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High content screening

Jelena Belic, Marina Koch, Peter Ulz, Martina Auer, Teresa Gerhalter, Sumitra Mohan, Katja Fischereder, Edgar Petru, Thomas Bauernhofer, Jochen B Geigl, Michael R Speicher, Ellen Heitzer
Recent progress in the analysis of cell-free DNA fragments (cell-free circulating tumor DNA, ctDNA) now allows monitoring of tumor genomes by non-invasive means. However, previous studies with plasma DNA from patients with cancer demonstrated highly variable allele frequencies of ctDNA. Comprehensive genome-wide analysis of tumor genomes is greatly facilitated when plasma DNA has increased amounts of ctDNA. In order to develop a fast and cost-effective pre-screening method for the identification of plasma samples suitable for further extensive qualitative analysis, we adapted the recently described FAST-SeqS method...
2016: Advances in Experimental Medicine and Biology
Wei Li, Lorenzo Katin-Grazzini, Sanalkumar Krishnan, Chandra Thammina, Rania El-Tanbouly, Huseyin Yer, Emily Merewitz, Karl Guillard, John Inguagiato, Richard J McAvoy, Zongrang Liu, Yi Li
When subjected to shade, plants undergo rapid shoot elongation, which often makes them more prone to disease and mechanical damage. Shade-tolerant plants can be difficult to breed; however, they offer a substantial benefit over other varieties in low-light areas. Although perennial ryegrass (Lolium perenne L.) is a popular species of turf grasses because of their good appearance and fast establishment, the plant normally does not perform well under shade conditions. It has been reported that, in turfgrass, induced dwarfism can enhance shade tolerance...
2016: Frontiers in Plant Science
Y Xiao, Y Y DU, C Gao, W Kong
OBJECTIVE: To study the change of microRNA during the early stage of high phosphorus induced vascular smooth muscle cell (VSMC) calcification and its related mechanism. METHODS: The in vitro calcification model was created through stimulating VSMC cell line A7r5 with high Pi (2.6 mmol/L) for 7 d. The calcification was validated through ocresolphthalein complexone colorimetry to detect the cellular calcium content, real-time PCR to measure the calcification-related gene expression and alizarin red staining to observe the formation of calcium nodules...
October 18, 2016: Beijing da Xue Xue Bao. Yi Xue Ban, Journal of Peking University. Health Sciences
Fang Mao, Mamoun Taher, Oleksandr Kryshtal, Adam Kruk, Aleksandra Czyrska-Filemonowicz, Mikael Ottosson, Anna Mikaela Andersson, Urban Wiklund, Ulf Jansson
A combinatorial approach is applied to rapidly deposit and screen Ag-Al thin films to evaluate the mechanical, tribological and electrical properties as a function of chemical composition. Ag-Al thin films with large continuous composition gradients (6-60 at.% Al) were deposited by a custom-designed combinatorial magnetron sputtering system. X-ray diffraction (XRD), energy dispersive X-ray spectroscopy (EDX), scanning and transmission electron microscopy (SEM & TEM), X-ray photoelectron spectroscopy (XPS), nanoindentation, and four-point electrical resistance screening were employed to characterize the chemical composition, structure and physical properties of the films in a rapid and time-efficient way...
October 17, 2016: ACS Applied Materials & Interfaces
Ronald J Maggiore, Kathryn E Callahan, Janet A Tooze, Ira R Parker, Tina Hsu, Heidi D Klepin
The number of older adults with cancer is growing, necessitating more collaborative training in both geriatric principles and cancer care. We administered a web-based survey to U.S. geriatrics program directors (PDs) addressing cancer-specific training and perspectives on optimal training content and roles for geriatricians in cancer care. Of 140 PDs contacted, 67 (48%) responded. Topics considered very important in training included cancer screening (79%) and cancer-related pain management (70%). Respondents strongly agreed that some of the geriatrician's roles in cancer care included assessing functional status (64%) and assessing physical/cognitive function for goals of care (64%)...
October 17, 2016: Gerontology & Geriatrics Education
P Sneha, C George Priya Doss
The pace of anti-diabetic drug discovery is very slow in spite of increasing rate of prevalence of Type 2 Diabetes which remains a major public health concern. Though extensive research steps are taken in the past decade, yet craves for better of new treatment strategies to overcome the current scenario. One such general finding is the evolution of gliptins which discriminately inhibits DPP4 (Dipeptidyl peptidase-4) enzyme. Although the mechanism of action of gliptin is highly target oriented and accurate, still its long-term use stands unknown...
October 12, 2016: Life Sciences
Laura R Matthews, Linda OʼDwyer, Erica OʼNeill
OBJECTIVE: To examine rates of intrauterine device (IUD) insertion failure with and without prior misoprostol administration. Additional outcomes included difficulty of insertion, subjective pain, expulsion, and complications. DATA SOURCES: Systematic searches were performed in PubMed MEDLINE, Cochrane Central Register of Controlled Trials,, and Cumulative Index to Nursing and Allied Health Literature for articles with the following keywords: "misoprostol," "intrauterine devices," and "IUDs...
October 6, 2016: Obstetrics and Gynecology
Kostas Ioannidis, Eleni Melliou, Paraskevi Alizoti, Prokopios Magiatis
BACKGROUND: Recently published studies have demonstrated the strong anti-inflammatory properties of Scots pine (Pinus sylvestris L.) heartwood extracts, related to its stilbene content. In order to find alternative sources of Pinus heartwood extracts rich in stilbenes, a large number of samples was investigated, using a new developed high throughput screening method based on qNMR. RESULTS: The new method enabled us to measure the levels of pinosylvin, pinosylvin monomethyl ether and pinosylvin dimethylether in heartwood extracts in only 45 seconds per sample...
October 14, 2016: Journal of the Science of Food and Agriculture
Ningning Liu, Lailai Li, Xin Zhu, Zhiqiang Ling, Jianguo Feng, Ying Hu, Yi Wang, Lijun Mou, Yi Wang
Chronic kidney disease (CKD) is a worldwide health problem with growing prevalence in developing countries. Renal tubular epithelial-mesenchymal transition (EMT) is a critical step and key factor in the development of this condition. Renal tubulointerstitial fibrosis is a basic pathological change at the later stages of the disease. Therefore, blocking the development of EMT could be a critical factor in curing CKD. We have established a cell-based high-content screening (HCS) method to identify inhibitors of EMT in human proximal tubular epithelial (HK-2) cells by automatic acquisition and processing of dual-fluorescent labeled images...
October 10, 2016: Molecules: a Journal of Synthetic Chemistry and Natural Product Chemistry
Concha Domingo, Eric Lalanne, María M Catalá, Eva Pla, Juan L Reig-Valiente, Manuel Talón
Salinity is a complex trait that affects growth and productivity in many crops, including rice. Mutation induction, a useful tool to generate salt tolerant plants, enables the analysis of plants with similar genetic background, facilitating the understanding of the salt tolerance mechanisms. In this work, we generated three salt tolerant mutant lines by irradiation of a salt-sensitive cultivar plants and screened M2 plants at seedling stage in the presence of high salinity. These three lines, SaT20, SaS62, and SaT58, showed different responses to salinity, but exhibited similar phenotype to wild type plants, except SaT20 that displayed shorter height when grown in the absence of salt...
2016: Frontiers in Plant Science
Wararut Buncharoen, Kanokporn Saenphet, Supap Saenphet, Chatchote Titaram
ETHNOPHARMACOLOGICAL RELEVANCE: Traditional medicine has used Uvaria rufa Blume as an ethnomedicinal plant for treating fever, skin allergies, intestinal ulcers and prostate disorders including BPH. However, no scientific evidence supports the traditional use. AIM OF THE STUDY: This study aimed to evaluate the therapeutic potential of U. rufa on BPH using in vitro and in vivo models. MATERIALS AND METHODS: In vitro studies screened the efficacy of a 5α-reductase (5αR) inhibition and antioxidant activity of petroleum ether, ethyl acetate, ethanol and aqueous extracts from the stem of U...
October 9, 2016: Journal of Ethnopharmacology
Harrison Liu, Steven Chen, Kevin Huang, Jeffrey Kim, Han Mo, Raffael Iovine, Julie Gendre, Pauline Pascal, Qiang Li, Yaping Sun, Zhiqiang Dong, Michelle Arkin, Su Guo, Bo Huang
Drug discovery in whole-organisms such as zebrafish is a promising approach for identifying biologically-relevant lead compounds. However, high content imaging of zebrafish at cellular resolution is challenging due to the difficulty in orienting larvae en masse such that the cell type of interest is in clear view. We report the development of the multi-pose imaging method, which uses 96-well round bottom plates combined with a standard liquid handler to repose the larvae within each well multiple times, such that an image in a specific orientation can be acquired...
2016: PloS One
W Joon Chung, Jennifer L Goeckeler-Fried, Viktoria Havasi, Annette Chiang, Steven M Rowe, Zackery E Plyler, Jeong S Hong, Marina Mazur, Gary A Piazza, Adam B Keeton, E Lucile White, Lynn Rasmussen, Allan M Weissman, R Aldrin Denny, Jeffrey L Brodsky, Eric J Sorscher
Small molecules that correct the folding defects and enhance surface localization of the F508del mutation in the Cystic Fibrosis Transmembrane conductance Regulator (CFTR) comprise an important therapeutic strategy for cystic fibrosis lung disease. However, compounds that rescue the F508del mutant protein to wild type (WT) levels have not been identified. In this report, we consider obstacles to obtaining robust and therapeutically relevant levels of F508del CFTR. For example, markedly diminished steady state amounts of F508del CFTR compared to WT CFTR are present in recombinant bronchial epithelial cell lines, even when much higher levels of mutant transcript are present...
2016: PloS One
Ophélie I Berthuy, Sinan K Muldur, François Rossi, Pascal Colpo, Loïc J Blum, Christophe A Marquette
Microarray technology was developed in the early 1990s to measure the transcription levels of thousands of genes in parallel. The basic premise of high-density arraying has since been expanded to create cell microarrays. Cells on chip are powerful experimental tools for high-throughput and multiplex screening of samples or cellular functions. Miniaturization increases assay throughput while reducing both reagent consumption and cell population heterogeneity effect, making these systems attractive for a wide range of assays, from drug discovery to toxicology, stem cell research and therapy...
October 12, 2016: Lab on a Chip
Maximilian Billmann, Michael Boutros
RNA interference (RNAi) is a potent tool for perturbation of gene function in model organisms and human cells. In Drosophila, efficient RNAi enables screening approaches for components of cellular processes in vivo and in vitro. In cultured cells, measuring the effect of depleting gene products on a genome-wide scale can systematically associate gene function with diverse processes, such as cell growth and proliferation, signaling and trafficking. Here, we describe methods for RNAi experiments in cultured Drosophila cells with a focus on genome-wide loss-of-function screening...
2016: Methods in Molecular Biology
Stefan Vordenbäumen, Angelika Lueking, Petra Budde, Hans-Dieter Zucht, Heike Goehler, Ralph Brinks, Rebecca Fischer-Betz, Jutta Richter, Ellen Bleck, Jacqueline Detert, Hans-Eckhard Langer, Anne Sörgel, Gerd-Rüdiger Burmester, Peter Schulz-Knappe, Matthias Schneider
BACKGROUND: The aim was to identify novel diagnostic autoantibody candidates for rheumatoid arthritis (RA) by comprehensive screening for autoreactivity. METHOD: We incubated 5892 recombinant proteins coupled to fluorescent beads, with patients' sera for the detection of IgG-autoantibodies in three independent patient cohorts: A (n = 72 patients with established RA); B/B- (n = 116 patients with early RA (B) and n = 51 CCP-negative patients with early RA from B (B-)); and C (n = 184 patients with early seronegative RA), in comparison to matched healthy controls...
October 12, 2016: Arthritis Research & Therapy
Lian Li, Yan Li, Danyang Feng, Linghua Xu, Fengxin Yin, Hengchang Zang, Chunhui Liu, Fengshan Wang
Chondroitin sulfate (CS) plays important roles in the complement system. However, the CS structure is complicated due to different sources and the number and positions of sulfate groups. The objective of this study was to prepare different low molecular weight chondroitin sulfates (LMWCSs) and to investigate the biological activity in anti-complement capacity. A series of LMWCSs was prepared from different sources and characterized by ultraviolet-visible (UV) spectroscopy, high-performance liquid chromatography (HPLC), size exclusion chromatography-multiangle laser light scattering (SEC-MALLS) and nuclear magnetic resonance (NMR) spectroscopy...
October 11, 2016: International Journal of Molecular Sciences
Sudip Mondal, Evan Hegarty, Chris Martin, Sertan Kutal Gökçe, Navid Ghorashian, Adela Ben-Yakar
Next generation drug screening could benefit greatly from in vivo studies, using small animal models such as Caenorhabditis elegans for hit identification and lead optimization. Current in vivo assays can operate either at low throughput with high resolution or with low resolution at high throughput. To enable both high-throughput and high-resolution imaging of C. elegans, we developed an automated microfluidic platform. This platform can image 15 z-stacks of ∼4,000 C. elegans from 96 different populations using a large-scale chip with a micron resolution in 16 min...
October 11, 2016: Nature Communications
Martin R Schmuck, Thomas Temme, Katharina Dach, Denise de Boer, Marta Barenys, Farina Bendt, Axel Mosig, Ellen Fritsche
Current developmental neurotoxicity (DNT) testing in animals faces major limitations, such as high cost and time demands as well as uncertainties in their methodology, evaluation and regulation. Therefore, the use of human-based 3D in vitro systems in combination with high-content image analysis (HCA) might contribute to DNT testing with lower costs, increased throughput and enhanced predictivity for human hazard identification. Human neural progenitor cells (hNPCs) grown as 3D neurospheres mimic basic processes of brain development including hNPC migration and differentiation and are therefore useful for DNT hazard identification...
October 8, 2016: Archives of Toxicology
Susan E Leggett, Jea Yun Sim, Jonathan E Rubins, Zachary J Neronha, Evelyn Kendall Williams, Ian Y Wong
Single cells respond heterogeneously to biochemical treatments, which can complicate the analysis of in vitro and in vivo experiments. In particular, stressful perturbations may induce the epithelial-mesenchymal transition (EMT), a transformation through which compact, sensitive cells adopt an elongated, resistant phenotype. However, classical biochemical measurements based on population averages over large numbers cannot resolve single cell heterogeneity and plasticity. Here, we use high content imaging of single cell morphology to classify distinct phenotypic subpopulations after EMT...
October 10, 2016: Integrative Biology: Quantitative Biosciences From Nano to Macro
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