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RNA Probe (rna + probe)
Selected AbstractsSensitive and Specific Digoxigenin-labelled RNA Probes for Routine Detection of Citrus tristeza virus by Dot-blot HybridizationJOURNAL OF PHYTOPATHOLOGY, Issue 6 2006L. Barbarossa Abstract A non-radioactive dot-blot hybridization assay for the successful detection of Citrus tristeza virus (CTV) RNA in total nucleic acid extracts of infected citrus was developed. Two digoxigenin (DIG)-labelled minus-sense riboprobes, complementary to the coat protein gene sequence of a Chinese and an Apulian CTV isolate were synthesized. Several citrus tissues were evaluated as optimal virus source and leaf petioles were found appropriate material for reliable detection. The hybridization assay showed a detection limit corresponding to 0.2 mg of fresh infected tissue. The riboprobes allowed CTV detection in isolates from different geographical areas, grown in the screenhouse or in the field, resulting in similar hybridization patterns. The infected trees were tested during different seasons with positive results, although from July to August most of the samples gave a weaker hybridization signal, compared to other seasons. The high sensitivity and reliability of the molecular hybridization assay described make it a good alternative to serological methods for CTV detection. [source] Functional association of human Ki-1/57 with pre-mRNA splicing eventsFEBS JOURNAL, Issue 14 2009Gustavo C. Bressan The cytoplasmic and nuclear protein Ki-1/57 was first identified in malignant cells from Hodgkin's lymphoma. Despite studies showing its phosphorylation, arginine methylation, and interaction with several regulatory proteins, the functional role of Ki-1/57 in human cells remains to be determined. Here, we investigated the relationship of Ki-1/57 with RNA functions. Through immunoprecipitation assays, we verified the association of Ki-1/57 with the endogenous splicing proteins hnRNPQ and SFRS9 in HeLa cell extracts. We also found that recombinant Ki-1/57 was able to bind to a poly-U RNA probe in electrophoretic mobility shift assays. In a classic splicing test, we showed that Ki-1/57 can modify the splicing site selection of the adenoviral E1A minigene in a dose-dependent manner. Further confocal and fluorescence microscopy analysis revealed the localization of enhanced green fluorescent protein,Ki-1/57 to nuclear bodies involved in RNA processing and or small nuclear ribonucleoprotein assembly, depending on the cellular methylation status and its N-terminal region. In summary, our findings suggest that Ki-1/57 is probably involved in cellular events related to RNA functions, such as pre-mRNA splicing. Structured digital abstract ,,MINT-7041074: Ki-1/57 (uniprotkb:Q5JVS0) physically interacts (MI:0915) with SF2P32 (uniprotkb:Q07021) by two hybrid (MI:0018) ,,MINT-7041232: Ki-1/57 (uniprotkb:Q5JVS0) physically interacts (MI:0915) with SFRS9 (uniprotkb:Q13242) by pull down (MI:0096) ,,MINT-7041203: P80-Coilin (uniprotkb:P38432) and Ki-1/57 (uniprotkb:Q5JVS0) colocalize (MI:0403) by fluorescence microscopy (MI:0416) ,,MINT-7041217: SMN (uniprotkb:Q16637) and Ki-1/57 (uniprotkb:Q5JVS0) colocalize (MI:0403) by fluorescence microscopy (MI:0416) ,,MINT-7041189: SC-35 (uniprotkb:Q01130) and Ki-1/57 (uniprotkb:Q5JVS0) colocalize (MI:0403) by fluorescence microscopy (MI:0416) ,,MINT-7041169: NPM (uniprotkb:P06748) and Ki-1/57 (uniprotkb:Q5JVS0) colocalize (MI:0403) by fluorescence microscopy (MI:0416) ,,MINT-7041249: Ki-1/57 (uniprotkb:Q5JVS0) physically interacts (MI:0915) with SFRS9 (uniprotkb:O60506) by pull down (MI:0096) ,,MINT-7041065: Ki-1/57 (uniprotkb:Q5JVS0) physically interacts (MI:0915) with SFRS9 (uniprotkb:Q13242) by two hybrid (MI:0018) ,,MINT-7041069: Ki-1/57 (uniprotkb:Q5JVS0) physically interacts (MI:0915) with YB1 (uniprotkb:P67809) by two hybrid (MI:0018) ,,MINT-7041079: Ki-1/57 (uniprotkb:Q5JVS0) physically interacts (MI:0915) with HNRPQ (uniprotkb:O60506) by two hybrid (MI:0018) ,,MINT-7041087: Ki-1/57 (uniprotkb:Q5JVS0) physically interacts (MI:0218) with HNRPQ3 (uniprotkb:O60506-1), HNRPQ2 (uniprotkb:O60506-2) and HNRPQ-1 (uniprotkb:O60506-3) by anti bait coimmunoprecipitation (MI:0006) [source] Increase in activity, glycosylation and expression of cytokinin oxidase/dehydrogenase during the senescence of barley leaf segments in the darkPHYSIOLOGIA PLANTARUM, Issue 4 2007Klaus Conrad The aim of the study was to elucidate relations between senescence and cytokinin oxidase/dehydrogenase (CKX, EC 1.4.3.18/1.5.99.12). Segments derived from first foliage leaves of Hordeum vulgare L. cv. Alexis were put with their bases into water and kept in darkness. Their senescence was characterized, e.g. by a 60% decline in chlorophyll within 5 days. During this time, the in vitro activity of CKX increased fast and markedly, e.g. 14-fold. Application of 10,4M kinetin (Kin), which slightly retarded the loss of chlorophyll, multiplied the enhancement of CKX activity strongly. Both in the presence and in the absence of Kin, the proportion of glycosylated to non-glycosylated CKX increased during senescence. By hybridization with an antisense RNA probe derived from a fragment of the CKX gene Zmckx1 of maize, an increase of the corresponding transcript of the barley gene Hvckx1 in segments incubated without Kin was shown. The content of base and riboside cytokinins slowly declined in such segments, which argues against triggering but for facilitating senescence processes by CKX. [source] Human papillomavirus infection and premalignant lesions of the oral cavity: A cross-sectional study in Allahabad, North IndiaASIA-PACIFIC JOURNAL OF CLINICAL ONCOLOGY, Issue 2 2009Sharmistha DEBANTH Abstract Aim: To assess the role of human papillomavirus (HPV) in premalignant lesions of the oral cavity using the second-generation Hybrid Capture assay kit (Digene Corporation) and to study the correlation between this technique and morphological changes (koilocytosis) on histopathology in those lesions. Methods: A hospital-based cross-sectional study was undertaken including 92 patients with premalignant lesions of the oral cavity (the study group) and a control group of 35 patients with no oral disease. All the participants were interviewed regarding possible risk factors. Oral exfoliated cells in the saliva were tested for HPV DNA using an HPV RNA probe of 13 high-risk HPV genotypes (16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59 and 68). Simultaneously biopsy specimens of the lesions were examined under a light microscope for evidence of koilocytosis, an empirical marker for HPV infection. Pearson's ,2 test using SPSS V.16 was applied for statistical analysis. Results: HPV DNA was detected in 44.6% of the study group (41 out of 92), and 14.3% of the controls (five out of 35). The association was independent of the influence of betel quid and tobacco chewing, two established causal factors for oral pre-cancers. Out of the total 92 participants in the study group there was evidence of koilocytosis on the histological sections of 42 individuals (45.6%). Conclusion: The results support a strong association between HPV infection and oral premalignant lesions, particularly oral lichen planus and squamous papilloma. Koilocytosis on histology is a good predictor of HPV infection. [source] Epstein-Barr virus-associated haemophagocytic lymphohistiocytosis in Wiskott-Aldrich syndromeACTA PAEDIATRICA, Issue 7 2003S Pasic A patient with Wiskott-Aldrich syndrome who developed Epstein-Barr virus-associated haemophagocytic lymphohistiocytosis (EBV-HLH) is described in this study. At 4 mo of age the patient developed fever associated with bicytopenia and splenomegaly. Analysis of a bone marrow specimen revealed extensive haemophagocytosis, and in situ hybridization for EBV of the bone marrow specimen using an EBV-encoded RNA probe was positive. Diagnosis of EBV-HLH was established and immunotherapy with HLH-94 protocol was started. HLH has been described in patients with other well-defined primary immunodeficiencies such as X-linked lymphoproliferative syndrome, Chediak-Higashi syndrome and Griscelli disease. Also, HLH was reported recently in severe combined immunodeficiency and DiGeorge syndrome. Conclusion: The possibility of an underlying primary immunodeficiency should be considered in paediatric patients who present with HLH during infancy. [source] Predictive value of clinical and microbiological parameters for the treatment outcome of scaling and root planingJOURNAL OF CLINICAL PERIODONTOLOGY, Issue 7 2005P. F. Brochut Abstract Objectives: To compare the clinical and microbiological outcome of non-surgical periodontal therapy after 6 months with data obtained after hygienic phase or 6 weeks after completion of non-surgical therapy, in order to evaluate the value of clinical and microbiological parameters to predict treatment success. Material and Methods: Clinical and microbiological data were available from 271 sites in 10 systemically healthy non-smokers with moderate-to-advanced chronic periodontal disease (24,32 sites per individual). Subgingival plaque samples were tested for the presence of Porphyromonas gingivalis, Actinobacillus actinomycetemcomitans, Tannerella forsythensis and Treponema denticola using RNA probes. Results: Stepwise multiple linear regression analysis revealed a significant impact of the number of sites with visible plaque index >1 after hygienic phase on the bleeding tendency of a subject at month 6 (p<0.01). Furthermore, an association could be demonstrated between the number of residual pockets (PD>3 mm) 6 months after therapy and the number of bleeding sites and suppurating sites after hygienic phase (p=0.016). Six weeks after therapy, the mean total bacterial loads had a significant impact on the bleeding tendency of a subject at month 6 (p<0.01). Although the average numbers of sites with persisting P. gingivalis, A. actinomycetemcomitans, T. forsythensis and T. denticola seemed to be very similar 6 weeks and 6 months after therapy, large variations were noted between subjects, and therefore the microbiological status of a subject at week 6 could not predict the status at month 6. Conclusions: The present study showed a limited potential of microbiological tests, performed after hygienic phase or shortly after non-surgical periodontal therapy, to predict the clinical outcome 6 months later, but confirmed the importance of an establishment of perfect oral hygiene before non-surgical therapy. [source] | |||||||||||||