Comparison of two different real time PCR assays and chemistry for detection of norovirus genogroup II. Method of choice? Toplak Nataša, Zimšek Mijovski Janet, Kovac Minka, Poljšak-Prijatelj MatejaNoroviruses (Caliciviridae) are one of the main cause of acute non-bacterial gastroenteritis in humans. Noroviruses can not be propagated in cell cultures. Thus Real Time PCR molecular methods for caliciviral detection have been recently introduced. The aim of the study was to compare two different Real Time PCR assays for detection of norovirus strains (genogroup II) and the comparison between different RT-PCR reagents and Real Time PCR instruments was made. |  | |
Comparison of two different real time PCR assays and chemistry for detection of norovirus genogroup II. Method of choice? Toplak Nataša, Zimšek Mijovski Janet, Kovac Minka, Poljšak-Prijatelj MatejaNoroviruses (Caliciviridae) are one of the main cause of acute non-bacterial gastroenteritis in humans. Noroviruses can not be propagated in cell cultures. Thus Real Time PCR molecular methods for caliciviral detection have been recently introduced. The aim of the study was to compare two different Real Time PCR assays for detection of norovirus strains (genogroup II) and the comparison between different RT-PCR reagents and Real Time PCR instruments was made. |  | |
qPCRas a primary screen in drug discovery GauravJaggi, Frank Boeckler, Andreas Joerger and Alan FershtWe report the use of qPCR technique to follow the thermal unfolding of proteins by the binding of the dye SYPRO Orange, and exploit its potential as a robust and high-throughput primary screen for small molecule drug discovery. |  | |
Targeting Inflammatory Cytokines Using Adenoviruses: gene delivery of biological therapies in ovarian cancer Michael A. Salako, Hagen Kulbe, Iain A. McNeish, Frances R. BalkwillConstitutive TNF-alpha expression is characteristic of the malignant ovarian surface epithelium. Adenoviral mutants hold great promise as gene therapy vectors but their efficacy is hindered by an inflammatory cascade orchestrated by TNF-alpha. We found that delivering TNF-alpha shRNA to ovarian cancer cells using oncolytic adenoviruses could reduce the inflammatory cascade generated by adenoviruses and also had direct anti-tumour activity on the cancer cells. |  | |
Efficient downregulation of the lung liquid clearing gammaENaC subunit by RNAi Nihal Yueksekdag, Marei Drechsel, Christa Schmidt, and Josef RoseneckerCF is caused by mutations in the gene encoding for CFTR. CFTR functions as chloride channel on the apical membrane of epithelia thereby regulating the transport of chloride and also sodium ions indirectly. It seems that the regulation of ENaC fails due to the mutated CFTR protein. And it is assumed that ENaC plays a role in the pathogenesis of chronic lung disease in CF-patients. |  | |
Hyperbaric Bioreactors use with Yarrowia Lipolytica Cultures: Cellular Adaptation to Hyperbaric Conditions Marlene Lopes, Nelma Gomes, Manuel Mota, Isabel BeloIn this work, a pressurized bioreactor was used for Y. lipolytica batch cultivation under increased air pressure from 1 bar to 6 bar. Cell growth was strongly enhanced by the pressure rise. The increase of oxygen availability caused the induction of the antioxidant enzyme SOD. The pre-growth of Y. lipolytica under increased pressure conditions did not affect the lipase production ability of the cells. |  | |
Improving the efficiency of 384 "mini-tube" technology using mosquito nanlolitre pipetting Joby Jenkins, Wayne Bowen, Rob Lewis & Chloe Milburn384-well plate “mini-tube” consumables have been developed by automation companies such as REMP and The Automation Partnership (TAP) to hold compound samples divided into single-use storage tubes. Each tube holds 40 -75 µL, and is filled, sealed, stored and then thawed just once before use. However, when accessed by conventional pipetting technology, the tube’s practical working volume is as little as 20 µL, which means high compound wastage. The seal must also be pierced prior to pipetting to p |  | |
Application of genetic programming in analysis of quantitative gene expression profiles for identification of nodal status in bladder cancer Anirban P. Mitra, Arpit A. Almal, Ben George, David W. Fry, Peter F. Lenehan, Vincenzo Pagliarulo, Richard J. Cote, Ram H. Datar, William P. WorzelNodal involvement in bladder cancer is an independent indicator of prognosis. This study employed an iterative machine learning process called genetic programming on quantitative expression values of 70 genes to classify primary urothelial carcinoma samples into those associated with or without nodal metastasis. The generated rules showed a strong predilection for ICAM1, MAP2K6 and KDR resulting in gene expression motifs that cumulatively suggested a pattern ICAM1>MAP2K6>KDR for node positive ca |  | |
How to Perform High Throughput siRNA Transfection Mark Hewitson, Hanno Hermann and Neil BennWe demonstrate the optimization of siRNA transfection process for time and costs in a high throughput application with two independent measurements of the RNAi effect: The visual monitoring of Eg5 expression and the quantitative PCR measurement of GAPDH mRNA after transfection of Hela cells. The results show, that 1 wash cycle is sufficient for the removal of any remnants of the transfection complex from disposable tips, making the tips re-usable. |  | |
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