383 research outputs found

    MicroRNA markers for forensic body fluid identification obtained from microarray screening and quantitative RT-PCR confirmation

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    MicroRNAs (miRNAs) are non-protein coding molecules with important regulatory functions; many have tissue-specific expression patterns. Their very small size in principle makes them less prone to degradation processes, unlike messenger RNAs (mRNAs), which were previously proposed as molecular tools for forensic body fluid identification. To identify suitable miRNA markers for forensic body fluid identification, we first screened total RNA samples derived from saliva, semen, vaginal secretion, and venous and menstrual blood for the expression of 718 human miRNAs using a microarray platform. All body fluids could be easily distinguished from each other on the basis of complete array-based miRNA expression profiles. Results from quantitative reverse transcription PCR (RT-PCR; TaqMan) assays for microarray candidate markers confirmed strong over-expression in the targeting body fluid of several miRNAs for venous blood and several others for semen. However, no candidate markers from array experiments for other body fluids such as saliva, vaginal secretion, or menstrual blood could be confirmed by RT-PCR. Time-wise degradation of venous blood and semen stains for at least 1 year under lab conditions did not significantly affect the detection sensitivity of the identified miRNA markers. The detection limit of the TaqMan assays tested for selected venous blood and semen miRNA markers required only subpicogram amounts of total RNA per single RT-PCR test, which is considerably less than usually needed for reliable mRNA RT-PCR detection. We therefore propose the application of several stable miRNA markers for the forensic identification of blood stains and several others for semen stain identification, using commercially available TaqMan assays. Additional work remains necessary in search for suitable miRNA markers for other forensically relevant body fluids

    Towards a dynamic earthquake risk framework for Switzerland

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    Scientists from different disciplines at ETH Zurich are developing a dynamic, harmonised, and user-centred earthquake risk framework for Switzerland, relying on a continuously evolving earthquake catalogue generated by the Swiss Seismological Service (SED) using the national seismic networks. This framework uses all available information to assess seismic risk at various stages and facilitates widespread dissemination and communication of the resulting information. Earthquake risk products and services include operational earthquake (loss) forecasting (OE(L)F), earthquake early warning (EEW), ShakeMaps, rapid impact assessment (RIA), structural health monitoring (SHM), and recovery and rebuilding efforts (RRE). Standardisation of products and workflows across various applications is essential for achieving broad adoption, universal recognition, and maximum synergies. In the Swiss dynamic earthquake risk framework, the harmonisation of products into seamless solutions that access the same databases, workflows, and software is a crucial component. A user-centred approach utilising quantitative and qualitative social science tools like online surveys and focus groups is a significant innovation featured in all products and services. Here we report on the key considerations and developments of the framework and its components. This paper may serve as a reference guide for other countries wishing to establish similar services for seismic risk reduction.</p

    Evidence for the Complexity of MicroRNA-Mediated Regulation in Ovarian Cancer: A Systems Approach

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    MicroRNAs (miRNAs) are short (∼22 nucleotides) regulatory RNAs that can modulate gene expression and are aberrantly expressed in many diseases including cancer. Previous studies have shown that miRNAs inhibit the translation and facilitate the degradation of their targeted messenger RNAs (mRNAs) making them attractive candidates for use in cancer therapy. However, the potential clinical utility of miRNAs in cancer therapy rests heavily upon our ability to understand and accurately predict the consequences of fluctuations in levels of miRNAs within the context of complex tumor cells. To evaluate the predictive power of current models, levels of miRNAs and their targeted mRNAs were measured in laser captured micro-dissected (LCM) ovarian cancer epithelial cells (CEPI) and compared with levels present in ovarian surface epithelial cells (OSE). We found that the predicted inverse correlation between changes in levels of miRNAs and levels of their mRNA targets held for only ∼11% of predicted target mRNAs. We demonstrate that this low inverse correlation between changes in levels of miRNAs and their target mRNAs in vivo is not merely an artifact of inaccurate miRNA target predictions but the likely consequence of indirect cellular processes that modulate the regulatory effects of miRNAs in vivo. Our findings underscore the complexities of miRNA-mediated regulation in vivo and the necessity of understanding the basis of these complexities in cancer cells before the therapeutic potential of miRNAs can be fully realized
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