|siRNA Screening: Development of Hit Stratification Strategies|
Žaklina Strezoska, Annaleen Vermeulen, Emily M. Anderson, Anja Smith, Devin Leake
This poster compares different approaches to hit stratification and validation after an initial screen. Standard siRNA reagents deconvoluted from a pooled set of four were compared to a pooled set of four specificity enhanced reagents. High confidence hits were similar. To explore the validity of low confidence hits, a chimeric approach was used whereby a gene-specific seed sequence was introduced into a non-targeting siRNA scaffold. This work resulted in new hit stratification strategies.
|Identification of microRNA targets using microRNA modulation techniques and gene expression arrays|
Emily M. Anderson, Maren Mayer, Kevin Sullivan, Barbara Robertson, Žaklina Strezoska, Annaleen Vermeulen, and Devin Leake
By examining the overlap of messages down-regulated by miRNA mimics and up-regulated by miRNA inhibitors, we robustly identify miRNA-regulated messages, many of which have canonical seed matches and some which are not identied by standard target prediction programs.
|Specificity and functionality of microRNA inhibitors|
Barbara Robertson, Andrew Dalby, Jon Karpilow, Anastasia Khvorova, Devin Leake and Annaleen Vermeulen
Our findings indicate that features important for natural miRNA target recognition also appear to be important for inhibitor specificity. Understanding the specificity of inhibitors allows for better interpretation of inhibitor activity in endogenous systems.
|Alternative miRNA design for therapeutic RNAi applications|
Anja van Brabant Smith, Barb Robertson, Annaleen Vermeulen, Christina Yamada, Angela Reynolds, Anastasia Khvorova, Devin Leake
For in vivo applications, the design of miRNA inhibitors and miRNA mimics must be optimized for stability and potency. However, stabilized miRNA mimic molecules can lose functionality compared to standard miRNA mimic molecules due, in part, to the activity of the stabilized passenger strand acting as a miRNA inhibitor. We discuss how mismatches affect the activity of the stabilized miRNA mimics, perhaps by generating a passenger strand that is less functional as an inhibitor molecule.
|Safety Of A Sublingual Tablet of House Dust Mite Allergen Extracts In An Environmental Exposure Chamber Study|
Michel Roux, Agnès Viatte, Robert K. Zeldin
In an environmental exposure chamber (EEC) study, treatment with HDM sublingual tablets in patients with HDM-associated allergic rhinitis was generally well tolerated regardless of dose. While asthma and related symptoms were more common during the peri-EEC challenge periods, rates did not differ between active and placebo treatment.
|Safety Of The 300 IR and 500 IR Doses Of A House Dust Mite Allergen Extracts Sublingual Tablet In Adults with Allergic Rhinitis|
Hélène Nguyen, Michel Roux, Josiane Cognet-Sicé, Robert K. Zeldin
In a randomized, DBPC study in patients with HDM-associated allergic rhinitis, treatment with HDM sublingual tablets at doses of 300IR and 500IR was associated with a favorable safety profile. There was no appreciable difference in tolerability between the tested doses.
|An Alternative to Direct Probe: Pyroprobe Coupled to High Resolution Time-of-Flight Mass Spectrometry|
Jonathan D. Byer, Joe Binkley, Karen Sam, Jeff Patrick
Direct Pyroprobe is not a substitute for direct probe, but there is some overlap, as well as additional functionality. Direct probes coupled to LECO Pegasus instruments provide unique features:
– Time-of-flight mass spectra which shows little-to-no spectral skewing
– Automatic peak finding and spectral deconvolution for better identification
– Pegasus HT has a very low maintenance source which is beneficial for direct analysis techniques
|A Reduced Workflow Solution for the Analysis of Gamma-Hydroxybutyrate (GHB) in Human Hair Samples Via an Automated Bead Mill as a Precursor to GCxGC-TOFMS and GC-HRT|
Brittany M. Watt, David E. Alonso, Joe Binkley, Jeff Patrick, Frank A. Kero, Victor Vandell, Elena Gairloch, Brad Nolt, Tom Enzweller, Rhys Jones, Lee Williams, Karen S. Scott
- The Biotage Bead Ruptor 24 was effective in pre-treating hair samples prior to the extraction procedure.
- The enhanced chromatographic resolution of GCxGC-TOFMS and high resolving power of GC-HRT minimized interferences and facilitated accurate analysis of GHB in hair.
|A FastGC Proton-Transfer-Reaction Quadrupole Ion Guide Time-of-Flight (PTR-QiTOF) Mass Spectrometer|
Alfons Jordan1, Lukas Märk1, Jens Herbig1, Christian Lindinger1, Rene Gutmann1, Lukas Fischer1, Eugen Hartungen1, Simone Jürschik1, Gernot Hanel1, Philipp Sulzer1, Tilmann D. Märk1,2
In order to identify and quantify isomeric compounds independently in real-time, we introduce a novel approach where we integrate a fast GC column (“fastGC”) into a PTR-QiTOF instrument. The outstanding sensitivity of the new PTR-QiTOF (up to 4,700 cps/ppbv) is crucial for applications where time per analysis is limited, e.g. nosespace in food and flavor research. Particularly, in these fields the chemical environment is normally very complex so that a high selectivity is essential as well.