50 research outputs found

    The Digital MIQE Guidelines Update: Minimum Information for Publication of Quantitative Digital PCR Experiments for 2020

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    Digital PCR (dPCR) has developed considerably since the publication of the Minimum Information for Publication of Digital PCR Experiments (dMIQE) guidelines in 2013, with advances in instrumentation, software, applications, and our understanding of its technological potential. Yet these developments also have associated challenges; data analysis steps, including threshold setting, can be difficult and preanalytical steps required to purify, concentrate, and modify nucleic acids can lead to measurement error. To assist independent corroboration of conclusions, comprehensive disclosure of all relevant experimental details is required. To support the community and reflect the growing use of dPCR, we present an update to dMIQE, dMIQE2020, including a simplified dMIQE table format to assist researchers in providing key experimental information and understanding of the associated experimental process. Adoption of dMIQE2020 by the scientific community will assist in standardizing experimental protocols, maximize efficient utilization of resources, and further enhance the impact of this powerful technology

    Spectroscopy for amateur astronomers: recording, processing, analysis and interpretation

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    This accessible guide presents the astrophysical concepts behind astronomical spectroscopy, covering both the theory and the practical elements of recording, processing, analysing and interpreting your spectra. It covers astronomical objects, such as stars, planets, nebulae, novae, supernovae, and events such as eclipses and comet passages. Suitable for anyone with only a little background knowledge and access to amateur-level equipment, the guide's many illustrations, sketches and figures will help you understand and practise this scientifically important and growing field of amateur astronomy, up to the level of Pro-Am collaborations. Accessible to non-academics, it benefits many groups from novices and learners in astronomy clubs, to advanced students and teachers of astrophysics. This volume is the perfect companion to the Spectral Atlas for Amateur Astronomers, which provides detailed commented spectral profiles of more than 100 astronomical objects

    Interferon-Mediated Long Non-Coding RNA Response in Macrophages in the Context of HIV.

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    Interferons play a critical role in the innate immune response against a variety of pathogens, such as HIV-1. Recent studies have shown that long non-coding genes are part of a reciprocal feedforward/feedback relationship with interferon expression. They presumably contribute to the cell type specificity of the interferon response, such as the phenotypic and functional transition of macrophages throughout the immune response. However, no comprehensive understanding exists today about the IFN-lncRNA interplay in macrophages, also a sanctuary for latent HIV-1. Therefore, we completed a poly-A+ RNAseq analysis on monocyte-derived macrophages (MDMs) treated with members of all three types of IFNs (IFN-alpha, IFN-epsilon, IFN-gamma or IFN-lambda) and on macrophages infected with HIV-1, revealing an extensive non-coding IFN and/or HIV-1 response. Moreover, co-expression correlation with mRNAs was used to identify important (long) non-coding hub genes within IFN- or HIV-1-associated gene clusters. This study identified and prioritized IFN related hub lncRNAs for further functional validation

    Replication competent virus as an important source of bias in HIV latency models utilizing single round viral constructs

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    The central memory T cell (TCM) model forms a unique HIV-1 latency model based on primary cells that closely resemble in vivo TCM. The virus employed in this model is based on an engineered vector incapable of replication after initial infection. We show that despite this strategy, replication competent viral particles are released into the culture medium due to recombination between overlapping sequences of the env deleted HIV genome that is co-transfected with intact env. This finding emphasizes the need for careful data analysis and interpretation if similar constructs are employed and urges for additional caution during laboratory work
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