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Journal of Biomolecular Screening

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https://www.readbyqxmd.com/read/27932699/a-systematic-investigation-of-the-best-buffers-for-use-in-screening-by-maldi-mass-spectrometry
#1
Jessica Chandler, Carl Haslam, Neil Hardy, Melanie Leveridge, Peter Marshall
Matrix-assisted laser desorption/ionization-mass spectrometry (MALDI-MS) offers a label-free alternative for the screening of biochemical targets in both 1536- and 6144-assay formats, as well as potentially providing increased sensitivity, reproducibility, and the simultaneous detection of multiple assay components within a specified m/z range. Ion suppression effects are one of the principal limitations reported for MS analysis. Within MALDI-MS screening, it has been identified that certain biochemical components incorporated into the assay (e...
December 8, 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27932697/htra3-isoform-specific-elisas-for-early-detection-of-preeclampsia
#2
Yao Wang, Ying Li, Jonathan Hyett, Fabricio da Silva Costa, Guiying Nie
Preeclampsia is a serious disorder of human pregnancy occurring after 20 weeks of gestation. It can be divided into subtypes of early onset (<34 weeks of gestation) and late onset (>34 weeks). Presymptomatic detection to identify those at high risk is important for managing this disease. HtrA3, a serine protease with high expression in the developing placenta, exists in long (HtrA3-L) and short (HtrA3-S) isoforms. They are identical, except HtrA3-S lacks the C-terminal PDZ domain. We have previously shown by Western blot analysis that serum HtrA3 levels at the end of the first trimester are significantly higher in women who later develop preeclampsia than in controls...
December 8, 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/28139961/functional-comparison-of-neuronal-cells-differentiated-from-human-induced-pluripotent-stem-cell-derived-neural-stem-cells-under-different-oxygen-and-medium-conditions
#3
Kazuto Yamazaki, Kazuyuki Fukushima, Michiko Sugawara, Yoshikuni Tabata, Yoichi Imaizumi, Yasuharu Ishihara, Masashi Ito, Kappei Tsukahara, Jun Kohyama, Hideyuki Okano
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% O2) 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...
December 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/28139960/development-of-an-in-vitro-model-to-screen-cyp1b1-targeted-anticancer-prodrugs
#4
Zhiying Wang, Yao Chen, Laura M Drbohlav, Judy Qiju Wu, Michael Zhuo Wang
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. KLE cells demonstrated superior stability of CYP1B1 expression relative to transiently transfected cells and did not express any appreciable amount of cognate CYP1A1 or CYP1A2, which would have compromised the specificity of the screening assay...
December 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27647668/one-step-seeding-of-neural-stem-cells-with-vitronectin-supplemented-medium-for-high-throughput-screening-assays
#5
Sheng Dai, Rong Li, Yan Long, Steve Titus, Jinghua Zhao, Ruili Huang, Menghang Xia, Wei Zheng
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). We developed a simplified method with one-step seeding of neural stem cells in assay plates by supplementing the medium with a recombinant human vitronectin (VTN), thus avoiding plate precoating...
December 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27455994/rethinking-nuclear-receptors-as-potential-therapeutic-targets-for-retinal-diseases
#6
Mayur Choudhary, Goldis Malek
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...
December 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27269812/a-platform-to-enable-the-pharmacological-profiling-of-small-molecules-in-gel-based-electrophoretic-mobility-shift-assays
#7
Timothy L Foley, Dorjbal Dorjsuren, Thomas S Dexheimer, Michael D Burkart, William C Wight, Anton Simeonov
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. Throughputs of greater than 1000 samples per day were achieved when this system was paired with a quantitative laser-based imaging system, yielding data of remarkable quality...
December 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27650791/high-throughput-platform-for-identifying-molecular-factors-involved-in-phenotypic-stabilization-of-primary-human-hepatocytes-in-vitro
#8
Jing Shan, David J Logan, David E Root, Anne E Carpenter, Sangeeta N Bhatia
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...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27650790/slas-europe-high-content-screening-conference-in-dresden-a-glimpse-of-the-future
#9
EDITORIAL
Maria Montoya, Thierry Dorval, Marc Bickle
No abstract text is available yet for this article.
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27461835/development-of-a-high-throughput-gene-expression-screen-for-modulators-of-ras-mapk-signaling-in-a-mutant-ras-cellular-context
#10
Bryan Severyn, Thi Nguyen, Michael D Altman, Lixia Li, Kumiko Nagashima, George N Naumov, Sriram Sathyanarayanan, Erica Cook, Erick Morris, Marc Ferrer, Bill Arthur, Yair Benita, Jim Watters, Andrey Loboda, Jeff Hermes, D Gary Gilliland, Michelle A Cleary, Pamela M Carroll, Peter Strack, Matt Tudor, Jannik N Andersen
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 2.0 reagent system and performed both primary and confirmatory gene expression-based high-throughput screens (GE-HTSs) using KRAS mutant colon cancer cells (SW837) and leveraging a highly annotated chemical library...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27412535/development-of-a-3d-tissue-culture-based-high-content-screening-platform-that-uses-phenotypic-profiling-to-discriminate-selective-inhibitors-of-receptor-tyrosine-kinases
#11
Tijmen H Booij, Maarten J D Klop, Kuan Yan, Csaba Szántai-Kis, Balint Szokol, Laszlo Orfi, Bob van de Water, Gyorgy Keri, Leo S Price
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. Image analysis tools were developed to process 3D image data to measure over 800 phenotypic parameters...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27358388/identification-of-novel-inhibitors-of-the-type-i-interferon-induction-pathway-using-cell-based-high-throughput-screening
#12
Zoe O Gage, Andri Vasou, David W Gray, Richard E Randall, Catherine S Adamson
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. We developed and executed a cell-based high-throughput screen (HTS) targeting components that participate in and/or regulate the IRF3 and nuclear factor (NF)-κB branches of the IFN induction pathway...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27256155/a-novel-automated-high-content-analysis-workflow-capturing-cell-population-dynamics-from-induced-pluripotent-stem-cell-live-imaging-data
#13
Maximilian Kerz, Amos Folarin, Ruta Meleckyte, Fiona M Watt, Richard J Dobson, Davide Danovi
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. Evaluations cannot be easily integrated with other biological data sets including analysis of endpoint images. Here, we present a workflow that incorporates a novel CellProfiler-based image analysis pipeline enabling segmentation of single-channel images with a robust R-based software solution to reduce the dimension of time to a single data point...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27233291/assay-establishment-and-validation-of-a-high-throughput-screening-platform-for-three-dimensional-patient-derived-colon-cancer-organoid-cultures
#14
Karsten Boehnke, Philip W Iversen, Dirk Schumacher, María José Lallena, Rubén Haro, Joaquín Amat, Johannes Haybaeck, Sandra Liebs, Martin Lange, Reinhold Schäfer, Christian R A Regenbrecht, Christoph Reinhard, Juan A Velasco
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. Single cells were embedded in an extracellular matrix by an automated workflow and subsequently self-organized into organoid structures within 4 days of culture before being exposed to compound treatment...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27226477/reproducibility-of-uniform-spheroid-formation-in-384-well-plates-the-effect-of-medium-evaporation
#15
Viswanath Das, Tomáš Fürst, Soňa Gurská, Petr Džubák, Marián Hajdúch
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...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27112173/ranking-differential-drug-activities-from-dose-response-synthetic-lethality-screens
#16
Rajarshi Guha, Lesley A Mathews Griner, Jonathan M Keller, Xiaohu Zhang, David Fitzgerald, Antonella Antignani, Ira Pastan, Craig J Thomas, Marc Ferrer
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...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27112172/in-vivo-chemical-screen-in-zebrafish-embryos-identifies-regulators-of-hematopoiesis-using-a-semiautomated-imaging-assay
#17
Guruchandar Arulmozhivarman, Martin Stöter, Marc Bickle, Martin Kräter, Manja Wobus, Gerhard Ehninger, Friedrich Stölzel, Michael Brand, Martin Bornhäuser, Nona Shayegi
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. Successful HSCT depends critically on the abundance of engraftment-competent HSPCs, which are currently difficult to obtain in large numbers. Therefore, finding compounds that enhance either the number or the activity of HSPCs could improve prognosis for patients undergoing HSCT and is of great clinical interest...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27044685/a-multiplexed-high-content-screening-approach-using-the-chromobody-technology-to-identify-cell-cycle-modulators-in-living-cells
#18
Kenji Schorpp, Ina Rothenaigner, Julia Maier, Bjoern Traenkle, Ulrich Rothbauer, Kamyar Hadian
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. The CCC technology employs intracellularly functional single-domain antibodies coupled to a fluorescent protein (chromobodies) to visualize the cell cycle-dependent redistribution of the proliferating cell nuclear antigen (PCNA) in living cells...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/26950929/single-cell-phenotype-classification-using-deep-convolutional-neural-networks
#19
Oliver Dürr, Beate Sick
Deep learning methods are currently outperforming traditional state-of-the-art computer vision algorithms in diverse applications and recently even surpassed human performance in object recognition. Here we demonstrate the potential of deep learning methods to high-content screening-based phenotype classification. We trained a deep learning classifier in the form of convolutional neural networks with approximately 40,000 publicly available single-cell images from samples treated with compounds from four classes known to lead to different phenotypes...
October 2016: Journal of Biomolecular Screening
https://www.readbyqxmd.com/read/27601436/matrix-based-activity-pattern-classification-as-a-novel-method-for-the-characterization-of-enzyme-inhibitors-derived-from-high-throughput-screening
#20
Douglas S Auld, Marta Jimenez, Kimberley Yue, Scott Busby, Yu-Chi Chen, Scott Bowes, Greg Wendel, Thomas Smith, Ji-Hu Zhang
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. Here we describe a novel method for determining the MOI of a compound without the need for curve fitting of the enzyme inhibition data...
September 6, 2016: Journal of Biomolecular Screening
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