We develop a flexible and computationally efficient approach for analyzing high-throughput chemical genetic screens. In such screens, a library of genetic mutants is phenotyped in a large number of stresses. Typically, interactions between genes and stresses are detected by grouping the mutants and …
Read A simple high throughput assay to evaluate water consumption in the fruit fly, Scientific Reports on DeepDyve, the largest online rental service for scholarly research with thousands of academic publications available at your fingertips.
Dr. Linda Baum and CDMD colleagues published a paper on a high throughput screen to find known drugs that might increase sugars on muscle cells as a possible new therapeutic intervention in DMD. Click for article ...
TY - JOUR. T1 - The establishment of a novel high-throughput screening system using RNA-guided genome editing to identify chemicals that suppress aldosterone synthase expression. AU - Ito, Ryo. AU - Morita, Masanobu. AU - Nakano, Taichi. AU - Sato, Ikuko. AU - Yokoyama, Atsushi. AU - Sugawara, Akira. N1 - Funding Information: This work was supported by JSPS KAKENHI Grant Numbers JP26893012 (RI), JP16K19550 (RI), 19K22793 (AS), 20H04099 (AS), and the Platform Project for Supporting Drug Discovery and Life Science Research from Japan Agency for Medical Research and Development (AMED) . PY - 2021/1/1. Y1 - 2021/1/1. N2 - Aldosterone is synthesized in the adrenal by the aldosterone synthase CYP11B2. Although the control of CYP11B2 expression is important to maintain the mineral homeostasis, its overexpression induced by the depolarization-induced calcium (Ca2+) signaling activation has been reported to increase the synthesis of aldosterone in primary aldosteronism (PA). The drug against PA focused ...
Miltenyi Biotec is a global provider of products and services that advance biomedical research and cellular therapy. The companys innovative tools support research at every level, from basic research to translational research to clinical application. This integrated portfolio enables scientists and clinicians to obtain, analyze, and utilize the cell. Miltenyi Biotecs technologies cover techniques of sample preparation, cell isolation, cell sorting, flow cytometry, cell culture, molecular analysis, and preclinical imaging. Their more than 25 years of expertise spans research areas including immunology, stem cell biology, neuroscience, and cancer, and clinical research areas like hematology, graft engineering, and apheresis. In their commitment to the scientific community, Miltenyi Biotec also offers comprehensive scientific support, consultation, and expert training. Today, Miltenyi Biotec has more than 2,000 employees in 25 countries - all dedicated to helping researchers and clinicians around ...
High‐content flow (HCF) screening systems, such as the iQue Screener and HTFC Screening System from IntelliCyt, have facilitated the implementation of flow cytometry assays for high‐throughput screening
Type: Heart. Grant holder: Dr Paul Welsh, Dr Preiss, Professors Woodward, Chalmers and Sattar; Cardiovascular Research Centre, University of Glasgow. Amount Awarded: £77,630 over two years. Year: 2013 - Finished. Cutting-edge technology is being harnessed to give a more detailed analysis of the chemical make-up of blood samples than has yet been possible. The research team has developed the ability to measure the levels of more than 100 different components in blood, covering a broad range of fats and sugars. The researchers will test the new technologys ability to predict heart attacks, thereby improving drug prescription and prevention. This new technology is being applied in a major international trial of diabetes patients, although this will be of particular interest in Scotland where rising diabetes rates threaten to reverse current declining heart attack trends.. ...
High-throughput cell-based screens have become an important experimental tool for the analysis of many cellular processes. Whole genome sequences and methods for gene silencing by RNA interference (RNAi) have enabled loss-of-function analysis in ex vivo and in vivo, opening new avenues for functional analysis that were previously unfeasible [1, 2]. Different experimental methods to assess phenotypic changes are being used, from single-channel homogenous readouts to multi-channel cytometry and imaging, producing large data sets that need to be analyzed to extract phenotypically relevant information. RNAi screening has found a broad user-base as a genetic method to dissect many different cellular processes, such as cell survival, signaling pathways and other cellular phenotypes in a high-throughput manner [3-6].. High-throughput screens are mostly performed using 96- to 384-well plates and produce large data sets that need to be normalized, summarized and ranked to generate a list of significant ...
Cytotoxicity tests for high-throughput drug discovery.: Despite theoretical obstacles associated with performing cell-based assays in high-density formats (micr
Included among the quantitative high throughput screens (qHTS) conducted in support of the US Tox21 program are those being evaluated for the detection of genotoxic compounds. One such screen is based on the induction of increased cytotoxicity in seven isogenic chicken DT40 cell lines deficient in DNA repair pathways compared to the parental DNA repair-proficient cell line. To characterize the utility of this approach for detecting genotoxic compounds and identifying the type(s) of DNA damage induced, we evaluated nine of 42 compounds identified as positive for differential cytotoxicity in qHTS (actinomycin D, adriamycin, alachlor, benzotrichloride, diglycidyl resorcinol ether, lovastatin, melphalan, trans-1,4-dichloro-2-butene, tris(2,3-epoxypropyl)isocyanurate) and one non-cytotoxic genotoxic compound (2-aminothiamine) for (1) clastogenicity in mutant and wild-type cells; (2) the comparative induction of γH2AX positive foci by melphalan; (3) the extent to which a 72-hr exposure duration ...
DasGupta et al. [3] developed a high-throughput assay based on the known ability of canonical Wnt signaling to activate transcription of luciferase reporter constructs in transfected cells. Improving on the widely used construct TOP-Flash [13], they generated two new reporters each containing multiple TCF-binding sites upstream of a different minimal promoter. Because only the TCF sites were common between the reporters, off-target effects unrelated to β-catenin/TCF signaling were minimized. Reporters with mutated TCF-binding sites also served as specificity controls. The authors first validated the behavior of these reporters in transfection assays of Drosophila cell lines. Then they scaled up the transfections to incorporate approximately 22,000 double-stranded RNAs (dsRNAs), so as to induce RNAi [3], and tested the individual effects on Wingless-induced signaling. The library of dsRNA sequences, previously used in other high-throughput RNAi screens, is directed at all known open reading ...
Understanding the tens of thousands of proteins that compose the ...The bulk of these so far enigmatic proteins may now be open to study ...The scientists have developed the first widely applicable high-through...Most studies of enzymes rely on substrate assays which allow research...One major hurdle in protein research though is that developing conve...,New,high-throughput,screening,technique,makes,probing,puzzling,proteins,possible,biological,biology news articles,biology news today,latest biology news,current biology news,biology newsletters
TY - JOUR. T1 - Application of a high throughput method of biomarker discovery to improvement of the EarlyCDT®-Lung Test. AU - Macdonald, Isabel K.. AU - Murray, Andrea. AU - Healey, Graham F.. AU - Parsy-Kowalska, Celine B.. AU - Allen, Jared. AU - McElveen, Jane. AU - Robertson, Chris. AU - Sewell, Herbert F.. AU - Chapman, Caroline J.. AU - Robertson, John F.R.. PY - 2012/12/13. Y1 - 2012/12/13. N2 - The National Lung Screening Trial showed that CT screening for lung cancer led to a 20% reduction in mortality. However, CT screening has a number of disadvantages including low specificity. A validated autoantibody assay is available commercially (EarlyCDT®-Lung) to aid in the early detection of lung cancer and risk stratification in patients with pulmonary nodules detected by CT.Recent advances in high throughput (HTP) cloning and expression methods have been developed into a discovery pipeline to identify biomarkers that detect autoantibodies. The aim of this study was to demonstrate the ...
in Journal of pharmacy & pharmaceutical sciences : a publication of the Canadian Society for Pharmaceutical Sciences, Societe canadienne des sciences pharmaceutiques (2013), 16(2), 217-30. PURPOSE: ATPases, which constitute a major category of ion transporters in the human body, have a variety of significant biological and pathological roles. However, the lack of high throughput assays for ... [more ▼]. PURPOSE: ATPases, which constitute a major category of ion transporters in the human body, have a variety of significant biological and pathological roles. However, the lack of high throughput assays for ATPases has significantly limited drug discovery in this area. We have recently found that the genetic deletion of the ATP dependent calcium pump PMCA4 (plasma membrane calcium/calmodulin dependent ATPase, isoform 4) results in infertility in male mice due to a selective defect in sperm motility. In addition, recent discoveries in humans have indicated that a single nucleotide polymorphism ...
Next, an unlabeled compound was analyzed using the kinetic probe competition assay. The reaction contained fluorescent tracer 178 and the indicated concentrations of staurosporine, an unlabelled ABL-binder. The reaction was started by adding Tb-labelled ABL resulting in a signal increase for the trace without competitor (Fig. 3, grey). In contrast, the signal in presence of the competitor is lower the higher its concentration, and the initial signal increase is followed by a decrease in signal over time. The latter indicates that due to slower binding kinetics the competitor equilibrates more slowly than the tracer. ...
Quantitative high resolution melting: two methods to determine SNP allele frequencies from pooled samples. . Biblioteca virtual para leer y descargar libros, documentos, trabajos y tesis universitarias en PDF. Material universiario, documentación y tareas realizadas por universitarios en nuestra biblioteca. Para descargar gratis y para leer online.
Gene-expression analysis is ubiquitously used both in life sciences and in medical research [1-3]. Microarray and reverse transcription quantitative real-time polymerase chain reaction (RT-qPCR) analyses are commonly used to measure transcript abundance. Microarrays allow the massive parallel analysis of thousands of genes but still require significant time and financial expenses. RT-qPCR is used as a conventional routine method for expression profiling of a moderate number of genes and is highly suitable when only a small amount of sample is available [1, 4]. High throughput thermal cyclers and PCR platforms help to overcome the limitation of RT-qPCR and to perform simultaneous expression analysis of tens and hundreds of genes for one or more samples depending on the platform [1].. In research practice these two technologies are able to solve a broad spectrum of questions supplementing each other. In clinical diagnostics RT-qPCR is still more popular with the advantage of speed and a relatively ...
Gentaur molecular products has all kinds of products like :search , MarkerGene \ MarkerGene™ Multiple Drug Resistance Microtiterplate Assay Kit, High-throughput assay system based on measurement of efflux of green fluorescent dye known to bind to MDR transporters ABCG2 and ABCB1 \ M1580 for more molecular products just contact us
EN) - High-throughput assay setup for the Investigator 24plex GO! Kit from Bode Buccal 2 Assembled Cassette samples using the STAR Q Punch ...
Within the framework of virtual organization CSLabGrid, high-throughput molecular screening has been performed for new antituberculosis compounds. Using the FlexX program installed on the Institute of Food Biotechnology and Genomics (IFBG) Cluster and models of four promising ligand binding sites on the surface of FtsZ protein from Mycobacterium tuberculosis, virtual screening has been done for the database containing 2886 compounds synthesized in the Institute of Organic Chemistry of the NAS of Ukraine. Based on the LE and ΔG scores, the docking scores of CCDC Gold, and the results of molecular dynamics, a group of Mycobacterial FtsZ inhibitors has been selected. In vitro validation have revealed 6 compounds with the highest inhibition of GTPase activity of FtsZ. Also, based on in vitro experiment, three of selected compounds demonstrste strong inhibition of FtsZ polymerization together with inhibition of its GTPase activity ...
Use NucleoSpin 8/96 Blood QuickPure kits to isolate genomic DNA from whole blood, serum, plasma, and body fluids in flexible 8-well strip or high-throughput 96-well plate formats. Use animal or human blood samples, fresh or frozen.
The overuse and misuse of antibiotics have resulted in the emergence of antibiotic-resistant pathogens along with the perturbation of healthy human microbiota via killing putatively beneficial commensal bacteria. Hence, targeting infectious bacterial pathogens is important for reducing the evolution of antibiotic-resistant bacteria and preserving the endogenous human microbiome. Cell lytic enzymes including bacteriophage endolysins, autolysins, and peptidoglycan targeting enzymes are useful antibiotic alternatives, and genetic information of numerous cell lytic enzymes is currently available. However, the identification of their antimicrobial function and specificity has been limited due to time-intensive protocols to identify their specific targets. Here, we developed Lego-like sandwich microchip for rapidly accessing the function of diverse genes that are suggestive of encoding cell lytic enzymes. This approach can be used to quantify antimicrobial activity in a high-throughput manner by ...
Radiotherapy (RT) is one of the most commonly used treatments for cancer. Approximately 50% of all cancer patients are treated with RT. For many indications, radiotherapy is combined with other treatment modalities, such as surgery and/or chemotherapy [1-4]. The biological basis for the therapeutic effects of RT is that the applied ionizing radiation (IR) causes lethal double-strand breaks in the cellular DNA leading to tumor cell death. However, IR-induced DNA damage also triggers DNA damage response (DDR) signaling pathways in cells. These can result either in cell cycle arrest and DNA damage repair or in cell death. Differences in the functioning of these processes in different cells or under different conditions determine the final effect of a certain dose of IR [5]. Cancer cells are generally more vulnerable to DNA damage than healthy cells [6].. Despite its broad use and implementation of improved methods, clinical success of radiotherapy is variable. While survival rates after RT are high ...
We developed a platform combining fluorogen-activating protein (FAP) technology with high-throughput flow cytometry to detect real-time protein trafficking to and from the plasma membrane in living cells. The hybrid platform facilitates drug discovery for trafficking receptors such as G protein-coupled receptors and was validated with the β2-adrenergic receptor (β2AR) system. When a chemical library containing ∼1200 off-patent drugs was screened against cells expressing FAP-tagged β2ARs, all 33 known β2AR-active ligands in the library were successfully identified, together with a number of compounds that might regulate receptor internalization in a nontraditional manner. Results indicated that the platform identified ligands of target proteins regardless of the associated signaling pathway; therefore, this approach presents opportunities to search for biased receptor modulators and is suitable for screening of multiplexed targets for improved efficiency. The results revealed that ligands ...
To assess the quantitative ability of this approach, we performed a second experiment where we mixed fully methylated and nonmethylated DNA (the mixtures contained 0, 1, 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 95, 99, and 100% of methylated DNA) and estimated the corresponding relative methylation (see Fig. 3b ). The means and standard deviations shown are derived from six replicates of the PCR, transcription, cleavage, and MS measurement steps. In these mixing experiments we found a direct correlation between the estimated relative methylation based on the signal intensities of the two cleavage products and the relative amount of methylated template DNA. As depicted in Fig. 3b , the linear relationship between the actual and estimated relative methylation spans the entire range in these mixtures. The average standard deviation of relative methylation was ≈5% for mixtures between 10% and 90%. Outside of this range, quantitative data can be analyzed, but limitations in the dynamic range of ...
Current molecular biology laboratories rely heavily on the purification and manipulation of nucleic acids. Yet, commonly used centrifuge- and column-based protocols require specialised equipment, often use toxic reagents and are not economically scalable or practical to use in a high-throughput manner. Although it has been known for some time that magnetic beads can provide an elegant answer to these issues, the development of open-source protocols based on beads has been limited. In this article, we provide step-by-step instructions for an easy synthesis of functionalised magnetic beads, and detailed protocols for their use in the high-throughput purification of plasmids, genomic DNA and total RNA from different sources, as well as environmental TNA and PCR amplicons. We also provide a bead-based protocol for bisulfite conversion, and size selection of DNA and RNA fragments. Comparison to other methods highlights the capability, versatility and extreme cost-effectiveness of using magnetic ...
Flow cytometry allows high-content, multiparameter analysis of single cells, making it a promising tool for drug discovery and profiling of intracellular signaling. To add high-throughput capacity to flow cytometry, we developed a cell-based multiplexing technique called fluorescent cell barcoding ( …
In this study, we identified a novel class of CRAC channel blockers using high-throughput chemical library screen and asked how inhibition of Ca2+ signaling during TCR stimulation influenced differentiation and effector functions of T cells in vitro and in vivo. Using a combination of NFAT translocation as readout and a cell line harboring amplified CRAC currents, we identified a novel class of immunomodulators, compound 5, and its analogs. A more potent analog of the lead compound, compound 5D, blocked CRAC currents generated by WT Orai1 and a constitutively active mutant of Orai1 without affecting Orai1 and STIM1 translocation, indicating that the blocking mechanism directly involves the pore-forming subunit, Orai1. Further studies of CRAC current inhibition showed block by extracellular, but not intracellular, application of compound 5D, suggesting that the binding site is accessible only from the extracellular face of Orai1. Comprehensive mutational analysis of all the residues with possible ...
The process of washing suspension cells - pipetting, centrifuging, and dispensing - can be very disruptive and lead to loss of reagents and cells and cause unnecessary strain. In addition, the procedure is overly time consuming and is subject to many error-prone steps. Overall, these limitations make it unfeasible to perform many cellular assays with suspension cells, including high throughput assays.. The DropArray platform lets these cells be retained, allowing entire assays to be performed directly on the proprietary, wall-less microtiter plate - the DropArray Microplate. Its unique design allows researchers to culture as little as 250 cells (384-format) or 750 cells (96-well format) or as many as 10,000 cells (384-well format) or 40,000 cells (96-well format) per assay. In addition, plate washing is performed using gentle, low-velocity liquid exchange that keeps suspension cells within the well. Overall, this enables fully automatable, plate-based assays with suspension cells that were ...
PubMed comprises more than 30 million citations for biomedical literature from MEDLINE, life science journals, and online books. Citations may include links to full-text content from PubMed Central and publisher web sites.
We have developed a novel high-throughput screening platform for the discovery of small-molecules catalysts for bond-forming reactions. The method employs an in vitro selection for bond-formation using amphiphilic DNA-encoded small molecules charged with reaction substrate, which enables selections to be con
Computational and statistical methods development for high-throughput, high-content biological data. Modeling and integration of data from high throughput assays for biomarker discovery, clinical outcome prediction and disease classification.
From the brief descriptions of the project I have been able to find, the proposed NIH research plan will only deal with mutations in single genes. Unless the high-throughput analyses that go unnamed consist of karyotyping or in situ probes for duplications, this study wont even skim the surface of the genetics of cancer. I dont think you can even do either of these in a high-throughput manner. The epigenetic effects may be picked up by the gene expression analysis, but it will be difficult to distinguish between mutations to regulatory regions (cis effects), mutations to transcription factors that control the expression of the gene (trans effects), and epigenetic effects.. Without more detail, its hard to judge the merits of this project. The research will definitely result in some worthwhile discoveries, but it may be wiser to spend the money on more detailed analyses. Just because high-throughput works for genome sequencing, doesnt mean that Francis Collins needs to apply it to every ...
Skaga, Erlend; Kulesskiy, Evgeny; Brynjulfsen, Marit; Sandberg, Cecilie Jonsgar; Kyttala, Aija; Langmoen, Iver Arne; Laakso, Aki; Gaal-Paavola, Em-Lia; Perola, Markus; Wennerberg, Krister; Vik-Mo, Einar ...
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A critical unmet need in implementing personalized medicine is the ability to sort through the millions of single nucleotide polymorphisms (SNPs) present in the human genome and to pinpoint which of these DNA variations are causative in disease. A key under-studied function of SNPs is their ability to generate or disrupt genomic binding sites for transcription factors involved in cancer. Toward this goal, we are inventing the SNP-SNAP (Specificity and Affinity for Proteins) microarray as a prototype high throughput device to evaluate SNP function. The SNP-SNAP arrays will be used to display a quarter-million prostate cancer- related SNPs as double-stranded DNA molecules and to assay transcription factors (i.e., drug targets) for their binding to these SNP DNA sequences. The resulting data will be correlated with prostate cancer incidence. The million-plus data points from the SNP-SNAP arrays will be analyzed using SNP-Sequence Specificity Landscapes, creating a prostate cancer molecular ...
The Spindle Assembly Checkpoint (SAC) works to maintain the genomic balance by monitoring the correctness of attachments between the chromosomes and microtubules during early cell division (mitosis). Importantly, chromosome missegregation and genomic instability is caused by defects in SAC function, which can lead to cell transformation and cancer. Moreover, malfunction of SAC can confer resistance to microtubule-targeting drugs (MTAs) such as paclitaxel. The identification and functional characterization of novel SAC regulating biomolecules and analysis of their expression profiles in tumor cells may in the future facilitate improved cancer diagnosis and predict patients response to MTA therapy. The microRNA- (miRNA) and protein phosphatase-mediated regulation of mitotic signaling were investigated in my thesis. First, to identify new miRNAs that regulate SAC signaling, a cell-based high-throughput screen (HTS) was performed to test the ability of 810 different pre-miRNAs to override a drug ...
The CeMM Library of Unique Drugs (CLOUD) is the first condensed set of FDA-approved drugs representing all clinical compounds. Its potential was shown in a combinatorial high throughput screen at the CeMM chemical screening platform, published in Nature Chemical Biology: by testing all CLOUD compounds in combination with each other, a pair of hitherto unrelated drugs proved to be highly effective against multiple prostate cancer cell lines known for their resistance to therapy.
In this study, we present a quantitative high-throughput assay with reduced costs for methylation analysis. On the MALDI-TOF MS, mass spectrum information can be used to determine methylated and unmethylated DNAs to assess the degree of methylation for each CpG island independently, and to estimate the average methylation for the entire target region (5, 6, 13, 14). Using this method, both hypermethylation and hypomethylation can be detected in samples.. Recently, the company developed a product that showed robustness of (C/T) cleavage with the MassARRAY system for methylation analysis based on a study assessing a set of ,400 candidate genes in 59 different cancer cell lines (6). In the MassCLEAVE kit (Sequenom, Inc.), the T-cleavage reagent contains dCTP, rUTP, rGTP, and rATP, and the C-cleavage reagent contains a mixture of dTTP, rCTP, rGTP, and rATP, resulting in a unique cleavage 3′ of rCTP or rUTP only, respectively. In EpiTYPER two cleavage reactions, the reverse strand is cleaved by ...
Video, Wyss researchers have created a high-throughput platform to generate an Adeno-associated virus 2 (AAV2) library containing 200,000 variants, each carrying a distinct mutation in the virus capsid protein.
The voltage-sensitive L-type Ca2+-channel (LTCC) CaV1.2 is a crucial component for controlling intracellular activity and thereby essential in the cardiovascular and neuronal system. It is widely expressed in vascular smooth muscle tissue and the heart muscle 1-3. The opening of the channels leads to an increase of intracellular calcium, which act as second messenger, and thereby affects a variety of cellular processes 4 including heart muscle contraction and CaV1.2 is therefore an important target in e.g. safety pharmacology screening. CaV1.2 channels are known to require a large depolarization for their activation and once activated they display a long-lasting current flow, which typically can be blocked by low micromolar concentrations of e.g. dihydropyridines, phenylalkylamines and benzothiazepines 5,6.. In these studies, currents from HEK-hCaV1.2 were recorded on the high-throughput platform Qube 384 in both single-hole and multi-hole mode. Success rates, IV characteristics and the ...
Love J, Mancia F, Shapiro L, Punta M, Rost B, Girvin M, Wang D-N, Zhou M, Hunt JF, Szyperski T, et al. The New York Consortium on Membrane Protein Structure (NYCOMPS): a high-throughput platform for structural genomics of integral membrane proteins. J Struct Funct Genomics. 2010 ;11(3):191-9. ...
Love J, Mancia F, Shapiro L, Punta M, Rost B, Girvin M, Wang D-N, Zhou M, Hunt JF, Szyperski T, et al. The New York Consortium on Membrane Protein Structure (NYCOMPS): a high-throughput platform for structural genomics of integral membrane proteins. J Struct Funct Genomics. 2010 ;11(3):191-9. ...
TY - JOUR. T1 - Evaluation of polymeric gene delivery nanoparticles by nanoparticle tracking analysis and high-throughput flow cytometry.. AU - Shmueli, Ron B.. AU - Bhise, Nupura S.. AU - Green, Jordan J.. PY - 2013. Y1 - 2013. N2 - Non-viral gene delivery using polymeric nanoparticles has emerged as an attractive approach for gene therapy to treat genetic diseases(1) and as a technology for regenerative medicine(2). Unlike viruses, which have significant safety issues, polymeric nanoparticles can be designed to be non-toxic, non-immunogenic, non-mutagenic, easier to synthesize, chemically versatile, capable of carrying larger nucleic acid cargo and biodegradable and/or environmentally responsive. Cationic polymers self-assemble with negatively charged DNA via electrostatic interaction to form complexes on the order of 100 nm that are commonly termed polymeric nanoparticles. Examples of biomaterials used to form nanoscale polycationic gene delivery nanoparticles include polylysine, ...
The technology provides numerous systems and methods to detect stress in stem cells by measuring potency factors (and markers) and prioritized differentiation factors (and markers). Potency factors can be thought of as brake pedals for differentiation. That is, their expression and activity is associated with toti- or pluri-potency. Differentiation factors can be thought of as accelerator pedals for differentiation. That is, their expression and activity is associated with normal differentiation that occurs when potency-maintaining extracellular growth factors are removed. Under stress conditions, however, potency factors can decrease and differentiation factors can increase even in the presence of potency-maintaining extracellular growth factors. This type of differentiation is referred to as compensatory or prioritized differentiation. That is, under compensatory or prioritized differentiation, stem cells can be induced by stress from a toti- or pluripotent state into a more ...
To screen for chemical toxicity in human or mouse stem cell lines (undifferentiated or differentiated) by quantitative high throughput screening (qHTS) at NCGC, or by using lower throughput assays at NIEHS. Initially, the project focuses on fostering collaborations with stem cell technology providers and assessing control compounds and subsets of NTP chemicals using various assay approaches. Stem cell technology platforms and model systems shown to be useful for in vitro toxicology screening will be employed with larger sets of chemicals for hazard identification and chemical prioritization for toxicity testing. Data have been generated on a library of 80 predominantly developmental neurotoxicants evaluated for effects on neurite outgrowth in a human stem cell-derived neural cell population, cytotoxicity in different neural populations derived from human stem cells, and effects on the beating of human stem cell derived cardiomyocytes. Dose-response analysis has been carried out on the data from ...
Drugs currently available for leishmaniasis treatment often show parasite resistance, highly toxic side effects and costs that are prohibitive commonly for patients from the tropical endemic countries. There is an urgent need for new drugs as a treatment solution for this neglected disease. Here the authors describe the development and implementation of an automated high-throughput viability screening assay for the discovery of new drugs against Leishmania. Assay validation was done with Leishmania promastigote forms, and included the screening of 4000 compounds with known pharmacological properties. In an attempt to find new compounds with leishmanicidal properties, 26,500 structurally diverse chemical compounds were screened. A cut-off of 70% growth inhibition in the primary screening led to the identification of 567 active compounds. Cellular toxicity and selectivity were responsible for the exclusion of 78% of the pre-selected compounds. The activity of the remaining 124 compounds was ...
Patients with hereditary diffuse gastric cancer (HDGC), a cancer predisposition syndrome associated with germline mutations of the CDH1 (E-cadherin) gene, have few effective treatment options. Despite marked differences in natural history, histopathology, and genetic profile to patients afflicted by sporadic gastric cancer, patients with HDGC receive, in large, identical systemic regimens. The lack of a robust preclinical in vitro system suitable for effective drug screening has been one of the obstacles to date which has hampered therapeutic advances in this rare disease. In order to identify therapeutic leads selective for the HDGC subtype of gastric cancer, we compared gene expression profiles and drug phenotype derived from an oncology library of 1912 compounds between gastric cancer cells established from a patient with metastatic HDGC harboring a c.1380delA CDH1 germline variant and sporadic gastric cancer cells. Unsupervised hierarchical cluster analysis shows select gene expression alterations
The WHO gold standard method for malaria diagnosis is microscopy. It is considered inexpensive and field adapted, even though it is time-consuming and requires specifically trained personnel. Molecular detection methods achieve much higher detection sensitivities [1, 26], and they are better adapted to automation of the process and objective reading of results by machines. This potential makes them a valuable option for large-scale epidemiologic studies. Unfortunately few efforts have been spent on developing the high-throughput approaches needed for such studies.. The 18S rRNA gene is the most frequently cited marker for malaria detection. It is composed of highly conserved regions which can be targeted for a qualitative detection of Plasmodium spp., and of variable zones allowing species identification [4-7]. However, the 18S rRNA genes in Plasmodium spp. also have unusual properties, such as the existence of three stage-specific A-, S- and O-types, as well as copy number and strain-specific ...
We present a novel automated methodology that compensates for the effect of cell morphology on flow cytometry data, and thereby enables a quantitative analysis of high‐throughput flow cytometry data. The algorithm normalizes the effect of the physical characteristics of cell size and cell granularity on the fluorescence intensity, thereby enabling the analysis of fluorescence intensities (protein abundance) in the presence of different morphological characteristics of cells in a population. In contrast to traditional gating, which discards the large majority of cells, the regression model retains all cells and thereby provides more accurate statistics, higher consistency across replicates and the ability to handle biological samples that contain far fewer cells (at least 10‐fold), allowing for faster and cheaper data acquisition. This is relevant when one is looking for rare cells (e.g., stem cells), or when performing high‐throughput screens where only a few hundred cells per experimental ...
To understand the function of cellular protein networks, spatial and temporal context is essential. Fluorescence correlation spectroscopy (FCS) is a single-molecule method to study the abundance, mobility and interactions of fluorescence-labeled biomolecules in living cells. However, manual acquisition and analysis procedures have restricted live-cell FCS to short-term experiments of a few proteins. Here, we present high-throughput (HT)-FCS, which automates screening and time-lapse acquisition of FCS data at specific subcellular locations and subsequent data analysis. We demonstrate its utility by studying the dynamics of 53 nuclear proteins. We made 60,000 measurements in 10,000 living human cells, to obtain biophysical parameters that allowed us to classify proteins according to their chromatin binding and complex formation. We also analyzed the cell-cycle-dependent dynamics of the mitotic kinase complex Aurora B/INCENP and showed how a rise in Aurora concentration triggers two-step complex ...
In collaboration with Dr. Lisa DeLouise, the Kobie Lab has developed patented nanowell array technology called microbubbles, which allow for the long term culture and functional interrogation of B cells at the single cell level. This high-throughput platform enables the rapid screening of millions of individual B cells to obtain precise resolution of cellular diversity and isolation of rare B cells with desired functional profiles. This platform is particularly advantageous for high-content single cell analysis and therapeutic monoclonal antibody development.. « back to all projects. ...
In contrast to upstream optimization, which focuses mainly on preservation of cells and proteins as they grow, downstream optimization focuses primarily on achieving maximum yield from each lot, and ensuring that the resulting product remains as concentrated and high-quality as possible. These improvements may be achieved by implementing additional filtration and purification steps, introducing new high-throughput methods, and/or re-engineering existing processes to streamline the pipeline from cell harvest to final product.. Chromatographic separation methods are often used to isolate mAbs (or other desired proteins) from the fermentation broth after harvesting. Chromatographic separation is highly selective, resulting in a significantly higher flow and purity rates than conventional filtration techniques. In fact, recent advances have helped further integrate chromatographic separation into the production process, allowing for fine-tuned adjustment of elution conditions, thus effectively ...
The virologic synapse (VS), which is formed between a virus-infected and uninfected cell, plays a central role in the transmission of certain viruses, such as HIV and HTLV-1. During VS formation, HTLV-1-infected T-cells polarize cellular and viral proteins toward the uninfected T-cell. This polarization resembles anterior-posterior cell polarity induced by immunological synapse (IS) formation, which is more extensively characterized than VS formation and occurs when a T-cell interacts with an antigen-presenting cell. One measure of cell polarity induced by both IS or VS formation is the repositioning of the microtubule organizing center (MTOC) relative to the contact point with the interacting cell. Here we describe an automated, high throughput system to score repositioning of the MTOC and thereby cell polarity establishment. The method rapidly and accurately calculates the angle between the MTOC and the IS for thousands of cells. We also show that the system can be adapted to score anterior-posterior
Assessment of autoantibody responses in cancer patients, 978-3-639-85018-5, Cancer insistently remains among the leading causes of death worldwide, and identification of novel biomarkers that could aid in early diagnosis of this disease is of great importance. Circulating antibodies found in cancer patients blood have been described to have promising biomarker qualities. This work describes approaches that were successfully applied to identify novel antigens eliciting antibody formation in several types of cancer. One of the approaches, T7 phage-display SEREX approach that was elaborated within this study, resulted in the identification of 1064 antigens, which were further used for the development of an antigen microarray. This high-throughput platform enabled systematic comparison of antibody responses in cancer patients and healthy controls, and the results revealed that signatures of autoantibody responses holds great promise to be used for cancer diagnostics. Apart from the experimental part, the
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The combined effect of multiple mutations on protein function is hard to predict; thus, the ability to functionally assess a vast number of protein sequence variants would be practically useful for protein engineering. Here we present a high-throughput platform that enables scalable assembly and parallel characterization of barcoded protein variants with combinatorial modifications. We demonstrate this platform, which we name CombiSEAL, by systematically characterizing a library of 948 combination mutants of the widely used Streptococcus pyogenes Cas9 (SpCas9) nuclease to optimize its genome-editing activity in human cells. The ease with which the editing activities of the pool of SpCas9 variants can be assessed at multiple on- and off-target sites accelerates the identification of optimized variants and facilitates the study of mutational epistasis. We successfully identify Opti-SpCas9, which possesses enhanced editing specificity without sacrificing potency and broad targeting range. This ...
Innate Immunity has long been regarded as the non-specific arm of immune response, acting immediately and in a generic way to defend the host from infections. In the post genomic era, our knowledge of the innate immune system is enriched by findings on the specificity of innate immune reactions as well as to novel functions that do not strictly correlate with immunological defense and surveillance, immune modulation or inflammation. The advent of high-throughput platforms for genome and proteome-wide profiling, together with the enormous quantity of raw genetic information that has accumulated in the databases, have stirred new expectations in biomedical research, and led scientists to revisit established biological systems from a global and integrative perspective. Innate Immunity research now faces the challenge of integrating isolated biochemical pathways into complex gene and protein regulatory circuits. This volume collects topics on natural killer cells, mast cells, phagocytes, toll-like ...
Centrosome amplification (CA) is a hallmark of virtually all types of cancers including solid tumors and hematological malignancies. Cancer cells with extra centrosomes use centrosome clustering (CC) to allow for successful division. Because normal cells do not rely on this mechanism, CC is regarded as a promising target to selectively eradicate cells harboring supernumerary centrosomes. To identify novel inhibitors of CC, we developed a cell-based high-throughput screen that reports differential drug cytotoxicity for isogenic cell populations with different centrosome contents. We identified CP-673451 and crenolanib, two chemically related compounds originally developed for inhibition of PDGFR-β, as robust inhibitors of CC with selective cytotoxicity for cells with extra centrosomes. We demonstrate that these compounds induce mitotic spindle multipolarity by activation of the actin severing protein cofilin, leading to destabilization of the cortical actin network, and provide evidence that ...
Forward transfection is a widely used method that works well for adherent cell types; however, if one is using suspension cells or a high-throughput format, reverse transfection, where cells are added to the plated transfection reagent complex, is a more appropriate approach. This method can be used to increase the throughput and reproducibility of a CRISPR-Cas9 screen by optimizing gene editing efficiency as demonstrated in our recently published application note: Optimization of reverse transfection of Dharmacon Edit-R synthetic crRNA and tracrRNA components with DharmaFECT transfection reagent in a Cas9-expressing cell line.. An un-cleavable ubiquitin moiety fused to EGFP allows constitutive degradation of the EGFP protein, while disruption of the proteasome components by functional protein knockout leads to accumulation of EGFP and detectable fluorescence. This functional knockout data shows that determining optimal transfection conditions prior to arrayed screening leads to high gene ...
Here we describe a rapid high-throughput method for performing RNA interference (RNAi) in moss, in which phenotyping is performed within 1 week after transformation. The moss Physcomitrella patens is a great plant model system for reverse genetic studies due to its amenability to homologous recombination as well as RNAi. Our lab has developed a rapid RNAi assay to screen for growth phenotypes in moss protonemal tissue. Here we describe how we have recently further facilitated this assay by modifying the PEG-mediated transformation protocol allowing for transformations to be carried out in a semiautomated fashion in a 96-well plate format ...
Cellular senescence plays an important role in organismal aging and age-related diseases. However, it is challenging to isolate low numbers of senescent cells from small volumes of biofluids for downstream analysis. Furthermore, there is no technology that could selectively remove senescent cells in a high-throughput manner. In this work, we developed a novel microfluidic chip platform, termed senescence chip, for ultrahigh-throughput isolation and removal of senescent cells. The core component of our senescence chip is a slanted and tunable 3D micropillar array with a variety of shutters in the vertical direction for rapid cell sieving, taking advantage of the characteristic cell size increase during cellular senescence. The 3D configuration achieves high throughput, high recovery rate, and device robustness with minimum clogging. We demonstrated proof-of-principle applications in isolation and enumeration of senescent mesenchymal stem cells (MSCs) from undiluted human whole blood, and ...
Video articles in JoVE about retinoblastoma protein include In Vivo Detection and Analysis of Rb Protein SUMOylation in Human Cells, Workflow for High-content, Individual Cell Quantification of Fluorescent Markers from Universal Microscope Data, Supported by Open Source Software, Analysis of Cell Cycle Position in Mammalian Cells, Induction of Protein Deletion Through In Utero Electroporation to Define Deficits in Neuronal Migration in Transgenic Models, Whole Mount Dissection and Immunofluorescence of the Adult Mouse Cochlea, In vitro Coculture Assay to Assess Pathogen Induced Neutrophil Trans-epithelial Migration, An In Vitro Model for Studying Cellular Transformation by Kaposi Sarcoma Herpesvirus, A Protein Preparation Method for the High-throughput Identification of Proteins Interacting with a Nuclear Cofactor Using LC-MS/MS Analysis, Genome-wide Analysis using ChIP to Identify Isoform-specific Gene Targets, Assessing Replication and Beta Cell Function in Adenovirally-transduced
Sigma-Aldrich offers abstracts and full-text articles by [Lily Mahapatra, Chengjian Mao, Neal Andruska, Chen Zhang, David J Shapiro].
The mucin MUC16 and the glycosylphosphatidylinositol anchored glycoprotein mesothelin likely facilitate the peritoneal metastasis of ovarian tumors. The biochemical basis and the kinetics of the binding between these two glycoproteins are not clearly understood. Here we have addressed this deficit and provide further evidence supporting the role of the MUC16-mesothelin interaction in facilitating cell-cell binding under conditions that mimic the peritoneal environment. In this study we utilize recombinant-Fc tagged human mesothelin to measure the binding kinetics of this glycoprotein to MUC16 expressed on the ovarian tumor cell line OVCAR-3. OVCAR-3 derived sublines that did not express MUC16 showed no affinity for mesothelin. In a flow cytometry-based assay mesothelin binds with very high affinity to the MUC16 on the OVCAR-3 cells with an apparent Kd of 5-10 nM. Maximum interaction occurs within 5 mins of incubation of the recombinant mesothelin with the OVCAR-3 cells and significant binding is
Our main objectives are to identify key actors of RNA degradation pathways in plants, and to determine their impact on genome expression, development or stress response. Our studies currently focus on new co-factors of the RNA exosome, on enzymes that adenylate or uridylate RNAs and on new factors associated to P-bodies and decapping activators. Finally, we address the roles of RNA degradation pathways during viral infections.. Our main experimental strategies include forward and reverse genetics in the model plant Arabidopsis thaliana, protein biochemistry approaches coupled to mass spectrometry analyses, and new high-throughput techniques based on Illumina and Nanopore sequencing to identify 3 modifications of transcripts.. Key funding of our current research includes the NetRNA LabEx (2011-2028) and the ANR grants 3modRN (2015-2021) and URIVir (2021-2025).. ...
As a valuable aid in the construction of AOPs, toxicogenomics can be used to identify toxicity pathways or perturbed biology in response to different classes of substances. This knowledge can then be used to derive dose-response, identifying appropriate biological models pertinent to assessing the adverse outcome, and establishment of high throughput assays for screening or prioritizing.. ...
Using a novel screening platform to rapidly evaluate the cellular effects of 1,000 chemical compounds, a team led by UC San Francisco scientists has identified eight drugs that may stimulate nervous system repair in multiple sclerosis (MS).. All eight compounds have previously been approved by the U.S. Food and Drug Administration (FDA) for the treatment of other conditions. One of the most promising agents is an antihistamine, though the scientists caution that MS patients should not use the drug until clinical trials have established whether it can safely and effectively treat MS, and if it does, what the proper dosages and treatment regimens would be. Because of the drugs emergence as a clear front-runner in the new study, a Phase 2 clinical trial to evaluate its effectiveness in MS is already underway at UCSF.. A major unmet need in the development of therapeutics for repair in MS has been the ability to screen compounds in a high-throughput manner, said Jonah Chan, PhD, the Debbie and ...
In this project, we develop single-cell manipulation surfaces on which cells are rapidly adhered or released on/from desired positions at a single-cell level. On the surfaces coated with photo-responsive materials, multiple cells can be freely patterned and arrayed in a light-guided manner, and accordingly, the properties and interactions of individual cells are accurately investigated in a high-throughput manner. Furthermore, only the desired cells are selectively obtained by light irradiation. This technology is expected to be applied to isolation of cancer cells from blood samples for assessment of cancer therapy and purification of functional cells for cell therapy and tissue engineering.. ...
Previous efforts toward preparing multicellular aggregates (spheroids) have been made in traditional rocker-plate [1], porous foam block [2], and microarray chip cultures [3] in order to maintain liver-specific functions in vitro. These approaches all employ static culture methods and thus physiological flow conditions could not be simulated. Furthermore, the ability to analyze cell viability and function in a high-throughput manner is hindered due to the opaque substrates used in all three systems. To effectively coalesce otherwise monolayer liver cells seeded into microfluidic channels,we present a high-density hepatic spheroid trapping array chip for rapid three-dimensional spheroid self-assembly. Microfabricated 3-D bioreactors address many of the issues associated with traditional liver cell culture such as providing more in vivo-like spatial cell-cell interactions [4]. A platform that closely mimics the in vivo architecture of the human liver has yet to be achieved. We aim to bring hepatic ...
Mass-spectrometry-based proteomics enables the high-throughput identification and quantification of proteins, including sequence variants and post-translational modifications (PTMs) in biological samples. However, most workflows require that such variations be included in the search space used to analyze the data, and doing so remains challenging with most analysis tools. In order to facilitate the search for known sequence variants and PTMs, the Proteomics Standards Initiative (PSI) has designed and implemented the PSI extended FASTA format (PEFF). PEFF is based on the very popular FASTA format but adds a uniform mechanism for encoding substantially more metadata about the sequence collection as well as individual entries, including support for encoding known sequence variants, PTMs, and proteoforms. The format is very nearly backward compatible, and as such, existing FASTA parsers will require little or no changes to be able to read PEFF files as FASTA files, although without supporting any of the
RNA sequencing (RNASeq) provides the ability to comprehensively assay the transcriptome in a high-throughput manner. Current there are a variety of library preparation methodologies for measuring and sequencing the transcriptome depending on (a) the sample source and (b) outcomes of interest. Beyond protocol selection, the requisite computational tools and resources are significant considerations in processing, analyzing and reporting the experimental results. While there are many resources readily available to effectively perform RNA-seq experiments, optimal protocols and analysis tools for the cancer domain remain to be developed.. We have developed and characterized a set of protocols and analysis procedures that comprise an RNA-seq pipeline that can effectively be used in a cancer research setting. The analysis pipeline consists of a sequence of functions and tools to process and clean the raw data, generate quality control and summary metrics, and perform secondary analyses that include ...
Methods are provided for detecting biomolecular interactions. The use of labels is not required and the methods can be performed in a high-throughput manner. The invention also relates to optical devices.
TY - JOUR. T1 - YAP and TAZ regulate cell volume. AU - Perez-Gonzalez, Nicolas A.. AU - Rochman, Nash D.. AU - Yao, Kai. AU - Tao, Jiaxiang. AU - Le, Minh Tam Tran. AU - Flanary, Shannon. AU - Sablich, Lucia. AU - Toler, Ben. AU - Crentsil, Eliana. AU - Takaesu, Felipe. AU - Lambrus, Bram. AU - Huang, Jessie. AU - Fu, Vivian. AU - Chengappa, Pragati. AU - Jones, Tia M.. AU - Holland, Andrew J.. AU - An, Steven. AU - Wirtz, Denis. AU - Petrie, Ryan J.. AU - Guan, Kun Liang. AU - Sun, Sean X.. N1 - Funding Information: This work has been funded in part by National Institutes of Health grants R01GM114674 and U54CA210172. The authors declare no competing financial interests. Publisher Copyright: © 2019 Perez-Gonzalez et al.. PY - 2019/10/7. Y1 - 2019/10/7. N2 - How mammalian cells regulate their physical size is currently poorly understood, in part due to the difficulty in accurately quantifying cell volume in a high-throughput manner. Here, using the fluorescence exclusion method, we demonstrate ...
TY - JOUR. T1 - Introducing discrete frequency infrared technology for high-throughput biofluid screening. AU - Hughes, Caryn. AU - Clemens, Graeme. AU - Bird, Benjamin. AU - Dawson, Timothy. AU - Ashton, Katherine M.. AU - Jenkinson, Michael D.. AU - Brodbelt, Andrew. AU - Weida, Miles. AU - Fotheringham, Edeline. AU - Barre, Matthew. AU - Rowlette, Jeremy. AU - Baker, Matthew J.. PY - 2016/2/4. Y1 - 2016/2/4. N2 - Accurate early diagnosis is critical to patient survival, management and quality of life. Biofluids are key to early diagnosis due to their ease of collection and intimate involvement in human function. Large-scale mid-IR imaging of dried fluid deposits offers a high-throughput molecular analysis paradigm for the biomedical laboratory. The exciting advent of tuneable quantum cascade lasers allows for the collection of discrete frequency infrared data enabling clinically relevant timescales. By scanning targeted frequencies spectral quality, reproducibility and diagnostic potential ...
Colorectal tumor (CRC) is the third most common cancer and the fourth leading cause of cancer death in the world. as FGFR EGFR CD44 EpCAM CA IX PPARγ and COX-2) overexpressed by cancerous cells have also been shown to be effective. This review aims to put forth an overview of drug delivery technologies that have been and may be developed for the treatment of CRC. [19] found that treatment with a combination of Irinotecan (Camptosar Pharmacia) a potent inhibitor of topoisomerase I 5 and Leucovorin resulted in significantly longer progression-free survival (median 7 4.3 months; = 0.004) greater confirmed response (39% 21% < 0.001) and longer overall survival (median 14.8 12.6 months; = 0.04) than 5-FU/Leucovorin alone. LY335979 An extensive body of data shows that Fluoropyrimidines Irinotecan and Oxaliplatin have emerged as cornerstones of chemotherapy for CRC. However these traditional pharmaceutical therapeutic regimens are usually accompanied by severe mucositis myelosuppression and cumulative ...
Data Availability StatementThe datasets used and/or analyzed through the present research are available through the corresponding writer on reasonable demand. no factor between organizations B and C (P 0.05). Tumor mass in organizations B, C and D was considerably less than that in group A (P 0.05), which in group D was significantly less than that in organizations B and C (P 0.05), whereas there MK-1775 cost is no factor between organizations B and C (P 0.05). Weighed against organizations C and B, mice in group D got considerably lower IL-6 level (P 0.05), but significantly higher IL-12 level (P 0.05). There is no factor in IL-6 and IL-12 amounts between organizations B and C (P 0.05). To conclude, erlotinib coupled with cisplatin KRT20 can inhibit the tumor development of mice with LLC, and inhibition of IL-6 known level and upregulation of IL-12 level could be among its therapeutic systems. (10), there is absolutely no factor in effectiveness between erlotinib and chemotherapy MK-1775 cost ...
Protein interactions with low molecular weight ligand have great implication in biology for both allosteric regulation and enzymatic activity. Another important...
An awful lot of drug discovery comes down (sooner or later) to screening compound collections. This has been true for a long time now, and it doesnt look like its going away, either. So with that in mind, whats in your collection? Did you buy a bunch of stuff from the vendors to fill it out? If so, your chemical
Protein-protein interactions (PPIs) play a critical role in all cellular processes, ranging from cellular division to apoptosis. Elucidating and analyzing PPIs is thus essential to understanding the underlying mechanisms in biology. Indeed, this has been a major focus of research in recent years, providing a wealth of experimental data about protein associations [1-9]. Current PPI networks have been constructed using a number of techniques, such as yeast-two-hybrid (Y2H), co-immunopurification or coaffinity purification, followed by mass spectroscopy and curation of published low-throughput experiments [10-16]. Despite this tremendous push, the current coverage of PPIs is still rather poor (for example, , 10% of interactions in humans) [17]. Additionally, despite considerable improvements in high-throughput (HTP) techniques, they are still prone to spurious errors and systematic biases, yielding a significant number of false-positives and false-negatives [18-21]. This limitation impedes our ...
TY - JOUR. T1 - Probabilistic modeling of personalized drug combinations from integrated chemical screen and molecular data in sarcoma.. AU - Berlow, Noah. AU - Rikhi, R. AU - Geltzeiler, M. AU - Abraham, J. AU - Svalina, M N. AU - Davis, L E. AU - Wise, E. AU - Mancini, M. AU - Noujaim, J. AU - Mansoor, A. AU - Quist, M J. AU - Matlock, Kevin. AU - Goros, M W. AU - Hernandez, B S. AU - Doung, Y C. AU - Thway, K. AU - Tsukahara, T. AU - Nishio, J. AU - Huang, E T. AU - Airhart, S. AU - Bult, C J. AU - Gandour-Edwards, R. AU - Maki, R G. AU - Jones, R L. AU - Michalek, J E. AU - Milovancev, M. AU - Ghosh, Souparno. AU - Pal, Ranadip. AU - Keller, C. PY - 2019/6/17. Y1 - 2019/6/17. M3 - Article. SP - 593. JO - Default journal. JF - Default journal. ER - ...
Stem cell platform recreates the human intestinal epithelium. Altis Biosystems, Inc. is a preclinical contract research organization whose patent-pending stem cell platform recreates the human intestinal epithelium in a high-throughput format for drug screening and microbiome research. Altis provides a variety of services for customers, including toxicology, permeability, ELISA, transport, custom cell isolation, and more. In addition to providing services utilizing its platform, Altis is able to ship its platform to customers as well.. ...
High-throughput screening of cellular effects of RNA interference (RNAi) libraries is now being increasingly applied to explore the role of genes in specific cell biological processes and disease states. However, the technology is still limited to specialty laboratories, due to the requirements for robotic infrastructure, access to expensive reagent libraries, expertise in high-throughput screening assay development, standardization, data analysis and applications. In the future, alternative screening platforms will be required to expand functional large-scale experiments to include more RNAi constructs, allow combinatorial loss-of-function analyses (e.g. genegene or gene-drug interaction), gain-of-function screens, multi-parametric phenotypic readouts or comparative analysis of many different cell types. Such comprehensive perturbation of gene networks in cells will require a major increase in the flexibility of the screening platforms, throughput and reduction of costs. As an alternative for ...
Workstation, apparatuses and methods for the high-throughput synthesis, screening and/or characterization of combinatorial libraries. The invention relates to an array, which permits various high-throughput methods for synthesis, screening and/or characterization in the same array, without requiring sample transfer from the array. In a preferred embodiment, the synthesis, screening, and/or characterization steps are carried out in a highly parallel fashion, where more than one compound is synthesized, screened, and/or characterized at the same time. The invention may be practiced at the microscale. The array may comprise thermal channels, for regulating the temperature of the wells in the array. The wells of the array may comprise a membrane, which is used in various screening and characterization methods. The invention also relates to a covered array, comprising the array and an array cover, as well as an apparatus comprising the array, which comprises the array, an array cover and a stage. The array,
Fernández did not need to go on a hiking trip to find the answers. Instead, together with a number of other PhD fellows, she conducted a two-week clinical study at the ACTA (Academic Centre for Dentistry Amsterdam) research labs - one of the scientific partners in the TiFN Oral Health project. Sixty-one volunteers agreed to abstain from all oral hygiene activities during the trial period, and had their saliva tested regularly. Fernández developed and optimised an automated high-throughput scratch assay connected to a mathematical model. The aim was to measure metabolite levels in saliva over time, and link them to re-epithelialization - an essential component of wound healing - in the gums, she explains. Setting up the testing procedure was not easy, she stresses: I was the only one in our project team working with high-throughput microscopy, so I had to learn all about it and start up the procedure by myself.. Fernández succeeded in identifying the metabolite signatures of 10 ...
PubMed Identifiers (PMIDs) point to literature references that support an interaction. A PMID may be used to support more than one interaction. The lpr score (lowest PMID re-use) is the lowest number of distinct interactions (RIGIDs: see column 35) that any one PMID (supporting the interaction in this row) is used to support. A value of one indicates that at least one of the PMIDs supporting this interaction has never been used to support any other interaction. This likely indicates that only one interaction was described by that reference and that the present interaction is not derived from high throughput methods. The hpr score (highest PMID re-use) is the highest number of interactions (RIGIDs: see column 35) that any one PMID (supporting the interaction in this row) is used to support. A high value (e.g. greater than 50) indicates that one PMID describes at least 50 other interactions and it is more likely that high-throughput methods were used. The np score (number PMIDs) is the total ...
PubMed Identifiers (PMIDs) point to literature references that support an interaction. A PMID may be used to support more than one interaction. The lpr score (lowest PMID re-use) is the lowest number of distinct interactions (RIGIDs: see column 35) that any one PMID (supporting the interaction in this row) is used to support. A value of one indicates that at least one of the PMIDs supporting this interaction has never been used to support any other interaction. This likely indicates that only one interaction was described by that reference and that the present interaction is not derived from high throughput methods. The hpr score (highest PMID re-use) is the highest number of interactions (RIGIDs: see column 35) that any one PMID (supporting the interaction in this row) is used to support. A high value (e.g. greater than 50) indicates that one PMID describes at least 50 other interactions and it is more likely that high-throughput methods were used. The np score (number PMIDs) is the total ...