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ABSTRACT: PCR is a well-understood and established laboratory technique often used in molecular diagnostics. Huge experience has been accumulated over the last years regarding the design of PCR assays and their set-up, including in-depth troubleshooting to obtain the optimal PCR assay for each purpose. Here we report a PCR troubleshooting that came up with a surprising result never observed before. With this report we hope to sensitize the reader to this peculiar problem and to save troubleshooting efforts in similar situations, especially in time-critical and ambitious diagnostic settings.
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ABSTRACT: This study investigates the relation between overweight/obesity and self-rated health (SRH), and whether this relation varies by social factors. Data was taken from the German Health Interview and Examination Survey for Children and Adolescents (KiGGS, baseline 2003‒2006). For the definition of overweight and obesity, body mass index was calculated based on standardized height and weight measurements. SRH of adolescents (n = 6813, 11‒17 years) was raised with the question: "How would you describe your health in general?" The response categories were "very good", "good", "fair", "poor", and "very poor". We dichotomized these responses into: "very good/good" vs. "fair/poor/very poor". Socio-economic status (SES) in the family of origin and adolescents' school type were analyzed as modifying factors. Prevalence and age-adjusted odds ratios with 95% confidence intervals were calculated by binary logistic regression models. We found that overweight and obese boys and obese girls reported fair to very poor SRH more often than their normal weight peers, and that these differences were more apparent in early than late adolescence. In addition, the relation between obesity and SRH was similarly strong in all sub-groups, but there was seldom a relation between overweight and SRH. In summary, the results show that obesity is linked to poor SRH regardless of SES and school type, while the relation between overweight and SRH varies by social factors among adolescents.
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ABSTRACT: The HERV-K(HML-2) family contains the most recently integrated and best preserved endogenized proviral sequences in the human genome. All known elements have nevertheless been subjected to mutations or deletions that render expressed particles non-infectious. Moreover, these post-insertional mutations hamper the analysis of the general biological properties of this ancient virus family. The expression of consensus sequences and sequences of elements with reverted post-insertional mutations has therefore been very instrumental in overcoming this limitation. We investigated the particle morphology of a recently reconstituted HERV-K113 element termed oriHERV-K113 using thin-section electron microscopy (EM) and could demonstrate that strong overexpression by substitution of the 5'LTR for a CMV promoter and partial codon optimization altered the virus assembly type and morphology. This included a conversion from the regular C-type to an A-type morphology with a mass of cytoplasmic immature cores tethered to the cell membrane and the membranes of vesicles. Overexpression permitted the release and maturation of virions but reduced the envelope content. A weaker boost of virus expression by Staufen-1 was not sufficient to induce these morphological alterations.
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