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BioAssay: AID 743365

qHTS profiling of inhibitors of Plasmodium falciparum (DD2) proliferation (acoustic dispense)

Plasmodium falciparum is a virulent malarial parasite that kills 1-2 million people worldwide each year. Transmitted by mosquitoes, the sporozoite form of the parasite enters the blood stream to infect hepatocytes, where they become merozoites that infect red blood cells (erythrocytes). Within erythrocytes, merozoites begin a 72-hour cycle of growth and division, after which the parasite is more ..
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 Tested Compounds
 Tested Compounds
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Inactive(127)
 
 
 Tested Substances
 Tested Substances
All(127)
 
 
Inactive(127)
 
 
 Related BioAssays
 Related BioAssays
AID: 743365
Data Source: NCGC (MMAcousticdd2)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: Other
BioAssay Version:
Deposit Date: 2014-04-01
Modify Date: 2014-04-02

Data Table ( Complete ):           View All Data
Tested Compounds:
Related Experiments
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AIDNameTypeComment
743367qHTS profiling of inhibitors of Plasmodium falciparum strainsSummarydepositor-specified cross reference
743322qHTS profiling for inhibitors of Plasmodium falciparum (3D7) proliferationConfirmatorysame project related to Summary assay
743323qHTS profiling for inhibitors of Plasmodium falciparum (DD2) proliferationConfirmatorysame project related to Summary assay
743324qHTS profiling for inhibitors of Plasmodium falciparum (HB3) proliferationConfirmatorysame project related to Summary assay
743325qHTS profiling of Chloroquine Analogs as inhibitors of Plasmodium falciparum (3D7) proliferationConfirmatorysame project related to Summary assay
743326qHTS profiling of Chloroquine Analogs as inhibitors of Plasmodium falciparum (DD2) proliferationConfirmatorysame project related to Summary assay
743327qHTS profiling of Chloroquine Analogs as inhibitors of Plasmodium falciparum (HB3) proliferationConfirmatorysame project related to Summary assay
743344qHTS profiling of the MIPE4 collection as inhibitors of Plasmodium falciparum (3D7) proliferationConfirmatorysame project related to Summary assay
743345qHTS profiling of the MIPE4 collection as inhibitors of Plasmodium falciparum (DD2) proliferationConfirmatorysame project related to Summary assay
743346qHTS profiling of the MIPE4 collection as inhibitors of Plasmodium falciparum (DD2) proliferation (Rep 2)Confirmatorysame project related to Summary assay
743347qHTS profiling of the MIPE4 collection as inhibitors of Plasmodium falciparum (HB3) proliferationConfirmatorysame project related to Summary assay
743364qHTS profiling of inhibitors of Plasmodium falciparum (3D7) proliferation (acoustic dispense)Confirmatorysame project related to Summary assay
743366qHTS profiling of inhibitors of Plasmodium falciparum (HB3) proliferation (acoustic dispense)Confirmatorysame project related to Summary assay
Description:
Plasmodium falciparum is a virulent malarial parasite that kills 1-2 million people worldwide each year. Transmitted by mosquitoes, the sporozoite form of the parasite enters the blood stream to infect hepatocytes, where they become merozoites that infect red blood cells (erythrocytes). Within erythrocytes, merozoites begin a 72-hour cycle of growth and division, after which the parasite is released to infect new red blood cells. Gametocytes, which form within erythrocytes as well, circulate in the bloodstream and are ingested by the mosquito, thereby completing the life cycle. To profile malarial isolates for differential chemical responses and to identify inhibitors of malarial growth, an assay that measures P. falciparum growth and replication within erythrocytes was employed. This assay uses a DNA binding fluorescent dye (SYBR Green) to detect parasite DNA following cell lysis. Because mature erythrocytes lack DNA, only Plasmodium-derived DNA is detected.
Protocol
The SYBR Green viability assay was adapted from a method described previously (Plouffe et al., 2008). For screening 3 ul culture medium was dispensed into 1536 well black clear bottom plates (Aurora Biotechnologies) using a Multidrop Combi (Thermo Fisher Scientific Inc.), 23 nL of compounds in DMSO were added by an acoustic dispenser (EDC Biosystems), and 5 ul of erythrocytes infected with P. falciparum (0.3% parasitemia, 2.5% hematocrit final concentration) were added. The plates were incubated at 37 C in a humidified incubator in 5% CO2 for 72 h, and 2 ul lysis buffer (20 mM Tris HCl, 10 mM EDTA, 0.16% saponin, 1.6% triton X, 10X SYBR Green I supplied as 10,000X final concentration by Invitrogen) was added to each well. The plates were mixed for 25 sec with gentle shaking and incubated overnight at room temperature in the dark. The following morning, fluorescence intensity at 485(14) nm excitation and 535(25) nm emission wavelengths was measured on an EnVision (Perkin Elmer) plate reader. Plate reads were normalized relative to the control inhibitor (0.29 uM artemisinin) and vehicle (DMSO) wells present on each plate and then corrected by an algorithm using vehicle-only control plates at the beginning and end of the compound plate stack.
Comment
Compound Ranking:
1. Compounds are first classified as having full titration curves, partial modulation, partial curve (weaker actives), single point activity (at highest concentration only), or inactive. See data field "Curve Description". For this assay, apparent inhibitors are ranked higher than compounds that showed apparent activation.
2. For all inactive compounds, PUBCHEM_ACTIVITY_SCORE is 0. For all active compounds, a score range was given for each curve class type given above. Active compounds have PUBCHEM_ACTIVITY_SCORE between 40 and 100. Inconclusive compounds have PUBCHEM_ACTIVITY_SCORE between 1 and 39. Fit_LogAC50 was used for determining relative score and was scaled to each curve class' score range.
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
1PhenotypeIndicates type of activity observed: inhibitor, activator, fluorescent, cytotoxic, inactive, or inconclusive.String
2Potency*Concentration at which compound exhibits half-maximal efficacy, AC50. Extrapolated AC50s also include the highest efficacy observed and the concentration of compound at which it was observed.FloatμM
3EfficacyMaximal efficacy of compound, reported as a percentage of control. These values are estimated based on fits of the Hill equation to the dose-response curves.Float%
4Analysis CommentAnnotation/notes on a particular compound's data or its analysis.String
5Activity_ScoreActivity score.Integer
6Curve_DescriptionA description of dose-response curve quality. A complete curve has two observed asymptotes; a partial curve may not have attained its second asymptote at the highest concentration tested. High efficacy curves exhibit efficacy greater than 80% of control. Partial efficacies are statistically significant, but below 80% of control.String
7Fit_LogAC50The logarithm of the AC50 from a fit of the data to the Hill equation (calculated based on Molar Units).Float
8Fit_HillSlopeThe Hill slope from a fit of the data to the Hill equation.Float
9Fit_R2R^2 fit value of the curve. Closer to 1.0 equates to better Hill equation fit.Float
10Fit_InfiniteActivityThe asymptotic efficacy from a fit of the data to the Hill equation.Float%
11Fit_ZeroActivityEfficacy at zero concentration of compound from a fit of the data to the Hill equation.Float%
12Fit_CurveClassNumerical encoding of curve description for the fitted Hill equation.Float
13Excluded_PointsWhich dose-response titration points were excluded from analysis based on outlier analysis. Each number represents whether a titration point was (1) or was not (0) excluded, for the titration series going from smallest to highest compound concentrations.String
14Max_ResponseMaximum activity observed for compound (usually at highest concentration tested).Float%
15Activity at 0.0000338500 uM (3.385e-05μM**)% Activity at given concentration.Float%
16Activity at 0.0000927659 uM (9.27659e-05μM**)% Activity at given concentration.Float%
17Activity at 0.0001693500 uM (0.00016935μM**)% Activity at given concentration.Float%
18Activity at 0.0002761802 uM (0.00027618μM**)% Activity at given concentration.Float%
19Activity at 0.0004467900 uM (0.00044679μM**)% Activity at given concentration.Float%
20Activity at 0.0008521697 uM (0.00085217μM**)% Activity at given concentration.Float%
21Activity at 0.00150 uM (0.00150088μM**)% Activity at given concentration.Float%
22Activity at 0.00257 uM (0.00257289μM**)% Activity at given concentration.Float%
23Activity at 0.00570 uM (0.00569606μM**)% Activity at given concentration.Float%
24Activity at 0.00773 uM (0.00773354μM**)% Activity at given concentration.Float%
25Activity at 0.023 uM (0.022589μM**)% Activity at given concentration.Float%
26Activity at 0.031 uM (0.0308383μM**)% Activity at given concentration.Float%
27Activity at 0.068 uM (0.068144μM**)% Activity at given concentration.Float%
28Activity at 0.104 uM (0.104288μM**)% Activity at given concentration.Float%
29Activity at 0.205 uM (0.205107μM**)% Activity at given concentration.Float%
30Activity at 0.327 uM (0.326898μM**)% Activity at given concentration.Float%
31Activity at 0.621 uM (0.621234μM**)% Activity at given concentration.Float%
32Activity at 1.094 uM (1.09414μM**)% Activity at given concentration.Float%
33Activity at 1.876 uM (1.87564μM**)% Activity at given concentration.Float%
34Activity at 4.152 uM (4.15244μM**)% Activity at given concentration.Float%
35Activity at 5.632 uM (5.63194μM**)% Activity at given concentration.Float%
36Activity at 16.49 uM (16.4932μM**)% Activity at given concentration.Float%
37Activity at 23.19 uM (23.1939μM**)% Activity at given concentration.Float%
38Activity at 49.83 uM (49.827μM**)% Activity at given concentration.Float%
39Activity at 100.00 uM (100μM**)% Activity at given concentration.Float%
40Compound QCNCGC designation for data stage: 'qHTS', 'qHTS Verification', 'Secondary Profiling'String

* Activity Concentration. ** Test Concentration.

Data Table (Concise)
Data Table ( Complete ):     View All Data
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