scholarly journals High-Throughput Analysis of Antimalarial Susceptibility Data by the WorldWide Antimalarial Resistance Network (WWARN)In VitroAnalysis and Reporting Tool

2013 ◽  
Vol 57 (7) ◽  
pp. 3121-3130 ◽  
Author(s):  
Charles J. Woodrow ◽  
Sabina Dahlström ◽  
Richard Cooksey ◽  
Jennifer A. Flegg ◽  
Hervé Le Nagard ◽  
...  
Keyword(s):  
High Throughput ◽  
Artemisinin Combination Therapy ◽  
Primary Data ◽  
Enzyme Linked Immunosorbent Assay ◽  
Data Sets ◽  
High Throughput Analysis ◽  
Susceptibility Data ◽  
Drug Concentrations ◽  
Reporting Tool

ABSTRACTAssessment ofin vitrosusceptibility is a fundamental component of antimalarial surveillance studies, but wide variations in the measurement of parasite growth and the calculation of inhibitory constants make comparisons of data from different laboratories difficult. Here we describe a Web-based, high-throughputin vitroanalysis and reporting tool (IVART) generating inhibitory constants for large data sets. Fourteen primary data sets examining laboratory-determined susceptibility to artemisinin derivatives and artemisinin combination therapy partner drugs were collated from 11 laboratories. Drug concentrations associated with half-maximal inhibition of growth (IC50s) were determined by a modified sigmoidEmaxmodel-fitting algorithm, allowing standardized analysis of 7,350 concentration-inhibition assays involving 1,592 isolates. Examination of concentration-inhibition data revealed evidence of apparent paradoxical growth at high concentrations of nonartemisinin drugs, supporting amendment of the method for calculating the maximal drug effect in each assay. Criteria for defining more-reliable IC50s based on estimated confidence intervals and growth ratios improved correlation coefficients for the drug pairs mefloquine-quinine and chloroquine-desethylamodiaquine in 9 of 11 and 8 of 8 data sets, respectively. Further analysis showed that maximal drug inhibition was higher for artemisinins than for other drugs, particularly in ELISA (enzyme-linked immunosorbent assay)-based assays, a finding consistent with the earlier onset of action of these drugs in the parasite life cycle. This is the first high-throughput analytical approach to apply consistent constraints and reliability criteria to large, diverse antimalarial susceptibility data sets. The data also illustrate the distinct biological properties of artemisinins and underline the need to apply more sensitive approaches to assessingin vitrosusceptibility to these drugs.

scholarly journals Correlation of In Vitro Activity, Serum Levels, and In Vivo Efficacy of Posaconazole against Rhizopus microsporus in a Murine Disseminated Infection

2009 ◽  
Vol 53 (12) ◽  
pp. 5022-5025 ◽  
Author(s):  
M. Mar Rodríguez ◽  
F. Javier Pastor ◽  
Enrique Calvo ◽  
Valentina Salas ◽  
Deanna A. Sutton ◽  
...  
Keyword(s):  
Serum Levels ◽  
Rhizopus Microsporus ◽  
In Vitro Susceptibility ◽  
In Vivo Efficacy ◽  
Susceptibility Data ◽  
Microdilution Method ◽  
Drug Concentrations ◽  
Broth Microdilution Method

ABSTRACT A broth microdilution method was used to evaluate the in vitro activities of seven antifungal agents against 15 clinical strains of Rhizopus microsporus. Amphotericin B (AMB) and posaconazole (POS) were the most active drugs. In a model of disseminated R. microsporus infection in immunosuppressed mice, we studied the efficacy of POS administered once or twice daily against four of the strains previously tested in vitro and compared it with that of liposomal AMB (LAMB). LAMB was the most effective treatment for the two strains with intermediate susceptibility to POS. For the two POS-susceptible strains, LAMB and POS at 20 mg/kg of body weight twice a day orally showed similar efficacies. The in vivo efficacy of POS administered twice a day orally correlated with the in vitro susceptibility data and the serum drug concentrations.

scholarly journals High-Throughput Screening of Novel Peptide Inhibitors of an Integrin Receptor from the Hexapeptide Library by Using a Protein Microarray Chip

2004 ◽  
Vol 9 (8) ◽  
pp. 687-694 ◽  
Author(s):  
Yoonsuk Lee ◽  
Dong-Ku Kang ◽  
Soo-Ik Chang ◽  
Moon Hi Han ◽  
In-Cheol Kang
Keyword(s):  
High Throughput ◽  
High Throughput Screening ◽  
Competitive Inhibition ◽  
Protein Microarray ◽  
Dependent Manner ◽  
High Throughput Analysis ◽  
Microarray Chip ◽  
New Drug

Protein microarray is an emerging technology that makes high-throughput analysis possible for protein-protein interactions and analysis of proteome and biomarkers in parallel. The authors investigated the application of a novel protein microarray chip, Proteo Chip, in new drug discovery. Integrin αvβ3 microarray immobilized on the Proteo Chip was employed to screen new active peptides against the integrin from multiple hexapeptide sublibraries of a positional scanning synthetic peptide combinatorial library (PS-SPCL). The integrin αvβ3-vitronectin interaction was successfully demonstrated on the integrin microarray in a dose-dependent manner andwas inhibited not only by the syntheticRGDpeptide but also by various integrin antagonists on the integrin microarray chip. Novel peptide ligands with high affinity to the integrin were also identified from the peptide libraries with this chip-based screening system by a competitive inhibition assay in a simultaneous and highthroughput fashion. The authors have confirmed antiangiogenic functions of the novel peptides thus screened through an in vitro and in vivo angiogenesis assay. These results provide evidence that the Proteo Chip is a promising tool for highthroughput screening of lead molecules in new drug development.

scholarly journals Fluorometric High-Throughput Assay for Measuring Chlamydial Neutralizing Antibody

2012 ◽  
Vol 19 (11) ◽  
pp. 1864-1869 ◽  
Author(s):  
Timothy Southern ◽  
Leah Bess ◽  
Jillian Harmon ◽  
Lacey Taylor ◽  
Harlan Caldwell
Keyword(s):  
High Throughput ◽  
Neutralizing Antibodies ◽  
Chlamydial Infection ◽  
Transmitted Disease ◽  
Neutralizing Antibody ◽  
Human Populations ◽  
High Throughput Analysis ◽  
Content Type ◽  
Fluorometric Analysis

ABSTRACTChlamydia trachomatisis an obligate intracellular mucosotropic pathogen that causes human infections of global importance.C. trachomatiscauses trachoma, the leading cause of preventable blindness worldwide, and is the most common cause of bacterial sexually transmitted disease. Although oculogenital infections are treatable with antibiotics, a vaccine is needed to controlC. trachomatisinfection. Ideally, a vaccine would provide coverage against most, if not all, naturally occurring antigenically distinct serovariants. The development of a subunit vaccine to prevent oculogenital disease could be advanced by identifying chlamydial antigens that elicit pan-neutralizing antibodies, particularly among infected human populations of known risk factors. There is currently no objective high-throughputin vitroassay to screen human sera for neutralization to aid in identification of these antigens. This report describes an objective, high-throughputin vitroassay that measuresC. trachomatis-neutralizing antibodies. Antibody-mediated neutralization of chlamydial infection was performed in a 96-well microtiter format, and neutralization was quantified by immunostaining fixed cells followed by automated fluorometric analysis. This report shows that fluorometric analysis ofC. trachomatisinfection directly correlates to labor-intensive manual inclusion counts. Furthermore, this report shows that fluorometry can be used to identifyC. trachomatis serovar- and serocomplex-specific neutralization. This objective, high-throughput analysis of serum neutralization is amenable to epidemiological studies of human chlamydial infection, human clinical vaccine trials, and preclinical animal model experiments ofChlamydiainfection.

scholarly journals A Propagated Skeleton Approach to High Throughput Screening of Neurite Outgrowth for In Vitro Parkinson’s Disease Modelling

Cells ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 931
Author(s):  
Justus Schikora ◽  
Nina Kiwatrowski ◽  
Nils Förster ◽  
Leonie Selbach ◽  
Friederike Ostendorf ◽  
...  
Keyword(s):  
Parkinson’S Disease ◽  
Parkinson's Disease ◽  
Neurite Outgrowth ◽  
High Throughput ◽  
High Throughput Screening ◽  
Image Data ◽  
Large Data ◽  
Therapeutic Effects ◽  
Data Sets

Neuronal models of neurodegenerative diseases such as Parkinson’s Disease (PD) are extensively studied in pathological and therapeutical research with neurite outgrowth being a core feature. Screening of neurite outgrowth enables characterization of various stimuli and therapeutic effects after lesion. In this study, we describe an autonomous computational assay for a high throughput skeletonization approach allowing for quantification of neurite outgrowth in large data sets from fluorescence microscopic imaging. Development and validation of the assay was conducted with differentiated SH-SY5Y cells and primary mesencephalic dopaminergic neurons (MDN) treated with the neurotoxic lesioning compound Rotenone. Results of manual annotation using NeuronJ and automated data were shown to correlate strongly (R2-value 0.9077 for SH-SY5Y cells and R2-value 0.9297 for MDN). Pooled linear regressions of results from SH-SY5Y cell image data could be integrated into an equation formula (y=0.5410·x+1792; y=0.8789·x+0.09191 for normalized results) with y depicting automated and x depicting manual data. This automated neurite length algorithm constitutes a valuable tool for modelling of neurite outgrowth that can be easily applied to evaluate therapeutic compounds with high throughput approaches.

scholarly journals An in vitro assay for clonogenic, high‐throughput analysis of intestinal stem cells

2012 ◽  
Vol 26 (S1) ◽  
Author(s):  
Adam David Gracz ◽  
Michael J Johnston ◽  
Fengchao Wang ◽  
Ian A Williamson ◽  
Yuli Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
High Throughput ◽  
In Vitro Assay ◽  
Intestinal Stem Cells ◽  
High Throughput Analysis ◽  
Vitro Assay ◽  
Throughput Analysis
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