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https://www.readbyqxmd.com/read/28548903/microenvironment-of-a-tumor-organoid-system-enhances-hepatocellular-carcinoma-malignancy-related-hallmarks
#1
Yang Wang, Kazuki Takeishi, Zhao Li, Eduardo Cervantes-Alvarez, Alexandra Collin de l'Hortet, Jorge Guzman-Lepe, Xiao Cui, Jiye Zhu
Organ-like microenviroment and 3-dimensional (3D) cell culture conformations have been suggested as promising approaches to mimic in a micro-scale a whole organ cellular functions and interactions present in vivo. We have used this approach to examine biologic features of hepatocellular carcinoma (HCC) cells. In this study, we demonstrate that hepatocellular carcinoma (HCC) cells, fibroblasts, endothelial cells and extracellular matrix can generate organoid-like spheroids that enhanced numerous features of human HCC observed in vivo...
May 26, 2017: Organogenesis
https://www.readbyqxmd.com/read/28548572/analytical-validation-of-a-reference-laboratory-elisa-for-the-detection-of-feline-leukemia-virus-p27-antigen
#2
Jesse S Buch, Genevieve H Clark, Roberta Cahill, Brendon Thatcher, Peter Smith, Ramaswamy Chandrashekar, Christian M Leutenegger, Thomas P O'Connor, Melissa J Beall
Feline leukemia virus (FeLV) is an oncogenic retrovirus of cats. Immunoassays for the p27 core protein of FeLV aid in the detection of FeLV infections. Commercial microtiter-plate ELISAs have rapid protocols and visual result interpretation, limiting their usefulness in high-throughput situations. The purpose of our study was to validate the PetChek FeLV 15 ELISA, which is designed for the reference laboratory, and incorporates sequential, orthogonal screening and confirmatory protocols. A cutoff for the screening assay was established with 100% accuracy using 309 feline samples (244 negative, 65 positive) defined by the combined results of FeLV PCR and an independent reference p27 antigen ELISA...
May 1, 2017: Journal of Veterinary Diagnostic Investigation
https://www.readbyqxmd.com/read/28548214/the-performance-of-malbac-and-mda-methods-in-the-identification-of-concurrent-mutations-and-aneuploidy-screening-to-diagnose-beta-thalassaemia-disorders-at-the-single-and-multiple-cell-levels
#3
WeiQiang Liu, HuiMin Zhang, Dan Hu, SiJia Lu, XiaoFang Sun
AIM: To select an optimal whole-genome amplification (WGA) method to improve the efficiency of the preimplantation genetic diagnosis and screening (PGD/PGS) of beta-thalassaemia disorders. METHODS: Fifty-seven fibroblast samples with defined beta-thalassaemia variations and forty-eight single-blastomere samples were amplified from single-, two-, and five-cell samples by multiple annealing and looping-based amplification cycles (MALBAC) and the multiple displacement amplification (MDA) method...
May 26, 2017: Journal of Clinical Laboratory Analysis
https://www.readbyqxmd.com/read/28546429/structural-studies-unravel-the-active-conformation-of-apo-ror%C3%AE-t-nuclear-receptor-and-a-common-inverse-agonism-of-two-diverse-classes-of-ror%C3%AE-t-inhibitors
#4
Xiang Li, Marie Anderson, Delphine Collin, Ingo Muegge, John Wan, Debra Brennan, Stanley Kugler, Donna Terenzio, Charles Kennedy, Siqi Lin, Mark E Labadia, Brian Cook, Robert Hughes, Neil A Farrow
The nuclear receptor retinoid acid-related orphan receptor γt (RORγt) is a master regulator of the Th17/IL-17 pathway that plays crucial roles in the pathogenesis of autoimmunity. RORγt has recently emerged as a highly promising target for treatment of a number of autoimmune diseases. Through high-throughput screening we previously identified several classes of inverse agonists for RORγt. Here, we report the crystal structures for the ligand binding domain of RORγt in both apo and ligand-bound states. We show that apo RORγt adopts an active conformation capable of recruiting coactivator peptides and present a detailed analysis of the structural determinants that stabilize helix 12 (H12) of RORγt in the active state in the absence of a ligand...
May 25, 2017: Journal of Biological Chemistry
https://www.readbyqxmd.com/read/28545524/an-unsupervised-learning-approach-for-tracking-mice-in-an-enclosed-area
#5
Jakob Unger, Mike Mansour, Marcin Kopaczka, Nina Gronloh, Marc Spehr, Dorit Merhof
BACKGROUND: In neuroscience research, mouse models are valuable tools to understand the genetic mechanisms that advance evidence-based discovery. In this context, large-scale studies emphasize the need for automated high-throughput systems providing a reproducible behavioral assessment of mutant mice with only a minimum level of manual intervention. Basic element of such systems is a robust tracking algorithm. However, common tracking algorithms are either limited by too specific model assumptions or have to be trained in an elaborate preprocessing step, which drastically limits their applicability for behavioral analysis...
May 25, 2017: BMC Bioinformatics
https://www.readbyqxmd.com/read/28544957/zebrafish-as-a-visual-and-dynamic-model-to-study-the-transport-of-nanosized-drug-delivery-systems-across-the-biological-barriers
#6
Ye Li, Xiaoqing Miao, Tongkai Chen, Xiang Yi, Ruibing Wang, Haitao Zhao, Simon Ming-Yuen Lee, Xueqing Wang, Ying Zheng
With the wide application of nanotechnology to drug delivery systems, a simple, dynamic and visual in vivo model for high-throughput screening of novel formulations with fluorescence markers across biological barriers is desperately needed. In vitro cell culture models have been widely used, although they are far from a complimentary in vivo system. Mammalian animal models are common predictive models to study transport, but they are costly and time consuming. Zebrafish (Danio rerio), a small vertebrate model, have the potential to be developed as an "intermediate" model for quick evaluations...
May 10, 2017: Colloids and Surfaces. B, Biointerfaces
https://www.readbyqxmd.com/read/28543988/absolute-minimal-sampling-of-homonuclear-2d-nmr-tocsy-spectra-for-high-throughput-applications-of-complex-mixtures
#7
Alexandar Hansen, Da-Wei Li, Cheng Wang, Rafael Bruschweiler
Modern applications of 2D NMR spectroscopy to diagnostic screening, metabolomics, quality control, and other high-throughput applications are often limited by the time-consuming sampling requirements along the indirect time domain t1. 2D Total Correlation Spectroscopy (TOCSY) provides unique spin-connectivity information for the analysis of a large number of compounds in complex mixtures, but standard methods typically require >100 t1-increments for an accurate spectral reconstruction, rendering these experiments ineffective for high-throughput applications...
May 23, 2017: Angewandte Chemie
https://www.readbyqxmd.com/read/28543938/high-throughput-assay-for-enantiomeric-excess-determination-in-1-2-and-1-3-diols-and-direct-asymmetric-reaction-screening
#8
Elena G Shcherbakova, Valentina Brega, Vincent M Lynch, Tony D James, Pavel Anzenbacher
A simple and efficient method for determination of the yield, enantiomeric/diasteriomeric excess (ee/de), and absolute configuration of crude chiral diols without the need of work-up and product isolation in a high throughput setting is described. This approach utilizes a self-assembled iminoboronate ester formed as a product by dynamic covalent self-assembly of a chiral diol with an enantiopure fluorescent amine such as tryptophan methyl ester or tryptophanol and 2-formylphenylboronic acid. The resulting diastereomeric boronates display different photophysical properties and allow for fluorescence-based ee determination of molecules containing a 1,2- or 1,3-diol moiety...
May 23, 2017: Chemistry: a European Journal
https://www.readbyqxmd.com/read/28543037/naringenin-responsive-riboswitch-based-fluorescent-biosensor-module-for-escherichia-coli-co-cultures
#9
Yu Xiu, Sungho Jang, J Andrew Jones, Nicholas A Zill, Robert J Linhardt, Qipeng Yuan, Gyoo Yeol Jung, Mattheos A G Koffas
The ability to design and construct combinatorial synthetic metabolic pathways has far exceeded our capacity for efficient screening and selection of the resulting microbial strains. The need for high-throughput rapid screening techniques is of upmost importance for the future of synthetic biology and metabolic engineering. Here we describe the development of a RNA riboswitch-based biosensor module with dual fluorescent reporters, and demonstrate a high-throughput flow cytometry-based screening method for identification of naringenin over producing Escherichia coli strains in co-culture...
May 20, 2017: Biotechnology and Bioengineering
https://www.readbyqxmd.com/read/28542706/modulatory-features-of-the-novel-spider-toxin-%C3%AE-trtx-df1a-isolated-from-the-venom-of-the-spider-davus-fasciatus
#10
Fernanda C Cardoso, Zoltan Dekan, Jennifer J Smith, Jennifer R Deuis, Irina Vetter, Volker Herzig, Paul F Alewood, Glenn F King, Richard J Lewis
BACKGROUND AND PURPOSE: Naturally occurring dysfunction in NaV channels results in complex disorders such as chronic pain, making these channels an attractive target for new therapies. In the pursuit of novel NaV modulators, we investigated spider venoms for new inhibitors of NaV channels. EXPERIMENTAL APPROACH: We used high-throughput screens to identify a NaV modulator in venom of the spider Davus fasciatus. Further characterization of this venom peptide was undertaken using fluorescent and electrophysiological assays, molecular modeling and a rodent pain model...
May 24, 2017: British Journal of Pharmacology
https://www.readbyqxmd.com/read/28542603/a-conformational-switch-high-throughput-screening-assay-and-allosteric-inhibition-of-the-flavivirus-ns2b-ns3-protease
#11
Matthew Brecher, Zhong Li, Binbin Liu, Jing Zhang, Cheri A Koetzner, Adham Alifarag, Susan A Jones, Qishan Lin, Laura D Kramer, Hongmin Li
The flavivirus genome encodes a single polyprotein precursor requiring multiple cleavages by host and viral proteases in order to produce the individual proteins that constitute an infectious virion. Previous studies have revealed that the NS2B cofactor of the viral NS2B-NS3 heterocomplex protease displays a conformational dynamic between active and inactive states. Here, we developed a conformational switch assay based on split luciferase complementation (SLC) to monitor the conformational change of NS2B and to characterize candidate allosteric inhibitors...
May 25, 2017: PLoS Pathogens
https://www.readbyqxmd.com/read/28542462/a-human-monocytic-nf-%C3%AE%C2%BAb-fluorescent-reporter-cell-line-for-detection-of-microbial-contaminants-in-biological-samples
#12
Claire Battin, Annika Hennig, Patrick Mayrhofer, Renate Kunert, Gerhard J Zlabinger, Peter Steinberger, Wolfgang Paster
Sensing of pathogens by innate immune cells is essential for the initiation of appropriate immune responses. Toll-like receptors (TLRs), which are highly sensitive for various structurally and evolutionary conserved molecules derived from microbes have a prominent role in this process. TLR engagement results in the activation of the transcription factor NF-κB, which induces the expression of cytokines and other inflammatory mediators. The exquisite sensitivity of TLR signalling can be exploited for the detection of bacteria and microbial contaminants in tissue cultures and in protein preparations...
2017: PloS One
https://www.readbyqxmd.com/read/28541033/synthesis-and-screening-of-phase-change-chalcogenide-thin-film-materials-for-data-storage
#13
Samuel Guérin, Brian Elliott Hayden, Dan Hewak, Chris Vian
A combinatorial synthetic methodology based on evaporation sources in ultra-high vacuum has been used to directly synthesise compositional gradient thin film libraries of the amorphous phases of GeSbTe alloys at room temperature over a wide compositional range. An optical screen is described which allows the rapid parallel mapping of the amorphous to crystalline phase transition temperature, and the optical contrast associated with the phase change, on such libraries. The results are shown to be consistent with the literature for compositions where published data are available along the Sb2Te3-GeTe tie line...
May 25, 2017: ACS Combinatorial Science
https://www.readbyqxmd.com/read/28540517/development-of-microtiter-plate-culture-method-for-rapid-screening-of-%C3%AE%C2%B5-poly-l-lysine-producing-strains
#14
Yong-Juan Liu, Xu-Sheng Chen, Jun-Jie Zhao, Long Pan, Zhong-Gui Mao
ε-Poly-L-lysine (ε-PL) produced by Streptomyces albulus possesses a broad spectrum of antimicrobial activity and is widely used as a food preservative. To extensively screen ε-PL-overproducing strain, we developed an integrated high-throughput screening assay using ribosome engineering technology. The production protocol was scaled down to 24- and 48-deep-well microtiter plates (MTPs). The microplate reader assay was used to monitor ε-PL production. A good correlation was observed between the fermentation results obtained in both 24-(48)-deep-well MTPs and conventional Erlenmeyer flasks...
May 24, 2017: Applied Biochemistry and Biotechnology
https://www.readbyqxmd.com/read/28540424/screening-and-optimizing-of-inhibitors-for-ammonia-oxidizing-bacteria-in-sediments-of-malodorous-river
#15
Jianhua Wang, Yan He, Jin Zhu, Huimin Guan, Minsheng Huang
The proper use of selective ammonia-oxidizing archaea (AOA) and/or ammonia-oxidizing bacteria (AOB) inhibitors is critical to distinguish AOA and AOB contribution. In this research, three inhibitors including ampicillin, dicyandiamide (DCD), and allylthiourea (ATU) were examined mainly focusing on inhibiting dosage, adaptability, and effects. The results showed that the optimized inhibitory dosage of ampicillin, DCD, and ATU was separately 1.5 g L(-1), 1 mM, and 25 μM. Among the three inhibitors, ATU exhibited the strongest and persistent inhibition effects and resulted in up to 90% inhibition in the AOB-enriched culture...
May 24, 2017: Applied Microbiology and Biotechnology
https://www.readbyqxmd.com/read/28539608/development-and-validation-of-a-high-throughput-whole-blood-thrombolysis-plate-assay
#16
T Bonnard, L S Law, Z Tennant, C E Hagemeyer
The objective of this work was to develop a high throughput assay for testing in vitro the thrombolytic activity using citrated whole blood samples, and to overcome the limitations of currently available techniques. We successfully developed a method that involves forming halo shaped, tissue factor induced, whole blood clots in 96 well plates, and then precisely measuring the thrombolysis process with a spectrophotometer plate reader. We here describe the implementation of this novel method, which we refer to as halo assay, and its validation with plasmin, urokinase and tissue plasminogen activator at different doses...
May 24, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28538186/an-engineered-virus-library-as-a-resource-for-the-spectrum-wide-exploration-of-virus-and-vector-diversity
#17
Wenli Zhang, Jun Fu, Jing Liu, Hailong Wang, Maren Schiwon, Sebastian Janz, Lukas Schaffarczyk, Lukas von der Goltz, Eric Ehrke-Schulz, Johannes Dörner, Manish Solanki, Philip Boehme, Thorsten Bergmann, Andre Lieber, Chris Lauber, Andreas Dahl, Andreas Petzold, Youming Zhang, A Francis Stewart, Anja Ehrhardt
Adenoviruses (Ads) are large human-pathogenic double-stranded DNA (dsDNA) viruses presenting an enormous natural diversity associated with a broad variety of diseases. However, only a small fraction of adenoviruses has been explored in basic virology and biomedical research, highlighting the need to develop robust and adaptable methodologies and resources. We developed a method for high-throughput direct cloning and engineering of adenoviral genomes from different sources utilizing advanced linear-linear homologous recombination (LLHR) and linear-circular homologous recombination (LCHR)...
May 23, 2017: Cell Reports
https://www.readbyqxmd.com/read/28537730/discovery-of-small-molecules-as-multi-toll-like-receptor-agonists-with-proinflammatory-and-anticancer-activities
#18
Lei Zhang, Varun Dewan, Hang Hubert Yin
Therapies based on activation of multiple Toll-like receptors (TLRs) may offer superior therapeutic profiles than that of single TLR activation. To discover new small molecules that could activate multiple TLRs, we performed a cell-based high-throughput screening of a small-molecule library based on TLR3-mediated NF-κB activation. Subsequent structural optimization and counter screening of other TLRs produced the first small molecule (CU-CPT24e) capable of simultaneously activating TLRs 3, 8, and 9. Biochemical studies demonstrated that CU-CPT24e could induce a strong immune response via the production of various cytokines in human monocytic THP-1 cells...
May 24, 2017: Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28537726/hot-spotting-with-thermal-scanning-a-ligand-and-structure-independent-assessment-of-target-ligandability
#19
Molly Chilton, Ben Clennell, Fredrik Edfeldt, Stefan Geschwindner
Evaluating the ligandability of a protein target is a key component when defining hit-finding strategies or to prioritize amongst drug targets. Computational as well as biophysical approaches based on nuclear magnetic resonance (NMR) fragment screening are powerful approaches, but suffer from specific constraints that limit their usage. Here, we demonstrate the applicability of high-throughput thermal scanning (HTTS) as a simple and generic biophysical fragment screening method to reproduce assessments from NMR-based screening...
May 24, 2017: Journal of Medicinal Chemistry
https://www.readbyqxmd.com/read/28536964/biosynthesis-of-heliotropin-by-a-novel-strain-of-serratia-liquefaciens
#20
Mingtao Zhao, Pu Zheng, Pengcheng Chen, Siqin Liu
The present study has been conducted towards isolation of bacteria capable of producing heliotropin via microbial conversion. Strain ZMT-1 capable of synthesizing heliotropin efficiently was obtained by enrichment culture of soil samples and a high-throughput screening method, and identified as Serratia liquefaciens. Heliotropin was identified by gas chromatography and gas chromatography-mass spectrometry analysis. In addition, the culture medium was optimized to improve heliotropin yield by experimental designs...
May 23, 2017: Applied Biochemistry and Biotechnology
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