SABIIA - Sistema Aberto e Integrado de Informação em Agricultura (Sabiia)

Paiva,Marcelo Henrique Santos; Guedes,Duschinka Ribeiro Duarte; Leal,Walter Soares; Ayres,Constância Flávia Junqueira.

Abstract Tissue samples from mosquitoes artificially infected with Zika virus and shown to be positive by RT-qPCR were reexamined by RT-PCR. Using these samples we compared the two methods employed in virus RNA detection for vector competence studies. Results demonstrated that, albeit useful, RT-PCR gave false negatives with low viral loads (< 106 RNA copies/ml).

Frequencies of BCR-ABL1 fusion transcripts among Sudanese chronic myeloid leukaemia patients

Palavras-chave: ZIKV; RT-PCR; Aedes aegypti; Vector competence.

Ano: 2017

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572017000400597

Osman,Emad-Aldin I.; Hamad,Kamal; Elmula,Imad M. Fadl; Ibrahim,Muntaser E..

The incidence of one or other rearrangement in chronic myeloid leukemia (CML) patients varies in different reported series. In this study we report the frequencies of BCR-ABL1 fusion transcript variants studied in 43 CML patients from Sudan. The study includes 46 Sudanese patients, three of which negative for the BCR-ABL1 fusion transcript. More than half of 43 positive patients showed b2a2 fusion transcript (53.5%), while (41.9%) showed b3a2 transcript and the remaining (4.6%) coexpression of b3a2/ b2a2 and b3a2/b2a2/e19a2. We detected neither coexpression of p210/p190 nor e1a2 alone. Male patients showed a tendency to express b2a2, while female tende to express b3a2 (p = 0.017). Moreover, a single nucleotide polymorphism was detected in BCR exon 13 in...

Anoxia- and hypoxia-induced expression of LDH-A* in the Amazon Oscar, Astronotus crassipinis

Palavras-chave: BCR-ABL; Chronic myeloid leukemia; Ph chromosome; Sudanese; RT-PCR.

Ano: 2010

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572010000200005

Almeida-Val,Vera Maria Fonseca; Oliveira,Alice Reis; Silva,Maria de Nazaré Paula da; Ferreira-Nozawa,Monica Stropa; Araújo,Roziete Mendes; Val,Adalberto Luis; Nozawa,Sérgio Ricardo.

Adaptation or acclimation to hypoxia occurs via the modulation of physiologically relevant genes, such as erythropoietin, transferrin, vascular endothelial growth factor, phosphofructokinase and lactate dehydrogenase A. In the present study, we have cloned, sequenced and examined the modulation of the LDH-A gene after an Amazonian fish species, Astronotus crassipinis (the Oscar), was exposed to hypoxia and anoxia. In earlier studies, we have discovered that adults of this species are extremely tolerant to hypoxia and anoxia, while the juveniles are less tolerant. Exposure of juveniles to acute hypoxia and anoxia resulted in increased LDH-A gene expression in skeletal and cardiac muscles. When exposed to graded hypoxia juveniles show decreased LDH-A...

Development of a quantitative competitive reverse transcriptase polymerase chain reaction for the quantification of growth hormone gene expression in pigs

Palavras-chave: Amazon; Fishes; Gene expression; RT-PCR; LDH-A; Hypoxia; Anoxia.

Ano: 2011

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572011000200025

Franco,Maurício Machaim; Almeida,Juliana Franco; Souza,Guilherme Rocha Lino de; Antunes,Robson Carlos; Goulart,Luiz Ricardo.

After the advent of the genome projects, followed by the discovery of DNA polymorphisms, basic understanding of gene expression is the next focus to explain the association between polymorphisms and the level of gene expression, as well as to demonstrate the interaction among genes. Among the various techniques for the investigation of transcriptional profiling involving patterns of gene expression, quantitative PCR is the simplest analytical laboratory technique. The objective of this work was to analyze two strategies of a competitive PCR technique for the quantification of the pig growth hormone (GH) gene expression. A pair of primers was designed targeting exons 3 and 5, and two competitive PCR strategies were performed, one utilizing a specific...

Identification and detection of a novel human endogenous retrovirus-related gene, and structural characterization of its related elements

Palavras-chave: Pig; GH gene; Gene expression; RT-PCR.

Ano: 2003

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572003000100003

Liang,Qiaoyi; Ding,Jiayi; Zheng,Shu.

Up-regulation of human endogenous retroviruses (HERVs) is associated with many diseases, including cancer. In this study, an H family HERV (HERV-H)-related gene was identified and characterized. Its spliced transcript lacks protein-coding capacity and may belong to the emerging class of noncoding RNAs (ncRNAs). The 1.3-kb RNA consisting of four exons is transcribed from an Alu element upstream of a 5.0-kb structurally incomplete HERV-H element. RT-PCR and quantitative RT-PCR results indicated that expression of this HERV-related transcript was negatively associated with colon, stomach, and kidney cancers. Its expression was induced upon treatment with DNA methylation and histone deacetylation inhibitors. A BLAT search using long terminal repeats (LTRs)...

In silicio expression analysis of PKS genes isolated from Cannabis sativa L.

Palavras-chave: Endogenous retrovirus; Noncoding RNA; RT-PCR; Quantitative real-time PCR; Transcription.

Ano: 2009

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572009000400005

Flores-Sanchez,Isvett J.; Linthorst,Huub J.M.; Verpoorte,Robert.

Cannabinoids, flavonoids, and stilbenoids have been identified in the annual dioecious plant Cannabis sativa L. Of these, the cannabinoids are the best known group of this plant's natural products. Polyketide synthases (PKSs) are responsible for the biosynthesis of diverse secondary metabolites, including flavonoids and stilbenoids. Biosynthetically, the cannabinoids are polyketide substituted with terpenoid moiety. Using an RT-PCR homology search, PKS cDNAs were isolated from cannabis plants. The deduced amino acid sequences showed 51%-73% identity to other CHS/STS type sequences of the PKS family. Further, phylogenetic analysis revealed that these PKS cDNAs grouped with other non-chalcone-producing PKSs. Homology modeling analysis of these cannabis PKSs...

Crude alkaloid extract of Rhazya stricta inhibits cell growth and sensitizes human lung cancer cells to cisplatin through induction of apoptosis

Palavras-chave: Cannabis sativa; Homology modeling; Polyketide synthases; RT-PCR.

Ano: 2010

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572010000400017

Elkady,Ayman I..

There is an urgent need to improve the clinical management of non-small cell lung cancer (NSCLC), one of the most frequent causes of cancer-related deaths in men and women worldwide. Rhazya stricta, an important medicinal plant used in traditional Oriental medicine, possesses anti-oxidant, anti-carcinogenic and free radical scavenging properties. This study was done to explore the potential anticancer activity of a crude alkaloid extract of R. stricta (CAERS) against the NSCLC line A549. CAERS markedly suppressed the growth of A549 cells and considerably enhanced the anti-proliferative potential of cisplatin. CAERS-mediated inhibition of A549 cell growth correlated with the induction of apoptosis that was accompanied by numerous morphological changes, DNA...

Considerações estatísticas na análise de dados de expressão gênica gerados pela técnica de RT-qPCR.

Palavras-chave: Apoptosis; Lung cancer; Medicinal plant; RT-PCR; Western blot.

Ano: 2013

URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1415-47572013000100003

Infoteca-e

LOBO, A. M. B. O.; LOBO, R. N. B..

A transcriptase reversa (RT) seguida de quantificação utilizando a técnica da reação em cadeia da polimerase (qPCR) em tempo real ou, simplesmente reverse transcription-qPCR (RT-qPCR), é um método in vitro para amplificação enzimática de sequências definidas de DNA complementar obtido a partir do RNA. A técnica de RT-qPCR é uma das mais utilizadas em experimentos de quantificação da expressão gênica por ser considerado um dos métodos mais sensível e específico de quantificação gênica dos que outros utilizados para esta finalidade. No entanto, para obter resultados, pela técnica de RT-qPCR, que possam ser adequadamente analisados é necessário que o experimento tenha um desenho experimental com tratamentos, ensaios, unidade experimental e delineamento bem...

Tipo: Documentos (INFOTECA-E)

Palavras-chave: RT-qPCR; Expressão gênica; Expressão gênica relativa; RT-PCR; Genetic processes.; Genética animal; Genética molecular; Gene expression; Genetics..

Ano: 2014

URL: http://www.infoteca.cnptia.embrapa.br/infoteca/handle/doc/1007398

Diagnóstico de mastite subclínica caprina por meio da detecção de Staphylococcus aureus utilizando a técnica de RT-PCR.

Infoteca-e

MONTEIRO, J. P.; SOUZA, V. de.

A presente publicação descreve os procedimentos necessários para a identificação e confirmação molecular de estirpes de S. aureus causadoras de mastite subclínica, provenientes de amostras de leite de cabra, por meio da técnica de RT-PCR.

Tipo: Comunicado Técnico (INFOTECA-E)

Palavras-chave: RT-PCR; PCR.; Leite de cabra; Caprino; Teste de mamite; Staphylococcus aureus; Diagnóstico; Infecção; Doença animal; Goat milk; Mastitis; Diagnostic techniques; Animal diseases..

Ano: 2015

URL: http://www.infoteca.cnptia.embrapa.br/infoteca/handle/doc/1047787

Les Toxi-Infections Alimentaires Collectives (TIAC) virales associées à la consommation d'huîtres en France, hiver 2000-2001

Zidane, Mohamed; Le Guyader, Soizick; Pommepuy, Monique.

During the winter 2000/2001, several outbreaks associated with the consumption of shellfish were declared in France. Following these events, a close cooperation was organized between administrations and laboratories (DGAL, DPMA, InVS, AFSSA, Ifremer, CHU-Dijon and CNC) to gather competences and to manage the situation. This report synthesizes the information collected during these outbreaks. From the outset, the epidemiologic investigations directed the research towards viral contamination and shellfish as contamination source. Several outbreaks were investigated. Oysters and stools samples from ill patients were analysed by RT-PCR for viruses (enterovirus, rotavirus, norovirus and astrovirus). The genic amplification was done by using the same primers in...

Tipo: Text

Palavras-chave: Épidémie; Gastro-entérites; Huîtres; Contamination virale; RT-PCR; Norovirus; TIAC.

Ano: 2004

URL: http://archimer.ifremer.fr/doc/00068/17941/15476.pdf

Differential gene expression in Litopenaeus vannamei shrimp in response to diet changes

Genotoxicity and activation of cellular defenses in transplanted zebra mussels Dreissena polymorpha along the Seine river

Chavez-calvillo, G.; Perez-rueda, E.; Lizama, G.; Zuniga Aguilar, J. J.; Gaxiola, G.; Cuzon, Gerard; Arena-ortiz, L..

We evaluated changes in gene expression of Litopenaeus vannamei shrimp in response to a change in diet by comparing specimens which were fed with an animal protein diet against those which underwent a change from an animal protein diet to a plant protein diet. In addition, we also compared specimens which were fed with a plant protein diet with another group which underwent a change from plant to animal protein diet. Results showed that, in the case of hepatopancreas tissue, a total of six genes were differentially expressed and exhibited a high degree of homology with sequences which codify for proteins related to metabolism, transcription regulation, and rRNA subunits. In the case of muscle tissue, four differentially expressed sequences were recorded...

Tipo: Text

Palavras-chave: Differential display; RT-PCR; Litopenaeus vannamei; Protein diet; Alternative protein sources.

Ano: 2010

URL: http://archimer.ifremer.fr/doc/00002/11298/7853.pdf

Chatel, Annelle; Faucet-marquis, Virginie; Gourlay-france, Catherine; Pfohl-leszkowicz, Annie; Hubert, Francoise.

The aim of the present study was to confirm the relevance of studying DNA adduct formation in a field study. In that context, freshwater mussels Dreissena polymorpha, collected in a reference station, were transplanted in different sites with a pollution gradient. After one and two months, mussels were collected and DNA adduct formation was analyzed using the 32P post labelling technique on both gills and digestive glands. In addition, the expression of genes involved in the detoxification system (catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST), HSP70, aryl hydrocarbon receptor (AHR), P glycoprotein (PgP), metallothionein (MT)) was assessed by RT-PCR. DNA adducts were observed at amount comparable to data from literature....

Tipo: Text

Palavras-chave: Biomarker; Pollution; DNA adducts; Detoxification; Dreissena polymorpha; RT-PCR.

Ano: 2015

URL: https://archimer.ifremer.fr/doc/00195/30639/32450.pdf

Contamination de l'environnement littoral par les rotavirus du groupe A. Détection par amplification enzymatique en chaîne et analyse du polymorphisme de restriction de séquences virales dans les eaux de surfaces et les coquillages

TRANSCRIPTOS DE FUSIÓN DEL GEN BCR/ABL EN PACIENTES CON LEUCEMIA MIELOIDE CRÓNICA

Dubois, Eric.

Les rotavirus font partie des principaux virus entériques humains susceptibles d'être rencontrés dans l'environnement. Leur mise en évidence par culture cellulaire dans des prélèvements est longue et peu sensible, ce qui explique le manque de données sur la contamination du littoral. La RT-seminested PCR a été développée pour pallier ces inconvénients. Après une mise au point méthodologique cette technique a été utilisée lors d'études in vitro et in situ. Les études in vitro sur le devenir des rotavirus en eau de mer, suivi par RT-PCR quantitative et par multiplication en culture cellulaire, montrent que dans des conditions proches de celles du milieu naturel (eau de mer non stérile et à une température relativement basse), la détection d'ARN viral peut...

Tipo: Text

Palavras-chave: Rotavirus; Contamination; Coquillages; Eaux de stations d'épuration; Echantillons cliniques; RT-PCR; RFLP; Epidémiologie moléculaire.

Ano: 1992

URL: http://archimer.ifremer.fr/doc/00442/55376/56894.pdf

International Journal of Morphology

Artigas,Carmen Gloria; Melo,Angélica; Roa,Juan Carlos; Roa,Iván; Quijada,Ingrid; Vittini,Cecilia; Cabrera,María Elena; Risueño,Concepción.

La anormalidad citogenética más común en la leucemia mieloide crónica (LMC) es el cromosoma Philadelphia, producida por la t(9;22), cuya expresión molecular es el gen de fusión BCR-ABL, que codifica proteínas con actividad tirosinquinasa. Según el punto de ruptura de los genes BCR o ABL se produce una proteína de fusión de 210-kD(p210) o 190-kD(p190). La presencia de este gen de fusión en pacientes con LMC tiene implicancia diagnóstica. Con el propósito de detectar transcriptos de fusión del gen BCR/ABL en pacientes con leucemia mieloide crónica, procedentes de la IX Región de Chile, se estudiaron 14 muestras de sangre de 11 pacientes con LMC. A 2 de ellos, se les realizó seguimiento durante su tratamiento con Gleevec. Se aplicó la técnica de reacción en...

Tipo: Journal article

Palavras-chave: BCR/ABL; RT-PCR; Leucemia mieloide crónica; Chile.

Ano: 2003

URL: http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-95022003000300004

Virus de Distemper Canino: detección molecular de diferentes aislamientos provenientes de perros de la provincia de santa fe, argentina, entre los años 2000 y 2010

InVet

Pinotti,M.; Gollan,A; Canavesio,M.; Passeggi,C.; Larrateguy,M.V.; Paz,M.E.; Formentini,E..

En la ciudad de Santa Fe, Argentina, durante los años 2000 y 2010, se aislaron 13 cepas autóctonas del virus Distemper Canino. Las muestras se tomaron de caninos con signos clínicos de la enfermedad y consistieron en hisopados nasales, oculares, faríngeos (tonsilas) y conjuntivales. Los aislamientos se realizaron en células MDCK, las cuales mostraron efecto citopatogénico característico. Los virus provenientes de la cosecha, se detectaron por RT-PCR de una región que codifica la nucleoproteína NP asociada al RNA (posición 769 a la 1055 del genoma) obteniéndose en todos los casos un producto de 287 pb comparable con el obtenido con cepas vacunales (cepas Ondersterpoort y Lederle), lo que indica presencia viral en todas las muestras procesadas. Las cepas...