|Nitric Oxide Decreases the Expression and Activity of the Ubiquitin-Conjugating Enzyme UbcH10|
Nick D. Tsihlis, PhD, Chris S. Oustwani, BA, Ashley K. Vavra, MD, Qun Jiang, MD and Melina R. Kibbe, MD
Nitric oxide (NO) has been shown to limit the formation of neointimal hyperplasia in animal models of arterial injury. Ubiquitination proceeds via formation of thioester bonds and NO can act to disrupt those bonds. We report that NO decreases the activity and expression of UbcH10 in vitro, and decreases the expression of UbcH10 following arterial injury in vivo. Therefore, UbcH10 may be a promising therapeutic target for inhibiting neointimal hyperplasia.
|High-Throughput Multiplexed Assay for Analysis of Hematopoietic Stem Cells Differentiation and Hematopoietic Toxicity|
Oksana Sirenko*1, Pierre Turpin1, Yen-Wen Chen1, Jayne Hesley1, Juan L. Almara2, Daniel Zimmerman2, David Novo2, H. Roger Tang1, and Evan F. Cromwell1
Hematopoietic stem cells (HSCs) give rise to all the blood cell types and are important for cell therapy and drug development. During the development of lymphoid and myeloid lineages, HSC differentiate into committed hematopoietic progenitors. Monitoring the expansion and differentiation of HSCs into lineage-commited hematopoietic progenitors is important for research of hematopoiesis and developing therspeutic processes with HSCs
|The Transcreener® ADP2 Universal Kinase Assay from BellBrook Labs is readily performed on BMG LABTECH microplate readers using different assay formats|
The Transcreener® ADP2 FI assay kit from BellBrook Labs is a simple one-step competitive red fluorescence immunoassay based on the detection of ADP. In this application note we show that this assay is compatible with four different microplate readers from BMG LABTECH. With the PHERAstar FS and Plus, as well as the POLARstar and FLUOstar Omegas comparable standard curves and EC50 values were obtained.
|Predicting hepatotoxicity: Reactive metabolite trapping using glutathione and freshly isolated hepatocytes|
Birks, V., Webber, G., Geoffroy, S., Cole, R., and Wood, S.
This poster presents our results to date using clozapine (a compound known to be associated with GSH-adduct formation) as substrate and using stable isotope GSH (GSH13C2,15N) to enhance specificity. In addition, all analyses have been conducted using an Waters Acquity UPLC-MS/MS. Results we have obtained in hepatocytes are compared against findings using human liver microsomes (HLM).
|In-silico Prediction of Human Intestinal Absorption and human oral bioavailability|
Human intestinal absorption (HIA) and human oral bioavailability are two important ADME properties in drug design. We built some in-silico models for classification of human intestinal absorption (HIA) and human oral bioavailability by Kohonen’s self-organizing Neural Network (KohNN), and several quantitative models for prediction of HIA and bioavailability of several subset compounds datasets by Support Vector Machine (SVM).
|Label-Free High Throughput Homogeneous ADME Assays|
David Ricketts, Patricia De Pril, Anne Van Hoonacker, Patrick Englebienne
SoPRano is a novel label-free screening platform using specifically derivatised gold nanoparticles in solution form. It enables SPR-based screening in high throughput using simple homogenous assay protocols on a standard absorbance plate-reader. SoPRano is flexible and can be applied to many affinity screening applications.
|Homology modeling and docking studies of Comamonas testosteroni B-356 biphenyl-2, 3-dioxygenase involved in degradation of polychlorinated biphenyls|
M. Saqib Baig and N. Manickam
The three-dimensional model of a-subunit of Biphenyl dioxygenase (BphA) from Comamonas testosteroni B-356 has been constructed. The kinetic parameters of biphenyl compounds were well matched with the docking results. The binding properties of these biphenyl compounds along with identification of critical active site residues could be used for further site-directed mutagenesis experiments in order to identify their role in activity and substrate specificity.
|Anti-inflammatory activity of Dalbergia sissoo Roxb. heart-wood|
Ohm Devani, S. K. Lahiri, D. D. Santani, M. B. Shah
The heart-wood of Dalbergia sissoo Roxb. is reported to be used in traditional medicinal practices, of which anti-inflammatory is one. The present work evaluates the effect of the heart-wood extract on the carrageenan induced rat paw edema. The drug was found to be active at lowest of 3 doses tried. the drug can be said to posses anti-inflamatory activity but needs further evaluation of discover the pathway of the same.
|Necessity of quantum mechanics for predicting binding free energy|
Ting Zhou, Danzhi Huang, Amedeo Caflisch
We demonstrate the necessity of quantum mechanics (QM) for predicting binding free energy by comparing the results of the linear interaction energy model with continuum model (LIECE) and the equivalent model with QM (QMLIECE).