European Journal of Molecular Cancer最新文献

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Preclinical In-House Validation of Commercially Available Fluorescence In-Situ Hybridization Probes Used in Diagnosis of Haematological Malignancies 用于血液恶性肿瘤诊断的市售荧光原位杂交探针的临床前内部验证

European Journal of Molecular Cancer Pub Date : 2020-03-31 DOI: 10.31487/j.ejmc.2020.01.01

D. Shetty, E. Talker, H. Jain

{"title":"Preclinical In-House Validation of Commercially Available Fluorescence In-Situ Hybridization Probes Used in Diagnosis of Haematological Malignancies","authors":"D. Shetty, E. Talker, H. Jain","doi":"10.31487/j.ejmc.2020.01.01","DOIUrl":"https://doi.org/10.31487/j.ejmc.2020.01.01","url":null,"abstract":"World Health Organization states the importance of conventional cytogenetics and FISH in hematological\u0000malignancy for accurate diagnosis, treatment and monitoring response to therapy. Most FISH probes,\u0000however, are Analyte- Specific reagents (not FDA approved) and thus an elaborate validation procedure\u0000prior to diagnostic use becomes essential. This study focuses on validating FISH probes by assessing the\u0000analyte- sensitivity, specificity, accuracy, precision and determining normal reference ranges (cut-offs).\u0000Eight probes from two different manufacturers each were validated using cytogenetically normal peripheral\u0000blood (negative controls) and leukemia positive bone marrow samples (positive controls) to determine the\u0000most suitable probe for use in a diagnostic set-up. Both the controls were cytogenetically defined before\u0000initiating the validation procedure. Alongside this, the probe constructs were studied to understand signal\u0000co-localization, size and intensity. Accuracy was determined by metaphase FISH, precision by standard\u0000deviation or inter-observer variability and analyte specificity and sensitivity using standard formulae. The\u0000cut-off or the normal reference range was derived by BETAINV function in Microsoft Excel. Based on\u0000performance characteristics and qualitative data most relevant probes were suggested for diagnostic use.\u0000Although validation procedures may differ between test centres, it should be a mandate pre-clinical practice.\u0000A validated FISH probe surges dependability on generated reports and this study presents the most\u0000rudimentary yet essential parameters in a FISH probe validation.","PeriodicalId":377302,"journal":{"name":"European Journal of Molecular Cancer","volume":"286 4 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2020-03-31","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129683932","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2

The WRAP53α gene undergoes both transcriptional and posttranscriptional regulation in response to DNA damage WRAP53α基因在DNA损伤时经历转录和转录后调控

European Journal of Molecular Cancer Pub Date : 2018-09-01 DOI: 10.31487/j.ejmc.2018.01.004

D. Reisman

{"title":"The WRAP53α gene undergoes both transcriptional and posttranscriptional regulation in response to DNA damage","authors":"D. Reisman","doi":"10.31487/j.ejmc.2018.01.004","DOIUrl":"https://doi.org/10.31487/j.ejmc.2018.01.004","url":null,"abstract":"The Wrap53α mRNA transcript regulates expression of the p53 tumor suppressor gene by binding to the 5'untranslated region of the p53 mRNA transcript. The binding of Wrap53α mRNA, which we demonstrate here is induced in response to a variety of DNA damaging agents, stimulates translation of the p53 mRNA, which increases the levels of active p53 protein in the cell. This allows the cell to respond to DNA damage through a p53-mediated cell cycle arrest or apoptosis. In order to determine whether the Wrap53α gene is regulated at the transcriptional and/or post-transcriptional level we carried out two sets of experiments. In one, we cloned a region of the Wrap53α gene predicted to carry the promoter and transcriptional regulatory elements of Wrap53α and tested for alterations in its activity. In addition, we carried out a series of experiments designed to measure the stability of the Wrap53α mRNA. Our results indicate that while there is a clear transcriptional response to treatment of cells with agents that damage DNA, some treatments also give rise to a post-transcriptional response leading to changes in mRNA stability. © 2018 David Reisman. Hosting by Science Repository. All rights reserved.","PeriodicalId":377302,"journal":{"name":"European Journal of Molecular Cancer","volume":"44 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2018-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"123791826","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 2