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Pesquisa : D13.444.600.723 [Categoria DeCS]
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  1 / 3798 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


  2 / 3798 MEDLINE  
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[PMID]:28302536
[Au] Autor:Mendez-Pena JE; Sadow PM; Nose V; Hoang MP
[Ad] Endereço:Massachusetts General Hospital, Boston, MA 02114, USA.
[Ti] Título:RNA chromogenic in situ hybridization assay with clinical automated platform is a sensitive method in detecting high-risk human papillomavirus in squamous cell carcinoma.
[So] Source:Hum Pathol;63:184-189, 2017 May.
[Is] ISSN:1532-8392
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Detection of active human papillomavirus (HPV) is clinically important because its presence has been shown to correlate with favorable clinical outcomes and better response to treatment in oropharyngeal squamous cell carcinomas. Using a clinical automated platform, we compared the performance of commercially available HPV DNA and RNA in situ hybridization (ISH) probes in archival tissues of 57 squamous cell carcinomas. Importantly, a clinical automated platform gives (1) consistent and reproducible results for HPV ISH and (2) better standardization across clinical laboratories. Compared with polymerase chain reaction results, RNA ISH exhibited 93% concordance versus 81% of DNA ISH. RNA ISH was more sensitive than DNA ISH (100% versus 88%) and more specific (87% versus 74%). When only accounting for 2+-3+ positivity, sensitivity was 92% for RNA ISH versus 73% for DNA ISH, highlighting the ease of interpretation. p16 exhibited 96% sensitivity, whereas specificity was only 55%. In 3 cases, both RNA and DNA ISH were positive, whereas polymerase chain reaction results were negative, suggesting that ISH methods might be a more sensitive method. Performing on a clinical automated platform, RNA ISH is sensitive in determining high-risk HPV status in formalin-fixed, paraffin-embedded tissues and has the potential of being a standalone clinical test.
[Mh] Termos MeSH primário: Carcinoma de Células Escamosas/virologia
Neoplasias de Cabeça e Pescoço/virologia
Hibridização In Situ
Técnicas de Diagnóstico Molecular
Papillomaviridae/genética
Infecções por Papillomavirus/virologia
RNA Viral/genética
[Mh] Termos MeSH secundário: Adulto
Idoso
Idoso de 80 Anos ou mais
Automação Laboratorial
Biomarcadores Tumorais/análise
Biópsia
Carcinoma de Células Escamosas/química
Carcinoma de Células Escamosas/patologia
Sondas de DNA de HPV
Feminino
Fixadores
Formaldeído
Neoplasias de Cabeça e Pescoço/química
Neoplasias de Cabeça e Pescoço/patologia
Testes de DNA para Papilomavírus Humano
Seres Humanos
Imuno-Histoquímica
Hibridização In Situ/normas
Masculino
Meia-Idade
Técnicas de Diagnóstico Molecular/normas
Infecções por Papillomavirus/patologia
Inclusão em Parafina
Reação em Cadeia da Polimerase
Valor Preditivo dos Testes
Sondas RNA
Reprodutibilidade dos Testes
Fixação de Tecidos/métodos
Adulto Jovem
[Pt] Tipo de publicação:COMPARATIVE STUDY; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Biomarkers, Tumor); 0 (DNA Probes, HPV); 0 (Fixatives); 0 (RNA Probes); 0 (RNA, Viral); 1HG84L3525 (Formaldehyde)
[Em] Mês de entrada:1709
[Cu] Atualização por classe:170911
[Lr] Data última revisão:
170911
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170318
[St] Status:MEDLINE


  3 / 3798 MEDLINE  
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[PMID]:27987409
[Au] Autor:Zouari M; Campuzano S; Pingarrón JM; Raouafi N
[Ad] Endereço:University of Tunis El Manar, Faculty of Science, Department of Chemistry, Laboratoire de Chimie Analytique et Electrochimie (LR99ES15), Sensors & Biosensors Group, Rue Béchir Salem Belkheria, 2092 Tunis El-Manar, Tunis, Tunisia.
[Ti] Título:Competitive RNA-RNA hybridization-based integrated nanostructured-disposable electrode for highly sensitive determination of miRNAs in cancer cells.
[So] Source:Biosens Bioelectron;91:40-45, 2017 May 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:A new method for the detection of miRNAs making use of a competitive RNA/RNA hybridization configuration is described in this work. A biotinylated miRNA (biotin-miRNA) of identical sequence to that of the target miRNA is mixed with the samples to be analyzed allowing competition to be accomplished with the target miRNA for a thiolated RNA probe assembled onto a gold nanoparticles (AuNPs) modified screen-printed electrode. After labeling the hybridized biotin-miRNA with streptavidin-HRP conjugates, amperometric detection at -0.20V was carried out using the H O /hydroquinone (HQ) system. The decrease in the amperometric response was proportional to the concentration of model target miRNA-21 in the 100 fM to 25.0 pM range. The integrated sensor provided a very low detection limit (25 fM, 0.25 attomol in 10µL sample) for miRNA-21 without any amplification step, a complete discrimination against single nucleotide mismatched sequences under practical conditions and high storage stability. The usefulness of the developed method was demonstrated by determining the endogenous levels of the mature target miRNA in total RNA (RNA ) extracted from cancerous and non-cancerous cells.
[Mh] Termos MeSH primário: Técnicas Eletroquímicas/métodos
Ouro/química
Nanopartículas Metálicas/química
MicroRNAs/análise
Hibridização de Ácido Nucleico/métodos
Sondas RNA/química
[Mh] Termos MeSH secundário: Neoplasias da Mama/genética
Linhagem Celular
Eletrodos
Feminino
Peroxidase do Rábano Silvestre/química
Seres Humanos
Peróxido de Hidrogênio/química
Hidroquinonas/química
Limite de Detecção
Células MCF-7
MicroRNAs/genética
Sondas RNA/genética
Estreptavidina/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Hydroquinones); 0 (MIRN21 microRNA, human); 0 (MicroRNAs); 0 (RNA Probes); 7440-57-5 (Gold); 9013-20-1 (Streptavidin); BBX060AN9V (Hydrogen Peroxide); EC 1.11.1.- (Horseradish Peroxidase); XV74C1N1AE (hydroquinone)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170531
[Lr] Data última revisão:
170531
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161218
[St] Status:MEDLINE


  4 / 3798 MEDLINE  
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[PMID]:27956502
[Au] Autor:Masuda I; Igarashi T; Sakaguchi R; Nitharwal RG; Takase R; Han KY; Leslie BJ; Liu C; Gamper H; Ha T; Sanyal S; Hou YM
[Ad] Endereço:Department of Biochemistry and Molecular Biology, Thomas Jefferson University, 233 South 10th Street, Philadelphia, PA 19107, USA.
[Ti] Título:A genetically encoded fluorescent tRNA is active in live-cell protein synthesis.
[So] Source:Nucleic Acids Res;45(7):4081-4093, 2017 Apr 20.
[Is] ISSN:1362-4962
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Transfer RNAs (tRNAs) perform essential tasks for all living cells. They are major components of the ribosomal machinery for protein synthesis and they also serve in non-ribosomal pathways for regulation and signaling metabolism. We describe the development of a genetically encoded fluorescent tRNA fusion with the potential for imaging in live Escherichia coli cells. This tRNA fusion carries a Spinach aptamer that becomes fluorescent upon binding of a cell-permeable and non-toxic fluorophore. We show that, despite having a structural framework significantly larger than any natural tRNA species, this fusion is a viable probe for monitoring tRNA stability in a cellular quality control mechanism that degrades structurally damaged tRNA. Importantly, this fusion is active in E. coli live-cell protein synthesis allowing peptidyl transfer at a rate sufficient to support cell growth, indicating that it is accommodated by translating ribosomes. Imaging analysis shows that this fusion and ribosomes are both excluded from the nucleoid, indicating that the fusion and ribosomes are in the cytosol together possibly engaged in protein synthesis. This fusion methodology has the potential for developing new tools for live-cell imaging of tRNA with the unique advantage of both stoichiometric labeling and broader application to all cells amenable to genetic engineering.
[Mh] Termos MeSH primário: Aptâmeros de Nucleotídeos/química
Biossíntese de Proteínas
Sondas RNA/química
RNA de Transferência/química
RNA de Transferência/metabolismo
[Mh] Termos MeSH secundário: Aptâmeros de Nucleotídeos/metabolismo
Compostos de Benzil/química
Escherichia coli/genética
Corantes Fluorescentes
Imidazolinas/química
Microscopia de Fluorescência
Ribossomos/metabolismo
Spinacia oleracea/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (3,5-difluoro-4-hydroxybenzylidene imidazolinone); 0 (Aptamers, Nucleotide); 0 (Benzyl Compounds); 0 (Fluorescent Dyes); 0 (Imidazolines); 0 (RNA Probes); 9014-25-9 (RNA, Transfer)
[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:161214
[St] Status:MEDLINE
[do] DOI:10.1093/nar/gkw1229


  5 / 3798 MEDLINE  
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[PMID]:27889163
[Au] Autor:Rohde A; Hammerl JA; Appel B; Dieckmann R; Al Dahouk S
[Ad] Endereço:German Federal Institute for Risk Assessment, Diedersdorfer Weg 1, 12277 Berlin, Germany; Free University Berlin, Department of Biology, Chemistry and Pharmacy, Takustr. 3, 14195 Berlin, Germany. Electronic address: alexander.rohde@bfr.bund.de.
[Ti] Título:Differential detection of pathogenic Yersinia spp. by fluorescence in situ hybridization.
[So] Source:Food Microbiol;62:39-45, 2017 Apr.
[Is] ISSN:1095-9998
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Yersinia enterocolitica, Y. pseudotuberculosis and Y. pestis are pathogens of major medical importance, which are responsible for a considerable number of infections every year. The detection of these species still relies on cultural methods, which are slow, labour intensive and often hampered by the presence of high amounts of accompanying flora. In this study, fluorescence in situ hybridization (FISH) was used to develop a fast, sensitive and reliable alternative to detect viable bacteria in food. For this purpose, highly specific probes targeting the 16S and 23S ribosomal RNA were employed to differentially detect each of the three species. In order to enable the differentiation of single nucleotide polymorphisms (SNPs), suitable competitor oligonucleotides and locked nucleic acids (LNAs) were used. Starved cells still showed a strong signal and a direct viable count (DVC) approach combined with FISH optimized live/dead discrimination. Sensitivity of the FISH test was high and even a single cell per gram of spiked minced pork meat could be detected within a day, demonstrating the applicability to identify foodborne hazards at an early stage. In conclusion, the established FISH tests proved to be promising tools to compensate existing drawbacks of the conventional cultural detection of these important zoonotic agents.
[Mh] Termos MeSH primário: Inocuidade dos Alimentos/métodos
Yersinia enterocolitica/isolamento & purificação
Yersinia pestis/isolamento & purificação
Yersinia pseudotuberculosis/isolamento & purificação
[Mh] Termos MeSH secundário: Bactérias/genética
Carga Bacteriana
Microbiologia de Alimentos
Hibridização in Situ Fluorescente
Polimorfismo de Nucleotídeo Único/imunologia
Sondas RNA
RNA Ribossômico 16S
RNA Ribossômico 23S
Carne Vermelha/microbiologia
Sensibilidade e Especificidade
Yersinia enterocolitica/genética
Yersinia pestis/genética
Yersinia pseudotuberculosis/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA Probes); 0 (RNA, Ribosomal, 16S); 0 (RNA, Ribosomal, 23S)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170828
[Lr] Data última revisão:
170828
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161128
[St] Status:MEDLINE


  6 / 3798 MEDLINE  
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[PMID]:27836600
[Au] Autor:Kaplan M; Kilic T; Guler G; Mandli J; Amine A; Ozsoz M
[Ad] Endereço:Gediz University, Department of Biomedical Engineering, 35665 Seyrek, Turkey.
[Ti] Título:A novel method for sensitive microRNA detection: Electropolymerization based doping.
[So] Source:Biosens Bioelectron;92:770-778, 2017 Jun 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:In the proposed study, for the first time, sensitive electrochemical detection of a breast cancer biomarker microRNA (miRNA), mir-21 was achieved via electropolymerized polypyrrole (PPy) modified pencil graphite electrodes (PPy/PGE). The detection of hybridization of electrochemically doped probe miRNA, antimir-21, with its complementary target, mir-21 was monitored by either electrochemical impedance spectroscopy (EIS) via comparison of charge transfer resistance (R ) values before and after hybridization or by electrochemical reduction signal of an hybridization indicator, Meldola's blue (MDB). The study covers all the optimization steps for hybridization procedure and electropolymerization of pyrrole as well as detection from real samples of breast cancer cell line, MCF-7. The designed sensor shows a high selectivity and a low detection limit of 0.17nM thanks to electrical conductivity and porous structure of PPy.
[Mh] Termos MeSH primário: Técnicas Eletroquímicas/métodos
Grafite/química
MicroRNAs/análise
Polímeros/química
Pirróis/química
[Mh] Termos MeSH secundário: Técnicas Biossensoriais/métodos
Neoplasias da Mama/genética
Espectroscopia Dielétrica/métodos
Eletrodos
Feminino
Seres Humanos
Limite de Detecção
Células MCF-7
MicroRNAs/genética
Hibridização de Ácido Nucleico/métodos
Polimerização
Sondas RNA/química
Sondas RNA/genética
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (MIRN21 microRNA, human); 0 (MicroRNAs); 0 (Polymers); 0 (Pyrroles); 0 (RNA Probes); 30604-81-0 (polypyrrole); 7782-42-5 (Graphite)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170321
[Lr] Data última revisão:
170321
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161113
[St] Status:MEDLINE


  7 / 3798 MEDLINE  
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[PMID]:27825885
[Au] Autor:McVey C; Huang F; Elliott C; Cao C
[Ad] Endereço:Institute for Global Food Security, School of Biological Sciences, Queen's University Belfast, 18-30 Malone Road, Belfast BT9 5BN, UK.
[Ti] Título:Endonuclease controlled aggregation of gold nanoparticles for the ultrasensitive detection of pathogenic bacterial DNA.
[So] Source:Biosens Bioelectron;92:502-508, 2017 Jun 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The development of an ultrasensitive biosensor for the low-cost and on-site detection of pathogenic DNA could transform detection capabilities within food safety, environmental monitoring and clinical diagnosis. Herein, we present an innovative approach exploiting endonuclease-controlled aggregation of plasmonic gold nanoparticles (AuNPs) for label-free and ultrasensitive detection of bacterial DNA. The method utilizes RNA-functionalized AuNPs which form DNA-RNA heteroduplex structures through specific hybridization with target DNA. Once formed, the DNA-RNA heteroduplex is susceptible to RNAse H enzymatic cleavage of the RNA probe, allowing the target DNA to liberate and hybridize with another RNA probe. This continuously happens until all of the RNA probes are cleaved, leaving the nanoparticles unprotected and thus aggregated upon exposure to a high electrolytic medium. The assay is ultrasensitive, allowing the detection of target DNA at femtomolar level by simple spectroscopic analysis (40.7 fM and 2.45fM as measured by UV-vis and dynamic light scattering (DLS), respectively). The target DNA spiked food matrix (chicken meat) is also successfully detected at a concentration of 1.2pM (by UV-vis) or 18.0fM (by DLS). In addition to the ultra-high sensitivity, the total analysis time of the assay is less than 3h, thus demonstrating its practicality for food analysis.
[Mh] Termos MeSH primário: Campylobacter jejuni/isolamento & purificação
Colorimetria/métodos
DNA Bacteriano/análise
Análise de Alimentos/métodos
Ouro/química
Nanopartículas Metálicas/química
Aves Domésticas/microbiologia
[Mh] Termos MeSH secundário: Animais
Infecções por Campylobacter/microbiologia
Galinhas
Sondas de DNA/química
Seres Humanos
Nanopartículas Metálicas/ultraestrutura
Hibridização de Ácido Nucleico/métodos
Sondas RNA/química
Ribonuclease H/química
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Probes); 0 (DNA, Bacterial); 0 (RNA Probes); 7440-57-5 (Gold); EC 3.1.26.4 (Ribonuclease H)
[Em] Mês de entrada:1703
[Cu] Atualização por classe:170331
[Lr] Data última revisão:
170331
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161110
[St] Status:MEDLINE


  8 / 3798 MEDLINE  
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[PMID]:27762072
[Au] Autor:Sánchez Barreiro F; Vieira FG; Martin MD; Haile J; Gilbert MT; Wales N
[Ad] Endereço:Centre for GeoGenetics, Natural History Museum of Denmark, University of Copenhagen, Øster Voldgade 5-7, 1350, Copenhagen K, Denmark.
[Ti] Título:Characterizing restriction enzyme-associated loci in historic ragweed (Ambrosia artemisiifolia) voucher specimens using custom-designed RNA probes.
[So] Source:Mol Ecol Resour;17(2):209-220, 2017 Mar.
[Is] ISSN:1755-0998
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Population genetic studies of nonmodel organisms frequently employ reduced representation library (RRL) methodologies, many of which rely on protocols in which genomic DNA is digested by one or more restriction enzymes. However, because high molecular weight DNA is recommended for these protocols, samples with degraded DNA are generally unsuitable for RRL methods. Given that ancient and historic specimens can provide key temporal perspectives to evolutionary questions, we explored how custom-designed RNA probes could enrich for RRL loci (Restriction Enzyme-Associated Loci baits, or REALbaits). Starting with genotyping-by-sequencing (GBS) data generated on modern common ragweed (Ambrosia artemisiifolia L.) specimens, we designed 20 000 RNA probes to target well-characterized genomic loci in herbarium voucher specimens dating from 1835 to 1913. Compared to shotgun sequencing, we observed enrichment of the targeted loci at 19- to 151-fold. Using our GBS capture pipeline on a data set of 38 herbarium samples, we discovered 22 813 SNPs, providing sufficient genomic resolution to distinguish geographic populations. For these samples, we found that dilution of REALbaits to 10% of their original concentration still yielded sufficient data for downstream analyses and that a sequencing depth of ~7m reads was sufficient to characterize most loci without wasting sequencing capacity. In addition, we observed that targeted loci had highly variable rates of success, which we primarily attribute to similarity between loci, a trait that ultimately interferes with unambiguous read mapping. Our findings can help researchers design capture experiments for RRL loci, thereby providing an efficient means to integrate samples with degraded DNA into existing RRL data sets.
[Mh] Termos MeSH primário: Ambrosia/classificação
Ambrosia/genética
DNA Antigo/isolamento & purificação
DNA de Plantas/genética
DNA de Plantas/isolamento & purificação
Sondas RNA
[Mh] Termos MeSH secundário: Polimorfismo de Nucleotídeo Único
Análise de Sequência de DNA
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA, Ancient); 0 (DNA, Plant); 0 (RNA Probes)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170407
[Lr] Data última revisão:
170407
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:161021
[St] Status:MEDLINE
[do] DOI:10.1111/1755-0998.12610


  9 / 3798 MEDLINE  
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[PMID]:27634438
[Au] Autor:Kiani SJ; Taheri T; Rafati S; Samimi-Rad K
[Ti] Título:PUF Proteins: Cellular Functions and Potential Applications.
[So] Source:Curr Protein Pept Sci;18(3):250-261, 2017.
[Is] ISSN:1875-5550
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:RNA-binding proteins play critical roles in the regulation of gene expression. Among several families of RNA-binding proteins, PUF (Pumilio and FBF) proteins have been the subject of extensive investigations, as they can bind RNA in a sequence-specific manner and they are evolutionarily conserved among a wide range of organisms. The outstanding feature of these proteins is a highly conserved RNA-binding domain, which is known as the Pumilio-homology domain (PUM-HD) that mostly consists of eight tandem repeats. Each repeat recognizes an RNA base with a simple three-letter code that can be programmed in order to change the sequence-specificity of the protein. Using this tailored architecture, researchers have been able to change the specificity of the PUM-HD and target desired transcripts in the cell, even in subcellular compartments. The potential applications of this versatile tool in molecular cell biology seem unbounded and the use of these factors in pharmaceutics might be an interesting field of study in near future.
[Mh] Termos MeSH primário: Proteínas de Ligação a RNA/química
Proteínas de Ligação a RNA/metabolismo
[Mh] Termos MeSH secundário: Animais
Regulação da Expressão Gênica
Seres Humanos
MicroRNAs
Biossíntese de Proteínas
Conformação Proteica
Domínios Proteicos
Sondas RNA
Proteínas de Ligação a RNA/genética
Transdução de Sinais
[Pt] Tipo de publicação:JOURNAL ARTICLE; REVIEW
[Nm] Nome de substância:
0 (MicroRNAs); 0 (PUM1 protein, human); 0 (Puf-A protein, human); 0 (RNA Probes); 0 (RNA-Binding Proteins)
[Em] Mês de entrada:1705
[Cu] Atualização por classe:170515
[Lr] Data última revisão:
170515
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160917
[St] Status:MEDLINE
[do] DOI:10.2174/1389203717666160914172908


  10 / 3798 MEDLINE  
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[PMID]:27321444
[Au] Autor:Adinolfi B; Pellegrino M; Giannetti A; Tombelli S; Trono C; Sotgiu G; Varchi G; Ballestri M; Posati T; Carpi S; Nieri P; Baldini F
[Ad] Endereço:Istituto di Fisica Applicata "Nello Carrara", Consiglio Nazionale delle Ricerche, Via Madonna del Piano 10, 50019 Sesto Fiorentino (Fi), Italy. Electronic address: b.adinolfi@ifac.cnr.it.
[Ti] Título:Molecular beacon-decorated polymethylmethacrylate core-shell fluorescent nanoparticles for the detection of survivin mRNA in human cancer cells.
[So] Source:Biosens Bioelectron;88:15-24, 2017 Feb 15.
[Is] ISSN:1873-4235
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:One of the main goals of nanomedicine in cancer is the development of effective drug delivery systems, primarily nanoparticles. Survivin, an overexpressed anti-apoptotic protein in cancer, represents a pharmacological target for therapy and a Molecular Beacon (MB) specific for survivin mRNA is available. In this study, the ability of polymethylmethacrylate nanoparticles (PMMA-NPs) to promote survivin MB uptake in human A549 cells was investigated. Fluorescent and positively charged core PMMA-NPs of nearly 60nm, obtained through an emulsion co-polymerization reaction, and the MB alone were evaluated in solution, for their analytical characterization; then, the MB specificity and functionality were verified after adsorption onto the PMMA-NPs. The carrier ability of PMMA-NPs in A549 was examined by confocal microscopy. With the optimized protocol, a hardly detectable fluorescent signal was obtained after incubation of the cells with the MB alone (fluorescent spots per cell of 1.90±0.40 with a mean area of 1.04±0.20µm ), while bright fluorescent spots inside the cells were evident by using the MB loaded onto the PMMA-NPs. (27.50±2.30 fluorescent spots per cell with a mean area of 2.35±0.16µm ). These results demonstrate the ability of the PMMA-NPs to promote the survivin-MB internalization, suggesting that this complex might represent a promising strategy for intracellular sensing and for the reduction of cancer cell proliferation.
[Mh] Termos MeSH primário: Corantes Fluorescentes/química
Proteínas Inibidoras de Apoptose/genética
Nanopartículas/química
Polimetil Metacrilato/química
Sondas RNA/química
RNA Mensageiro/análise
RNA Mensageiro/genética
[Mh] Termos MeSH secundário: Células A549
Técnicas Biossensoriais/métodos
Seres Humanos
Nanopartículas/ultraestrutura
Imagem Óptica/métodos
Sondas RNA/genética
Espectrometria de Fluorescência/métodos
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (BIRC5 protein, human); 0 (Fluorescent Dyes); 0 (Inhibitor of Apoptosis Proteins); 0 (RNA Probes); 0 (RNA, Messenger); 9011-14-7 (Polymethyl Methacrylate)
[Em] Mês de entrada:1702
[Cu] Atualização por classe:170821
[Lr] Data última revisão:
170821
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:160621
[St] Status:MEDLINE



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