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Pesquisa : D13.444.735.635 [Categoria DeCS]
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Id: biblio-1021917
Autor: Sánchez, Evelyn; Tricon, David; Mora, Roxana; Quiroz, Daniela; Decroocq, Véronique; Prieto, Humberto.
Título: A fast and efficient protocol for small RNA extraction in Japanese plum and other Prunus species
Fonte: Electron. j. biotechnol;30:103-109, nov. 2017. ilus, tab, graf.
Idioma: en.
Projeto: FONDEF.
Resumo: Background: Small ribonucleic acids represent an important repertoire of mobile molecules that exert key roles in several cell processes including antiviral defense. Small RNA based repertoire includes both small interfering RNA (siRNA) and microRNA (miRNA) molecules. In the Prunus genus, sharka disease, caused by the Plum pox virus (PPV), first occurred on European plum (Prunus domestica) and then spread over among all species in this genus and thus classified as quarantine pathogen. Next-generation sequencing (NGS) was used for the study of siRNA/miRNA molecules; however, NGS relies on adequate extraction protocols. Currently, knowledge of PPV-Prunus interactions in terms of siRNA populations and miRNA species is still scarce, and siRNA/miRNA extraction protocols are limited to species such as peach, almond, and sweet cherry. Results: We describe a reliable procedure for siRNA/miRNA purification from Prunus salicina trees, in which previously used protocols did not allow adequate purification. The procedure was based on a combination of commercially available RNA purification kits and specific steps that yielded high quality purifications. The resulting molecules were adequate for library construction and NGS, leading to the development of a pipeline for analysis of both siRNAs and miRNAs in the PPV­P. salicina interactions. Results showed that PPV infection led to altered siRNA profiles in Japanese plum as characterized by decreased 24-nt and increased 21- and 22-nt siRNAs. Infections showed miR164 and miR160 generation and increased miR166, miR171, miR168, miR319, miR157, and miR159. Conclusion: We propose this protocol as a reliable and reproducible small RNA isolation procedure for P. salicina and other Prunus species.
Descritores: RNA de Plantas/isolamento & purificação
MicroRNAs/isolamento & purificação
RNA Interferente Pequeno/isolamento & purificação
Prunus domestica/genética
-Doenças das Plantas/virologia
Vírus Eruptivo da Ameixa/fisiologia
Interações Hospedeiro-Patógeno
Sequenciamento de Nucleotídeos em Larga Escala
Reação em Cadeia da Polimerase em Tempo Real
Prunus domestica/imunologia
Prunus domestica/virologia
Responsável: CL1.1 - Biblioteca Central


  2 / 16 LILACS  
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Texto completo SciELO Saúde Pública
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Id: lil-761805
Autor: Irrazábal, Gabriela.
Título: Religión y salud: la intervención pública de agentes religiosos católicos formados en bioética en el debate parlamentario sobre la muerte digna en la Argentina / Religion and health: the public intervention of Catholic religious agents trained in bioethics in the parliamentary debate on death with dignity in Argentina
Fonte: Salud colect;11(3):331-349, jul.-sep. 2015.
Idioma: es.
Resumo: Desde una perspectiva sociológica, este trabajo aborda una de las aristas de la intervención pública de ciertos sectores del catolicismo en la elaboración y sanción de leyes de salud. En particular se hace foco en el debate en comisiones parlamentarias sobre la llamada ley de "muerte digna" (Ley 26742) en el cual se convocó a un grupo de expertos en bioética para asesorar a los senadores sobre los alcances y límites de la ley. La mayoría de los expertos invitados pregonan la perspectiva de la bioética personalista, un desarrollo teológico de la bioética del catolicismo contemporáneo. En el debate no participaron representantes de otros credos consolidando la ampliamente estudiada imbricación entre el catolicismo y lo político en Argentina.

This paper discusses from a sociological perspective one of Catholicism's fronts of public intervention in the development and enactment of health legislation. In particular we analyze the debate in parliamentary committees on the so-called "death with dignity" law (No. 26742), for which a group of bioethics experts was convened to counsel senators regarding the scope and limits of the law. The majority of the invited experts advocated a personalist bioethics perspective, which is a theological bioethics development of contemporary Catholicism. In the debate no representatives of other faiths were present, reinforcing the widely studied overlap between Catholicism and politics in Argentina.
Descritores: Mesembryanthemum/fisiologia
RNA de Plantas/genética
Salinidade
Análise de Sequência de RNA
-Arabidopsis/efeitos dos fármacos
Arabidopsis/crescimento & desenvolvimento
Genes de Plantas
Mesembryanthemum/genética
Raízes de Plantas/crescimento & desenvolvimento
Cloreto de Sódio/farmacologia
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


  3 / 16 LILACS  
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Texto completo SciELO Brasil
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Id: lil-719257
Autor: Rêgo, MJBM.; Santos, PB.; Carvalho-Junior, LB.; Stirling, J.; Beltrão, EIC..
Título: Evaluation of Parkia pendula lectin mRNA differentially expressed in seedlings / Avaliação de mRNA da lectina de Parkia pendula diferencialmente expressa em plântulas
Fonte: Braz. j. biol;74(2):489-492, 5/2014. tab, graf.
Idioma: en.
Resumo: Parkia pendula (Willd.) Walp. (Fabaceae) is a neotropical species of the genus Parkia more abundantly distributed in Central to South America. From the seeds of P. pendula a glucose/mannose specific lectin (PpeL) was isolated that has been characterised and used as a biotechnological tool but until now this is the first manuscript to analyse P. pendula mRNA expression in seedlings. For this porpoise a Differential display reverse transcription polimerase chain reaction (DDRT-PCR) was used to evaluate the expression of P. pendula lectin mRNAs in non-rooted seedlings. No bands were observed in the agarose gel, indicating the absence of mRNA of PpeL seedlings. our findings confirm that lectins mRNAs are differently regulated among species even if they are grouped in the same class.

Parkia pendula (Willd.) Walp. (Fabaceae) é a espécie neotropical do gênero Parkia mais abundantemente distribuída na América Central a do Sul. Das sementes de P. pendula foi isolada uma lectina glicose/manose específica (Ppel) que foi caracterizada e usada como ferramenta biotecnológica, porém até o momento esse é o primeiro artigo a analisar a expressão do mRNA nas plântulas de P. pendula. Para esse propósito uma reação de PCR diferencial de transcriptase reversa (DDRT-PCR) foi utilizada para avaliar a expressão do mRNA da lectina de P. pendula em plântulas não enraizadas. Nenhuma banda foi observada no gel de agarose, indicando a ausência de mRNA das plântulas de PpeL. Nossos achados confirmam que os mRNAs de lectinas são regulados de forma diferentes entre as espécies, mesmo que sejam agrupadas na mesma classe.
Descritores: Fabaceae/genética
Lectinas de Plantas/genética
RNA Mensageiro/análise
RNA de Plantas/análise
-Fabaceae/química
Lectinas de Plantas/análise
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Plântulas
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


  4 / 16 LILACS  
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Texto completo SciELO Brasil
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Id: lil-719244
Autor: Rêgo, MJBM.; Santos, PB.; Carvalho-Junior, LB.; Stirling, J.; Beltrão, EIC..
Título: Evaluation of Parkia pendula lectin mRNA differentially expressed in seedlings / Avaliação de mRNA da lectina de Parkia pendula diferencialmente expressa em plântulas
Fonte: Braz. j. biol;74(2):489-492, 5/2014. tab, graf.
Idioma: en.
Resumo: Parkia pendula (Willd.) Walp. (Fabaceae) is a neotropical species of the genus Parkia more abundantly distributed in Central to South America. From the seeds of P. pendula a glucose/mannose specific lectin (PpeL) was isolated that has been characterised and used as a biotechnological tool but until now this is the first manuscript to analyse P. pendula mRNA expression in seedlings. For this porpoise a Differential display reverse transcription polimerase chain reaction (DDRT-PCR) was used to evaluate the expression of P. pendula lectin mRNAs in non-rooted seedlings. No bands were observed in the agarose gel, indicating the absence of mRNA of PpeL seedlings. our findings confirm that lectins mRNAs are differently regulated among species even if they are grouped in the same class.

Parkia pendula (Willd.) Walp. (Fabaceae) é a espécie neotropical do gênero Parkia mais abundantemente distribuída na América Central a do Sul. Das sementes de P. pendula foi isolada uma lectina glicose/manose específica (Ppel) que foi caracterizada e usada como ferramenta biotecnológica, porém até o momento esse é o primeiro artigo a analisar a expressão do mRNA nas plântulas de P. pendula. Para esse propósito uma reação de PCR diferencial de transcriptase reversa (DDRT-PCR) foi utilizada para avaliar a expressão do mRNA da lectina de P. pendula em plântulas não enraizadas. Nenhuma banda foi observada no gel de agarose, indicando a ausência de mRNA das plântulas de PpeL. Nossos achados confirmam que os mRNAs de lectinas são regulados de forma diferentes entre as espécies, mesmo que sejam agrupadas na mesma classe.
Descritores: Fabaceae/genética
Lectinas de Plantas/genética
RNA Mensageiro/análise
RNA de Plantas/análise
-Fabaceae/química
Lectinas de Plantas/análise
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Plântulas
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


  5 / 16 LILACS  
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Texto completo SciELO Chile
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Id: lil-591904
Autor: Rai, Vineeta; Ghosh, Jayadri Sekhar; Dey, Nrisingha.
Título: Isolation of total RNA from hard bamboo tissue rich in polyphenols and polysaccharides for gene expression studies
Fonte: Electron. j. biotechnol;13(5):22-23, Sept. 2010. ilus, tab.
Idioma: en.
Projeto: Department of Science and technology.
Resumo: RNA isolation from hard and woody internodal bamboo (Bambusa balcooa) tissue is very difficult due to the presence of secondary metabolites, polysaccharides, and polyphenolics. These compounds often co-precipitate with isolated RNA and hinder downstream applications. We have developed an efficient, cost effective and reproducible RNA isolation method from hard tissue of bamboo internode. This protocol includes an additional organic solvent refinement steps to remove endogenous phenolic compounds and acidic phenol (pH 4.2) to critically stabilize RNA in extraction buffer. In addition to these, two 2M Lithium chloride washing steps were introduced to eliminate DNA and polysaccharides contamination. The RNA isolated from the present protocol was found to be superior, when compared to total RNA extracted by other available protocols. The A260/A280 absorption ratio of the isolated RNA was found ranging between 1.89-1.97. The integrity of 28S and 18S rRNA was highly satisfactory when analyzed in agarose denaturing gel. RNA was further used for RT PCR, northern hybridization, cDNA library and subtractive hybridization without any further refinement.
Descritores: RNA de Plantas/isolamento & purificação
Bambusa/genética
-Northern Blotting
Compostos Fenólicos
Reação em Cadeia da Polimerase
Polissacarídeos
Responsável: CL1.1 - Biblioteca Central


  6 / 16 LILACS  
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Texto completo SciELO Chile
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Id: lil-531920
Autor: Rubio-Piña, Jorge A; Vázquez-Flota, Felipe A.
Título: Isolation of functional total RNA from Argemone mexicana tissues
Fonte: Electron. j. biotechnol;11(4):15-16, Oct. 2008. ilus, tab.
Idioma: en.
Resumo: RNA extraction from recalcitrant plant tissues is frequently complicated by the presence of secondary metabolites, polysaccharides and polyphenols. These compounds may co precipitate with RNA, often rendering it unsuitable for either cDNA synthesis or hybridization in northern blot analyses and therefore, interfering with the gene analysis expression in such tissues. We have developed an efficient RNA extraction method from A. mexicana tissues. The procedure includes the use of polyvinylpolypyrrolidone (PVPP), to remove secondary metabolites, proteins and polyphenols, and a two-step precipitation with LiCl, to eliminate polysaccharides, and thus increasing RNA yield. The quality of the resulting RNA was evaluated spectrophotometrically and by agarose gel electrophoresis. Moreover, the RNA obtained by this method, could be used directly for both RT-PCR and northern blot analysis, without any further purification.
Descritores: Papaveraceae/anatomia & histologia
Papaveraceae/genética
RNA de Plantas
-Northern Blotting/métodos
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Responsável: CL1.1 - Biblioteca Central


  7 / 16 LILACS  
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Texto completo SciELO Chile
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Id: lil-531894
Autor: Vasanthaiah, Hemanth K. N; Katam, Ramesh; Sheikh, Mehboob B.
Título: Efficient protocol for isolation of functional RNA from different grape tissue rich in polyphenols and polysaccharides for gene expression studies
Fonte: Electron. j. biotechnol;11(3):42-51, July 2008. ilus, tab.
Idioma: en.
Resumo: Extraction of RNA from plant tissue containing high levels of polyphenols and polysaccharides is tedious and difficult in grapes. Although several protocols have been published for plant RNA isolation, most have failed to yield high quality RNA in sufficient quantity from mature and diseased grape tissue. We describe a protocol for isolating intact and high quality RNA from various grape tissues as evident by high A260/A280 absorbance ratio (1.8 to 1.9) and electrophoretic profile on denaturing formaldehyde agarose gel. On an average, 205 µg RNA per g of fresh tissue were obtained using this modified protocol. RNA quality was further assessed through RT-PCR, differential display RT-PCR and subtractive hybridization, and found to be suitable for molecular studies.
Descritores: RNA de Plantas
Vitis/genética
-Compostos Fenólicos/análise
Polissacarídeos/isolamento & purificação
Reação em Cadeia da Polimerase
Responsável: CL1.1 - Biblioteca Central


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Texto completo SciELO Chile
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Id: lil-499183
Autor: Bazzini, Ariel A; Mongelli, Vanesa C; Hopp, H. Esteban; Del Vas, Mariana; Asurmendi, Sebastian.
Título: A practical approach to the understanding and teaching of RNA silencing in plants
Fonte: Electron. j. biotechnol;10(2):178-190, Apr. 15, 2007. ilus, graf.
Idioma: en.
Projeto: FONCyT; . ANPCyT; . CONICET; . INTA.
Resumo: Gene silencing, also called RNA interference (RNAi) is a specific mechanism of RNA degradation involved in gene regulation, development and defense in eukaryotic organisms. It became an important subject in the teaching programs of molecular biology, genetics and biotechnology courses in the last years. The aim of this work is to provide simple and inexpensive assays to understand and teach gene silencing using plants as model systems. The use of transient and permanent transgenic plants for expressing reporter genes, like those derived from jellyfish green fluorescent protein (gfp) encoding gene, provides a nice, colorful and conclusive image of gene silencing. Three experimental approaches to evidence RNA silencing are depicted. In the first approach gene silencing is demonstrated after transient expression of reporter genes in non-transgenic plants. In the second, silencing is triggered against a reporter gene stably integrated into a transgenic plant. The third approach involves the triggering of RNA silencing against endogenous genes using viral vectors. In addition we illustrate systemic gene silencing showing how the silencing signal is spread over a plant and finally it is also demonstrated the suppression of gene silencing. The first group of experiments is recommended to be tough on undergraduate courses, the following two sections are recommended for graduate courses. Hopefully, it will help students to understand this important phenomenon and to unravel the importance of gene silencing as a key gene regulation mechanism and as a molecular and biotechnological tool.
Descritores: RNA de Plantas/genética
Inativação Gênica
Interferência de RNA
Ensino
-Biotecnologia/educação
Proteínas de Fluorescência Verde
Modelos Genéticos
Plantas Geneticamente Modificadas/genética
Interferência Viral
Responsável: CL1.1 - Biblioteca Central


  9 / 16 LILACS  
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Texto completo SciELO Chile
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Id: lil-481306
Autor: Le Provost, Gregoire; Herrera, Raúl; Paiva, Jorge A; Chaumeil, Philippe; Salin, Franck; Plomion, Christophe.
Título: A micromethod for high throughput RNA extraction in forest trees
Fonte: Biol. Res;40(3):291-297, 2007. ilus.
Idioma: en.
Projeto: Fundação para a Ciencia e a Tecnología; . Fondecyt. Enlace.
Resumo: A large quantity of high quality RNA is often required in the analysis of gene expression. However, RNA extraction from samples taken from woody plants is generally complex, and represents the main limitation to study gene expression, particularly in refractory species like conifers. Standard RNA extraction protocols are available but they are highly time consuming, and not adapted to large scale extraction. Here we present a high-throughput RNA extraction protocol. This protocol was adapted to a micro-scale by modifying the classical cetyltrimethylammonium (CTAB) protocol developed for pine: (i) quantity of material used (100-200 mg of sample), (ii) disruption of samples in microtube using a mechanical tissue disrupter, and (iii) the use of SSTE buffer. One hundred samples of woody plant tissues/organs can be easily treated in two working days. An average of 15 /ig of high quality RNA per sample was obtained. The RNA extracted is suitable for applications such as real time reverse transcription polymerase chain reaction, cDNA library construction or synthesis of complex targets for microarray analysis.
Descritores: Técnicas Genéticas
RNA de Plantas/isolamento & purificação
Árvores/genética
-Compostos de Cetrimônio
DNA Complementar/genética
Regulação da Expressão Gênica de Plantas
Análise em Microsséries
Reprodutibilidade dos Testes
Reação em Cadeia da Polimerase Via Transcriptase Reversa
RNA de Plantas/genética
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME


  10 / 16 LILACS  
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Texto completo SciELO Chile
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Id: lil-468185
Autor: Leitao, Louis; Maoret, Jean-José; Biolley, Jean-Philippe.
Título: Changes in PEP Carboxylase, Rubisco and Rubisco activase mRNA levels from maize (Zea mays) exposed to a chronic ozone stress
Fonte: Biol. Res;40(2):137-153, 2007. graf, tab.
Idioma: en.
Resumo: We quantified the ozone impact on levels of Zea mays L. cv. Chambord mRNAs encoding C4-phosphoenolpyruvate carboxylase (C4-PEPc), ribulose-l,5-bisphosphate carboxylase/oxygenase small and large subunits (Rubisco-SSU and Rubisco-LSU, respectively) and Rubisco activase (RCA) using real-time RT-PCR. Foliar pigment content, PEPc and Rubisco protein amounts were simultaneously determined. Two experiments were performed to study the ozone response of the 5th and the 10th leaf. For each experiment, three ozone concentrations were tested in open-top chambers: non-filtered air (NF, control) and non-filtered air containing 40 (+40) and 80 nL L-1 (+80) ozone. Regarding the 5th leaf, +40 atmosphere induced a loss in pigmentation, PEPc and Rubisco activase mRNAs. However, it was unable to notably depress carboxylase protein amounts and mRNAs encoding Rubisco. Except for Rubisco mRNAs, all other measured parameters from 5th leaf were depressed by +80 atmosphere. Regarding the 10th leaf, +40 atmosphere increased photosynthetic pigments and transcripts encoding Rubisco and Rubisco activase. Rubisco and PEPc protein amounts were not drastically changed, even if they tended to be increased. Level of C4-PEPc mRNA remained almost stable. In response to +80 atmosphere, pigments and transcripts encoding PEPc were notably decreased. Rubisco and PEPc protein amounts also declined to a lesser extent. Conversely, the level of transcripts encoding both Rubisco subunits and Rubisco activase that were not consistently disturbed tended to be slightly augmented. So, the present study suggests that maize leaves can respond differentially to a similar ozone stress.
Descritores: Ozônio/farmacologia
Fosfoenolpiruvato Carboxilase/metabolismo
Ribulose-Bifosfato Carboxilase/metabolismo
Zea mays/efeitos dos fármacos
Zea mays/enzimologia
-Fosfoenolpiruvato Carboxilase/efeitos dos fármacos
Folhas de Planta/efeitos dos fármacos
Folhas de Planta/enzimologia
Proteínas de Plantas/metabolismo
Reação em Cadeia da Polimerase Via Transcriptase Reversa
RNA Mensageiro/efeitos dos fármacos
RNA de Plantas/efeitos dos fármacos
Ribulose-Bifosfato Carboxilase/efeitos dos fármacos
Zea mays/genética
Tipo de Publ: Research Support, Non-U.S. Gov't
Responsável: BR1.1 - BIREME



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