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  1 / 1622 MEDLINE  
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[PMID]:29034805
[Au] Autor:Vural K; Kosova F; Kurt FÖ; Tuglu I
[Ad] Endereço:1 Department of Medical Pharmacology, Faculty of Medicine, Celal Bayar University, Manisa, Turkey.
[Ti] Título:In vitro investigation of the effect of matrix molecules on the behavior of colon cancer cells under the effect of geldanamycin derivative.
[So] Source:Tumour Biol;39(10):1010428317720569, 2017 Oct.
[Is] ISSN:1423-0380
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:The chaperone-binding drug, 17-allylamino-17-demethoxygeldanamycin, has recently come into clinical use. It is a derivative of geldanamycin, an ansamycin benzoquinone antibiotic with anti-carcinogenic effect. Understanding the effect of this drug on the cancer cells and their niche is important for treatment. We applied 17-allylamino-17-demethoxygeldanamycin to colon cancer cell line (Colo 205) on matrix molecules to investigate the relationship of apoptosis with terminal deoxynucleotidyl transferase dUTP nick end labeling immunocytochemistry and related gene expression. We used laminin and collagen I for matrix molecules and vascular endothelial growth factor for angiogenic structure. We also examined apoptosis-related signaling pathway including mitochondrial proteins, cytochrome c, Bcl-2, caspase-9, Apaf-1 expression using real-time polymerase chain reaction. There was clear effect of 17-allylamino-17-demethoxygeldanamycin that killed more cells on tissue culture plastic compared to matrix molecules. The IC value was 0.58 µg/mL for tissue culture plastic compared with 0.64 µg/mL for laminin and 0.75 µg/mL for collagen I. The analyses showed that more cells on matrix molecules underwent apoptosis compared to that on tissue culture plastic. Apoptosis-related gene expression was similar in which Bcl-2 expression decreased and proapoptotic gene expression of the cells on matrix molecules increased compared to that on tissue culture plastic. However, the application of 17-allylamino-17-demethoxygeldanamycin was more effective for the cells on collagen I compared to the cells on laminin. There was also a decrease in angiogenesis as shown by the vascular endothelial growth factor staining. This was more pronounced by coating of the tissue culture plastic with matrix molecules. Our results supported the anti-cancer effect of 17-allylamino-17-demethoxygeldanamycin, and this effect depended on matrix molecules. This effect occurs through apoptosis, and related genes were also altered. All these genes may serve for novel target under the effect of matrix substrate. However, correct interpretation of the results requires further studies.
[Mh] Termos MeSH primário: Anticarcinógenos/farmacologia
Benzoquinonas/farmacologia
Neoplasias do Colo/tratamento farmacológico
Neoplasias do Colo/metabolismo
Lactamas Macrocíclicas/farmacologia
[Mh] Termos MeSH secundário: Apoptose/efeitos dos fármacos
Fator Apoptótico 1 Ativador de Proteases/metabolismo
Caspase 9/metabolismo
Linhagem Celular Tumoral
Colágeno/metabolismo
Colo/efeitos dos fármacos
Colo/metabolismo
DNA Nucleotidilexotransferase/metabolismo
Seres Humanos
Proteínas Proto-Oncogênicas c-bcl-2/metabolismo
Rifabutina/farmacologia
Transdução de Sinais/efeitos dos fármacos
Fator A de Crescimento do Endotélio Vascular/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Anticarcinogenic Agents); 0 (Apoptotic Protease-Activating Factor 1); 0 (Benzoquinones); 0 (Lactams, Macrocyclic); 0 (Proto-Oncogene Proteins c-bcl-2); 0 (Vascular Endothelial Growth Factor A); 1W306TDA6S (Rifabutin); 3T006GV98U (quinone); 4GY0AVT3L4 (tanespimycin); 9007-34-5 (Collagen); EC 2.7.7.31 (DNA Nucleotidylexotransferase); EC 3.4.22.- (Caspase 9); Z3K3VJ16KU (geldanamycin)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171020
[Lr] Data última revisão:
171020
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:171017
[St] Status:MEDLINE
[do] DOI:10.1177/1010428317720569


  2 / 1622 MEDLINE  
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[PMID]:28698239
[Au] Autor:Gaspar I; Wippich F; Ephrussi A
[Ad] Endereço:Developmental Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg 69117, Germany.
[Ti] Título:Enzymatic production of single-molecule FISH and RNA capture probes.
[So] Source:RNA;23(10):1582-1591, 2017 Oct.
[Is] ISSN:1469-9001
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Arrays of singly labeled short oligonucleotides that hybridize to a specific target revolutionized RNA biology, enabling quantitative, single-molecule microscopy analysis and high-efficiency RNA/RNP capture. Here, we describe a simple and efficient method that allows flexible functionalization of inexpensive DNA oligonucleotides by different fluorescent dyes or biotin using terminal deoxynucleotidyl transferase and custom-made functional group conjugated dideoxy-UTP. We show that (i) all steps of the oligonucleotide labeling-including conjugation, enzymatic synthesis, and product purification-can be performed in a standard biology laboratory, (ii) the process yields >90%, often >95% labeled product with minimal carryover of impurities, and (iii) the oligonucleotides can be labeled with different dyes or biotin, allowing single-molecule FISH, RNA affinity purification, and Northern blot analysis to be performed.
[Mh] Termos MeSH primário: DNA Nucleotidilexotransferase/metabolismo
Hibridização in Situ Fluorescente/métodos
Sondas RNA/química
[Mh] Termos MeSH secundário: Animais
Biotina
Didesoxinucleotídeos/química
Didesoxinucleotídeos/metabolismo
Drosophila melanogaster/genética
Feminino
Corantes Fluorescentes/química
Sondas de Oligonucleotídeos/química
Oligonucleotídeos/química
Ovário/fisiologia
Sondas RNA/metabolismo
Nucleotídeos de Uracila/química
Nucleotídeos de Uracila/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Dideoxynucleotides); 0 (Fluorescent Dyes); 0 (Oligonucleotide Probes); 0 (Oligonucleotides); 0 (RNA Probes); 0 (Uracil Nucleotides); 6SO6U10H04 (Biotin); 84445-38-5 (2',3'-dideoxyuridine-5'-triphosphate); EC 2.7.7.31 (DNA Nucleotidylexotransferase)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171019
[Lr] Data última revisão:
171019
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170713
[St] Status:MEDLINE
[do] DOI:10.1261/rna.061184.117


  3 / 1622 MEDLINE  
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[PMID]:28521514
[Au] Autor:Lai CC; Huang PH; Yang AH; Chiang SC; Tang CY; Tseng KW; Huang CH
[Ad] Endereço:* Division of Cardiovascular Surgery, Department of Surgery, Taipei Veterans General Hospital, National Yang-Ming University School of Medicine, Taipei, Taiwan.
[Ti] Título:Baicalein Attenuates Lung Injury Induced by Myocardial Ischemia and Reperfusion.
[So] Source:Am J Chin Med;45(4):791-811, 2017.
[Is] ISSN:0192-415X
[Cp] País de publicação:Singapore
[La] Idioma:eng
[Ab] Resumo:Baicalein is an active component of Scutellaria baicalensis Georgi, which has traditionally been used to treat cardiovascular diseases in China. In this study, we investigated if treatment with baicalein can attenuate the lung injury induced by myocardial ischemia and reperfusion (I/R). Myocardial I/R, induced by a 40-min occlusion of the left anterior descending coronary artery and a 3-h reperfusion, significantly increased histological damage and the wet-to-dry weight ratio of lungs in rats. The terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL)-positive nuclei and caspase-3 activation was significantly increased in the lungs. Serum and bronchoalveolar lavage fluid levels of tumor necrosis factor-[Formula: see text] (TNF-[Formula: see text]), interleukin-1[Formula: see text] (IL-1[Formula: see text]), and interleukin-6 (IL-6) were significantly elevated, as were TNF-[Formula: see text] levels in the lung. Intravenous administration with baicalein at doses of 3, 10, and 30[Formula: see text]mg/kg for ten minutes before myocardial I/R significantly reduced histological damage, the wet-to-dry weight ratio, and apoptosis in the lung. Baicalein also significantly inhibited the increase in levels of TNF-[Formula: see text], IL-1[Formula: see text], and IL-6. Moreover, baicalein increased Bcl-2 and decreased p53, Bax, and cytochrome [Formula: see text] in lungs. Phosphorylation of the prosurvival kinases, including Akt and extracellular signal-regulated kinases 1 and 2 (ERK1/2), was increased, while the phosphorylation of the pro-apoptotic mitogen-activated protein kinases, including p38 and c-Jun N-terminal kinase (JNK), was decreased. In conclusion, treatment with baicalein attenuates the lung injury induced by myocardial I/R. The mechanisms might be related to the limiting of apoptosis, possibly via the inhibition of both the extrinsic and intrinsic pathways of apoptosis, including the inhibition of TNF-[Formula: see text] production and modulation of pro- and anti-apoptotic signaling elements.
[Mh] Termos MeSH primário: Apoptose/efeitos dos fármacos
Flavanonas/uso terapêutico
Pneumopatias/tratamento farmacológico
Pneumopatias/etiologia
Isquemia Miocárdica/complicações
Reperfusão Miocárdica/efeitos adversos
Fitoterapia
Scutellaria baicalensis/química
[Mh] Termos MeSH secundário: Animais
Apoptose/genética
Caspase 3/metabolismo
Citocinas/metabolismo
DNA Nucleotidilexotransferase/metabolismo
Nucleotídeos de Desoxiuracil/metabolismo
Flavanonas/administração & dosagem
Flavanonas/isolamento & purificação
Infusões Intravenosas
Pulmão/metabolismo
Pneumopatias/metabolismo
Pneumopatias/prevenção & controle
Masculino
Ratos Sprague-Dawley
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Cytokines); 0 (Deoxyuracil Nucleotides); 0 (Flavanones); 1173-82-6 (deoxyuridine triphosphate); 49QAH60606 (baicalein); EC 2.7.7.31 (DNA Nucleotidylexotransferase); EC 3.4.22.- (Caspase 3)
[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:170520
[St] Status:MEDLINE
[do] DOI:10.1142/S0192415X17500422


  4 / 1622 MEDLINE  
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[PMID]:28441732
[Au] Autor:Tauraite D; Jakubovska J; Dabuzinskaite J; Bratchikov M; Meskys R
[Ad] Endereço:Department of Molecular Microbiology and Biotechnology, Institute of Biochemistry, Life Sciences Center, Vilnius University, Sauletekio al. 7, Vilnius LT-10257, Lithuania. daiva.tauraite@bchi.vu.lt.
[Ti] Título:Modified Nucleotides as Substrates of Terminal Deoxynucleotidyl Transferase.
[So] Source:Molecules;22(4), 2017 Apr 22.
[Is] ISSN:1420-3049
[Cp] País de publicação:Switzerland
[La] Idioma:eng
[Ab] Resumo:The synthesis of novel modified nucleotides and their incorporation into DNA sequences opens many possibilities to change the chemical properties of oligonucleotides (ONs), and, therefore, broaden the field of practical applications of modified DNA. The chemical synthesis of nucleotide derivatives, including ones bearing thio-, hydrazino-, cyano- and carboxy groups as well as 2-pyridone nucleobase-containing nucleotides was carried out. The prepared compounds were tested as substrates of terminal deoxynucleotidyl transferase (TdT). The nucleotides containing 4-aminocytosine, 4-thiouracil as well as 2-pyridone, 4-chloro- and 4-bromo-2-pyridone as a nucleobase were accepted by TdT, thus allowing enzymatic synthesis of 3'-terminally modified ONs. The successful UV-induced cross-linking of 4-thiouracil-containing ONs to TdT was carried out. Enzymatic post-synthetic 3'-modification of ONs with various photo- and chemically-reactive groups opens novel possibilities for future applications, especially in analysis of the mechanisms of polymerases and the development of photo-labels, sensors, and self-assembling structures.
[Mh] Termos MeSH primário: Citosina/análogos & derivados
Citosina/química
DNA Nucleotidilexotransferase/química
Tiouracila/análogos & derivados
Tiouracila/química
[Mh] Termos MeSH secundário: Engenharia Genética
Mutagênese
Oligonucleotídeos/síntese química
Especificidade por Substrato
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Oligonucleotides); 59X161SCYL (Thiouracil); 8J337D1HZY (Cytosine); EC 2.7.7.31 (DNA Nucleotidylexotransferase)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170807
[Lr] Data última revisão:
170807
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170427
[St] Status:MEDLINE


  5 / 1622 MEDLINE  
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[PMID]:28369937
[Au] Autor:Huang Y; Cai J; Tang JF; Zhang HY; Wang ZW; Jian JC; Wu ZH; Lu YS
[Ad] Endereço:College of Fishery, Guangdong Ocean University, Zhanjiang, 524088, People's Republic of China.
[Ti] Título:Identification and expression analysis of terminal deoxynucleotidyl transferase in humphead snapper Lutjanus sanguineus.
[So] Source:J Fish Biol;90(5):2194-2199, 2017 May.
[Is] ISSN:1095-8649
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:A tdt gene was identified successfully from humphead snapper Lutjanus sanguineus, which contained 1710 bp encoding a protein of 463 amino acids. Results of quantitative real-time polymerase chain reaction (qRT-PCR) indicated that tdt mainly expressed in thymus and head kidney and the transcripts of tdt in these tissues were up-regulated significantly at 36 and 48 h after Vibrio harveyi infection. Meanwhile Tdt-producing cells were found in thymus and head kidney.
[Mh] Termos MeSH primário: DNA Nucleotidilexotransferase/metabolismo
Proteínas de Peixes/metabolismo
Regulação Enzimológica da Expressão Gênica/fisiologia
Perciformes/metabolismo
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Animais
DNA Nucleotidilexotransferase/genética
Proteínas de Peixes/genética
Perciformes/genética
Filogenia
Reação em Cadeia da Polimerase em Tempo Real
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Fish Proteins); EC 2.7.7.31 (DNA Nucleotidylexotransferase)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171010
[Lr] Data última revisão:
171010
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170404
[St] Status:MEDLINE
[do] DOI:10.1111/jfb.13259


  6 / 1622 MEDLINE  
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[PMID]:28248816
[Au] Autor:Dunlap JB; Cascio MJ; Stacey X; Click S; Troxell ML
[Ad] Endereço:*Department of Pathology, Oregon Health & Science University, Portland, OR †Department of Pathology, Stanford University Medical Center, Stanford, CA.
[Ti] Título:TdT-positive Infiltrate in Inflamed Pediatric Kidney: A Potential Diagnostic Pitfall.
[So] Source:Am J Surg Pathol;41(5):706-716, 2017 May.
[Is] ISSN:1532-0979
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:We encountered a patient with infantile nephrotic syndrome associated with a dense interstitial inflammatory infiltrate and prominent extramedullary hematopoiesis. Immunohistochemical analysis revealed numerous terminal deoxynucleotidyl transferase (TdT)-positive cells, which may raise concern for lymphoblastic lymphoma. Thus, we further characterized a group of pediatric kidneys with inflammation. TdT-positive nuclei were quantitated, and dual immunostains for TdT/CD79a, TdT/CD3, and TdT/CD43 were performed in a subset of cases; flow cytometry was performed in 1 case. TdT-positive nuclei were present in inflamed pediatric kidneys in 40 of 42 patients. TdT counts (average of 3 maximal high-power fields) ranged from 1 to >200, with a mean of 47. The presence and number of TdT-positive nuclei showed a strong association with younger patient age. Extramedullary hematopoiesis was identified in 11/42 patients, all under the age of 1. The presence of extramedullary hematopoiesis did not correlate with TdT count (P=0.158). Dual immunostaining and flow cytometric analysis in 1 case showed weak expression of B-cell markers and favored normal precursor B cells. Although TdT is a common marker of lymphoblastic lymphoma, we have demonstrated that TdT-positive cells may be part of the inflammatory milieu in infant kidneys. Together with cytologic, architectural, and clinical features, these data can help to avoid misinterpretation of involvement by lymphoblastic lymphoma/leukemia.
[Mh] Termos MeSH primário: DNA Nucleotidilexotransferase/análise
Rim/química
Nefrite/metabolismo
Síndrome Nefrótica/metabolismo
[Mh] Termos MeSH secundário: Adolescente
Biomarcadores/análise
Biópsia
Complexo CD3/análise
Antígenos CD79/análise
Criança
Pré-Escolar
Diagnóstico Diferencial
Citometria de Fluxo
Hematopoese Extramedular
Seres Humanos
Imuno-Histoquímica
Lactente
Recém-Nascido
Rim/patologia
Rim/cirurgia
Leucossialina/análise
Masculino
Nefrectomia
Nefrite/diagnóstico
Nefrite/cirurgia
Síndrome Nefrótica/diagnóstico
Síndrome Nefrótica/cirurgia
Leucemia-Linfoma Linfoblástico de Células Precursoras/diagnóstico
Leucemia-Linfoma Linfoblástico de Células Precursoras/metabolismo
Valor Preditivo dos Testes
Adulto Jovem
[Pt] Tipo de publicação:CASE REPORTS; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Biomarkers); 0 (CD3 Complex); 0 (CD79 Antigens); 0 (CD79A protein, human); 0 (Leukosialin); 0 (UN1 sialoglycoprotein, human); EC 2.7.7.31 (DNA Nucleotidylexotransferase)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:171116
[Lr] Data última revisão:
171116
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170302
[St] Status:MEDLINE
[do] DOI:10.1097/PAS.0000000000000828


  7 / 1622 MEDLINE  
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[PMID]:28051018
[Au] Autor:Zhang KD; Tong LR; Wang SM; Peng RY; Huang HD; Dong YC; Zhang XX; Li Q; Bai C
[Ad] Endereço:Department of Respiratory and Critical Care Medicine, Changhai Hospital, The Second Military Medical University, Shanghai 200433; Department of Respiratory Medicine, Yancheng First People's Hospital, Yancheng, Jiangsu 224000, China.
[Ti] Título:Apoptosis of Lewis Lung Carcinoma Cells Induced by Microwave via p53 and Proapoptotic Proteins .
[So] Source:Chin Med J (Engl);130(1):15-22, 2017 5th Jan 2017.
[Is] ISSN:0366-6999
[Cp] País de publicação:China
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Microwave therapy is a minimal invasive procedure and has been employed in clinical practice for the treatment of various types of cancers. However, its therapeutic application in non-small-cell lung cancer and the underlying mechanism remains to be investigated. This study aimed to investigate its effect on Lewis lung carcinoma (LLC) tumor in vivo. METHODS: Fifty LLC tumor-bearing C57BL/6 mice were adopted to assess the effect of microwave radiation on the growth and apoptosis of LLC tumor in vivo. These mice were randomly assigned to 10 groups with 5 mice in each group. Five groups were treated by single pulse microwave at different doses for different time, and the other five groups were radiated by multiple-pulse treatment of a single dose. Apoptosis of cancer cells was determined by terminal deoxynucleotidyl transferase dUTP nick-end labeling assay. Western blotting was applied to detect the expression of proteins. RESULTS: Single pulse of microwave radiation for 5 min had little effect on the mice. Only 15-min microwave radiation at 30 mW/cm2 significantly increased the mice body temperature (2.20 ± 0.82)°C as compared with the other groups (0.78 ± 0.29 °C, 1.24 ± 0.52 °C, 0.78 ± 0.42 °C, respectively), but it did not affect the apoptosis of LLC tumor cells significantly. Continous microwave radiation exposure, single dose microwave radiation once per day for up to seven days, inhibited cell division and induced apoptosis of LLC tumor cells in a dose- and duration-dependent manner. It upregulated the protein levels of p53, Caspase 3, Bax and downregulated Bcl-2 protein. CONCLUSIONS: Multiple exposures of LLC-bearing mice to microwave radiation effectively induced tumor cell apoptosis at least partly by upregulating proapoptotic proteins and downregulating antiapoptotic proteins. Continuous radiation at low microwave intensity for a short time per day is promising in treating non-small-cell lung cancer.
[Mh] Termos MeSH primário: Carcinoma Pulmonar de Lewis/terapia
Micro-Ondas
[Mh] Termos MeSH secundário: Animais
Apoptose/efeitos da radiação
Proteínas Reguladoras de Apoptose/metabolismo
Temperatura Corporal/efeitos da radiação
Carcinoma Pulmonar de Lewis/metabolismo
Carcinoma Pulmonar de Lewis/patologia
Caspase 3/metabolismo
Divisão Celular/efeitos da radiação
Linhagem Celular Tumoral
DNA Nucleotidilexotransferase/metabolismo
Imuno-Histoquímica
Masculino
Camundongos
Camundongos Endogâmicos C57BL
Proteína Supressora de Tumor p53/metabolismo
Proteína X Associada a bcl-2/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Apoptosis Regulatory Proteins); 0 (Tumor Suppressor Protein p53); 0 (bcl-2-Associated X Protein); EC 2.7.7.31 (DNA Nucleotidylexotransferase); EC 3.4.22.- (Caspase 3)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170313
[Lr] Data última revisão:
170313
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170105
[St] Status:MEDLINE
[do] DOI:10.4103/0366-6999.196587


  8 / 1622 MEDLINE  
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[PMID]:27668724
[Au] Autor:Liu Y; Xiong E; Li X; Li J; Zhang X; Chen J
[Ad] Endereço:State Key Laboratory of Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082, PR China.
[Ti] Título:Sensitive electrochemical assay of alkaline phosphatase activity based on TdT-mediated hemin/G-quadruplex DNAzyme nanowires for signal amplification.
[So] Source:Biosens Bioelectron;87:970-975, 2017 Jan 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Taking TdT-mediated hemin/G-quadruplex DNAzyme nanowires as NADH oxidase and HRP-mimicking DNAzyme, a novel DNA-based electrochemical method has been developed for sensitive and selective assay of alkaline phosphatase (AP) activity. The double-stranded DNA (dsDNA) probe consisted of thiol-functionalized DNA1 and 3'-phosphorylated DNA2, was immobilized on a gold nanoparticles (AuNPs) modified glassy carbon (GC) electrode. In the presence of AP, 3'-phosphoryl end of DNA2 was dephosphorylated. Terminal deoxynucletidyl transferase (TdT) catalyzed the sequential addition of deoxynucleotides (dTTPs) at 3'-OH end of DNA2 to extend DNA2 with a poly-T sequence. Then, G-rich DNA3 strand hybridized with the poly-T sequence of DNA2. Upon addition of hemin, the hemin/G-quadruplex DNAzyme was formed. In the presence of NADH, the hemin/G-quadruplex DNAzyme oxidased NADH to NAD , accompanied by the formation of H O which was further catalyzed by hemin/G-quadruplex DNAzyme (served as a HRP-mimicking DNAzyme) with the thionine (Thi) as electron transfer mediator, leading to the amplified electrochemical signal. Under optimized conditions, the response peak current was linear with the concentration of AP in the range from 0.1UL to 5UL with the detection limit of 0.03UL . Also, the developed biosensor possessed good selectivity, reproducibility and stability, and simple sensing structure, showing promising practical applications in AP activity assay.
[Mh] Termos MeSH primário: Fosfatase Alcalina/análise
DNA Catalítico/química
Técnicas Eletroquímicas/métodos
Quadruplex G
Hemina/química
Nanofios/química
[Mh] Termos MeSH secundário: Fosfatase Alcalina/sangue
Animais
Artemia/enzimologia
DNA Nucleotidilexotransferase/química
Ensaios Enzimáticos/métodos
Seres Humanos
Limite de Detecção
Nanofios/ultraestrutura
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Catalytic); 743LRP9S7N (Hemin); EC 2.7.7.31 (DNA Nucleotidylexotransferase); EC 3.1.3.1 (Alkaline Phosphatase)
[Em] Mês de entrada:1701
[Cu] Atualização por classe:170816
[Lr] Data última revisão:
170816
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160927
[St] Status:MEDLINE


  9 / 1622 MEDLINE  
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[PMID]:27592241
[Au] Autor:Shi K; Dou B; Yang J; Yuan R; Xiang Y
[Ad] Endereço:Key Laboratory of Luminescent and Real-Time Analytical Chemistry, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715, PR China.
[Ti] Título:Target-triggered catalytic hairpin assembly and TdT-catalyzed DNA polymerization for amplified electronic detection of thrombin in human serums.
[So] Source:Biosens Bioelectron;87:495-500, 2017 Jan 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Specific and sensitive detection of protein biomarkers is of great importance in biomedical and bioanalytical applications. In this work, a dual amplified signal enhancement approach based on the integration of catalytic hairpin assembly (CHA) and terminal deoxynucleotidyl transferase (TdT)-mediated in situ DNA polymerization has been developed for highly sensitive and label-free electrochemical detection of thrombin in human serums. The presence of the target thrombin leads to the unfolding and capture of a significant number of hairpin signal probes with free 3'-OH termini on the sensor electrode. Subsequently, TdT can catalyze the elongation of the signal probes and formation of many G-quadruplex sequence replicates with the presence of dGTP and dATP at a molar ratio of 6:4. These G-quadruplex sequences bind hemin and generate drastically amplified current response for sensitive detection of thrombin in a completely label-free fashion. The sensor shows a linear range of 0.5pM-10.0nM and a detection limit of 0.12pM for thrombin. Moreover, the developed sensor can selectively discriminate the target thrombin against other non-target proteins and can be employed to monitor thrombin in human serum samples.
[Mh] Termos MeSH primário: Aptâmeros de Nucleotídeos/metabolismo
Técnicas Biossensoriais/métodos
DNA Nucleotidilexotransferase/metabolismo
Quadruplex G
Trombina/análise
[Mh] Termos MeSH secundário: Aptâmeros de Nucleotídeos/química
Técnicas Eletroquímicas/métodos
Hemina/química
Hemina/metabolismo
Seres Humanos
Limite de Detecção
Polimerização
Trombina/metabolismo
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Aptamers, Nucleotide); 145563-68-4 (thrombin aptamer); 743LRP9S7N (Hemin); EC 2.7.7.31 (DNA Nucleotidylexotransferase); EC 3.4.21.5 (Thrombin)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:170816
[Lr] Data última revisão:
170816
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160905
[St] Status:MEDLINE


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[PMID]:27581387
[Au] Autor:Banovic F; Dunston S; Linder KE; Rakich P; Olivry T
[Ad] Endereço:1 Department of Small Animal Medicine & Surgery, The University of Georgia, College of Veterinary Medicine, Athens, GA, USA.
[Ti] Título:Apoptosis as a Mechanism for Keratinocyte Death in Canine Toxic Epidermal Necrolysis.
[So] Source:Vet Pathol;54(2):249-253, 2017 Mar.
[Is] ISSN:1544-2217
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:In humans and dogs, toxic epidermal necrolysis (TEN) is a life-threatening dermatosis characterized by sudden epidermal death resulting in extensive skin detachment. There is little information on the pathogenesis of keratinocyte cell death in canine TEN. We studied the occurrence of apoptosis in skin lesions of dogs with TEN to determine if apoptosis contributes to the pathogenesis of this disease. Immunostaining with antibodies to activated caspase-3 and the terminal deoxynucleotidyl-transferase (TdT)-mediated deoxyuridine triphosphate (dUTP) nick-end labeling technique revealed positive apoptotic keratinocytes in basal and suprabasal epidermal compartments in 17 biopsy specimens collected from 3 dogs with TEN and 16 from 3 dogs with erythema multiforme (EM). There was no significant difference in the number of positively stained epidermal cells between TEN and EM. These results suggest that apoptosis of epidermal keratinocytes and lymphocytic satellitosis represent one of the early steps in the pathogenesis of canine TEN, as in the human disease counterpart.
[Mh] Termos MeSH primário: Apoproteínas/fisiologia
Doenças do Cão/patologia
Queratinócitos/patologia
Síndrome de Stevens-Johnson/veterinária
[Mh] Termos MeSH secundário: Animais
Caspase 3/genética
Caspase 3/metabolismo
DNA Nucleotidilexotransferase/metabolismo
Nucleotídeos de Desoxiuracil/metabolismo
Cães
Regulação Enzimológica da Expressão Gênica
Marcação In Situ das Extremidades Cortadas
Síndrome de Stevens-Johnson/patologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Apoproteins); 0 (Deoxyuracil Nucleotides); 1173-82-6 (deoxyuridine triphosphate); EC 2.7.7.31 (DNA Nucleotidylexotransferase); EC 3.4.22.- (Caspase 3)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171030
[Lr] Data última revisão:
171030
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160902
[St] Status:MEDLINE
[do] DOI:10.1177/0300985816666609



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