Surgical plume resulting from routine LEEPs for HSIL of the cervix uteri has the risk of contamination with high-risk HPV. Further investigations of infectiousness of surgical plume are necessary for evaluation of potential hazards to involved healthcare professionals.
We present a disposable microarray hybridization chamber with an integrated micropump to speed up diffusion based reaction kinetics by generating convective flow. The time-to-result for the hybridization reaction was reduced from 60 min (standard protocol) down to 15 min for a commercially available microarray. The integrated displacement micropump is pneumatically actuated. It includes two active microvalves and is designed for low-cost, high volume manufacturing. The setup is made out of two microstructured polymer parts realized in polycarbonate (PC) separated by a 25 μm thermoplastic elastomer (TPE) membrane. Pump rate can be controlled between 0.3 μl s(-1) and 5.7 μl s(-1) at actuation frequencies between 0.2 Hz and 8.0 Hz, respectively.
The expression of genes involved in C4 photosynthesis in maize is under tight tissue-specific
and light-dependent control. There is strong evidence that this control is at least in part brought about by
DOF transcription factors binding to the respective promoters. We analyzed the interaction of DOF1 and
DOF2 proteins with a functional and a cryptic endogenous binding site derived from the maize
phosphoenolpyruvate carboxylase promoter (−300 bp region) in the nucleosomal context. Various DNA
fragments comprising this promoter region were reconstituted into mononucleosomes from purified
components, resulting in different positions of the DOF binding sites on the nucleosome surface. Binding
of recombinant transcription factors to the different types of nucleosomes was examined using
electrophoretic mobility shift assays. Changing the translational position of the binding site on the
nucleosome surface strongly affected the efficiency of the interaction with the DOF factors. Deletion of
individual recognition motifs revealed a positive impact of DOF protein binding to the main binding site
on interactions with the cryptic binding site. The addition of the chromosomal high-mobility group (HMG)
protein HMGB5 to the binding reaction mixture facilitated nucleosome binding of the transcription factor
independent from the position of the recognition sites. The relevance of the data for the activation of the
promoter in vivo is discussed.
Summary
Background and objectives
Onychomycosis (OM) and tinea pedis (TP) are common fungal infections. Currently, diagnosis is based on direct microscopy and culture that have a low to moderate sensitivity and/or require up to 3–4 weeks until results are obtained. PCR techniques have emerged for the diagnosis of fungal infections, but little is known about their sensitivity and specificity in diagnosing. Here, we compared the diagnostic value of a DNA‐chip technology, that detects 56 fungal pathogens, in a single‐center prospective diagnostic study with microscopy and culture in suspected OM/TP.
Patients and methods
Microscopy, culture and DNA microarray assays were performed on scraping material from patients with suspected OM (n = 67) or TP (n = 73). To test whether swabs can be used as an alternative for scraping, PCR yields were compared in a further 13 patients with OM and 11 patients with TP.
Results
DNA microarrays had the highest sensitivity. Combination of DNA‐chip technology with microscopy further increased the sensitivity, and results from this combined laboratory diagnosis can be obtained within 24 hours. Comparison of sampling techniques (scraping, dry or wet swab) for DNA‐chip assays showed similar results in suspected OM or TP.
Conclusions
DNA‐chip technology shows high sensitivity for OM and TP diagnosis, especially when combined with microscopy.
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