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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.

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LC-MS Approaches to Profiling of Non-Radiolabelled Metabolites in Response to Recent Regulatory Changes
Richard Clayton, Caroline Anderson, Brian Morrison and Lindsay Corfield

Following publication of the FDA MIST guidelines and revision of ICH M3, there is increasing interest in obtaining metabolic profiling data at an early stage in the development of a drug. This has led to a requirement to estimate the relative abundance of metabolites in samples prior to the synthesis of the radiolabelled compound and from a wider range of studies.

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Fast PK and Semi Quantitative Analysis of Metabolites Using High Resolution MS
Rohan A. Thakur and Mike Koleto

It has become essential to eliminate poor candidates as early as possible in the drug development process. This realization led to the application of high throughput principles to the in vivo Lead Optimization process, wherein the NCE is first given to rodents to determine the PK profile in a rapid and limited study. This rapid primary screen is known as Fast PK and several critical decisions are based on this initial study.

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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

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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.

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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).

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In-silico Prediction of Human Intestinal Absorption and human oral bioavailability
Aixia Yan*

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).

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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.

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Mechanisms of beneficial effects of human placenta MSC transplantation in rats with experimental ischemic stroke
Yarygin K.N., Kholodenko I.V., Konieva A.A., Burunova V.V., Tairova R.T., Gubsky L.V., Cheglakov I.B., PirogovYu.A., Yarygin V.N., Skvortsova V.I.

Human MSC transplantation significantly reduced the volume of experimental ischemic lesion in rat model. It also greatly stimulated stem and progenitor cell proliferation in the subventricular region and hippocampus and cell migration from these areas towards the ischemia site. Beneficial effects of MSC in rats with brain ischemia are likely to be associated with enhancement of host neural stem and progenitor cell proliferation and migration towards the affected area.

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Scientific News
High Throughput Mass Spectrometry-Based Screening Assay Trends
Dr John Comley provides an insight into HT MS-based screening with a focus on future user requirements and preferences.
Non-Disease Proteins Kill Brain Cells
Scientists at the forefront of cutting-edge research into neurodegenerative diseases such as Alzheimer’s and Parkinson’s have shown that the mere presence of protein aggregates may be as important as their form and identity in inducing cell death in brain tissue.
Potential Treatment for Life-Threatening Viral Infections Revealed
The findings point to new therapies for Dengue, West Nile and Ebola.
Gut Microbes Signal to the Brain When They're Full
Don't have room for dessert? The bacteria in your gut may be telling you something.
Personalized Drug Screening for Multiple Myeloma Patients
A personalized method for testing the effectiveness of drugs that treat multiple myeloma may predict quickly and more accurately the best treatments for individual patients with the bone marrow cancer.
Nanocarriers May Carry New Hope for Brain Cancer Therapy
Berkeley lab researchers develop nanoparticles that can carry therapeutics across the brain blood barrier.
Cancer-Fighting Tomato Component Traced
The metabolic pathway associated with lycopene, the bioactive red pigment found in tomatoes, has been traced by researchers at the University of Illinois.
Batten Disease may Benefit from Gene Therapy
NIH-funded animal study suggests one-shot approach to injecting genes.
Shedding Light on “Dark” Cellular Receptors
UNC and UCSF labs create a new research tool to find homes for two orphan cell-surface receptors, a crucial step toward finding better therapeutics and causes of drug side effects.
Molecule Proves Key to Brain Repair After Stroke
Scientists found that a molecule known as growth and differentiation factor 10 (GDF10) plays a key role in repair mechanisms following stroke.

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