1,716 results match your criteria Journal of Biomolecular Screening [Journal]


Functional Comparison of Neuronal Cells Differentiated from Human Induced Pluripotent Stem Cell-Derived Neural Stem Cells under Different Oxygen and Medium Conditions.

J Biomol Screen 2016 Dec 19;21(10):1054-1064. Epub 2016 Aug 19.

2 Department of Physiology, School of Medicine, Keio University, Shinjuku-ku, Tokyo, Japan.

Because neurons are difficult to obtain from humans, generating functional neurons from human induced pluripotent stem cells (hiPSCs) is important for establishing physiological or disease-relevant screening systems for drug discovery. To examine the culture conditions leading to efficient differentiation of functional neural cells, we investigated the effects of oxygen stress (2% or 20% O) and differentiation medium (DMEM/F12:Neurobasal-based [DN] or commercial [PhoenixSongs Biologicals; PS]) on the expression of genes related to neural differentiation, glutamate receptor function, and the formation of networks of neurons differentiated from hiPSCs (201B7) via long-term self-renewing neuroepithelial-like stem (lt-NES) cells. Expression of genes related to neural differentiation occurred more quickly in PS and/or 2% O than in DN and/or 20% O, resulting in high responsiveness of neural cells to glutamate, N-methyl-d-aspartate (NMDA), α-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA), and ( S)-3,5-dihydroxyphenylglycine (an agonist for mGluR), as revealed by calcium imaging assays. Read More

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http://dx.doi.org/10.1177/1087057116661291DOI Listing
December 2016
45 Reads

Development of an In Vitro Model to Screen CYP1B1-Targeted Anticancer Prodrugs.

J Biomol Screen 2016 Dec;21(10):1090-1099

1 School of Pharmacy, Department of Pharmaceutical Chemistry, University of Kansas, Lawrence, KS, USA.

Cytochrome P450 1B1 (CYP1B1) is an anticancer therapeutic target due to its overexpression in a number of steroid hormone-related cancers. One anticancer drug discovery strategy is to develop prodrugs specifically activated by CYP1B1 in malignant tissues to cytotoxic metabolites. Here, we aimed to develop an in vitro screening model for CYP1B1-targeted anticancer prodrugs using the KLE human endometrial carcinoma cell line. Read More

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http://dx.doi.org/10.1177/1087057116675315DOI Listing
December 2016
34 Reads

High-Throughput Platform for Identifying Molecular Factors Involved in Phenotypic Stabilization of Primary Human Hepatocytes In Vitro.

J Biomol Screen 2016 Oct 4;21(9):897-911. Epub 2016 Aug 4.

Harvard-MIT Division of Health Sciences and Technology, MIT, Cambridge, MA, USA The Broad Institute of MIT and Harvard, Cambridge, MA, USA Department of Medicine, Brigham and Women's Hospital, Boston, MA, USA Institute for Medical Engineering and Science, MIT, Cambridge, MA, USA Department of Electrical Engineering and Computer Science, MIT, Cambridge, MA, USA David H. Koch Institute for Integrative Cancer Research, MIT, Cambridge, MA, USA Howard Hughes Medical Institute, Chevy Chase, MD, USA

Liver disease is a leading cause of morbidity worldwide and treatment options are limited, with organ transplantation being the only form of definitive management. Cell-based therapies have long held promise as alternatives to whole-organ transplantation but have been hindered by the rapid loss of liver-specific functions over a period of days in cultured hepatocytes. Hypothesis-driven studies have identified a handful of factors that modulate hepatocyte functions in vitro, but our understanding of the mechanisms involved remains incomplete. Read More

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http://dx.doi.org/10.1177/1087057116660277DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC6352718PMC
October 2016
21 Reads

SLAS Europe High-Content Screening Conference in Dresden: A Glimpse of the Future?

J Biomol Screen 2016 Oct;21(9):883-6

Max Planck Institute of Molecular Cell Biology and Genetics, Dresden, Germany

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http://dx.doi.org/10.1177/1087057116662825DOI Listing
October 2016
21 Reads

One-Step Seeding of Neural Stem Cells with Vitronectin-Supplemented Medium for High-Throughput Screening Assays.

J Biomol Screen 2016 Dec 26;21(10):1112-1124. Epub 2016 Sep 26.

1 National Center for Advancing Translational Sciences, National Institutes of Health, Bethesda, MD, USA.

Human neuronal cells differentiated from induced pluripotent cells have emerged as a new model system for the study of disease pathophysiology and evaluation of drug efficacy. Differentiated neuronal cells are more similar in genetics and biological content to human brain cells than other animal disease models. However, culture of neuronal cells in assay plates requires a labor-intensive procedure of plate precoating, hampering its applications in high-throughput screening (HTS). Read More

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http://dx.doi.org/10.1177/1087057116670068DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5285267PMC
December 2016
17 Reads

Matrix-Based Activity Pattern Classification as a Novel Method for the Characterization of Enzyme Inhibitors Derived from High-Throughput Screening.

J Biomol Screen 2016 Dec 27;21(10):1075-1089. Epub 2016 Sep 27.

1 Novartis Institutes for Biomedical Research, Center for Proteomic Chemistry, Cambridge, MA, USA.

One of the central questions in the characterization of enzyme inhibitors is determining the mode of inhibition (MOI). Classically, this is done with a number of low-throughput methods in which inhibition models are fitted to the data. The ability to rapidly characterize the MOI for inhibitors arising from high-throughput screening in which hundreds to thousands of primary inhibitors may need to be characterized would greatly help in lead selection efforts. Read More

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http://dx.doi.org/10.1177/1087057116667255DOI Listing
December 2016
14 Reads

Functional Characterization of Acetylcholine Receptors Expressed in Human Neurons Differentiated from Hippocampal Neural Stem/Progenitor Cells.

J Biomol Screen 2016 Dec 27;21(10):1065-1074. Epub 2016 Sep 27.

2 Department of Genomics-Based Drug Discovery, Doctoral Program in Clinical Sciences, Graduate School of Comprehensive Human Science, University of Tsukuba, Tsukuba, Ibaraki, Japan.

Neurotransmission mediated by acetylcholine receptors (AChRs) plays an important role in learning and memory functions in the hippocampus. Impairment of the cholinergic system contributes to Alzheimer's disease (AD), indicating the importance of AChRs as drug targets for AD. To improve the success rates for AD drug development, human cell models that mimic the target brain region are important. Read More

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http://journals.sagepub.com/doi/10.1177/1087057116665567
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http://dx.doi.org/10.1177/1087057116665567DOI Listing
December 2016
5 Reads

Development of a High-Throughput Gene Expression Screen for Modulators of RAS-MAPK Signaling in a Mutant RAS Cellular Context.

J Biomol Screen 2016 Oct 26;21(9):989-97. Epub 2016 Jul 26.

Oncology, Merck & Co. Inc., Boston, MA, USA.

The RAS-MAPK pathway controls many cellular programs, including cell proliferation, differentiation, and apoptosis. In colorectal cancers, recurrent mutations in this pathway often lead to increased cell signaling that may contribute to the development of neoplasms, thereby making this pathway attractive for therapeutic intervention. To this end, we developed a 26-member gene signature of RAS-MAPK pathway activity utilizing the Affymetrix QuantiGene Plex 2. Read More

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http://dx.doi.org/10.1177/1087057116658646DOI Listing
October 2016
39 Reads

Rethinking Nuclear Receptors as Potential Therapeutic Targets for Retinal Diseases.

J Biomol Screen 2016 Dec 28;21(10):1007-1018. Epub 2016 Jul 28.

1 Department of Ophthalmology, Duke University School of Medicine, Durham, NC, USA.

Collectively, retinal diseases, including age-related macular degeneration, retinitis pigmentosa, and diabetic retinopathy, result in severe vision impairment worldwide. The absence and/or limited availability of successful drug therapies for these blinding disorders necessitates further understanding their pathobiology and identifying new targetable signaling pathways. Nuclear receptors are transcription regulators of many key aspects of human physiology, as well as pathophysiology, with reported roles in development, aging, and disease. Read More

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http://dx.doi.org/10.1177/1087057116659856DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5586077PMC
December 2016
6 Reads

Combining Unique Multiplex Gateway Cloning and Bimolecular Fluorescence Complementation (BiFC) for High-Throughput Screening of Protein-Protein Interactions.

J Biomol Screen 2016 Dec 28;21(10):1100-1111. Epub 2016 Jul 28.

1 Department for Molecular Medicine, Ruđer Bošković Institute, Zagreb, Croatia.

Protein interaction networks are the basis for human metabolic and signaling systems. Interaction studies often use bimolecular fluorescence complementation (BiFC) to reveal the formation and cellular localization of protein complexes. However, large-scale studies were either far from native conditions in human cells or limited by laborious restriction/ligation cloning techniques. Read More

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http://dx.doi.org/10.1177/1087057116659438DOI Listing
December 2016
8 Reads

Erratum.

Authors:

J Biomol Screen 2016 Aug;21(7):766

The guest editors of the June 2016 special issue of the Journal of Biomolecular Screening (Vol. 21, No. 5) (JBS) on "Innovative Screening Methodologies to Identify New Compounds for the Treatment of Central Nervous System Disorders" were not acknowledged in the final published issue. Read More

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http://dx.doi.org/10.1177/1087057116659074DOI Listing
August 2016
5 Reads

Development of a 3D Tissue Culture-Based High-Content Screening Platform That Uses Phenotypic Profiling to Discriminate Selective Inhibitors of Receptor Tyrosine Kinases.

J Biomol Screen 2016 Oct 7;21(9):912-22. Epub 2016 Jul 7.

Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands OcellO B.V., Leiden, The Netherlands

3D tissue cultures provide a more physiologically relevant context for the screening of compounds, compared with 2D cell cultures. Cells cultured in 3D hydrogels also show complex phenotypes, increasing the scope for phenotypic profiling. Here we describe a high-content screening platform that uses invasive human prostate cancer cells cultured in 3D in standard 384-well assay plates to study the activity of potential therapeutic small molecules and antibody biologics. Read More

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5030728PMC
http://dx.doi.org/10.1177/1087057116657269DOI Listing
October 2016
52 Reads
5 Citations
2.423 Impact Factor

In Vitro Assays for the Discovery of PCSK9 Autoprocessing Inhibitors.

J Biomol Screen 2016 Dec 11;21(10):1034-1041. Epub 2016 Jul 11.

3 Department of Cardiometabolic Diseases, Merck Research Laboratories, Kenilworth, NJ, USA.

PCSK9 plays a significant role in regulating low-density lipoprotein (LDL) cholesterol levels and has become an important drug target for treating hypercholesterolemia. Although a member of the serine protease family, PCSK9 only catalyzes a single reaction, the autocleavage of its prodomain. The maturation of the proprotein is an essential prerequisite for the secretion of PCSK9 to the extracellular space where it binds the LDL receptor and targets it for degradation. Read More

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http://dx.doi.org/10.1177/1087057116657312DOI Listing
December 2016
10 Reads

Identification of Positive Allosteric Modulators of Glycine Receptors from a High-Throughput Screen Using a Fluorescent Membrane Potential Assay.

J Biomol Screen 2016 Dec 11;21(10):1042-1053. Epub 2016 Jul 11.

1 Neusentis (Pfizer Ltd.), Granta Park, Great Abington, Cambridgeshire, UK.

Glycine receptor 3 (GlyRα3) is a ligand-gated ion channel of the cys-loop family that plays a key role in mediating inhibitory neurotransmission and regulation of pain signaling in the dorsal horn. Potentiation of GlyRα3 function is therefore of interest as a putative analgesic mechanism with which to target new therapeutics. However, to date, positive allosteric modulators (PAMs) of this receptor with sufficient selectivity to enable target validation studies have not been described. Read More

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http://dx.doi.org/10.1177/1087057116657779DOI Listing
December 2016
16 Reads

Early Perspective.

J Biomol Screen 2016 Dec 20;21(10):1019-1033. Epub 2016 Jul 20.

1 Chemical Biology Consortium Sweden, Science for Life Laboratories, Stockholm, Sweden.

The cellular thermal shift assay (CETSA) was introduced in 2013 as a means to assess drug binding in complex environments such as cell lysates, live cells, and even tissues. The assay principle relies on the well-proven biophysical concept of ligand-induced thermal stabilization of proteins, which in CETSA applications is measured as a persistent presence of soluble protein at elevated temperatures. Given its recent development, we have just started to learn about the benefits and pitfalls of the method as it is applied to a growing number of protein target classes, the majority of which are intracellular soluble proteins. Read More

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http://dx.doi.org/10.1177/1087057116659256DOI Listing
December 2016
3 Reads

The Use of Nucleosome Substrates Improves Binding of SAM Analogs to SETD8.

J Biomol Screen 2016 Sep 1;21(8):786-94. Epub 2016 Jul 1.

Lilly Research Laboratories, Eli Lilly and Company, Lilly Corporate Center, Indianapolis, IN, USA.

SETD8 is the methyltransferase responsible for monomethylation of lysine at position 20 of the N-terminus of histone H4 (H4K20). This activity has been implicated in both DNA damage and cell cycle progression. Existing biochemical assays have utilized truncated enzymes containing the SET domain of SETD8 and peptide substrates. Read More

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http://dx.doi.org/10.1177/1087057116656596DOI Listing
September 2016
11 Reads

Identification of Novel Inhibitors of the Type I Interferon Induction Pathway Using Cell-Based High-Throughput Screening.

J Biomol Screen 2016 Oct 29;21(9):978-88. Epub 2016 Jun 29.

School of Biology, University of St Andrews, St Andrews, Fife, UK Biomedical Sciences Research Complex (BSRC), University of St Andrews, St Andrews, Fife, UK

Production of type I interferon (IFN) is an essential component of the innate immune response against invading pathogens. However, its production must be tightly regulated to avoid harmful effects. Compounds that modulate the IFN response are potentially valuable for a variety of applications due to IFN's beneficial and detrimental roles. Read More

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5030734PMC
http://dx.doi.org/10.1177/1087057116656314DOI Listing
October 2016
9 Reads

An Automatic Quality Control Pipeline for High-Throughput Screening Hit Identification.

J Biomol Screen 2016 Sep 16;21(8):832-41. Epub 2016 Jun 16.

Genomics Institute of the Novartis Research Foundation, San Diego, CA, USA

The correction or removal of signal errors in high-throughput screening (HTS) data is critical to the identification of high-quality lead candidates. Although a number of strategies have been previously developed to correct systematic errors and to remove screening artifacts, they are not universally effective and still require fair amount of human intervention. We introduce a fully automated quality control (QC) pipeline that can correct generic interplate systematic errors and remove intraplate random artifacts. Read More

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http://dx.doi.org/10.1177/1087057116654274DOI Listing
September 2016
21 Reads

Development of a High-Throughput Screening Strategy for Upregulators of the OPG/RANKL Ratio with the Potential for Antiosteoporosis Effects.

J Biomol Screen 2016 Aug 14;21(7):738-48. Epub 2016 Jun 14.

Institute of Medicinal Biotechnology, Peking Union Medical College and Chinese Academy of Medical Sciences, Beijing, China

The ratio between osteoprotegerin (OPG) and the receptor activator of NF-κB ligand (RANKL) in the bone microenvironment indicates the level of osteoclastogenesis, and upregulation of this ratio would improve osteoporosis. In this study, we established a novel high-throughput screening (HTS) system using two stably transfected monoclonal cell lines that either express firefly luciferase under the OPG promoter control or concurrently express firefly and renilla luciferases under control of the OPG and RANKL promoters, respectively. With this system, we can conveniently and rapidly detect the effects of compounds on the expression of OPG and RANKL through changes in firefly and renilla luciferase activities. Read More

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http://dx.doi.org/10.1177/1087057116654657DOI Listing
August 2016
3 Reads

A Graphene Oxide-Based Sensing Platform for the Determination of Methicillin-Resistant Staphylococcus aureus Based on Strand-Displacement Polymerization Recycling and Synchronous Fluorescent Signal Amplification.

J Biomol Screen 2016 Sep 10;21(8):851-7. Epub 2016 Jun 10.

Department of Microbiology, The Medicine School of Hunan University of Chinese Medicine, Changsha, Hunan, People's Republic of China

To develop new technology for detecting methicillin-resistant Staphylococcus aureus (MRSA), a novel fluorescent biosensor based on Klenow fragment (KF)-assisted target recycling amplification and synchronous fluorescence analysis was created. Carboxy-fluorescein (FAM)-labeled single-stranded DNA (ssDNA) containing a capture probe and a signal probe was adsorbed onto the surface of graphene oxide (GO) via π-stacking interactions, resulting in the fluorescence quenching of the dye. When target and primer were introduced, the fluorescence was restored due to P0 being completely released from the surface of the GO. Read More

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http://dx.doi.org/10.1177/1087057116653564DOI Listing
September 2016
72 Reads

Flow Cytometric Method for the Detection of Flavonoids in Cell Lines.

J Biomol Screen 2016 Sep 8;21(8):858-65. Epub 2016 Jun 8.

Department of Food Safety and Food Quality, Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium

Here, we describe an easy-to-use flow cytometric method using diphenylboric acid 2-amino ethyl ester (DPBA) stain for the detection of flavonoids in cells from human/animal origin. Flavonoid bioavailability and bioactivity depend on structure, conjugation and the cell type to which they are presented. We have studied cellular uptake of five flavonoids with different structures and conjugation forms. Read More

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http://dx.doi.org/10.1177/1087057116653220DOI Listing
September 2016
7 Reads

Identification of Potential Pharmacoperones Capable of Rescuing the Functionality of Misfolded Vasopressin 2 Receptor Involved in Nephrogenic Diabetes Insipidus.

J Biomol Screen 2016 Sep 8;21(8):824-31. Epub 2016 Jun 8.

The Scripps Research Institute Molecular Screening Center, Department of Molecular Therapeutics, Scripps Florida, Jupiter, FL, USA

Pharmacoperones correct the folding of otherwise misfolded protein mutants, restoring function (i.e., providing "rescue") by correcting their trafficking. Read More

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http://jbx.sagepub.com/content/early/2016/06/08/108705711665
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http://dx.doi.org/10.1177/1087057116653925DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5594746PMC
September 2016
60 Reads

Characterization of Kinetic Binding Properties of Unlabeled Ligands via a Preincubation Endpoint Binding Approach.

J Biomol Screen 2016 Aug 7;21(7):729-37. Epub 2016 Jun 7.

Pharmaceutical Research Division, Takeda Pharmaceutical Company Limited, Fujisawa, Kanagawa, Japan.

The dissociation rates of unlabeled drugs have been well studied by kinetic binding analyses. Since kinetic assays are laborious, we developed a simple method to determine the kinetic binding parameters of unlabeled competitors by a preincubation endpoint assay. The probe binding after preincubation of a competitor can be described by a single equation as a function of time. Read More

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http://dx.doi.org/10.1177/1087057116652065DOI Listing
August 2016
6 Reads

A Platform to Enable the Pharmacological Profiling of Small Molecules in Gel-Based Electrophoretic Mobility Shift Assays.

J Biomol Screen 2016 Dec 10;21(10):1125-1131. Epub 2016 Jul 10.

1 Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, USA.

We describe a polyacrylamide gel casting cassette that overcomes limitations of commercially available gel electrophoresis equipment. This apparatus molds a single polyacrylamide gel that can evaluate more than 200 samples in parallel, is loaded with a multichannel pipettor, and is flexible with respect to composition of the separating matrix. We demonstrate its use to characterize inhibitors of enzymes that modify protein and nucleic acid substrates. Read More

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http://dx.doi.org/10.1177/1087057116652895DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5269537PMC
December 2016
9 Reads

A Novel Automated High-Content Analysis Workflow Capturing Cell Population Dynamics from Induced Pluripotent Stem Cell Live Imaging Data.

J Biomol Screen 2016 Oct 2;21(9):887-96. Epub 2016 Jun 2.

Centre for Stem Cells and Regenerative Medicine, King's College London, Tower Wing, Guy's Hospital, London, UK.

Most image analysis pipelines rely on multiple channels per image with subcellular reference points for cell segmentation. Single-channel phase-contrast images are often problematic, especially for cells with unfavorable morphology, such as induced pluripotent stem cells (iPSCs). Live imaging poses a further challenge, because of the introduction of the dimension of time. Read More

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5030730PMC
http://dx.doi.org/10.1177/1087057116652064DOI Listing
October 2016
7 Reads

Small Molecules Revealed in a Screen Targeting Epithelial Scattering Are Inhibitors of Microtubule Polymerization.

J Biomol Screen 2016 Aug 31;21(7):671-9. Epub 2016 May 31.

Brigham Young University, Provo, UT, USA

Stimulation of cultured epithelial cells with scatter factor/hepatocyte growth factor (HGF) results in the detachment of cell-cell junctions and initiation of cell migration. Instead of coordinating collective cell behavior within a tissue, cells become solitary and have few cell-cell interactions. Since epithelial scattering is recapitulated in cancer progression and since HGF signaling drives cancer metastasis in many cases, inhibitors of HGF signaling have been proposed to act as anticancer agents. Read More

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http://dx.doi.org/10.1177/1087057116651850DOI Listing
August 2016
10 Reads

Transcriptional Inhibitors Identified in a 160,000-Compound Small-Molecule DUX4 Viability Screen.

J Biomol Screen 2016 Aug 31;21(7):680-8. Epub 2016 May 31.

Lillehei Heart Institute, University of Minnesota, Minneapolis, MN, USA Department of Pediatrics, University of Minnesota, Minneapolis, MN, USA

Facioscapulohumeral muscular dystrophy is a genetically dominant, currently untreatable muscular dystrophy. It is caused by mutations that enable expression of the normally silent DUX4 gene, which encodes a pathogenic transcription factor. A screen based on Tet-on DUX4-induced mouse myoblast death previously uncovered compounds from a 44,000-compound library that protect against DUX4 toxicity. Read More

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http://dx.doi.org/10.1177/1087057116651868DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5501317PMC
August 2016
12 Reads

Assay Establishment and Validation of a High-Throughput Screening Platform for Three-Dimensional Patient-Derived Colon Cancer Organoid Cultures.

J Biomol Screen 2016 Oct 27;21(9):931-41. Epub 2016 May 27.

Eli Lilly and Company, Lilly Research Laboratories, Quantitative Biology, Alcobendas, Madrid, Spain

The application of patient-derived three-dimensional culture systems as disease-specific drug sensitivity models has enormous potential to connect compound screening and clinical trials. However, the implementation of complex cell-based assay systems in drug discovery requires reliable and robust screening platforms. Here we describe the establishment of an automated platform in 384-well format for three-dimensional organoid cultures derived from colon cancer patients. Read More

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http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5030729PMC
http://dx.doi.org/10.1177/1087057116650965DOI Listing
October 2016
54 Reads

Reproducibility of Uniform Spheroid Formation in 384-Well Plates: The Effect of Medium Evaporation.

J Biomol Screen 2016 Oct 25;21(9):923-30. Epub 2016 May 25.

Institute of Molecular and Translational Medicine, Faculty of Medicine and Dentistry, Palacky University, Olomouc, Czech Republic

Spheroid cultures of cancer cells reproduce the spatial dimension-induced in vivo tumor traits more effectively than the conventional two-dimensional cell cultures. With growing interest in spheroids for high-throughput screening (HTS) assays, there is an increasing demand for cost-effective miniaturization of reproducible spheroids in microtiter plates (MPs). However, well-to-well variability in spheroid size, shape, and growth is a frequently encountered problem with almost every culture method that has prevented the transfer of spheroids to the HTS platform. Read More

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http://dx.doi.org/10.1177/1087057116651867DOI Listing
October 2016
5 Reads

Corrigendum.

Authors:

J Biomol Screen 2016 Jul 17;21(6):653. Epub 2016 May 17.

Pedró-Rosa, L.; Buckner, F.; Ranade, R. Read More

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http://dx.doi.org/10.1177/1087057116651590DOI Listing
July 2016
6 Reads

FRET-Protease-Coupled Peptidyl-Prolyl cis-trans Isomerase Assay: New Internally Quenched Fluorogenic Substrates for High-Throughput Screening.

J Biomol Screen 2016 Aug 16;21(7):701-12. Epub 2016 May 16.

Istituto di Biostrutture e Bioimmagini-CNR and CIRPEB, Napoli, Italy

In this work, a sensitive and convenient protease-based fluorimetric high-throughput screening (HTS) assay for determining peptidyl-prolyl cis-trans isomerase activity was developed. The assay was based on a new intramolecularly quenched substrate, whose fluorescence and structural properties were examined together with kinetic constants and the effects of solvents on its isomerization process. Pilot screens performed using the Library of Pharmacologically Active Compounds (LOPAC) and cyclophilin A (CypA), as isomerase model enzyme, indicated that the assay was robust for HTS, and that comparable results were obtained with a CypA inhibitor tested both manually and automatically. Read More

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http://dx.doi.org/10.1177/1087057116650402DOI Listing
August 2016
16 Reads

Mycobacterium Cytidylate Kinase Appears to Be an Undruggable Target.

J Biomol Screen 2016 Aug 4;21(7):695-700. Epub 2016 May 4.

Department of Medicine, Division of Allergy and Infectious Diseases, Center for Emerging and Reemerging Infectious Disease (CERID), University of Washington, Seattle, WA, USA Departments of Global Health and Microbiology, University of Washington, Seattle, WA, USA

New and improved drugs against tuberculosis are urgently needed as multi-drug-resistant forms of the disease become more prevalent. Mycobacterium tuberculosis cytidylate kinase is an attractive target for screening due to its essentiality and different substrate specificity to the human orthologue. However, we selected the Mycobacterium smegmatis cytidylate kinase for screening because of the availability of high-resolution X-ray crystallographic data defining its structure and the high likelihood of active site structural similarity to the M. Read More

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http://dx.doi.org/10.1177/1087057116646702DOI Listing
August 2016
7 Reads

Fluorometric High-Throughput Screening Assay for Secreted Phospholipases A2 Using Phospholipid Vesicles.

J Biomol Screen 2016 Aug 4;21(7):713-21. Epub 2016 May 4.

Department of Chemistry, University of Washington, Seattle, WA, USA

There is interest in developing inhibitors of human group III secreted phospholipase A2 (hGIII-sPLA2) because this enzyme plays a role in mast cell maturation. There are no potent inhibitors for hGIII-sPLA2 reported to date, so we adapted a fluorescence-based enzyme activity monitoring method to a high-throughput screening format. We opted to use an assay based on phospholipid substrate present in phospholipid vesicles since this matrix more closely resembles the natural substrate of hGIII-sPLA2, as opposed to phospholipid/detergent mixed micelles. Read More

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http://dx.doi.org/10.1177/1087057116646742DOI Listing
August 2016
15 Reads

Development of a Human Whole Blood Screening Platform to Monitor JAK/STAT Signaling Using High-Throughput Flow Cytometry.

J Biomol Screen 2016 Sep 3;21(8):866-74. Epub 2016 May 3.

Bristol-Myers Squibb Company, Princeton, NJ, USA.

Oral agents targeting Janus-associated kinases (JAKs) are promising new agents in clinical development. To better understand the relationship between JAK inhibition and biological outcome, compounds targeting JAKs were evaluated in peripheral human whole blood. To date, these analyses are low throughput and costly. Read More

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http://dx.doi.org/10.1177/1087057116645095DOI Listing
September 2016
19 Reads

Simultaneous High-Throughput Conformational and Colloidal Stability Screening Using a Fluorescent Molecular Rotor Dye, 4-(4-(Dimethylamino)styryl)-N-Methylpyridinium Iodide (DASPMI).

J Biomol Screen 2016 Sep 2;21(8):842-50. Epub 2016 May 2.

Analytical Science Department, Analytical and Formulation Development, Boehringer Ingelheim, Fremont, CA, USA

Technologies to improve the throughput for screening protein formulations are continuously evolving. The purpose of this article is to highlight novel applications of a molecular rotor dye, 4-(4-(dimethylamino)styryl)-N-methylpyridinium iodide (DASPMI) in screening for the conformational stability, colloidal stability, and subtle pretransition dynamics of protein structures during early formulation development. The measurement of the apparent unfolding temperature (Tm) for a monoclonal antibody in the presence of Tween 80 was conducted and data were compared to the results of differential scanning calorimetry (DSC) measurements. Read More

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http://jbx.sagepub.com/lookup/doi/10.1177/1087057116646553
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http://dx.doi.org/10.1177/1087057116646553DOI Listing
September 2016
5 Reads

Morphological Evaluation of Nonlabeled Cells to Detect Stimulation of Nerve Growth Factor Expression by Lyconadin B.

J Biomol Screen 2016 Sep 28;21(8):795-803. Epub 2016 Apr 28.

Division of Bioscience, Graduate School of Pharmaceutical Sciences, Nagoya University, Nagoya, Japan

The success of drug development is greatly influenced by the efficiency of drug screening methods. Recently, phenotype-based screens have raised expectations, based on their proven record of identifying first-in-class drugs at a higher rate. Although fluorescence images are the data most commonly used in phenotype-based cell-based assays, nonstained cellular images have the potential to provide new descriptive information about cellular responses. Read More

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http://dx.doi.org/10.1177/1087057116645500DOI Listing
September 2016
8 Reads

Ranking Differential Drug Activities from Dose-Response Synthetic Lethality Screens.

J Biomol Screen 2016 Oct 25;21(9):942-55. Epub 2016 Apr 25.

Division of Preclinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, MD, USA

Synthetic lethal screens are used to discover new combination treatments for cancer. In traditional high-throughput synthetic lethal screens, compounds are tested at a single dose, and hit selection is based on threshold activity values from the variance of the efficacy of the compounds tested. The limitation of the single-dose screening for synthetic lethal screens is that it does not allow for the robust detection of differential activities from compound collections with a broad range of potencies and efficacies. Read More

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http://dx.doi.org/10.1177/1087057116644890DOI Listing
October 2016
29 Reads

In Vivo Chemical Screen in Zebrafish Embryos Identifies Regulators of Hematopoiesis Using a Semiautomated Imaging Assay.

J Biomol Screen 2016 Oct 25;21(9):956-64. Epub 2016 Apr 25.

Medizinische Klinik und Poliklinik I, Universitätsklinikum Carl Gustav Carus, Technische Universität Dresden, Dresden, Germany

Hematopoietic stem and progenitor cells (HSPCs) generate all cell types of the blood and are crucial for homeostasis of all blood lineages in vertebrates. Hematopoietic stem cell transplantation (HSCT) is a rapidly evolving technique that offers potential cure for hematologic cancers, such as leukemia or lymphoma. HSCT may be autologous or allogenic. Read More

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http://dx.doi.org/10.1177/1087057116644163DOI Listing
October 2016
25 Reads

384-Well Multiplexed Luminex Cytokine Assays for Lead Optimization.

J Biomol Screen 2016 Jul 19;21(6):548-55. Epub 2016 Apr 19.

Bristol-Myers Squibb, Lawrenceville, NJ, USA

Cytokines serve as a major mechanism of communication between immune cells and are the functional molecules at the end of immune pathways. Abnormalities in cytokines are involved in a wide variety of diseases, including chronic inflammation, autoimmune diseases, and cancer. Cytokines are not only direct targets of therapeutics but also important biomarkers for assessing drug efficacy and safety. Read More

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http://dx.doi.org/10.1177/1087057116644164DOI Listing
July 2016
10 Reads

Development of a Multiplex Assay for Studying Functional Selectivity of Human Serotonin 5-HT2A Receptors and Identification of Active Compounds by High-Throughput Screening.

J Biomol Screen 2016 Sep 19;21(8):816-23. Epub 2016 Apr 19.

BioFarma Research Group, Centro Singular de Investigación en Medicina Molecular y Enfermedades Crónicas (CIMUS), Universidade de Santiago de Compostela, Santiago de Compostela, Spain

G protein-coupled receptors (GPCRs) exist as collections of conformations in equilibrium, and the efficacy of drugs has been proposed to be associated with their absolute and relative affinities for these different conformations. The serotonin 2A (5-HT2A) receptor regulates multiple physiological functions, is involved in the pathophysiology of schizophrenia, and serves as an important target of atypical antipsychotic drugs. This receptor was one of the first GPCRs for which the functional selectivity phenomenon was observed, with its various ligands exerting differential effects on the phospholipase A2 (PLA2) and phospholipase C (PLC) signaling pathways. Read More

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http://dx.doi.org/10.1177/1087057116644162DOI Listing
September 2016
10 Reads

A Quantitative Spectrophotometric Assay to Monitor the tRNA-Dependent Pathway for Lipid Aminoacylation In Vitro.

J Biomol Screen 2016 Aug 12;21(7):722-8. Epub 2016 Apr 12.

Burnett School of Biomedical Sciences, College of Medicine, University of Central Florida, Orlando, FL, USA

The transfer RNA (tRNA)-dependent pathway for lipid aminoacylation is a two-step pathway composed of (1) a tRNA aminoacylation step catalyzed by an aminoacyl-tRNA synthetase, forming a specific aa-tRNA, and (2) a tRNA-dependent transfer step in which the amino acid acylating the tRNA is transferred to an acceptor lipid. The latter step is catalyzed by a transferase located within the cytoplasmic membrane of certain bacteria. Lipid aminoacylation modifies the biochemical properties of the membrane and enhances resistance of some pathogens to various classes of antimicrobial agents and components of the innate immune response. Read More

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http://dx.doi.org/10.1177/1087057116642987DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC5938733PMC
August 2016
7 Reads

Development of an Enhanced Phenotypic Screen of Cytotoxic T-Lymphocyte Lytic Granule Exocytosis Suitable for Use with Synthetic Compound and Natural Product Collections.

J Biomol Screen 2016 Jul 5;21(6):556-66. Epub 2016 Apr 5.

Department of Molecular and Cell Biology, University of Connecticut at Storrs, Storrs, CT, USA

We previously developed an assay of cytotoxic T-lymphocyte lytic granule exocytosis based on externalization of LAMP-1/CD107A using nonphysiological stimuli to generate maximal levels of exocytosis. Here, we used polystyrene beads coated with anti-CD3 antibodies to stimulate cells. Light scatter let us distinguish cells that contacted beads from cells that had not, allowing comparison of signaling events and exocytosis from stimulated and unstimulated cells in one sample. Read More

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http://dx.doi.org/10.1177/1087057116643260DOI Listing
July 2016
40 Reads

A Multiplexed High-Content Screening Approach Using the Chromobody Technology to Identify Cell Cycle Modulators in Living Cells.

J Biomol Screen 2016 Oct 4;21(9):965-77. Epub 2016 Apr 4.

Assay Development and Screening Platform, Institute of Molecular Toxicology and Pharmacology, Helmholtz Zentrum München, German Research Center for Environmental Health, Neuherberg, Bavaria, Germany

Many screening hits show relatively poor quality regarding later efficacy and safety. Therefore, small-molecule screening efforts shift toward high-content analysis providing more detailed information. Here, we describe a novel screening approach to identify cell cycle modulators with low toxicity by combining the Cell Cycle Chromobody (CCC) technology with the CytoTox-Glo (CTG) cytotoxicity assay. Read More

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http://dx.doi.org/10.1177/1087057116641935DOI Listing
October 2016
40 Reads

Identification of Small-Molecule Frequent Hitters of Glutathione S-Transferase-Glutathione Interaction.

J Biomol Screen 2016 Jul 4;21(6):596-607. Epub 2016 Apr 4.

Helmholtz Zentrum München für Gesundheit und Umwelt (GmbH), Assay Development and Screening Platform, Institute of Molecular Toxicology and Pharmacology, Neuherberg, Germany

In high-throughput screening (HTS) campaigns, the binding of glutathione S-transferase (GST) to glutathione (GSH) is used for detection of GST-tagged proteins in protein-protein interactions or enzyme assays. However, many false-positives, so-called frequent hitters (FH), arise that either prevent GST/GSH interaction or interfere with assay signal generation or detection. To identify GST-FH compounds, we analyzed the data of five independent AlphaScreen-based screening campaigns to classify compounds that inhibit the GST/GSH interaction. Read More

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http://dx.doi.org/10.1177/1087057116639992DOI Listing
July 2016
22 Reads

Characterization of Early Cortical Neural Network Development in Multiwell Microelectrode Array Plates.

J Biomol Screen 2016 Jun 29;21(5):510-9. Epub 2016 Mar 29.

Office of Research and Development, National Health and Environmental Effects Research Laboratory, U.S. Environmental Protection Agency, Research Triangle Park, NC, USA

We examined neural network ontogeny using microelectrode array (MEA) recordings made in multiwell MEA (mwMEA) plates over the first 12 days in vitro (DIV). In primary cortical cultures, action potential spiking activity developed rapidly between DIV 5 and 12. Spiking was sporadic and unorganized at early DIV, and became progressively more organized with time, with bursting parameters, synchrony, and network bursting increasing between DIV 5 and 12. Read More

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http://dx.doi.org/10.1177/1087057116640520DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4904353PMC
June 2016
20 Reads

NMR Binding and Functional Assays for Detecting Inhibitors of S. aureus MnaA.

J Biomol Screen 2016 Jul 29;21(6):579-89. Epub 2016 Mar 29.

Structural Chemistry, Merck Research Laboratories, Kenilworth, NJ, USA

Nonessential enzymes in the staphylococcal wall teichoic acid (WTA) pathway serve as highly validated β-lactam potentiation targets. MnaA (UDP-GlcNAc 2-epimerase) plays an important role in an early step of WTA biosynthesis by providing an activated form of ManNAc. Identification of a selective MnaA inhibitor would provide a tool to interrogate the contribution of the MnaA enzyme in the WTA pathway as well as serve as an adjuvant to restore β-lactam activity against methicillin-resistant Staphylococcus aureus (MRSA). Read More

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http://dx.doi.org/10.1177/1087057116640199DOI Listing
July 2016
13 Reads

Bacterial Expression and HTS Assessment of Soluble Epoxide Hydrolase Phosphatase.

J Biomol Screen 2016 Aug 23;21(7):689-94. Epub 2016 Mar 23.

Institute of Pharmaceutical Chemistry, Goethe University Frankfurt, Frankfurt, Germany

Soluble epoxide hydrolase (sEH) is a bifunctional enzyme that possesses an epoxide hydrolase and lipid phosphatase activity (sEH-P) at two distinct catalytic domains. While the physiological role of the epoxide hydrolase domain is well understood, the consequences of the phosphatase activity remain unclear. Herein we describe the bacterial expression of the recombinant N-terminal domain of sEH-P and the development of a high-throughput screening protocol using a sensitive and commercially available substrate fluorescein diphosphate. Read More

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http://dx.doi.org/10.1177/1087057116637609DOI Listing
August 2016
5 Reads

Antigen Selection for Enhanced Affinity T-Cell Receptor-Based Cancer Therapies.

J Biomol Screen 2016 Sep 18;21(8):769-85. Epub 2016 Mar 18.

Immunocore Ltd, Abingdon, UK.

Evidence of adaptive immune responses in the prevention of cancer has been accumulating for decades. Spontaneous T-cell responses occur in multiple indications, bringing the study of de novo expressed cancer antigens to the fore and highlighting their potential as targets for cancer immunotherapy. Circumventing the immune-suppressive mechanisms that maintain tumor tolerance and driving an antitumor cytotoxic T-cell response in cancer patients may eradicate the tumor or block disease progression. Read More

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http://dx.doi.org/10.1177/1087057116637837DOI Listing
September 2016
4 Reads

Small-Activating RNA Can Change Nucleosome Positioning in Human Fibroblasts.

J Biomol Screen 2016 Jul 18;21(6):634-42. Epub 2016 Mar 18.

Institute of Medical Biology, Chinese Academy of Medical Sciences and Peking Union Medical College, Kunming, China

RNA activation (RNAa) is a mechanism of positive gene expression regulation mediated by small-activating RNAs (saRNAs), which target gene promoters and have been used as tools to manipulate gene expression. Studies have shown that RNAa is associated with epigenetic modifications at promoter regions; however, it is unclear whether these modifications are the cause or a consequence of RNAa. In this study, we examined changes in nucleosome repositioning and the involvement of RNA polymerase II (RNAPII) in this process. Read More

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http://dx.doi.org/10.1177/1087057116637562DOI Listing
July 2016
14 Reads