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BioAssay: AID 624012

Anti-Fungal Single Agent: S.cerevisiae growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-06_Inhibitor_Dose_DryPowder_Activity_Set4

Assay Overview: Fungal growth in glycerol-supplemented medium vs. fungal growth in glucose-supplemented medium. The goal of this assay is to identify drug-like molecules that selectively inhibit the growth of Saccharomyces cerevisiae when they are cultured under conditions that require mitochondrial respiration to support metabolic needs. ..more
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 Tested Compounds
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Active(8)
 
 
 Tested Substances
 Tested Substances
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Active(9)
 
 
 Related BioAssays
 Related BioAssays
AID: 624012
Data Source: Broad Institute (2148-06_Inhibitor_Dose_DryPowder_Activity_Set4)
BioAssay Type: Confirmatory, Concentration-Response Relationship Observed
Depositor Category: NIH Molecular Libraries Probe Production Network
Deposit Date: 2012-04-05
Hold-until Date: 2013-04-05
Modify Date: 2013-04-05

Data Table ( Complete ):           View Active Data    View All Data
BioActive Compounds: 8
Related Experiments
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AIDNameTypeComment
588528Broad Institute Reversing Antifungal Drug Resistance: Single Agent Probe Inhibitor Probe ProjectSummarydepositor-specified cross reference: Summary assay
1979Fluorescence Cell-Based Primary HTS of C.albicans growth in the presence of Fluconazole and compoundScreeningsame project related to Summary assay
588529Anti-Fungal Single Agent: CaCi2 Measured in Microorganism System Using Plate Reader - 2148-01_Inhibitor_Dose_CherryPick_ActivityConfirmatorysame project related to Summary assay
588530CaCi17 Retest at Dose Measured in Microorganism System Using Plate Reader - 2148-02_Inhibitor_Dose_CherryPick_ActivityConfirmatorysame project related to Summary assay
588634Antifungal Single Agent - Mammalian Cell Toxicity Measured in Cell-Based and Microorganism Combination System Using Plate Reader - 2148-03_Inhibitor_Dose_CherryPick_ActivityConfirmatorysame project related to Summary assay
602184Anti-Fungal Single Agent: S.cerevisiae growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-06_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
602185Anti-Fungal Single Agent: C.glabrata growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-05_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
602186Anti-Fungal Single Agent: C.glabrata growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-04_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
602187Anti-Fungal Single Agent: S.cerevisiae growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-07_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
602188Antifungal Single Agent - Mammalian Cell Toxicity Measured in Cell-Based and Microorganism Combination System Using Plate Reader - 2148-03_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
602189CaCi17 Retest at Dose Measured in Microorganism System Using Plate Reader - 2148-02_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
602190Anti-Fungal Single Agen: CaCi2 Measured in Microorganism System Using Plate Reader - 2148-01_Inhibitor_Dose_DryPowder_ActivityConfirmatorysame project related to Summary assay
602394Antifungal Single Agent - Mammalian Cell Toxicity Measured in Cell-Based and Microorganism Combination System Using Plate Reader - 2148-03_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatorysame project related to Summary assay
623897CaCi17 Retest at Dose Measured in Microorganism System Using Plate Reader - 2148-02_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatorysame project related to Summary assay
623899Anti-Fungal Single Agent: CaCi2 Measured in Microorganism System Using Plate Reader - 2148-01_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatorysame project related to Summary assay
623903Anti-Fungal Single Agent: C.glabrata growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-04_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatorysame project related to Summary assay
623904Anti-Fungal Single Agent: C.glabrata growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-05_Inhibitor_Dose_DryPowder_Activity_Set2Confirmatorysame project related to Summary assay
623965Anti-Fungal Single Agent: C.glabrata growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-04_Inhibitor_Dose_DryPowder_Activity_Set3Confirmatorysame project related to Summary assay
623967Anti-Fungal Single Agent: C.glabrata growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-05_Inhibitor_Dose_DryPowder_Activity_Set3Confirmatorysame project related to Summary assay
623969Anti-Fungal Single Agent: S.cerevisiae growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-06_Inhibitor_Dose_DryPowder_Activity_Set3Confirmatorysame project related to Summary assay
623971Anti-Fungal Single Agent: S.cerevisiae growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-07_Inhibitor_Dose_DryPowder_Activity_Set3Confirmatorysame project related to Summary assay
623981Anti-Fungal Single Agen: CaCi2 Measured in Microorganism System Using Plate Reader - 2148-01_Inhibitor_Dose_DryPowder_Activity_Set3Confirmatorysame project related to Summary assay
623982CaCi17 Retest at Dose Measured in Microorganism System Using Plate Reader - 2148-02_Inhibitor_Dose_DryPowder_Activity_Set3Confirmatorysame project related to Summary assay
624006Anti-Fungal Single Agen: CaCi2 Measured in Microorganism System Using Plate Reader - 2148-01_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatorysame project related to Summary assay
624013Anti-Fungal Single Agent: C.glabrata growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-05_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatorysame project related to Summary assay
624014Anti-Fungal Single Agent: C.glabrata growth, Glycerol Measured in Microorganism System Using Plate Reader - 2148-04_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatorysame project related to Summary assay
624025Anti-Fungal Single Agent: S.cerevisiae growth, Glucose Measured in Microorganism System Using Plate Reader - 2148-07_Inhibitor_Dose_DryPowder_Activity_Set4Confirmatorysame project related to Summary assay
Description:
Keywords: Saccaromyces cerevisiae, glycolysis, respiration


Assay Overview: Fungal growth in glycerol-supplemented medium vs. fungal growth in glucose-supplemented medium. The goal of this assay is to identify drug-like molecules that selectively inhibit the growth of Saccharomyces cerevisiae when they are cultured under conditions that require mitochondrial respiration to support metabolic needs.


Expected Outcome: reduction of signal
Protocol
1) Inoculate an overnight culture in YPD media at 30 degrees C (note: colonies of S. cerevisiae should appear pink)
2)Preparing the assay plates:
a.Fill a clear, flat bottom, 384-well plate with 25uL of CSM-glycerol media
b.Pin in compounds (positive control: Amphotericin B, 10 uM)
3)Prepare cells: Dilute O/N culture of S. cerevisiae 1:1000 (e.g. 15 uL into 15 mL) directly from culture tube into CSM-glycerol media.
a.Add 25ul of cells to assay plates (which were pinned-into).
b.Incubate 48hr at 30 degrees C, no agitation, in a humidified incubator to prevent evaporation.
c.Read OD600 of plates.
Comment
PRESENCE OF CONTROLS: Neutral control wells (NC; n=168) and positive control wells (PC; n=8) were included on every plate.
EXPECTED OUTCOME: Active compounds result in decreasing readout signal.
ACTIVE CONCENTRATION LIMIT:
For each sample, the highest valid tested concentration (Max_Concentration) was determined and the active concentration limit (AC_limit) was set to equal (2)(Max_Concentration).
NORMALIZATION:
The raw signals of the plate wells were normalized using the 'Neutral Controls Minus Inhibitors' method in Genedata Assay Analyzer (v7.0.3):
The median raw signal of the intraplate neutral control wells was set to a normalized activity value of 0.
The median raw signal of the intraplate positive control wells was set to a normalized activity value of -100.
Experimental wells values were scaled to this range.
PATTERN CORRECTION: No plate pattern correction algorithm from Genedata Condoseo (v.7.0.3) was applied.
MEASUREMENT USED TO DETERMINE ACTIVE CONCENTRATION (AC): AC50
AC values were calculated using the curve fitting strategies in Genedata Screener Condoseo (7.0.3).
AC values were calculated up to the active concentration limit described for each sample.
pAC was set to equal -1*log10(AC)
PUBCHEM_ACTIVITY_OUTCOME:
Activity_Outcome = 1 (inactive) when:
a) compound shows activity but in a direction opposite to the expected outcome
in these cases, values describing curve fitting parameters (Sinf, S0, Hill Slope, log_AC50, log_AC50_SE) are set to null
b) curve fit is constant where activity is > -30% and < 30% at all tested concentrations, or
c) AC > AC_limit
Activity_Outcome = 2 (active) when:
AC <= AC_limit
Activity_Outcome = 3 (inconclusive) when:
a) Curve fitting strategy resulted in a constant fit with activity >= -70% but <= -30%, or
b) The fit was deemed not valid due to poor fit quality.
PUBCHEM_ACTIVITY_SCORE:
If PUBCHEM_ACTIVITY_OUTCOME = 1 (inactive) or 3 (inconclusive),
then PUBCHEM_ACTIVITY_SCORE = 0
If PUBCHEM_ACTIVITY_OUTCOME = 2 (active)
then PUBCHEM_ACTIVITY_SCORE = (10)(pAC)
Scores relate to AC in this manner:
120 = 1 pM
90 = 1 nM
60 = 1 uM
30 = 1 mM
0 = 1 M
When the active concentration (AC) is calculated to be greater than the highest valid tested concentration (Max_Concentration), the PUBCHEM_ACTIVITY_SCORE is calculated using Max_Concentration as the basis.
When the active concentration (AC) is calculated to be less than the lowest tested concentration, the PUBCHEM_ACTIVITY_SCORE is calculated using the lowest tested concentration as the basis.
Note:
The individual dose data point columns ('Activity_at_xxuM') reported here represent the median of valid (unmasked) replicate observations at each concentration. These values are the inputs to a curve fitting algorithm.
All other data columns represent values which are derived during the curve fitting algorithm; this may sometimes include automatic further masking of some replicate data points.
Occasionally this results in perceived inconsistencies: for example, between the derived 'Maximal_Activity' and the apparent most active data point.
Categorized Comment - additional comments and annotations
From PubChem:
Assay Format: Organism-based
Result Definitions
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TIDNameDescriptionHistogramTypeUnit
OutcomeThe BioAssay activity outcomeOutcome
ScoreThe BioAssay activity ranking scoreInteger
1AC50_Qualifier'>','=', or '<'String
2AC50_uM*The concentration at which activity reaches 50% of the maximumFloatμM
3pAC50_MEqual to -1*log10(AC50)String
4Hill_SlopeThe slope at AC50Float
5S0_(%)The fitted activity value at zero concentrationFloat%
6Sinf_(%)The fitted activity value at infinite concentrationFloat%
7Num_PointsThe number of data points used to generate the plotInteger
8Max_Activity_(%)The maximum activity value observed, based on mean of replicates per concentrationFloat%
9Max_Activity_Conc_uMThe concentration at which the maximum activity is observedFloatμM
10Max_Concentration_uMMaximum valid test concentrationFloatμM
11Activity_at_0.000006uM_(%) (6e-06μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
12Activity_at_0.000012uM_(%) (1.2e-05μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
13Activity_at_0.000023uM_(%) (2.3e-05μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
14Activity_at_0.000046uM_(%) (4.6e-05μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
15Activity_at_0.0001uM_(%) (0.0001μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
16Activity_at_0.000195uM_(%) (0.000195μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
17Activity_at_0.00038uM_(%) (0.00038μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
18Activity_at_0.00075uM_(%) (0.00075μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
19Activity_at_0.0015uM_(%) (0.0015μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
20Activity_at_0.003uM_(%) (0.003μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
21Activity_at_0.006uM_(%) (0.006μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
22Activity_at_0.012uM_(%) (0.012μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
23Activity_at_0.0235uM_(%) (0.0235μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
24Activity_at_0.05uM_(%) (0.05μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
25Activity_at_0.1uM_(%) (0.1μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
26Activity_at_0.195uM_(%) (0.195μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
27Activity_at_0.38uM_(%) (0.38μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
28Activity_at_0.8uM_(%) (0.8μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
29Activity_at_1.6uM_(%) (1.6μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%
30Activity_at_3uM_(%) (3μM**)The average measured activity of all accepted replicates at the specified concentrationFloat%

* Activity Concentration. ** Test Concentration.
Additional Information
Grant Number: 1 R03 MH086456-01

Data Table (Concise)
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