|Potential Use of RNAi on Human Skin|
Christine Collin-Djangoné, Florent Sahuc, Béatrice Bertino, Anne de Brugerolle, Jean-Eric Baudouin, Peggy Sextius, Séverine Teluob, Rachid Boulgana, Dmitry Samarsky, Peggy Tailor, Peter Welch and Yann Mahé
We designed highly efficient StealthTM RNAi targeting human tyrosinase with optimum IC50 in the 10-100 pM range. By silencing tyrosinase in melanocytes, we achieved the reconstruction of an artificial epidermis showing a long term lasting (>30 days) inhibition of melanogenesis. Since skin pigmentation can phenotypically be monitored, the use of StealthTM RNAi against tyrosinase on reconstructed pigmented epidermis is a precious tool for the evaluation of delivery vehicles.
|A Novel Multiplexed Digital Gene Expression Technology|
Gary K. Geiss1,#, Roger Bumgarner2, Brian Birditt1, Timothy Dahl1, Naeem Dowidar1, Dwayne L. Dunaway1, H. Perry Fell1, Sean Ferree1, Renee D. George1, Tammy Grogan1, Jeffrey J. James1, Malini Maysuria1, Jeffrey D. Mitton1, Paola Oliveri4, Jennifer L. Osborn3, Tao Peng2, Amber L. Ratcliffe1, Philippa J. Webster1, Eric H. Davidson4, and Leroy Hood5
We describe a novel platform, the nCounter Analysis System, for sensitive, highly multiplexed, digital gene expression analysis based on NanoString’s novel molecular barcoding technology. Detection of individual mRNA molecules using an assigned sequence of six different fluorescent spots per probe are detected, and then the number of times that code sequence appears in a sample are counted.
|Biosynthesis of Very Long Chain Polyunsaturated Fatty Acids in Chicory|
Hattem Mekky1, Michael R. Davey1, J. Brian Power1 , Maged E. Mohamed2 and Colin M. Lazarus2
Very long chain polyunsaturated fatty acids (VLCPUFAs) are declining in reserves, since they are produced by oily fish. VLCPUFAs are important in human nutrition because they are responsible for the production of inflammatory mediators such as prostaglandins, the regulation of cholesterol synthesis and its transport, and the maintenance of cellular membranes. This research aims to produce VLCPUFAs in edible plants.
|Targeting Inflammatory Cytokines Using Adenoviruses: gene delivery of biological therapies in ovarian cancer|
Michael A. Salako, Hagen Kulbe, Iain A. McNeish, Frances R. Balkwill
Constitutive 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.
|Evolution of the Contact Phase of Vertebrate Blood Coagulation|
M. B. Ponczek, D. Gailani and R. F. Doolittle
Three proteases involved in the contact phase of vertebrate blood coagulation were searched in genomes of amphibians, birds and mammals. Several data sources were used during this bioinformatics analysis. Our survey confirmed that the vertebrate contact phase has occurred by way of a series of gene duplications during the course of several hundred million years of evolution, beginning with the appearance of fXII and a PK-fXI predecessor in amphibians.
|Bayesian Parameter Estimation for Systems Biological Models of Dynamic Cellular Networks|
Jarad Niemi, Lingchong You and Mike West
Dynamic cellular networks determine production of proteins in the cells in our body. The Rb-E2F network is a key example: it controls cell cycle, proliferation and degradation - a key network in almost all cancers. We develop statistical models of such single-cell processes - fine-time scale dynamic network models. Model estimation requires informative priors and custom Metropolis proposals.
|Hyperbaric Bioreactors use with Yarrowia Lipolytica Cultures: Cellular Adaptation to Hyperbaric Conditions|
Marlene Lopes, Nelma Gomes, Manuel Mota, Isabel Belo
In 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.
|Treatment of Highland Frogs from the Two Legged Stage with Homeopathically Prepared Thyroxin (10-11 – 10-21)|
Gerhard Lingg1, P. Christian Endler1,*, Michael Frass1, and Harald Lothaller2
For the experiment presented here, we used the same setup as for the initial multilaboratory study[1,2,3,4], but a special preparation process of the homeopathic drug involving dilution steps of 1: 50,000 and leading to thyroxin 10–11 – 10–21 was used.
|A high-throughput protein array-based approach |
Rachel Van Dyk, Claudia Kirisits, Paul Potter, Fook Tim Chew, Reinhard Hiller
Here we present a novel discovery-oriented high-throughput approach to the screening of allergen reactivates in crude biological extracts is presented using seafood allergens as an example. The CPGR workflow permits the effective screening of hundreds of putative allergens in parallel using minute amounts of patient serum and constitutes a cost-efficient allergen-specific antibody screening method for a routine diagnostic setting.