systems biology in small fish models
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Systems Biology in Small Fish Models. D.L. Villeneuve The McKim Conference on Predictive Toxicology Sept. 25-27, 2007, Duluth, Minnesota. Systems Biology in Small Fish Models. What is systems biology or the “systems perspective”? The systems perspective in ecotoxicology. - PowerPoint PPT PresentationTRANSCRIPT
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Systems Biology in Small Fish ModelsD.L. VilleneuveThe McKim Conference on Predictive ToxicologySept. 25-27, 2007, Duluth, Minnesota
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Systems Biology in Small Fish ModelsWhat is systems biology or the systems perspective?The systems perspective in ecotoxicology.An overview of a systems-oriented small fish research program.A case studyConclusions
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The Systems Perspective The experimenter can draw a boundary around it, heuristically The experimenter can conduct defined perturbations within the boundaries The experimenter can, by reasoning generate sensible explanations for the changed behavior. Learning is assessed by how well the understanding enables prediction of changed system behavior in response to defined perturbations. A subset of the world whose behavior, and whose interaction with the world, we believe can be sensibly described.What is a system?Brent 2004, Nature Biotech. 22: 1211-1214Kuipers, B. 1994. MIT Press, Cambridge
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www.oz.net/~geoffsi/bm2003-days/bm2003-20.htm Systems biology is the study of an organism, viewed as an integrated and interacting network of genes, proteins and biochemical reactions which give rise to life. Systems are comprised of parts which interact.
Interaction of these parts gives rise to "emergent properties".
Emergent properties cannot be attributed to any single parts of the system. Irreducible.
To understand systems, and to be able to fully understand a system's emergent properties, systems need be studied as a whole. http://www.systemsbiology.org/Intro_to_ISB_and_Systems_Biology/Why_Systems_MatterThe Systems Perspective
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The Systems PerspectiveNon-equilibrium thermodynamics (1930s-40s) dealt with integration quantitatively aimed to discover general principles >> descriptive established connection to molecular mechanisms Investigation of biological self-organization (1950s) examination of how structures, oscillations, and/or waves arise in a steady or homogenous environment Feedback regulation in metabolism (late 1950s) Systems theory in biology (1960s) search for general biological laws governing behavior and evolution of living matter in a way analogous to the relation of physical laws and non-living matter Metabolic control analysis (1970s)Approaches to characterize properties of networks of interacting chemical reactions Convergence with mainstream molecular biology high throughput, genome-scale, data-richWesterhoff and Palsson, 2004 Nature Biotech. 22:1249-1252Wolkenhauer. 2001 Briefings in Bioinformatics. 2:258-270
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The Systems Perspective Shift from reductionism to a holistic perspective Should reduce complexity rather than adding additional layers of complexity
Search for organizing principles over construction of predictive descriptions (models) that exactly describe the evolution of a system in space and time.
Identification of new concepts and hypotheses that provide a conceptual structure with logical coherence to rival chemistry and physics.Mesarovic et al. 2004. Syst. Biol. 1:19-27
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The Systems Perspective in Ecotoxicology Shift from reductionism to a holistic perspective Historically, ecotoxicology and ecological risk assessment was holistic in focus Apical/integrative endpoints: Survival, growth, reproduction Not feasible to test every chemical, let alone every chemical mixture Increased emphasis on subtle, chronic impacts (e.g., development, behavior, etc.) with long-term population implications. Reductionist in the sense that testing of the universe of chemicals was the paradigm.
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Search for organizing principles that underlie biological response to stressors.
Develop a conceptual structure with logical coherence that allows us to predict, with reasonable and quantitative certainty, integrated, ecologically-relevant impacts.
The Systems Perspective in Ecotoxicology
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USEPA Cincinnati, OHD. Bencic, M. Kostich, I. Knoebl, D. Lattier, J. Lazorchak, G. Toth, R. Wang, USEPA Duluth, MN, and Grosse Isle, MIG. Ankley, E Durhan, M Kahl, K Jensen, E Makynen, D. Martinovic, D. Miller, D. Villeneuve, USEPA Athens, GAT. Collette, D. Ekman, J. Kenneke, T. Whitehead, Q. Teng USEPA-RTP, NCM. Breen, R. ConollyUSEPA STAR ProgramN. Denslow (Univ. of Florida), E. Orlando, (Florida Atlantic University), K. Watanabe (Oregon Health Sciences Univ.), M. Sepulveda (Purdue Univ.)USACE Vicksburg, MSE. PerkinsOther partnersJoint Genome Institute, DOE (Walnut Creek, CA)Sandia, DOE (Albuquerque, NM)Pacific Northwest National Laboratory (Richland, WA)C. Tyler (Univ. Exeter, UK)Linkage of Exposure and Effects Using Genomics, Proteomics, and Metabolomics in Small Fish Models
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11HSDP45011.GnRHNeuronal SystemGABADopamineBrain
GonadotrophPituitaryD1 RD2 R??Y2 RNPYY1 RGnRH R?ActivinInhibinGABAA RGABAB RY2 RD2 RGnRH Circulating Sex Steroids / Steroid Hormone Binding GloblulinPACAPFollistatinActivinPAC1 RActivin RFSHbLHbGPaCirculating LH, FSH FSH RLH RCirculating LDL, HDLLDL RHDL ROuter mitochondrial membraneInner mitochondrial membraneStAR P450sccpregnenolone3bHSDP450c17androstenedionetestosteroneCholesterolestradiolGonad(Generalized, gonadal, steroidogenic cell)Androgen / Estrogen Responsive Tissues(e.g. liver, fatpad, gonads)ARER3456BloodBloodCompartment17HSDP450arom11-ketotestosteroneprogesterone17-hydroxyprogesterone17,20-P (MIS)20HSD
Fipronil* (-)Muscimol* (+)
Apomorphine (+)Haloperidol (-)
Trilostane (-)Ketoconazole (-)
Fadrozole (-)Prochloraz (-,-)
Vinclozolin (-)Flutamide (-)-trebolone (+)Ethynyl estradiol (+)78910111221Chemical Probes
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Figure 1. Conceptual Overview of ResearchIncreasing Ecological RelevanceIncreasing Diagnostic (Screening) UtilityPhase 1.Fathead minnow 21 d reproduction test
Phase 2.Zebrafish genomics proteomics, metabalomicsPopulation modelingPhase 3. Fathead minnow molecular markers metabolomicsSystems modelingMolecular
mRNA, protein, Enzyme assaysCellular
Metabolite profilesOrganFunctional andStructural change(Pathology)Individual
Altered reproductionor developmentPopulation
Decreased numbersof animalsLevels of BiologicalOrganizationWell characterized genome, low eco / regulatory relevanceComputational modelingPoorly characterized genomehigh eco / regulatory relevances Depict the flow of information
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A Case Study with FadrozoleAromatase
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OvaryBrain
Chart1
105.67550015471.16001622921.1600162292
815.77180800828.35022964268.3502296426
1521.150972877432.967819434432.9678194344
2641.115170762472.568430940372.5684309403
3915.6434244345181.0148544948181.0148544948
4684.63776393561.038190912961.0381909129
IC50=8.82 +/- 1.58 M
A
*B*
*C*
*D*
*E*
*F*
fadrozole (uM)
fmol/h/mg
Fig1
A. Brain
B. Ovary
&A
&F
Fig1
5211.7958042402279.2917918464279.2917918464
3781.5648140955501.617257516501.617257516
3876.3664579737398.1876749747398.1876749747
ANOVA p=0.0593
brain fmol/h/mg
fmol/h/mg
Fig2
92.567078269817.040929608717.0409296087
17.61433272994.21165942574.2116594257
64.923946586826.759560242726.7595602427
A
AB
B
ANOVA p=0.0199
fmol/h/mg
Fig3
A. Brain
B. Ovary
&A
&F
Fig3
0.52182581730.12865458090.1286545809
0.55631737610.13913094450.1391309445
0.72898982040.20469249330.2046924933
ANOVA p=0.6284
pg P450aromB mRNA / ug total RNA
Fig4
69.6607717278.57367495528.5736749552
115.855689717829.04734423229.047344232
55.328877924517.284678991917.2846789919
ANOVA (log y) p=0.1352
pg P450aromB mRNA / ug total RNA
Fig5
A. Brain
B. Ovary
&A
&F
Fig5
105.67550015471.16001622921.1600162292
815.77180800828.35022964268.3502296426
1521.150972877432.967819434432.9678194344
2641.115170762472.568430940372.5684309403
3915.6434244345181.0148544948181.0148544948
4684.63776393561.038190912961.0381909129
IC50=8.82 +/- 1.58 M
A
*B*
*C*
*D*
*E*
*F*
fadrozole (uM)
fmol/h/mg
Fig6
2.18327194960.14883373870.1488337387
2.90690721950.10371134010.1037113401
4.60666491750.28372638360.2837263836
5.8648370230.101131210.10113121
6.4694631780.07793391910.0779339191
IC50 = 6.93 +/- 0.80 M
A
B
*C*
*D*
*E*
fadrozole (uM)
fmol/h/mg
Fig7
&A
&F
Fig7
0.04976453920.0685840784
0.12802174660.1345654982
0.26077568050.2648337828
0.50247712390.48457595
11
Ovary
Brain
nM [3H]-AD
AAo/AAmax
Fig8
&A
&F
Fig8
6238.69672043581189.97519258399.27288579079.2728857907163.3639139103163.3639139103
14779.15642915641319.06598297956.179350241556.1793502415298.2195570378298.2195570378
12595.69169960471345.642559555619.193451356319.1934513563112.9876512721112.9876512721
11456.29847238541367.716055976960.919357945160.9193579451493.7705441699493.7705441699
10637.79617562231433.2218780045134.5068291497134.5068291497596.6096776993596.6096776993
Brain
Ovary
mg/ml microsomal protein
fmol/h/mg
A.
B.
&A
&F
3197.58059960179.721953274479.7219532744
2579.974640757951.926748744751.9267487447
3073.9914938098110.7724416262110.7724416262
2291.777651179429.612770045929.6127700459
1493.553444648227.334051897327.3340518973
a
a
ab
b
b
c
ANOVA (log x) p
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PregnenoloneProgesterone17a-OH-Pregnenolone17a-OH-ProgesteroneCYP17 (hydroxylase)CYP11A3b-HSD3b-HSDDHEAAndrostenedioneTestosteroneCYP17 (lyase)3b-HSD17b-HSDCYP1917b-estradiolCholesterol20b-HSD1720-dihydroxy-4-pregnen-3-oneCYP17 (hydroxylase)CYP17 (lyase)estroneCYP1917b-HSD11-OH-TestosteroneCYP11B111-Ketotestosterone11HSD11-deoxycorticosteroneCYP21corticosteroneCYP11B1aldosteroneCYP11B211-deoxycortisolCYP21cortisolCYP11B2A Case Study with Fadrozole
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25oC, 12 hRIAAndrostenedioneTestosterone17b-HSDCYP1917b-estradiolestroneCYP1917b-HSD
Chart1
0.31989224950.03820228980.0382022898
0.20519668820.02605756530.0260575653
0.15406868360.02440912480.0244091248
A
B
C
24 h-E2
Fadrozole (ug/L)
Ex vivo E2 production (ng/ml*12 h)
E2-RIA d1,16
Small Fish Comp Tox
Phase 3, Fadrozole
Ex vivo steroid production
24 h, d.16 females, E2
Extraction: Notebook 341:83
RIA: Notebook 341:85-90
DLV, NDM, JET
adjusted
E2 (ng/ml)log E2dpmdpmBo/BmBo/BmMeanSDCV
60.77815125041777.781750.930.10775106140.10451585460.1061334580.00228763662.1554340001
20.30102999572480.792664.480.19245805230.21459120420.20352462820.01565050187.6897336556
0.6666666667-0.17609125914608.974475.540.44888644420.43280921380.4408478290.01136831872.5787398599
0.2222222222-0.65321251386630.838484.190.69250414040.91581895610.80416154830.157907420519.6362809051
0.0740740741-1.130333768510303.59759.941.13503102361.06953646171.10228374270.04631164894.2014271877
0.024691358-1.607455023210961.211064.91.21427852781.22677353491.22052603140.00883530430.7238931442
MeanSDCVSigmoid fit; regression using GraphPad Prism
Bm9217.219148.469182.8352178.798099924223.7268566834
NS877.97889.07883.527.84888527120.8883653195
linearadj for dil
CT iddpmBo/Bmlog ng/mlng/mldil factorng/ml
580525117.730.510187889-0.27460080.53137265330.53333333330.2833987484
580546121.840.6311749825-0.43005850.37148518620.61538461540.2286062684
580564205.580.400281228-0.11684840.76410246370.50.3820512319
580587147.170.7547189135-0.58426740.26045494060.50.1302274703
580607565.580.8051339177-0.64890510.224437230.57142857140.1282498457
580626302.650.6529611179-0.45721570.34896695190.50.1744834759
580645166.540.5160690973-0.28246340.52183908020.50.2609195401
580664959.810.4911598126-0.24885560.56382509270.50.2819125464
580689853.261.0807807632-1.0940160.08053487710.50.0402674385
580709640.021.0550870765-1.0391030.0913896470.57142857140.0522226554
5807210390.81.1455499641-1.2635920.05450144290.53333333330.0290674362
580745956.390.6112396023-0.40509460.39346435990.50.19673218
580763764.210.3470997305-0.029176910.93502471390.50.467512357
580784729.250.4633792066-0.21025840.61622824440.50.3081141222
580804299.660.4116171033-0.13435810.73390847020.57142857140.4193762687
580824644.330.4531470368-0.19570950.63722161650.53333333330.3398515288
580846519.770.6791223131-0.48973850.32378855980.57142857140.1850220341
5808613511.71.52159304710.57142857140
580887453.060.7915761723-0.6313210.23371091730.53333333330.1246458226
580905139.150.5127688249-0.27807060.52714416060.53333333330.2811435523
5809211174.41.23996739490.53333333330
5809410215.21.1243915914-1.2018970.06282073310.57142857140.0358975618
5809610847.51.200578602-1.4760980.03341196360.57142857140.0190925506
5809811691.71.30229784020.50
583886036.970.6209488373-0.41726870.38258796110.57142857140.2186216921
583908790.110.9526798296-0.85737110.13887654390.57142857140.0793580251
583926065.930.624438282-0.42164160.3787550220.57142857140.2164314412
583946796.590.7124768731-0.53125110.29427197230.57142857140.1681554127
583964965.120.4917996244-0.24972620.56269596380.57142857140.3215405508
583988958.980.973027292-0.89000120.12882459920.57142857140.0736140567
584005236.030.5244420774-0.29359420.50863448310.57142857140.290648276
584024204.680.4001727853-0.1166780.76440232620.57142857140.4368013292
584044340.080.4164873848-0.14173360.72154994850.57142857140.4123142563
584065392.30.5432713423-0.31830250.48050454510.57142857140.2745740258
584085053.270.5024209829-0.26415910.54430321560.57142857140.3110304089
5841011336.51.25949912730.57142857140
584126282.380.6505187476-0.45417740.35141686490.57142857140.2008096371
5841410655.51.1774441626-1.3745880.04220967420.57142857140.0241198138
584167125.030.7520512235-0.5808960.26248470370.57142857140.1499912593
584186680.150.6984467995-0.51376960.30635882840.57142857140.1750621877
584207013.730.7386404782-0.56400140.27289689860.57142857140.1559410849
584226722.130.7035050483-0.5200660.30194928110.57142857140.1725424464
584244831.40.4756874513-0.22751850.59221785910.57142857140.3384102052
584265363.110.5397541845-0.31371920.48560237260.57142857140.2774870701
584284459.020.430818688-0.16320570.68674309280.57142857140.3924246244
584304406.40.4244784057-0.15377360.70182106610.57142857140.4010406092
584324334.960.4158704664-0.14079960.72310339410.52631578950.3805807337
584343477.20.31251735840.034193231.08191521920.52631578950.5694290627
QA3830.870.3551317187-0.043097130.90553005640.52631578950.4765947665
QA4305.260.4122918578-0.13538160.73218090760.52631578950.3853583724
QA4043.070.3807000939-0.085735130.82085201760.52631578950.4320273777
QA4607.430.4487008868-0.1893090.64668233830.52631578950.3403591254
Mean0.776311330.4085849105
SD0.1117027630.0587909279
CV14.388913140114.3889131401
E2-RIA d1,16
0
0
0
0
0
0
T-RIA d1,16
Small Fish Comp Tox
Phase 3, Fadrozole
Small Fish Comp Tox
Phase 3, Fadrozole
Ex vivo steroid production
24 h, d.16 females, T
Extraction: Notebook 341:83
RIA: Notebook 341:85-90
DLV, NDM, JET
T (ng/ml)log TdpmdpmBo/BmBo/BmMeanSDCV
60.7781512504769.79853.710.07044473850.10121092360.08582783110.021754978125.3472303948
20.30102999571043.31942.920.17072079830.13391649280.15231864560.02602457417.0856127958
0.6666666667-0.17609125911549.651706.360.35635175810.41380372260.38507774030.040624673710.5497330623
0.2222222222-0.65321251382536.682366.960.71821004740.65598844430.68709924590.219172590231.898243457
0.0740740741-1.13033376852819.152856.240.82176729590.83536498180.82856613890.00961501591.160440366
0.024691358-1.60745502323178.793212.420.95361608990.9659452940.95978069190.16645720917.3432545992
Sigmoid fit R^2 =0.9967; Regression conducted using GraphPad Prism
MeanSDCV
Bm3300.843309.783305.31410.51633345512.4199041377
NS568.75586.53577.6412.57235856952.1765041496
Bm-3SD2069.2909996350.546859040710.5=MDL
polynomialadj for dil
CT iddpmBo/Bmlog ng/mlng/mldil factorng/ml
580522359.220.6531508577-0.62443230.23744755390.53333333330.1266386954
580543268.070.98634732210.61538461540
580562808.770.8179618502-0.98558960.10337378070.50.0516868903
580582468.550.6932326858-0.69866390.20014101590.50.100070508
580602550.650.7233316347-0.75839320.17442422420.57142857140.0996709853
580622877.090.8430088684-1.0630150.08649380440.50.0432469022
580642774.530.8054090121-0.95040710.11209671870.50.0560483593
580661770.590.4373512925-0.26710980.54061762460.50.2703088123
580682149.750.5763563774-0.49298310.32137855960.50.1606892798
580702136.890.5716417309-0.48519990.3271900590.57142857140.186965748
580721979.330.5138781451-0.39127760.40618361460.53333333330.2166312611
580742437.260.6817613568-0.67686420.21044363750.50.1052218188
580762880.740.8443470068-1.0675280.08559965220.50.0427998261
580782669.960.7670722631-0.85402370.13995109470.50.0699755474
580802964.090.8749042223-1.1817730.06580016760.57142857140.0376000958
580822842.620.8303717092-1.0226350.09492158910.53333333330.0506248475
580842588.930.7373655904-0.78779450.16300671670.57142857140.0931466953
580863538.441.08546855010.57142857140
580882536.740.7182320442-0.74800460.17864686530.53333333330.0952783281
580902819.140.8217636298-0.99667260.10076910470.53333333330.0537435225
580922624.60.7504426855-0.81623170.15267513050.53333333330.0814267363
580942263.510.6180623023-0.56302920.27350848250.57142857140.1562905614
580962318.40.6381857043-0.59788840.25241293110.57142857140.1442359606
580982350.160.6498293415-0.61849030.24071862830.50.1203593142
583882755.50.7984323617-0.9317560.11701566360.57142857140.0668660935
583903101.520.9252878831-1.4647370.03429754230.57142857140.0195985956
583923102.530.9256581625-1.4676940.03406481230.57142857140.019465607
583942637.440.7551499998-0.82672560.14903023970.57142857140.085160137
583962473.930.6952050651-0.70246950.19839489820.57142857140.1133685133
583982636.570.7548310463-0.82600440.14927792860.57142857140.0853016735
584002720.090.785450586-0.89844920.1263428880.57142857140.072195936
584022247.380.6121488303-5.53E-010.27994441010.57142857140.1599682344
584042706.770.7805672974-0.88632460.12991981690.57142857140.0742398954
584063008.320.8911195269-1.2551990.05556495920.57142857140.0317514053
584082700.020.7780926578-0.8803220.13172797030.57142857140.0752731259
584103295.10.99625687860.57142857140
584123066.90.9125957319-1.3750210.04216761130.57142857140.0240957779
584143055.740.9085043279-1.3495690.04471271070.57142857140.0255501204
584162893.350.84897-1.0832530.08255568780.57142857140.0471746787
584183006.830.8905732732-1.2525650.05590298520.57142857140.031944563
584202674.50.7687366874-0.85794330.13869368910.57142857140.0792535366
584222728.380.78848981-0.90608230.12414170330.57142857140.0709381162
584242827.980.825004491-1.0063240.09855439580.57142857140.0563167976
584262351.990.6505002438-0.61969050.24005430570.57142857140.137173889
584282528.630.7152588106-0.7419650.18114860750.57142857140.10351349
584302415.060.6736225423-0.66168090.21793104430.57142857140.1245320253
584322404.070.6695934626-0.65426870.22168244360.52631578950.1166749703
584342106.030.5603280455-0.46663540.3414794710.52631578950.1797260374
1
1
1
QA1922.260.4929555261-0.35752440.43901119990.52631578950.2310585263
QA2003.220.5226365359-0.40541780.39317165440.52631578950.2069324497
QA1885.690.4795484791-0.33586390.46146216580.52631578950.2428748241
QA1955.110.5049987718-0.37695340.41980402670.52631578950.2209494878
Mean0.42836226170.225453822
SD0.02898635170.0152559746
CV6.76678463926.7667846392
T-RIA d1,16
0
0
0
0
0
0
T (log ng/ml)
Bo/Bm
forSAS
CT idfadtime pointtissueex_vivo_wtE2_ng_mlT_ng_ml
5805201y28.30.28339874840.1266386954
5805401y33.30.22860626840
5805601y30.10.38205123190.0516868903
5805801y43.30.13022747030.100070508
5806031y61.30.12824984570.0996709853
5806231y27.40.17448347590.0432469022
5806431y33.50.26091954010.0560483593
5806631y28.50.28191254640.2703088123
58068301y17.70.04026743850.1606892798
58070301y300.05222265540.186965748
58072301y24.40.02906743620.2166312611
58074301y13.80.196732180.1052218188
5807601y13.30.4675123570.0427998261
5807801y22.50.30811412220.0699755474
5808001y21.50.41937626870.0376000958
5808201y33.50.33985152880.0506248475
5808431y23.20.18502203410.0931466953
5808631blankblank00
5808831y30.20.12464582260.0952783281
5809031y26.70.28114355230.0537435225
58092301y43.100.0814267363
58094301y9.70.03589756180.1562905614
58096301y35.20.01909255060.1442359606
58098301y42.100.1203593142
58388016y19.70.21862169210.0668660935
58390016y8.40.07935802510.0195985956
58392016y12.80.21643144120.019465607
58394016y7.50.16815541270.085160137
58396316y9.10.32154055080.1133685133
58398316y7.30.07361405670.0853016735
58400316y15.40.2906482760.072195936
58402316y90.43680132920.1599682344
584043016y10.30.41231425630.0742398954
584063016y150.27457402580.0317514053
584083016y130.31103040890.0752731259
584103016y3.300
58412016y140.20080963710.0240957779
58414016y1.70.02411981380.0255501204
58416016y8.30.14999125930.0471746787
58418016y12.70.17506218770.031944563
58420316y6.40.15594108490.0792535366
58422316y11.90.17254244640.0709381162
58424316y7.20.33841020520.0563167976
58426316y13.10.27748707010.137173889
584283016y120.39242462440.10351349
584303016y8.80.40104060920.1245320253
584323016y15.70.38058073370.1166749703
584343016y9.60.56942906270.1797260374
figures
CT idfadtime pointtissueex_vivo_wtE2_ng_mlT_ng_mlE2_relT_rel24 h-E224 h-T24 h-wts
5808631blankblank00Fad ug/LMeanSDcountSECVFad ug/LMeanSDcountSECVFad ug/LMeanSDcountSECV
5805201y28.30.28339874840.12663869540.8859193962.113302355200.31989224950.108052392680.038202289833.777746346700.05992455130.039097735280.013823136965.2449361585028.2259.142639506383.232411196432.3919911651
5805401y33.30.228606268400.7146352212030.20519668820.068941837770.026057565333.59792903230.10163480070.077833236670.029418198276.5812851741332.971428571412.886131341674.870499841639.0827207067
5805601y30.10.38205123190.05168689031.19431224890.8625327885300.15406868360.069039430780.024409124844.8108136597300.1464775850.043882857980.015514933229.9587529836302712.706803576984.492533488247.0622354701
5805801y43.30.13022747030.1000705080.40709792291.6699417148
5807601y13.30.4675123570.04279982611.46146822180.7142285617
5807801y22.50.30811412220.06997554740.96318095451.1677275147
5808001y21.50.41937626870.03760009581.31099227750.627457277
5808201y33.50.33985152880.05062484751.06239375730.8448097881
5806031y61.30.12824984570.09967098530.40091576441.6632746191
5806231y27.40.17448347590.04324690220.54544452460.7216892121
5806431y33.50.26091954010.05604835930.81564820820.9353154611
5806631y28.50.28191254640.27030881230.88127345024.5108191286
5808431y23.20.18502203410.09314669530.5783886121.5543995448
5808831y30.20.12464582260.09527832810.38964939841.5899714902
5809031y26.70.28114355230.05374352250.87886953430.8968531483
58068301y17.70.04026743850.16068927980.12587813112.6815266249
58070301y300.05222265540.1869657480.16325076813.1200191556
58072301y24.40.02906743620.21663126110.09086633473.6150668861
58074301y13.80.196732180.10522181880.61499514381.7559049916
58092301y43.100.081426736301.3588209592Normality ok except 30 ug/LNormality ok except 3 ug/LLevene's test okNormality ok except 3 ug/LLevene's ok
58094301y9.70.03589756180.15629056140.11221766652.6081223478Levene's test okKW p=0.0076KW p=0.8063
58096301y35.20.01909255060.14423596060.05968431772.4069593759KW p=0.0005
58098301y42.100.120359314202.0085142321
58388016y19.70.21862169210.06686609351.41898851181.6724071518d. 16-E2d. 16-Td. 16-wts
58390016y8.40.07935802510.01959859560.51508212580.4901861272Fad ug/LMeanSDcountSECVFad ug/LMeanSDcountSECVFad ug/LMeanSDcountSECV
58392016y12.80.21643144120.0194656071.40477244360.486859911800.15406868360.069039430780.024409124844.810813659700.03998194660.02442835180.008636726361.0984532737010.63755.379308904181.901872902150.5692963958
58394016y7.50.16815541270.0851601371.09143148882.129964749930.25837312740.11688507980.04132511645.238868349930.09681458710.036347464580.012850769337.543376070339.9253.212364505281.135742362632.3663929995
58412016y140.20080963710.02409577791.30337737950.6026664511300.34267421510.163292342480.057732561347.6523575945300.08821386870.056150345980.019852145263.65251486013010.96253.957971161481.39935412436.1046400126
58414016y1.70.02411981380.02555012040.15655234580.6390414316
58416016y8.30.14999125930.04717467870.97353502191.1798994973
58418016y12.70.17506218770.0319445631.13626068290.7989746793
58396316y9.10.32154055080.11336851332.08699485982.8354925861
58398316y7.30.07361405670.08530167350.47780025742.1335047598
58400316y15.40.2906482760.0721959361.88648510021.80571338
58402316y90.43680132920.15996823442.83510781684.0010116513
58420316y6.40.15594108490.07925353661.01215302961.9822330645
58422316y11.90.17254244640.07093811621.11990602061.7742536858
58424316y7.20.33841020520.05631679762.19648923641.4085556697
58426316y13.10.27748707010.1371738891.80106082273.4308957038
584043016y10.30.41231425630.07423989542.67617173451.8568354379
584063016y150.27457402580.03175140531.78215338330.7941435558
584083016y130.31103040890.07527312592.01877761011.882677863
584103016y3.30000
584283016y120.39242462440.103513492.54707585762.5890057566
584303016y8.80.40104060920.12453202532.6029988693.1147064061
584323016y15.70.38058073370.11667497032.47020176192.9181913367Normality okLevene's test okNormality okLevene's test okNormality okLevene's test ok
584343016y9.60.56942906270.17972603743.69594293514.4951797619ANOVA p=0.0199ANOVA p=0.0246ANOVA p=0.8848
&A
&F
figures
00.03820228980.0382022898
00.02605756530.0260575653
00.02440912480.0244091248
A
B
C
24 h-E2
Fadrozole (ug/L)
Ex vivo E2 production (ng/ml)
00.01382313690.0138231369
00.02941819820.0294181982
00.01551493320.0155149332
A
AB
B
24 h-T
Fadrozole (ug/L)
Ex vivo T production (ng/ml)
00.02440912480.0244091248
00.0413251160.041325116
00.05773256130.0577325613
A
AB
B
d. 16-E2
Fadrozole (ug/L)
Ex vivo E2 production (ng/ml)
00.00863672630.0086367263
00.01285076930.0128507693
00.01985214520.0198521452
A
B
B
d. 16-T
Fadrozole (ug/L)
Ex vivo T production (ng/ml)
03.23241119643.2324111964
04.87049984164.8704998416
04.49253348824.4925334882
24 h-wts
Fadrozole (ug/L)
Ex vivo tissue wt (mg)
01.90187290211.9018729021
01.13574236261.1357423626
01.3993541241.399354124
d. 16-wts
Fadrozole (ug/L)
Ex vivo tissue wt (mg)
MBD0023823A.bin
MBD0022DDB4.bin
Chart2
0.05992455130.01382313690.0138231369
0.10163480070.02941819820.0294181982
0.1464775850.01551493320.0155149332
A
AB
B
24 h-T
Fadrozole (ug/L)
Ex vivo T production (ng/ml*12 h)
E2-RIA d1,16
Small Fish Comp Tox
Phase 3, Fadrozole
Ex vivo steroid production
24 h, d.16 females, E2
Extraction: Notebook 341:83
RIA: Notebook 341:85-90
DLV, NDM, JET
adjusted
E2 (ng/ml)log E2dpmdpmBo/BmBo/BmMeanSDCV
60.77815125041777.781750.930.10775106140.10451585460.1061334580.00228763662.1554340001
20.30102999572480.792664.480.19245805230.21459120420.20352462820.01565050187.6897336556
0.6666666667-0.17609125914608.974475.540.44888644420.43280921380.4408478290.01136831872.5787398599
0.2222222222-0.65321251386630.838484.190.69250414040.91581895610.80416154830.157907420519.6362809051
0.0740740741-1.130333768510303.59759.941.13503102361.06953646171.10228374270.04631164894.2014271877
0.024691358-1.607455023210961.211064.91.21427852781.22677353491.22052603140.00883530430.7238931442
MeanSDCVSigmoid fit; regression using GraphPad Prism
Bm9217.219148.469182.8352178.798099924223.7268566834
NS877.97889.07883.527.84888527120.8883653195
linearadj for dil
CT iddpmBo/Bmlog ng/mlng/mldil factorng/ml
580525117.730.510187889-0.27460080.53137265330.53333333330.2833987484
580546121.840.6311749825-0.43005850.37148518620.61538461540.2286062684
580564205.580.400281228-0.11684840.76410246370.50.3820512319
580587147.170.7547189135-0.58426740.26045494060.50.1302274703
580607565.580.8051339177-0.64890510.224437230.57142857140.1282498457
580626302.650.6529611179-0.45721570.34896695190.50.1744834759
580645166.540.5160690973-0.28246340.52183908020.50.2609195401
580664959.810.4911598126-0.24885560.56382509270.50.2819125464
580689853.261.0807807632-1.0940160.08053487710.50.0402674385
580709640.021.0550870765-1.0391030.0913896470.57142857140.0522226554
5807210390.81.1455499641-1.2635920.05450144290.53333333330.0290674362
580745956.390.6112396023-0.40509460.39346435990.50.19673218
580763764.210.3470997305-0.029176910.93502471390.50.467512357
580784729.250.4633792066-0.21025840.61622824440.50.3081141222
580804299.660.4116171033-0.13435810.73390847020.57142857140.4193762687
580824644.330.4531470368-0.19570950.63722161650.53333333330.3398515288
580846519.770.6791223131-0.48973850.32378855980.57142857140.1850220341
5808613511.71.52159304710.57142857140
580887453.060.7915761723-0.6313210.23371091730.53333333330.1246458226
580905139.150.5127688249-0.27807060.52714416060.53333333330.2811435523
5809211174.41.23996739490.53333333330
5809410215.21.1243915914-1.2018970.06282073310.57142857140.0358975618
5809610847.51.200578602-1.4760980.03341196360.57142857140.0190925506
5809811691.71.30229784020.50
583886036.970.6209488373-0.41726870.38258796110.57142857140.2186216921
583908790.110.9526798296-0.85737110.13887654390.57142857140.0793580251
583926065.930.624438282-0.42164160.3787550220.57142857140.2164314412
583946796.590.7124768731-0.53125110.29427197230.57142857140.1681554127
583964965.120.4917996244-0.24972620.56269596380.57142857140.3215405508
583988958.980.973027292-0.89000120.12882459920.57142857140.0736140567
584005236.030.5244420774-0.29359420.50863448310.57142857140.290648276
584024204.680.4001727853-0.1166780.76440232620.57142857140.4368013292
584044340.080.4164873848-0.14173360.72154994850.57142857140.4123142563
584065392.30.5432713423-0.31830250.48050454510.57142857140.2745740258
584085053.270.5024209829-0.26415910.54430321560.57142857140.3110304089
5841011336.51.25949912730.57142857140
584126282.380.6505187476-0.45417740.35141686490.57142857140.2008096371
5841410655.51.1774441626-1.3745880.04220967420.57142857140.0241198138
584167125.030.7520512235-0.5808960.26248470370.57142857140.1499912593
584186680.150.6984467995-0.51376960.30635882840.57142857140.1750621877
584207013.730.7386404782-0.56400140.27289689860.57142857140.1559410849
584226722.130.7035050483-0.5200660.30194928110.57142857140.1725424464
584244831.40.4756874513-0.22751850.59221785910.57142857140.3384102052
584265363.110.5397541845-0.31371920.48560237260.57142857140.2774870701
584284459.020.430818688-0.16320570.68674309280.57142857140.3924246244
584304406.40.4244784057-0.15377360.70182106610.57142857140.4010406092
584324334.960.4158704664-0.14079960.72310339410.52631578950.3805807337
584343477.20.31251735840.034193231.08191521920.52631578950.5694290627
QA3830.870.3551317187-0.043097130.90553005640.52631578950.4765947665
QA4305.260.4122918578-0.13538160.73218090760.52631578950.3853583724
QA4043.070.3807000939-0.085735130.82085201760.52631578950.4320273777
QA4607.430.4487008868-0.1893090.64668233830.52631578950.3403591254
Mean0.776311330.4085849105
SD0.1117027630.0587909279
CV14.388913140114.3889131401
E2-RIA d1,16
0
0
0
0
0
0
T-RIA d1,16
Small Fish Comp Tox
Phase 3, Fadrozole
Small Fish Comp Tox
Phase 3, Fadrozole
Ex vivo steroid production
24 h, d.16 females, T
Extraction: Notebook 341:83
RIA: Notebook 341:85-90
DLV, NDM, JET
T (ng/ml)log TdpmdpmBo/BmBo/BmMeanSDCV
60.7781512504769.79853.710.07044473850.10121092360.08582783110.021754978125.3472303948
20.30102999571043.31942.920.17072079830.13391649280.15231864560.02602457417.0856127958
0.6666666667-0.17609125911549.651706.360.35635175810.41380372260.38507774030.040624673710.5497330623
0.2222222222-0.65321251382536.682366.960.71821004740.65598844430.68709924590.219172590231.898243457
0.0740740741-1.13033376852819.152856.240.82176729590.83536498180.82856613890.00961501591.160440366
0.024691358-1.60745502323178.793212.420.95361608990.9659452940.95978069190.16645720917.3432545992
Sigmoid fit R^2 =0.9967; Regression conducted using GraphPad Prism
MeanSDCV
Bm3300.843309.783305.31410.51633345512.4199041377
NS568.75586.53577.6412.57235856952.1765041496
Bm-3SD2069.2909996350.546859040710.5=MDL
polynomialadj for dil
CT iddpmBo/Bmlog ng/mlng/mldil factorng/ml
580522359.220.6531508577-0.62443230.23744755390.53333333330.1266386954
580543268.070.98634732210.61538461540
580562808.770.8179618502-0.98558960.10337378070.50.0516868903
580582468.550.6932326858-0.69866390.20014101590.50.100070508
580602550.650.7233316347-0.75839320.17442422420.57142857140.0996709853
580622877.090.8430088684-1.0630150.08649380440.50.0432469022
580642774.530.8054090121-0.95040710.11209671870.50.0560483593
580661770.590.4373512925-0.26710980.54061762460.50.2703088123
580682149.750.5763563774-0.49298310.32137855960.50.1606892798
580702136.890.5716417309-0.48519990.3271900590.57142857140.186965748
580721979.330.5138781451-0.39127760.40618361460.53333333330.2166312611
580742437.260.6817613568-0.67686420.21044363750.50.1052218188
580762880.740.8443470068-1.0675280.08559965220.50.0427998261
580782669.960.7670722631-0.85402370.13995109470.50.0699755474
580802964.090.8749042223-1.1817730.06580016760.57142857140.0376000958
580822842.620.8303717092-1.0226350.09492158910.53333333330.0506248475
580842588.930.7373655904-0.78779450.16300671670.57142857140.0931466953
580863538.441.08546855010.57142857140
580882536.740.7182320442-0.74800460.17864686530.53333333330.0952783281
580902819.140.8217636298-0.99667260.10076910470.53333333330.0537435225
580922624.60.7504426855-0.81623170.15267513050.53333333330.0814267363
580942263.510.6180623023-0.56302920.27350848250.57142857140.1562905614
580962318.40.6381857043-0.59788840.25241293110.57142857140.1442359606
580982350.160.6498293415-0.61849030.24071862830.50.1203593142
583882755.50.7984323617-0.9317560.11701566360.57142857140.0668660935
583903101.520.9252878831-1.4647370.03429754230.57142857140.0195985956
583923102.530.9256581625-1.4676940.03406481230.57142857140.019465607
583942637.440.7551499998-0.82672560.14903023970.57142857140.085160137
583962473.930.6952050651-0.70246950.19839489820.57142857140.1133685133
583982636.570.7548310463-0.82600440.14927792860.57142857140.0853016735
584002720.090.785450586-0.89844920.1263428880.57142857140.072195936
584022247.380.6121488303-5.53E-010.27994441010.57142857140.1599682344
584042706.770.7805672974-0.88632460.12991981690.57142857140.0742398954
584063008.320.8911195269-1.2551990.05556495920.57142857140.0317514053
584082700.020.7780926578-0.8803220.13172797030.57142857140.0752731259
584103295.10.99625687860.57142857140
584123066.90.9125957319-1.3750210.04216761130.57142857140.0240957779
584143055.740.9085043279-1.3495690.04471271070.57142857140.0255501204
584162893.350.84897-1.0832530.08255568780.57142857140.0471746787
584183006.830.8905732732-1.2525650.05590298520.57142857140.031944563
584202674.50.7687366874-0.85794330.13869368910.57142857140.0792535366
584222728.380.78848981-0.90608230.12414170330.57142857140.0709381162
584242827.980.825004491-1.0063240.09855439580.57142857140.0563167976
584262351.990.6505002438-0.61969050.24005430570.57142857140.137173889
584282528.630.7152588106-0.7419650.18114860750.57142857140.10351349
584302415.060.6736225423-0.66168090.21793104430.57142857140.1245320253
584322404.070.6695934626-0.65426870.22168244360.52631578950.1166749703
584342106.030.5603280455-0.46663540.3414794710.52631578950.1797260374
1
1
1
QA1922.260.4929555261-0.35752440.43901119990.52631578950.2310585263
QA2003.220.5226365359-0.40541780.39317165440.52631578950.2069324497
QA1885.690.4795484791-0.33586390.46146216580.52631578950.2428748241
QA1955.110.5049987718-0.37695340.41980402670.52631578950.2209494878
Mean0.42836226170.225453822
SD0.02898635170.0152559746
CV6.76678463926.7667846392
T-RIA d1,16
0
0
0
0
0
0
T (log ng/ml)
Bo/Bm
forSAS
CT idfadtime pointtissueex_vivo_wtE2_ng_mlT_ng_ml
5805201y28.30.28339874840.1266386954
5805401y33.30.22860626840
5805601y30.10.38205123190.0516868903
5805801y43.30.13022747030.100070508
5806031y61.30.12824984570.0996709853
5806231y27.40.17448347590.0432469022
5806431y33.50.26091954010.0560483593
5806631y28.50.28191254640.2703088123
58068301y17.70.04026743850.1606892798
58070301y300.05222265540.186965748
58072301y24.40.02906743620.2166312611
58074301y13.80.196732180.1052218188
5807601y13.30.4675123570.0427998261
5807801y22.50.30811412220.0699755474
5808001y21.50.41937626870.0376000958
5808201y33.50.33985152880.0506248475
5808431y23.20.18502203410.0931466953
5808631blankblank00
5808831y30.20.12464582260.0952783281
5809031y26.70.28114355230.0537435225
58092301y43.100.0814267363
58094301y9.70.03589756180.1562905614
58096301y35.20.01909255060.1442359606
58098301y42.100.1203593142
58388016y19.70.21862169210.0668660935
58390016y8.40.07935802510.0195985956
58392016y12.80.21643144120.019465607
58394016y7.50.16815541270.085160137
58396316y9.10.32154055080.1133685133
58398316y7.30.07361405670.0853016735
58400316y15.40.2906482760.072195936
58402316y90.43680132920.1599682344
584043016y10.30.41231425630.0742398954
584063016y150.27457402580.0317514053
584083016y130.31103040890.0752731259
584103016y3.300
58412016y140.20080963710.0240957779
58414016y1.70.02411981380.0255501204
58416016y8.30.14999125930.0471746787
58418016y12.70.17506218770.031944563
58420316y6.40.15594108490.0792535366
58422316y11.90.17254244640.0709381162
58424316y7.20.33841020520.0563167976
58426316y13.10.27748707010.137173889
584283016y120.39242462440.10351349
584303016y8.80.40104060920.1245320253
584323016y15.70.38058073370.1166749703
584343016y9.60.56942906270.1797260374
figures
CT idfadtime pointtissueex_vivo_wtE2_ng_mlT_ng_mlE2_relT_rel24 h-E224 h-T24 h-wts
5808631blankblank00Fad ug/LMeanSDcountSECVFad ug/LMeanSDcountSECVFad ug/LMeanSDcountSECV
5805201y28.30.28339874840.12663869540.8859193962.113302355200.31989224950.108052392680.038202289833.777746346700.05992455130.039097735280.013823136965.2449361585028.2259.142639506383.232411196432.3919911651
5805401y33.30.228606268400.7146352212030.20519668820.068941837770.026057565333.59792903230.10163480070.077833236670.029418198276.5812851741332.971428571412.886131341674.870499841639.0827207067
5805601y30.10.38205123190.05168689031.19431224890.8625327885300.15406868360.069039430780.024409124844.8108136597300.1464775850.043882857980.015514933229.9587529836302712.706803576984.492533488247.0622354701
5805801y43.30.13022747030.1000705080.40709792291.6699417148
5807601y13.30.4675123570.04279982611.46146822180.7142285617
5807801y22.50.30811412220.06997554740.96318095451.1677275147
5808001y21.50.41937626870.03760009581.31099227750.627457277
5808201y33.50.33985152880.05062484751.06239375730.8448097881
5806031y61.30.12824984570.09967098530.40091576441.6632746191
5806231y27.40.17448347590.04324690220.54544452460.7216892121
5806431y33.50.26091954010.05604835930.81564820820.9353154611
5806631y28.50.28191254640.27030881230.88127345024.5108191286
5808431y23.20.18502203410.09314669530.5783886121.5543995448
5808831y30.20.12464582260.09527832810.38964939841.5899714902
5809031y26.70.28114355230.05374352250.87886953430.8968531483
58068301y17.70.04026743850.16068927980.12587813112.6815266249
58070301y300.05222265540.1869657480.16325076813.1200191556
58072301y24.40.02906743620.21663126110.09086633473.6150668861
58074301y13.80.196732180.10522181880.61499514381.7559049916
58092301y43.100.081426736301.3588209592Normality ok except 30 ug/LNormality ok except 3 ug/LLevene's test okNormality ok except 3 ug/LLevene's ok
58094301y9.70.03589756180.15629056140.11221766652.6081223478Levene's test okKW p=0.0076KW p=0.8063
58096301y35.20.01909255060.14423596060.05968431772.4069593759KW p=0.0005
58098301y42.100.120359314202.0085142321
58388016y19.70.21862169210.06686609351.41898851181.6724071518d. 16-E2d. 16-Td. 16-wts
58390016y8.40.07935802510.01959859560.51508212580.4901861272Fad ug/LMeanSDcountSECVFad ug/LMeanSDcountSECVFad ug/LMeanSDcountSECV
58392016y12.80.21643144120.0194656071.40477244360.486859911800.15406868360.069039430780.024409124844.810813659700.03998194660.02442835180.008636726361.0984532737010.63755.379308904181.901872902150.5692963958
58394016y7.50.16815541270.0851601371.09143148882.129964749930.25837312740.11688507980.04132511645.238868349930.09681458710.036347464580.012850769337.543376070339.9253.212364505281.135742362632.3663929995
58412016y140.20080963710.02409577791.30337737950.6026664511300.34267421510.163292342480.057732561347.6523575945300.08821386870.056150345980.019852145263.65251486013010.96253.957971161481.39935412436.1046400126
58414016y1.70.02411981380.02555012040.15655234580.6390414316
58416016y8.30.14999125930.04717467870.97353502191.1798994973
58418016y12.70.17506218770.0319445631.13626068290.7989746793
58396316y9.10.32154055080.11336851332.08699485982.8354925861
58398316y7.30.07361405670.08530167350.47780025742.1335047598
58400316y15.40.2906482760.0721959361.88648510021.80571338
58402316y90.43680132920.15996823442.83510781684.0010116513
58420316y6.40.15594108490.07925353661.01215302961.9822330645
58422316y11.90.17254244640.07093811621.11990602061.7742536858
58424316y7.20.33841020520.05631679762.19648923641.4085556697
58426316y13.10.27748707010.1371738891.80106082273.4308957038
584043016y10.30.41231425630.07423989542.67617173451.8568354379
584063016y150.27457402580.03175140531.78215338330.7941435558
584083016y130.31103040890.07527312592.01877761011.882677863
584103016y3.30000
584283016y120.39242462440.103513492.54707585762.5890057566
584303016y8.80.40104060920.12453202532.6029988693.1147064061
584323016y15.70.38058073370.11667497032.47020176192.9181913367Normality okLevene's test okNormality okLevene's test okNormality okLevene's test ok
584343016y9.60.56942906270.17972603743.69594293514.4951797619ANOVA p=0.0199ANOVA p=0.0246ANOVA p=0.8848
&A
&F
figures
00.03820228980.0382022898
00.02605756530.0260575653
00.02440912480.0244091248
A
B
C
24 h-E2
Fadrozole (ug/L)
Ex vivo E2 production (ng/ml)
00.01382313690.0138231369
00.02941819820.0294181982
00.01551493320.0155149332
A
AB
B
24 h-T
Fadrozole (ug/L)
Ex vivo T production (ng/ml)
00.02440912480.0244091248
00.0413251160.041325116
00.05773256130.0577325613
A
AB
B
d. 16-E2
Fadrozole (ug/L)
Ex vivo E2 production (ng/ml)
00.00863672630.0086367263
00.01285076930.0128507693
00.01985214520.0198521452
A
B
B
d. 16-T
Fadrozole (ug/L)
Ex vivo T production (ng/ml)
03.23241119643.2324111964
04.87049984164.8704998416
04.49253348824.4925334882
24 h-wts
Fadrozole (ug/L)
Ex vivo tissue wt (mg)
01.90187290211.9018729021
01.13574236261.1357423626
01.3993541241.399354124
d. 16-wts
Fadrozole (ug/L)
Ex vivo tissue wt (mg)
MBD0023823A.bin
MBD0022DDB4.bin
-
Chart1
14.361.86326518321.8632651832
5.9591.54101529611.5410152961
3.068750.50539036360.5053903636
a
b
b
z
Female 24 h
Fadrozole (ug/L-nominal)
VTG (mg/ml)
Fadrozole, Female Vtg
Vtg
Fadrozole 24 h and 7 d exposures
FHM exposures in support of array validation
DLV, KJ, MK, LD
VTG ELISAS conducted by KJ
24-hr Fadrozole Exposure
Fad ug/LSexIDVtg (mg/ml)TreatSexIDVtg (mg/ml)Female 24 h
0F219.60M100550
0F49.040M30.004Mean14.365.9593.06875
0F1419.60M130.003SD5.27010978474.35864946321.4294598131
0F167.440M150.003SE1.86326518321.54101529610.5053903636
0F2610.20M250.004CV36.699928862973.143974881346.5811751709
0F2813.60M270.003
0F3814.30M370.002
0F4021.10M390.006
5F68.545M50.003
5F81.325M70.008
5F189.965M170.002
5F205.095M190.004
5F301.475M290.012
5F320.7825M310.004Male 24 h
5F428.815M410.0050550
5F4411.75M430.005Mean0.0031250.0053750.01775
50F106.0750M90.005SD0.00172688820.00320434970.0032043497
50F122.9550M110.004SE0.00061054720.00113290870.0011329087
50F221.5750M210.004CV55.260422417159.615808787118.0526744919
50F242.7850M230.111
50F343.8650M330.004
50F361.6950M350.002
50F463.2050M450
50F482.4350M470.012
7-d Fadrozole Exposure
TreatSexIDVtg (mg/ml)TreatSexIDVtg (mg/ml)Female 7d
0F218.40M10.0100550
0F420.40M30.004Mean19.3755.970.03175
0F1424.90M130.004SD4.49563280152.89367125520.0425768213
0F1617.90M150.003SE1.58944621981.0937049310.0150531795
0F2618.90M250.010CV23.203266072148.4702052802134.100224651
0F2816.50M270.003
0F3826.00M370.002
0F4012.00M390.004
5F65.495M50.004
5F88.035M70.002
5F1810.15M170.003
5F208.225M190
5F302.335M290Male 7d
5F323.085M3100550
5F424.545M410.011Mean0.0050.00250.0035
5F44no sample5M430SD0.00316227770.00377964470.0065683222
50F100.00750M90SE0.0011180340.00133630620.0023222526
50F120.02150M110CV63.2455532034151.1857892037187.6663499196
50F220.13550M210.012
50F240.00850M230
50F340.02550M330
50F360.03150M350
50F460.01750M450
50F480.01050M470.016
&A
&F
Figures
Fadrozole 24 h and 7 d exposures
FHM exposures in support of array validation
DLV, KJ, MK, LDStatistics
VTG ELISAS conducted by KJ
Female, 24 h
Kolmogorov-Smirnov p=0.150 (normal)
Female 24 hLevene's test p= 0.0046 (non-trans); p=0.0013 (log trans) - violated homogeneity
0550Non-parametric one-way ANOVA p=0.0017
Mean14.365.9593.06875Welch's one-way ANOVA p=.0005
SD5.27010978474.35864946321.4294598131Dunnett's test = both treatments different from control
SE1.86326518321.54101529610.5053903636Fishers LSD = (0-A, 5-B, 50-B)
CV36.699928862973.143974881346.5811751709
Female, 7d
Non-transformed data was not normally distributed
Male 24 hLog trans Kolm-Smir p>0.150 (normal)
0550Log trans Levene's test p=0.1547
Mean0.0031250.0053750.01775Log trans one-way ANOVA p
-
BrainsGnRHneuronscGnRHneuronsdopaminergicneuronssGnRH releasecGnRH releasedopamine releaseGnRHPituitaryFSH gonadotrophsLH gonadotrophs TestisOvariesLeydigcellsSertoli cellsSperm-atocytesThecacellsGranulosacellsOocytesFSHLHLiverSperm mat.oocyte mat.steroidssteroidsSteroid metab.vitellogenesisBlood supplyBlood supply
-
state transitionactivated proteinFigure Key
-
Inner mitochondrial membraneFadrozole
-
Evidence for compensatory/feedback response to fadrozole.exposureexposurepost-exposurepost-exposure
Chart1
10.64145564170.14586154520.07006656710.07006656710.11942236750.11942236750.081457320.08145732
10.53099060980.20668239950.06325857470.06325857470.14333392830.14333392830.15518425060.1551842506
10.59229788610.30946009320.05299525550.05299525550.17950254390.17950254390.09471836430.0947183643
10.96481614430.87126656920.25900102660.25900102660.21613330920.21613330920.1263969650.126396965
10.89354557341.81310187730.44434440540.44434440540.2364491670.2364491670.21820686290.2182068629
12.47728187023.01296882721.03462874651.03462874650.18095712190.18095712190.48690591810.4869059181
11.95546663281.61361971330.33879169720.33879169720.23921811950.23921811950.21922348270.2192234827
11.67699964292.22416526890.37471963770.37471963770.1584301510.1584301510.26822528110.2682252811
#
#
#
#
#
*
*
*
*
0
3
30
Time point
Ex vivo E2 production (relative to control)
Sheet1
CT idfadtime pointE2_relT_rel
58052010.8859193962.113302355224 hMeanSDcountSE24 hMeanSDcountSE
58054010.71463522120010.337777463580.1194223675010.652449361680.230675684
58056011.19431224890.862532788530.64145564170.215515811370.0814573231.69604608631.2988538970.4909206261
58058010.40709792291.6699417148300.14586154520.19817817980.0700665671302.44436682170.732301818180.2589077907
58076011.46146822180.7142285617
58078010.96318095451.167727514748 hMeanSDcountSE48 hMeanSDcountSE
58080011.31099227750.627457277010.405409570780.1433339283012.599367829180.9190153094
58082011.06239375730.844809788130.53099060980.438927343780.155184250631.53146446483.201770441981.1319967956
58060310.40091576441.6632746191300.20668239950.178922268680.0632585747307.80939420214.031428156281.4253250936
58062310.54544452460.7216892121
58064310.81564820820.93531546114 dMeanSDcountSE4 dMeanSDcountSE
58066310.88127345024.5108191286010.507709864380.1795025439010.449878228180.1590559729
58084310.5783886121.554399544830.59229788610.267903990980.094718364330.832015360.477066830480.1686685954
58088310.38964939841.5899714902300.30946009320.140212266870.0529952555302.15938457480.574971068370.2173186368
58090310.87886953430.8968531483
580683010.12587813112.68152662498 dMeanSDcountSE8 dMeanSDcountSE
580703010.16325076813.1200191556010.571834986270.2161333092010.794780176170.3003986704
580723010.09086633473.615066886130.96481614430.357504604280.12639696531.74401530281.779858235180.6292749138
580743010.61499514381.7559049916300.87126656920.685252305870.2590010266306.85463325914.157058620971.5712204709
5809230101.3588209592
580943010.11221766652.60812234789 dMeanSDcountSE9 dMeanSDcountSE
580963010.05968431772.4069593759010.668779237780.236449167011.016475565780.3593783827
5809830102.008514232130.89354557340.617182209880.218206862931.79445136781.261639283780.4460568465
58100021.71605163110301.81310187731.256795768980.4443444054302.94205177442.207579791380.7804973202
58102021.42032194730
58104020.6783947327010 dMeanSDcountSE10 dMeanSDcountSE
58106021.21729454190010.51182403280.1809571219010.451297209280.1595576585
58124020.63376734877.412989289232.47728187021.37717790680.486905918131.09665294591.000009533480.3535567611
58126020.90284214630303.01296882722.926372010781.0346287465302.58386779341.393396890280.492640195
58128020.76660974590
58130020.66471790620.587010710812 dMeanSDcountSE12 dMeanSDcountSE
58108320.23043089490010.676611017880.2392181195010.878360812880.3105474435
58110320.6827012862031.95546663280.620057644980.219223482730.3785333640.42759575680.1511779293
58112320.45773391323.3446588395301.61361971330.958247625980.3387916972300.86425836840.984991247380.3482469952
58114321.54096400210
58132320.38149488448.907056878716 dMeanSDcountSE16 dMeanSDcountSE
58134320.45607100630010.448108136680.158430151010.610984532780.2160156531
58136320.3568149942031.67699964290.758655660780.268225281132.42145756260.909096919180.3214142981
58138320.14171389720302.22416526891.059867187480.3747196377302.20634251471.404392497180.4965277291
581163020.15506631010.8359186709
581183020.62121881258.817714348
581203020.14861856826.5094939192
581223020.07808292287.6837057603
581403020.063377617615.0685743617
581423020.12147300968.2488788577
581443020.21674783179.8324766934
581463020.24887412365.4783910057
58148041.16496360950.7923813134
58150040.66956030620.3969951719
58152041.80328490081.2091583299
58154040.77185131130.7644784932
58172040.79403646330.8050046186
58174041.52296182141.2786626575
58176040.19466568370.8710047534
58178041.07867590391.882314662
58156340.182561881.2172883485
58158340.8074837650.7046476779
58160340.88608337721.4939078698
58162340.47663255181.2780671875
58180340.55671932810.3281136705
58182340.62004184420.1234123508
58184340.29930646610.8640514933
58186340.90955387680.6466342815
581643040.44609545583.1722537914
581663040.47863427951.7795378312
581683040.19610397611.9234332756
581883040.30131003741.3352961447
581903040.14847686852.3892897782
581923040.17217317942.2018824255
581943040.42342685552.3139987772
58196080.73202679920.579926722
58198080.73362518870.1589241651
58200081.55964973340.4647927377
58220081.33942969551.8651909446
58222081.82584268881.6628373958
58224080.40044137390.298751698T relativeMeanMeanMeanSESESE
58226080.40898452061.9695763367E2 relativeMeanMeanMeanSESESEdays03300330
58204380.44528937150.2421837166days0330033024 h11.69604608632.44436682170.2306756840.49092062610.2589077907
58206380.78028570160.29497316124 h10.64145564170.14586154520.11942236750.081457320.070066567148 h11.53146446487.80939420210.91901530941.13199679561.4253250936
58208381.18373194321.554991816548 h10.53099060980.20668239950.14333392830.15518425060.06325857474 d10.832015362.15938457480.15905597290.16866859540.2173186368
58210380.51697747120.14449520374 d10.59229788610.30946009320.17950254390.09471836430.05299525558 d11.74401530286.85463325910.30039867040.62927491381.5712204709
58228381.11814589834.89333674488 d10.96481614430.87126656920.21613330920.1263969650.25900102669 d11.79445136782.94205177440.35937838270.44605684650.7804973202
58230380.94651675333.91411118099 d10.89354557341.81310187730.2364491670.21820686290.444344405410 d11.09665294592.58386779340.15955765850.35355676110.492640195
58232381.36088639490.990275057810 d12.47728187023.01296882720.18095712190.48690591811.034628746512 d10.3785333640.86425836840.31054744350.15117792930.3482469952
58234381.36669562061.917755541312 d11.95546663281.61361971330.23921811950.21922348270.338791697216 d12.42145756262.20634251470.21601565310.32141429810.4965277291
582123081.93346858519.223973655916 d11.67699964292.22416526890.1584301510.26822528110.3747196377
582143080.32106662428.1566457122
582163080.74046528964.9391328698
582363080.10347622221.7491969416
582383080.481341944614.444714409
582403080.85561036894.8809207543
582423081.66343695014.5878484711
58244090.85443928330.5797096217
58246092.31574762030.8797289579
58248091.4531346763.4279996944
58250090.79170699250.2628597465
58268090.41896184590.6753578084
58270090.27024054740.3193272718
58272090.59138998360.8187135305
58274091.3043790511.0363033688
58252391.20249771572.4785770542
58254390.22042378652.0895620606
58256390.13050298160.9811789697
58258391.30315236354.5499086065
58276390.47273218650.9356139639
58278391.94772477061.4507268925
58280390.74083824940.9411042543
58282391.13049253360.9289391411
582603092.1931362283.2176226678
582623092.00300553992.4116925427
582643090.01259521297.0803043862
582663092.58600693062.4101458099
582843091.40526757011.1641742142
582863093.68352025935.2303497615
582883090.11328477340.5140791555
582903092.50799850421.5080456576
582920101.04296163931.9913261875
582940100.51883735320.9579141865
582960101.19255587191.3145291014
582980101.10908591050.8018020423
583160101.86615290480.7712967566
583180100.32364233420.719988558
583200100.55967400410.8143563836
583220101.3870899820.6287867841
583003100.68417001280.5794224326
583023101.67760702140.2468632857
583043103.58975152080.5226964592
583063103.78846635913.3010115332
583243103.09026508531.3205705416
583263104.30193788261.4191121066
583283101.03755334561.1040036395
583303101.64850373380.2795435687
5830830103.46710133443.8767238411
5831030104.65330945652.2911520153
58312301004.7121391322
5831430100.97388580361.2281966404
5833230101.23558033650.9860718437
5833430108.78154413053.8236361429
5833630100.64643049391.4431188687
5833830104.34589906182.3099038627
583400120.84404803492.0154109771
583420120.91767509021.1817282552
583440120.20892916450.2751245338
583460121.75495531760
583640120.70495723890.7872604238
583660122.26061914932.5289195238
583680120.47736315910.8480798011
583700120.83145284540.3634764851
583483121.9113584970.5288914175
583503121.43915420750
583523121.58101310
583543122.79447193530.7918777121
583723120.99613634720
583743122.67502013580
583763122.32850902271.0292427575
583783121.91806981710.6782550245
5835630122.3538733481.9097834245
5835830121.95227943690.9631825806
5836030121.10956845120
5836230121.91585828590
5838030120.14811509180.1472218845
5838230122.92953443522.4897008793
5838430122.04849333991.4041781786
5838630120.45123531740
583880161.41898851181.6724071518
583900160.51508212580.4901861272
583920161.40477244360.4868599118
583940161.09143148882.1299647499
584120161.30337737950.6026664511
584140160.15655234580.6390414316
584160160.97353502191.1798994973
584180161.13626068290.7989746793
583963162.08699485982.8354925861
583983160.47780025742.1335047598
584003161.88648510021.80571338
584023162.83510781684.0010116513
584203161.01215302961.9822330645
584223161.11990602061.7742536858
584243162.19648923641.4085556697
584263161.80106082273.4308957038
5840430162.67617173451.8568354379
5840630161.78215338330.7941435558
5840830162.01877761011.882677863
58410301600
5842830162.54707585762.5890057566
5843030162.6029988693.1147064061
5843230162.47020176192.9181913367
5843430163.69594293514.4951797619
Sheet1
0.11942236750.11942236750.14333392830.14333392830.17950254390.17950254390.21613330920.21613330920.2364491670.2364491670.18095712190.18095712190.23921811950.23921811950.1584301510.158430151
0.081457320.081457320.15518425060.15518425060.09471836430.09471836430.1263969650.1263969650.21820686290.21820686290.48690591810.48690591810.21922348270.21922348270.26822528110.2682252811
0.07006656710.07006656710.06325857470.06325857470.05299525550.05299525550.25900102660.25900102660.44434440540.44434440541.03462874651.03462874650.33879169720.33879169720.37471963770.3747196377
a
a
a
a
a
a
a
a
b
b
b
a
a
ab
b
b
b
ab
b
ab
b
c
a
a
24 h
48 h
4 d
8 d
9 d
10 d
12 d
16 d
Fadrozole (ug/L)
E2 production (relative to control)
Sheet2
0.2306756840.2306756840.91901530940.91901530940.15905597290.15905597290.30039867040.30039867040.35937838270.35937838270.15955765850.15955765850.31054744350.31054744350.21601565310.2160156531
0.49092062610.49092062611.13199679561.13199679560.16866859540.16866859540.62927491380.62927491380.44605684650.44605684650.35355676110.35355676110.15117792930.15117792930.32141429810.3214142981
0.25890779070.25890779071.42532509361.42532509360.21731863680.21731863681.57122047091.57122047090.78049732020.78049732020.4926401950.4926401950.34824699520.34824699520.49652772910.4965277291
a
a
b
a
b
a
a
b
a
a
b
a
a
a
a
b
b
a
ab
b
a
a
ab
b
24 h
48 h
4 d
8 d
9 d
10 d
12 d
16 d
Fadrozole (ug/L)
T production (relative to control)
Sheet3
0.11942236750.11942236750.081457320.081457320.07006656710.0700665671
0.14333392830.14333392830.15518425060.15518425060.06325857470.0632585747
0.17950254390.17950254390.09471836430.09471836430.05299525550.0529952555
0.21613330920.21613330920.1263969650.1263969650.25900102660.2590010266
0.2364491670.2364491670.21820686290.21820686290.44434440540.4443444054
0.18095712190.18095712190.48690591810.48690591811.03462874651.0346287465
0.23921811950.23921811950.21922348270.21922348270.33879169720.3387916972
0.1584301510.1584301510.26822528110.26822528110.37471963770.3747196377
&F
*
*
*
*
#
#
#
#
#
0
3
30
Time point
E2 production (relative to control)
0.2306756840.2306756840.49092062610.49092062610.25890779070.2589077907
0.91901530940.91901530941.13199679561.13199679561.42532509361.4253250936
0.15905597290.15905597290.16866859540.16866859540.21731863680.2173186368
0.30039867040.30039867040.62927491380.62927491381.57122047091.5712204709
0.35937838270.35937838270.44605684650.44605684650.78049732020.7804973202
0.15955765850.15955765850.35355676110.35355676110.4926401950.492640195
0.31054744350.31054744350.15117792930.15117792930.34824699520.3482469952
0.21601565310.21601565310.32141429810.32141429810.49652772910.4965277291
&F
#
#
#
#
#
#
#
*
0
3
30
Time point
T production (relative to control)
0.07006656710.07006656710.11942236750.11942236750.081457320.08145732
0.06325857470.06325857470.14333392830.14333392830.15518425060.1551842506
0.05299525550.05299525550.17950254390.17950254390.09471836430.0947183643
0.25900102660.25900102660.21613330920.21613330920.1263969650.126396965
0.44434440540.44434440540.2364491670.2364491670.21820686290.2182068629
1.03462874651.03462874650.18095712190.18095712190.48690591810.4869059181
0.33879169720.33879169720.23921811950.23921811950.21922348270.2192234827
0.37471963770.37471963770.1584301510.1584301510.26822528110.2682252811
#
#
#
#
#
*
*
*
*
0
3
30
Time point
Ex vivo E2 production (relative to control)
0.07006656710.07006656710.11942236750.11942236750.081457320.08145732
0.06325857470.06325857470.14333392830.14333392830.15518425060.1551842506
0.05299525550.05299525550.17950254390.17950254390.09471836430.0947183643
0.25900102660.25900102660.21613330920.21613330920.1263969650.126396965
0.44434440540.44434440540.2364491670.2364491670.21820686290.2182068629
1.03462874651.03462874650.18095712190.18095712190.48690591810.4869059181
0.33879169720.33879169720.23921811950.23921811950.21922348270.2192234827
0.37471963770.37471963770.1584301510.1584301510.26822528110.2682252811
#
#
#
#
#
*
#
#
0
3
30
Time point
Ex vivo T production (relative to control)
Chart2
11.69604608632.44436682170.07006656710.07006656710.11942236750.11942236750.081457320.08145732
11.53146446487.80939420210.06325857470.06325857470.14333392830.14333392830.15518425060.1551842506
10.832015362.15938457480.05299525550.05299525550.17950254390.17950254390.09471836430.0947183643
11.74401530286.85463325910.25900102660.25900102660.21613330920.21613330920.1263969650.126396965
11.79445136782.94205177440.44434440540.44434440540.2364491670.2364491670.21820686290.2182068629
11.09665294592.58386779341.03462874651.03462874650.18095712190.18095712190.48690591810.4869059181
10.3785333640.86425836840.33879169720.33879169720.23921811950.23921811950.21922348270.2192234827
12.42145756262.20634251470.37471963770.37471963770.1584301510.1584301510.26822528110.2682252811
#
#
#
#
#
*
#
#
0
3
30
Time point
Ex vivo T production (relative to control)
Sheet1
CT idfadtime pointE2_relT_rel
58052010.8859193962.113302355224 hMeanSDcountSE24 hMeanSDcountSE
58054010.71463522120010.337777463580.1194223675010.652449361680.230675684
58056011.19431224890.862532788530.64145564170.215515811370.0814573231.69604608631.2988538970.4909206261
58058010.40709792291.6699417148300.14586154520.19817817980.0700665671302.44436682170.732301818180.2589077907
58076011.46146822180.7142285617
58078010.96318095451.167727514748 hMeanSDcountSE48 hMeanSDcountSE
58080011.31099227750.627457277010.405409570780.1433339283012.599367829180.9190153094
58082011.06239375730.844809788130.53099060980.438927343780.155184250631.53146446483.201770441981.1319967956
58060310.40091576441.6632746191300.20668239950.178922268680.0632585747307.80939420214.031428156281.4253250936
58062310.54544452460.7216892121
58064310.81564820820.93531546114 dMeanSDcountSE4 dMeanSDcountSE
58066310.88127345024.5108191286010.507709864380.1795025439010.449878228180.1590559729
58084310.5783886121.554399544830.59229788610.267903990980.094718364330.832015360.477066830480.1686685954
58088310.38964939841.5899714902300.30946009320.140212266870.0529952555302.15938457480.574971068370.2173186368
58090310.87886953430.8968531483
580683010.12587813112.68152662498 dMeanSDcountSE8 dMeanSDcountSE
580703010.16325076813.1200191556010.571834986270.2161333092010.794780176170.3003986704
580723010.09086633473.615066886130.96481614430.357504604280.12639696531.74401530281.779858235180.6292749138
580743010.61499514381.7559049916300.87126656920.685252305870.2590010266306.85463325914.157058620971.5712204709
5809230101.3588209592
580943010.11221766652.60812234789 dMeanSDcountSE9 dMeanSDcountSE
580963010.05968431772.4069593759010.668779237780.236449167011.016475565780.3593783827
5809830102.008514232130.89354557340.617182209880.218206862931.79445136781.261639283780.4460568465
58100021.71605163110301.81310187731.256795768980.4443444054302.94205177442.207579791380.7804973202
58102021.42032194730
58104020.6783947327010 dMeanSDcountSE10 dMeanSDcountSE
58106021.21729454190010.51182403280.1809571219010.451297209280.1595576585
58124020.63376734877.412989289232.47728187021.37717790680.486905918131.09665294591.000009533480.3535567611
58126020.90284214630303.01296882722.926372010781.0346287465302.58386779341.393396890280.492640195
58128020.76660974590
58130020.66471790620.587010710812 dMeanSDcountSE12 dMeanSDcountSE
58108320.23043089490010.676611017880.2392181195010.878360812880.3105474435
58110320.6827012862031.95546663280.620057644980.219223482730.3785333640.42759575680.1511779293
58112320.45773391323.3446588395301.61361971330.958247625980.3387916972300.86425836840.984991247380.3482469952
58114321.54096400210
58132320.38149488448.907056878716 dMeanSDcountSE16 dMeanSDcountSE
58134320.45607100630010.448108136680.158430151010.610984532780.2160156531
58136320.3568149942031.67699964290.758655660780.268225281132.42145756260.909096919180.3214142981
58138320.14171389720302.22416526891.059867187480.3747196377302.20634251471.404392497180.4965277291
581163020.15506631010.8359186709
581183020.62121881258.817714348
581203020.14861856826.5094939192
581223020.07808292287.6837057603
581403020.063377617615.0685743617
581423020.12147300968.2488788577
581443020.21674783179.8324766934
581463020.24887412365.4783910057
58148041.16496360950.7923813134
58150040.66956030620.3969951719
58152041.80328490081.2091583299
58154040.77185131130.7644784932
58172040.79403646330.8050046186
58174041.52296182141.2786626575
58176040.19466568370.8710047534
58178041.07867590391.882314662
58156340.182561881.2172883485
58158340.8074837650.7046476779
58160340.88608337721.4939078698
58162340.47663255181.2780671875
58180340.55671932810.3281136705
58182340.62004184420.1234123508
58184340.29930646610.8640514933
58186340.90955387680.6466342815
581643040.44609545583.1722537914
581663040.47863427951.7795378312
581683040.19610397611.9234332756
581883040.30131003741.3352961447
581903040.14847686852.3892897782
581923040.17217317942.2018824255
581943040.42342685552.3139987772
58196080.73202679920.579926722
58198080.73362518870.1589241651
58200081.55964973340.4647927377
58220081.33942969551.8651909446
58222081.82584268881.6628373958
58224080.40044137390.298751698T relativeMeanMeanMeanSESESE
58226080.40898452061.9695763367E2 relativeMeanMeanMeanSESESEdays03300330
58204380.44528937150.2421837166days0330033024 h11.69604608632.44436682170.2306756840.49092062610.2589077907
58206380.78028570160.29497316124 h10.64145564170.14586154520.11942236750.081457320.070066567148 h11.53146446487.80939420210.91901530941.13199679561.4253250936
58208381.18373194321.554991816548 h10.53099060980.20668239950.14333392830.15518425060.06325857474 d10.832015362.15938457480.15905597290.16866859540.2173186368
58210380.51697747120.14449520374 d10.59229788610.30946009320.17950254390.09471836430.05299525558 d11.74401530286.85463325910.30039867040.62927491381.5712204709
58228381.11814589834.89333674488 d10.96481614430.87126656920.21613330920.1263969650.25900102669 d11.79445136782.94205177440.35937838270.44605684650.7804973202
58230380.94651675333.91411118099 d10.89354557341.81310187730.2364491670.21820686290.444344405410 d11.09665294592.58386779340.15955765850.35355676110.492640195
58232381.36088639490.990275057810 d12.47728187023.01296882720.18095712190.48690591811.034628746512 d10.3785333640.86425836840.31054744350.15117792930.3482469952
58234381.36669562061.917755541312 d11.95546663281.61361971330.23921811950.21922348270.338791697216 d12.42145756262.20634251470.21601565310.32141429810.4965277291
582123081.93346858519.223973655916 d11.67699964292.22416526890.1584301510.26822528110.3747196377
582143080.32106662428.1566457122
582163080.74046528964.9391328698
582363080.10347622221.7491969416
582383080.481341944614.444714409
582403080.85561036894.8809207543
582423081.66343695014.5878484711
58244090.85443928330.5797096217
58246092.31574762030.8797289579
58248091.4531346763.4279996944
58250090.79170699250.2628597465
58268090.41896184590.6753578084
58270090.27024054740.3193272718
58272090.59138998360.8187135305
58274091.3043790511.0363033688
58252391.20249771572.4785770542
58254390.22042378652.0895620606
58256390.13050298160.9811789697
58258391.30315236354.5499086065
58276390.47273218650.9356139639
58278391.94772477061.4507268925
58280390.74083824940.9411042543
58282391.13049253360.9289391411
582603092.1931362283.2176226678
582623092.00300553992.4116925427
582643090.01259521297.0803043862
582663092.58600693062.4101458099
582843091.40526757011.1641742142
582863093.68352025935.2303497615
582883090.11328477340.5140791555
582903092.50799850421.5080456576
582920101.04296163931.9913261875
582940100.51883735320.9579141865
582960101.19255587191.3145291014
582980101.10908591050.8018020423
583160101.86615290480.7712967566
583180100.32364233420.719988558
583200100.55967400410.8143563836
583220101.3870899820.6287867841
583003100.68417001280.5794224326
583023101.67760702140.2468632857
583043103.58975152080.5226964592
583063103.78846635913.3010115332
583243103.09026508531.3205705416
583263104.30193788261.4191121066
583283101.03755334561.1040036395
583303101.64850373380.2795435687
5830830103.46710133443.8767238411
5831030104.65330945652.2911520153
58312301004.7121391322
5831430100.97388580361.2281966404
5833230101.23558033650.9860718437
5833430108.78154413053.8236361429
5833630100.64643049391.4431188687
5833830104.34589906182.3099038627
583400120.84404803492.0154109771
583420120.91767509021.1817282552
583440120.20892916450.2751245338
583460121.75495531760
583640120.70495723890.7872604238
583660122.26061914932.5289195238
583680120.47736315910.8480798011
583700120.83145284540.3634764851
583483121.9113584970.5288914175
583503121.43915420750
583523121.58101310
583543122.79447193530.7918777121
583723120.99613634720
583743122.67502013580
583763122.32850902271.0292427575
583783121.91806981710.6782550245
5835630122.3538733481.9097834245
5835830121.95227943690.9631825806
5836030121.10956845120
5836230121.91585828590
5838030120.14811509180.1472218845
5838230122.92953443522.4897008793
5838430122.04849333991.4041781786
5838630120.45123531740
583880161.41898851181.6724071518
583900160.51508212580.4901861272
583920161.40477244360.4868599118
583940161.09143148882.1299647499
584120161.30337737950.6026664511
584140160.15655234580.6390414316
584160160.97353502191.1798994973
584180161.13626068290.7989746793
583963162.08699485982.8354925861
583983160.47780025742.1335047598
584003161.88648510021.80571338
584023162.83510781684.0010116513
584203161.01215302961.9822330645
584223161.11990602061.7742536858
584243162.19648923641.4085556697
584263161.80106082273.4308957038
5840430162.67617173451.8568354379
5840630161.78215338330.7941435558
5840830162.01877761011.882677863
58410301600
5842830162.54707585762.5890057566
5843030162.6029988693.1147064061
5843230162.47020176192.9181913367
5843430163.69594293514.4951797619
Sheet1
0.11942236750.11942236750.14333392830.14333392830.17950254390.17950254390.21613330920.21613330920.2364491670.2364491670.18095712190.18095712190.23921811950.23921811950.1584301510.158430151
0.081457320.081457320.15518425060.15518425060.09471836430.09471836430.1263969650.1263969650.21820686290.21820686290.48690591810.48690591810.21922348270.21922348270.26822528110.2682252811
0.07006656710.07006656710.06325857470.06325857470.05299525550.05299525550.25900102660.25900102660.44434440540.44434440541.03462874651.03462874650.33879169720.33879169720.37471963770.3747196377
a
a
a
a
a
a
a
a
b
b
b
a
a
ab
b
b
b
ab
b
ab
b
c
a
a
24 h
48 h
4 d
8 d
9 d
10 d
12 d
16 d
Fadrozole (ug/L)
E2 production (relative to control)
Sheet2
0.2306756840.2306756840.91901530940.91901530940.15905597290.15905597290.30039867040.30039867040.35937838270.35937838270.15955765850.15955765850.31054744350.31054744350.21601565310.2160156531
0.49092062610.49092062611.13199679561.13199679560.16866859540.16866859540.62927491380.62927491380.44605684650.44605684650.35355676110.35355676110.15117792930.15117792930.32141429810.3214142981
0.25890779070.25890779071.42532509361.42532509360.21731863680.21731863681.57122047091.57122047090.78049732020.78049732020.4926401950.4926401950.34824699520.34824699520.49652772910.4965277291
a
a
b
a
b
a
a
b
a
a
b
a
a
a
a
b
b
a
ab
b
a
a
ab
b
24 h
48 h
4 d
8 d
9 d
10 d
12 d
16 d
Fadrozole (ug/L)
T production (relative to control)
Sheet3
0.11942236750.11942236750.081457320.081457320.07006656710.0700665671
0.14333392830.14333392830.15518425060.15518425060.06325857470.0632585747
0.17950254390.17950254390.09471836430.09471836430.05299525550.0529952555
0.21613330920.21613330920.1263969650.1263969650.25900102660.2590010266
0.2364491670.2364491670.21820686290.21820686290.44434440540.4443444054
0.18095712190.18095712190.48690591810.48690591811.03462874651.0346287465
0.23921811950.23921811950.21922348270.21922348270.33879169720.3387916972
0.1584301510.1584301510.26822528110.26822528110.37471963770.3747196377
&F
*
*
*
*
#
#
#
#
#
0
3
30
Time point
E2 production (relative to control)
0.2306756840.2306756840.49092062610.49092062610.25890779070.2589077907
0.91901530940.91901530941.13199679561.13199679561.42532509361.4253250936
0.15905597290.15905597290.16866859540.16866859540.21731863680.2173186368
0.30039867040.30039867040.62927491380.62927491381.57122047091.5712204709
0.35937838270.35937838270.44605684650.44605684650.78049732020.7804973202
0.15955765850.15955765850.35355676110.35355676110.4926401950.492640195
0.31054744350.31054744350.15117792930.15117792930.34824699520.3482469952
0.21601565310.21601565310.32141429810.32141429810.49652772910.4965277291
&F
#
#
#
#
#
#
#
*
0
3
30
Time point
T production (relative to control)
0.07006656710.07006656710.11942236750.11942236750.081457320.08145732
0.06325857470.06325857470.14333392830.14333392830.15518425060.1551842506
0.05299525550.05299525550.17950254390.17950254390.09471836430.0947183643
0.25900102660.25900102660.21613330920.21613330920.1263969650.126396965
0.44434440540.44434440540.2364491670.2364491670.21820686290.2182068629
1.03462874651.03462874650.18095712190.18095712190.48690591810.4869059181
0.33879169720.33879169720.23921811950.23921811950.21922348270.2192234827
0.37471963770.37471963770.1584301510.1584301510.26822528110.2682252811
#
#
#
#
#
*
*
*
*
0
3
30
Time point
Ex vivo E2 production (relative to control)
0.07006656710.07006656710.11942236750.11942236750.081457320.08145732
0.06325857470.06325857470.14333392830.14333392830.15518425060.1551842506
0.05299525550.05299525550.17950254390.17950254390.09471836430.0947183643
0.25900102660.25900102660.21613330920.21613330920.1263969650.126396965
0.44434440540.44434440540.2364491670.2364491670.21820686290.2182068629
1.03462874651.03462874650.18095712190.18095712190.48690591810.4869059181
0.33879169720.33879169720.23921811950.23921811950.21922348270.2192234827
0.37471963770.37471963770.1584301510.1584301510.26822528110.2682252811
#
#
#
#
#
*
#
#
0
3
30
Time point
Ex vivo T production (relative to control)
-
Aromatase A (CYP19A) gene expressionexposurepost-exposure
Chart1
1278.58003333332470.72842825.8473428571547.6822690753547.6822690753980.5975400569980.5975400569641.229579791641.229579791
975.9153754940.35622513618.9757428571124.2794187459124.27941874591023.62192927471023.62192927472105.39382987842105.3938298784
1345.78477142865617.206255525.0565142857304.2239148383304.2239148383689.9761769474689.97617694741089.5953607861089.595360786
1050.209352757.39706666672481.59025216.537382967216.537382967578.9286455984578.9286455984546.0749489434546.0749489434
982.90881849.9924252790.015125236.1028913768236.1028913768262.3420559405262.3420559405477.6438439085477.6438439085
1168.462351108.833952394.2108857143224.8358402555224.8358402555209.874045623209.874045623453.6921081955453.6921081955
a
a
a
a
b
c
a
b
b
a
b
b
a
ab
b
a
a
b
0
3
30
Time point (d)
CYP19A mRNA (copies/ng total RNA)
Fadrozole Phase 3 ovary, Arom-A
Document Name: Fadrozole Phase 3 ovaryArom-A8-3-07.sds
Plate Type: Absolute Quantification
User: dvillene
Document Information
Operator: JET
Run Date: FridayAugust 032007 17:35:10
Last Modified: FridayAugust 032007 18:58:42
Instrument Type: Applied Biosystems 7500 Real-Time PCR System
Comments:
Notebook 345p. 41
AromA mRNAlog copiesCtCtMeanSDCV
5.00E+077.698970004315.6515.4715.560.12727922060.8179898497
5.00E+066.698970004319.3919.2219.3050.12020815280.6226788542
500004.698970004326.6526.5826.6150.04949747470.1859758583
50003.698970004330.1730.1530.160.01414213560.0468903701
5002.698970004333.0833.3233.20.16970562750.5111615286
Fad (ug/L)TimeSample NameCtQtycopies/nglog ratio24 h
015105226.4448629.3972.586-0.1187999043Fad (ug/L)MeanSDcountSECV
015105427.2129426.68588.5336-0.336956655901278.58003333331341.54210040416547.6822690753104.9243743394
015105828.2714855.42297.1084-0.633812987532470.72842594.41722723227980.5975400569105.0061685142
015107624.32191599.633831.99260.4766967437302825.84734285711696.53400142537641.22957979160.036293387
015107827.9418309.57366.1914-0.5430197774
015108825.6580753.411615.06820.1014629481
315106025.4393170.411863.40820.1635800842
315106225.7177924.611558.49220.085976715
315106423.134125808251.60.8098102491
315106625.495104.521902.09040.1725032356
315108426.1459050.441181.0088-0.0344747844
315108627.1830058.01601.1602-0.3277376978
315109025.3796866.951937.3390.1804777031
3015106824.81138775.72775.5140.3366155042
3015107024.54165392.843307.85680.4128187822
3015107424.49170950.283419.00560.427171894
3015109226.3651124.551022.491-0.0970684241
3015109425.9964917.911298.35820.0066666071
3015109623.61301646.726032.93440.6738006849
3015109825.3896238.571924.77140.1776512388
025110025.28102887.62057.7520.100455864948 h
025110425.495106.291902.12580.0663020687Fad (ug/L)MeanSDcountSECV
025112426.3352082.441041.6488-0.195215849701632.8157452.10709498074226.053547490327.688801313
025112825.7476486.811529.7362-0.02832062363618.9072436.84670071674218.423350358470.5835544839
325110826.546628.19932.5638-0.24325861453017626.9649521103.4958004042410551.7479002021119.7227989065
325111226.546734.38934.6876-0.24227068643-out822.7836191.98708557833110.843795539523.3338493351
325113227.1830054.97601.0994-0.4339908731
325113634363.97.278-2.3509251163
3025111620.382.45E+06490001.4772589124
3025112023.03440708.228814.16440.732243979
3025114022.92474097.59481.950.7639604933
3025114424.59160587.273211.74540.2938039432
045114826.0861171.51223.430.09816696564 d
045115026.2853707.191074.14380.0416502659Fad (ug/L)MeanSDcountSECV
045115225.7675413.041508.26080.18906428370975.915375351.51527902858124.279418745936.0190328007
045115426.7639467.27789.3454-0.092145077734940.3562252895.240030245681023.621929274758.6038718341
045117226.4249175.29983.50580.00336476533013618.97574285715570.348485708172105.393829878440.9013760718
045117426.8736760.55735.211-0.1230001639
045117628.0117563.04351.2608-0.443782474
045117826.1957108.271142.16540.0683168397
345115626.1657982.431159.64860.0749142479
345115823.35357657.387153.14760.8650850264
345116024.3193969.733879.39460.5993517968
345116225.5884810.291696.20580.2400663838
345118022.92474412.419488.24820.9877738766
345118224.02231780.754635.6150.6766951994
345118424.23203117.564062.35120.6193653064
345118623.29372411.947448.23880.8826414321
3045116423.16405379.038107.58060.9194791147
3045116622.72538322.0610766.44121.0426600128
3045116822.4664034.8813280.69761.1338087279
3045118821.491.19E+06238001.3871647969
3045119022.42655338.3813106.76761.1280834388
3045119221.94889063.1917781.26381.260550465
3045119423.09424503.978490.07940.9394995917
085119626.0363200.191264.0038-0.02722723
085119826.3351931.661038.6332-0.11251340898 d
085120026.2654553.921091.0784-0.0911196515Fad (ug/L)MeanSDcountSECV
085122025.9666064.411321.2882-0.007978053201345.7847714286860.47517272528304.223914838363.9385428483
085122226.7140711.04814.2208-0.21823341735617.206251951.54733430678689.976176947434.7423122359
085122424.58161252.423225.04840.3795606271305525.05651428572882.798354329371089.59536078652.176812072
085122627.0233311.03666.2206-0.3053575524
385120423.81266382.035327.64060.5975593102
385120623.62300208.226004.16440.6494769655
385120823.45335474.786709.49560.6977142628
385121024.11218335.954366.7190.5111796362
385122823.74277471.55549.430.615272768
385123024.22203943.784078.87560.4815648505
385123222.9480286.919605.73820.8535551363
385123424.55164779.333295.58660.3889571187
3085121224.2206563.24131.2640.4871073389
3085121423.68288951.635779.03260.6328795347
3085121623.04437150.068743.00120.8126849284
3085123624.2207233.784144.67560.4885149348
3085123824.33189326.223786.52440.4492651501
3085124022.83502370.1310047.40260.8730781952
3085124225.29102174.762043.49520.1813980123
F10NTC35.250
F11NTC35.150
F12NTC35.120
H10QA- 5110025.3101176.852023.537Mean2179.5973333333
H11QA- 5110025.11114742.842294.8568SD140.1860959888
H12QA- 5110025.16111019.912220.3982CV6.4317428658
G10QA-Gonad25.4791005.311820.1062Mean2032.3144
G11QA-Gonad25.21107627.592152.5518SD184.3203468945
G12QA-Gonad25.23106214.262124.2852CV9.0694799434
&A
&F
Fadrozole Phase 3 ovary, Arom-A
547.6822690753547.6822690753
980.5975400569980.5975400569
641.229579791641.229579791
24 h
Fadro