBI) and fluorescence studies. P450 surface acetylation and protein cross-linking have contributed to the investigation of protein complexes formation involving P450 and its redox partner or other P450 enzymes. Finally, covalent immobilization on polymer surfaces or electrodes has benefited the areas of biocatalysis and biosensor design. This article is part of a Special Issue entitled: Cytochrome P450 biodiversity and biotechnology, edited by Erika Plettner, Gianfranco Gilardi, Luet Wong, Vlada Urlacher, Jared Goldstone.
[Mh] Termos MeSH primário: Reagentes para Ligações Cruzadas/química
Sistema Enzimático do Citocromo P-450/química
Proteínas Imobilizadas/química
Marcadores de Fotoafinidade/química
Coloração e Rotulagem/métodos
[Mh] Termos MeSH secundário: Acetilação
Regulação Alostérica
Biocatálise
Técnicas Biossensoriais
Domínio Catalítico
Sistema Enzimático do Citocromo P-450/metabolismo
Corantes Fluorescentes/química
Seres Humanos
Proteínas Imobilizadas/metabolismo
Isoenzimas/química
Isoenzimas/metabolismo
Modelos Moleculares
Oxirredução
Estrutura Secundária de Proteína
[Pt] Tipo de publicação:JOURNAL ARTICLE; RESEARCH SUPPORT, NON-U.S. GOV'T; REVIEW
[Nm] Nome de substância:
0 (Cross-Linking Reagents); 0 (Fluorescent Dyes); 0 (Immobilized Proteins); 0 (Isoenzymes); 0 (Photoaffinity Labels); 9035-51-2 (Cytochrome P-450 Enzyme System)
[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:170620
[St] Status:MEDLINE


  10 / 62903 MEDLINE  
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[PMID]:29202622
[Au] Autor:Laenkholm AV; Grabau D; Møller Talman ML; Balslev E; Bak Jylling AM; Tabor TP; Johansen M; Brügmann A; Lelkaitis G; Di Caterino T; Mygind H; Poulsen T; Mertz H; Søndergaard G; Bruun Rasmussen B
[Ad] Endereço:a Department of Surgical Pathology , Zealand University Hospital , Slagelse , Denmark.
[Ti] Título:An inter-observer Ki67 reproducibility study applying two different assessment methods: on behalf of the Danish Scientific Committee of Pathology, Danish breast cancer cooperative group (DBCG).
[So] Source:Acta Oncol;57(1):83-89, 2018 Jan.
[Is] ISSN:1651-226X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:INTRODUCTION: In 2011, the St. Gallen Consensus Conference introduced the use of pathology to define the intrinsic breast cancer subtypes by application of immunohistochemical (IHC) surrogate markers ER, PR, HER2 and Ki67 with a specified Ki67 cutoff (>14%) for luminal B-like definition. Reports concerning impaired reproducibility of Ki67 estimation and threshold inconsistency led to the initiation of this quality assurance study (2013-2015). The aim of the study was to investigate inter-observer variation for Ki67 estimation in malignant breast tumors by two different quantification methods (assessment method and count method) including measure of agreement between methods. MATERIAL AND METHODS: Fourteen experienced breast pathologists from 12 pathology departments evaluated 118 slides from a consecutive series of malignant breast tumors. The staining interpretation was performed according to both the Danish and Swedish guidelines. Reproducibility was quantified by intra-class correlation coefficient (ICC) and Lights Kappa with dichotomization of observations at the larger than (>) 20% threshold. The agreement between observations by the two quantification methods was evaluated by Bland-Altman plot. RESULTS: For the fourteen raters the median ranged from 20% to 40% by the assessment method and from 22.5% to 36.5% by the count method. Light's Kappa was 0.664 for observation by the assessment method and 0.649 by the count method. The ICC was 0.82 (95% CI: 0.77-0.86) by the assessment method vs. 0.84 (95% CI: 0.80-0.87) by the count method. CONCLUSION: Although the study in general showed a moderate to good inter-observer agreement according to both ICC and Lights Kappa, still major discrepancies were identified in especially the mid-range of observations. Consequently, for now Ki67 estimation is not implemented in the DBCG treatment algorithm.
[Mh] Termos MeSH primário: Neoplasias da Mama/patologia
Imuno-Histoquímica/normas
Antígeno Ki-67/metabolismo
[Mh] Termos MeSH secundário: Biomarcadores/metabolismo
Conferências de Consenso como Assunto
Dinamarca
Feminino
Seres Humanos
Patologia Clínica/normas
Guias de Prática Clínica como Assunto
Reprodutibilidade dos Testes
Coloração e Rotulagem/métodos
Coloração e Rotulagem/normas
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Biomarkers); 0 (Ki-67 Antigen)
[Em] Mês de entrada:1802
[Cu] Atualização por classe:180201
[Lr] Data última revisão:
180201
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171206
[St] Status:MEDLINE
[do] DOI:10.1080/0284186X.2017.1404127



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