|Critical factors for successful RNAi experiments in primary cells and hard to transfect cell lines|
Markus Zumbansen1, Nicole Spottke1, Sheila Offizier1, Allison St. Amand2, Devin Leake2, Ludger Altrogge1, Meike Weigel1, Sandra Domzalski1, Dietmar Lenz, and Herbert Müller-Hartmann1
The amaxa Nucleofector® Technology is a well established method for effective, non-viral transfection of any nucleic acid substrate into hard-to-transfect cells, especially suspension and primary cells. With the
Nucleofector® 96-well Shuttle® system high throughput applications such as siRNA-library screenings have become amenable for the first time in these cell types. This renders target validation and identification possible
in cell types highly relevant for biomedical research. Here we dis
|Highly Efficient High-Throughput Transfection|
Markus Zumbansen1, Allison St. Amand2, Devin Leake2, Ludger Altrogge1, and Herbert Müller-Hartmann1
Successful RNAi experiments and large-scale siRNA screens require efficient delivery of highly functional and specific nucleic acids including siRNA oligonucleotides, shRNA vectors, or micro RNAs into an appropriate cell system. Cell types relevant for immunological research, such as primary T cells and several suspension cell lines, are poorly accessible using reagent-based transfection approaches.
|Automated High Throughput Nucleofection®|
Claudia Merz, Andreas Schroers, Eric Willimann
Using primary cells for RNAi based applications such as target identification or – validation, requires a highly efficient transfection technology in combination with a reliable and robust automation system. To accomplish these requirements we integrated the amaxa 96-well Shuttle® in a Tecan Freedom EVO® cell transfection workstation which is based on Tecan’s Freedom EVO® liquid handling platform and include all the necessary components and features for unattended cell transfection.
|DOCKING STUDIES OF A NEW HETEROCYCLIC METHYLTHIOMORPHOLIN PHENOLS DERIVATIVES AS ANTIHYPERTENSIVE DRUGS WITH ACE TARGET.CASE FESCDIPINE II |
1V. Vázquez,1O. Vázquez, 1,2M.A.Balboa, 1A. Ma. Velázquez, 1V.H. Abrego, 1B.Camacho, 1L. Martínez, 1L.A. Torres, 1A. Ramírez, 1I.Martínez, 3R.López-Castañares, 1S.Díaz-Barriga, 4J. Bocanegra, 4Froylan Gómez-Laguna, 1F. Olvera, 1E.Angeles.
As part of the Drug Design in Medicinal Chemistry Program of the UNAM, new methylthiomorpholinphenol compounds with cardiovascular effect, were studied. One of them, FESCDIPINE II, was an excellent antihypertensive drug, that has low toxicity and preliminary studies indicate that the Angiotensin-converting enzyme (ACE) system is the biological target of this compound.
|Use of gamma scinitigraphy to understand inhaled device/formulation variables on delivery efficiency and|
Peter Scholes and Karen Jones
Systemic delivery of both small molecules and macromolecules via inhaled therapies is an area of significant ongoing research1. The pulmonary route offers the physiological benefits of a highly vascularised, large surface area for absorption which can promote high bioavailability and a rapid onset of action. For biomolecules such as peptides, proteins and nucleic acid derivatives, inhaled drug delivery can also provide a viable alterative to intravenous administration.
|Quantifying the Impact of a Drug on Gastric Emptying: Measuring the Pharmacodynamic Effect in Clinical Trials|
A Rankin, M Paterson, A Connor
Many drug classes are known to alter the rate of gastric emptying. Whilst there is no specific regulatory guidance requiring the impact of drugs on GE to be measured, it is important to fully understand the mode of action and the relationship between the pharmacokinetic profile and the pharmacodynamic response.
|Building Flexibility ino Phase I Protocols and Early Clinical Development Programs|
Lloyd Stevens and Gareth King
The transition of a drug candidate into Phase I and other early drug development programs is undergoing considerable examination and change. This has largely been brought about by commercial and scientific drivers to reduce attrition rates coupled with an evolving regulatory environment, all of which encourage the pharmaceutical industry to build both scientific focus and flexibility into the drug development program.
|Evaluation of Human Regional Bioavailability to Assess Whether Modified Release Development is Feasible|
A Connor, G King and K Jones
Many modified release (MR) oral formulations rely on bioavailability from the distal regions of the gastrointestinal (GI) tract (i.e. the ileum and colon). Therefore, by assessing the bioavailability of a drug following delivery to the distal intestines, it is possible to determine whether MR formulation development is achievable.
|Diagnostic Significance of microRNA Tumor Biomarkers in Fine Needle Aspirates (FNA) of Pancreatic Cancer|
Martina Doleshal 1, Anna E. Szafranska 1, Hayward S. Edmunds 2, Gregory J. Tsongalis 2, Murray Korc 2 and Emmanuel Labourier 1
Collection of pancreatic fine needle aspirates (FNAs) in RNARetain™ allows stabilization and protection of RNA in tissues containing high levels of nucleases; Differential miRNA expression in pancreatic FNAs accurately classifies benign and malignant tissue; Combinations of miRNA and mRNA expression signatures improve the separation between normal tissue and chronic pancreatitis; microRNAs are suitable analytes for molecular characterization of fine needle aspirates from pancreatic tissues.