p-1 

P.O. Box 331, Millwood, VA 22646 (540) 837-1602 

May 24, 2013 

C O 

IIII III III IIIIIIIIII III III § if 

4013000004 1 — 

=* 2m 

o o 

** 

o 

Re: Testing Consent Order for Ethylene Dichloride; 

Request for Modification of Enforceable Consent 

Agreement and Amendment to Study Plan (Pocket No. OPPT2003-0010) 

Dear Sirs: 

The HAP Task Force requests a modification of the Enforceable Consent Agreement 

for Ethylene Dichloride ("EDC"), 68 Fed. Reg. 33125 (June 3, 2003), as amended, 74 Fed. 

Reg. 36196 (July 22,2009), and further amended by Addendum Modification provided by 

letter dated November 23, 2011 from Maria J. Doa (the "ECA"). Six copies of this 

submission are enclosed. 

Pursuant to the ECA and the Study Plan for an extended one-generation drinking 

water reproductive toxicity study of ethylene dichloride in rats, submitted July 2, 2012, and 

amended July 16, 2012 and August 7, 2012, the experimental start date was August 14, 2012, 

and the final report, unless a second generation study was triggered, was due June 28, 2013. 

As the study was about to begin, however, WIL discovered that it was unable to obtain the 

appropriate glass water bottles with teflon stoppers. It is not possible to use plastic bottles 

because EDC dissolves them. 

The problem with the stoppers was dealt with last fall, but an issue regarding the 

delivered target doses to the animals then arose. In pre-testing, the formulated 

concentrations resulted in significantly lower concentrations that did not meet the expected 

drinking water target doses determined by the PBPK modeling from Tier 1 studies. Stability, 

solubility, and volatile properties were addressed by WIL to determine why the formulated 

Task Force 

BY HAND 

Document Control Office 

Office of Pollution Prevention and Toxics 

Room 6428 

U.S. Environmental Protection Agency 

1200 Pennsylvania Ave., N.W. 

Washington, DC 20460 

CONTAINS NO CBI 
p. 2 

Document Control Office 

May 24,2013 -

Page 2 

concentrations were lower than expected. WIL now believes that the issue has been 

resolved with an improved analytical methodology. 

As the issues appear to have been satisfactorily resolved, the Task Force seeks EPA 

approval of a modification of the ECA and amendment to the Study Plan to provide for an 

experimental start date of July 8, 2013, and a final report deadline (unless a second 

generation study is triggered) of July 21, 2014. We also seek approval of the enclosed 

further addendum to Appendix C to the Study Plan. This consists of Protocol Amendment 2, 

which includes clarification of several procedures contained within the initial protocol, 

revision of the statistical analysis outlined in the protocol to align with the recommendations 

of EPA reviewers, and the revised study schedule. Finally, we enclose the CV of Dr. 

Sawhney Coder, emergency contact for the study, as an addendum to Appendix A to the 

Study Plan. 

The study will be conducted pursuant to the protocol as amended by Protocol 

Amendments 1 and 2. 

We look forward to hearing from EPA. 

Sincerely, 

Peter E. Voytek, Ph.D. 

Manager 

Enclosures 

cc: John E. Schaeffer 

W. Caffey Norman, Esq. 

4819-6034-2036.1. 
p. 3 

|EWIL R E SEARCH 

PROTOCOL ATTACHMENT 2 

Study Number: WIL-417005 

Sponsor: HAP Task Force 

Title of Studv: 

An Extended One-Generation Drinking Water Reproductive Toxicity Study of Ethylene 

Dichloride in Rats 

Protocol Modifications: 

1) 1 Objective: 

The following is added to this section: 

Animals will not be paired more than once. If warranted, Fi animals may be mated to 

produce a second generation (see Section 7.16). 

2) The following personnel is added to the protocol: 

2.8 Contributing Scientist (Statistical Analysis): 

Les Freshwater, MS 

BioSTAT Consultants, Inc. 

528 West Centre Avenue 

Portage, MI 49024 

Tel: (269) 329-7976 

Email: LesFreshwater@biostat.net 

3) 3 Study Schedule: 

The following dates are updated in the protocol: 

Proposed Experimental Starting 

(Animal Receipt) Date: 25 June 2013 

Proposed Experimental Start Date: 8 July 2013 
p. 4 

Page 2 of 14 

WIL-417005 

Protocol Attachment 2 

Proposed Experimental 

Completion/Termination Date: 29 April 2014 

Proposed Audited Draft Report Date: 27 May 2014 

4) 6.3 Drinking Water: 

The second sentence of this paragraph is replaced with the following: 

Reverse osmosis-treated tap water (with test substance added during the treatment 

period for the animals assigned to Groups 2-4) will be available ad libitum to all 

animals and will be provided from standard glass water bottles with metal sipper 

tubes and no plastic or rubber fittings. 

5) 7.4.4 Treatment Regimen: 

This section is replaced with the following: 

Untreated or test substance-treated drinking water formulations will be offered ad 

libitum via standard glass water bottles with metal sipper tubes and no plastic or 

rubber fittings, and supplied for the duration of the study, as noted in Sections 7.4.4.1 

and 7.4.4.2. Fresh vehicle and test substance drinking water formulations will be 

provided at least weekly. 

6) 7.5.1 Preparation of Test Substance-Treated Drinking Water: 

The third sentence is replaced with the following: 

Test substance formulations will be prepared at least weekly. 

7) 7.5.3 Stability of Test Substance-Treated Drinking Water: 

This section is replaced with: 

Stability of the formulated test substance in drinking water was assessed in a 

concurrent study (WIL-417006; Stanton, Draft) after 8 and 15 days of room 

temperature storage at concentrations bracketing those used in the current study and 

will not be assessed in this study. 
Page 3 of 14 

WIL-417005 

Protocol Attachment 2 

8) 7.5.4 Homogeneity and Concentration of Test Substance in the Drinking Water: 

As per study director notification dated 4 October 2012, the requirement for 

homogeneity assessment is removed as the test substance is soluble in the water to a 

concentration of 8 mg/mL, which is higher than the expected highest concentration in 

the vehicle. 

9) 7.11 Assignment of Fi Animals to Cohorts: 

The first sentence of paragraph 3 is replaced with the following. 

Assignment of same-sex littermates to a particular Cohort will be avoided. 

10) 7.14.3.1 Males: 

This section is replaced with the following: 

Recorded individually on a twice-weekly basis, beginning 1 week prior to the 

initiation of test substance administration, until euthanasia. 

11) 7.14.3.2 Females: 

This section is replaced with the following: 

Recorded individually on a twice-weekly basis, beginning 1 week prior to the 

initiation of test substance administration, until evidence of copulation is observed, 

and on gestation days 0, 3, 7, 10, 14, 17 and 20 and lactation days 1, 4, 7, 10, 14, 17 

and 21. 

Females with no evidence of mating or that fail to deliver will continue to be weighed 

on a twice-weekly basis until the scheduled euthanasia. 

12) 7.14,4.1 Males: 

This section is replaced with the following: 

Recorded individually on a twice-weekly basis on the days of body weight collection, 

beginning 1 week prior to the initiation of test substance administration, until 

euthanasia. 
Page 4 of 14 

WIL-417005 

Protocol Attachment 2 

13) 7.14.4.2 Females: 

This section is replaced with the following: 

Recorded individually on a twice-weekly basis on the days of body weight collection, 

beginning 1 week prior to the initiation of test substance administration, until the start 

of the mating period and on gestation days 0, 3, 7, 10, 14, 17 and 20 and lactation 

days 1,4, 7, 10, 14, 17 and 21. 

For females with no evidence of mating or that fail to deliver, individual food 

consumption will continue to be weighed on a twice-weekly basis until the scheduled 

euthanasia. 

14) 7.14.5.1 Males: 

This section is replaced with the following: 

Recorded (by weight) individually on a twice-weekly basis on the days of body 

weight collection, beginning 1 week prior to the initiation of test substance 

administration, until euthanasia. 

15) 7.14.5.2 Females: 

This section is replaced with the following: 

Recorded (by weight) individually on a twice-weekly basis on the days of body 

weight collection, beginning 1 week prior to the initiation of test substance 

administration, until the start of the mating period and on gestation days 0, 3, 7, 10, 

14, 17 and 20 and lactation days 1,4, 7, 10, 14, 17 and 21. 

For females with no evidence of mating or that fail to deliver, individual water 

consumption will continue to be weighed on a twice-weekly basis until the scheduled 

euthanasia. 

16) 7.14.6.2 Body Weights: 

This section is replaced with the following: 

Each pup will be weighed on PND 1, 4 (before culling), 7, 14 and 21, on a twiceweekly basis following weaning, and on the day of euthanasia. 
Page 5 of 14 

WIL-417005 

Protocol Attachment 2 

17) 7.14.6.3 Food Consumption: 

This section is replaced with the following: 

Cage food consumption will be recorded on PND 21, 24 and 28 (the days of body 

weight collection). Individual food consumption will be recorded on a twice-weekly 

basis on the days of body weight collection, beginning on PND 28 (when the animals 

are individually housed) until the day prior to euthanasia. 

18) 7.15 Developmental Neurotoxicity (Fi Cohort 2A): 

The first sentence of the second paragraph is replaced with the following: 

The functional observational battery tests will be counterbalanced across testing 

times, sexes, and treatment groups. Motor activity and auditory startle response tests 

will be counterbalanced with respect to cage assignment, testing times, sexes, and 

treatment groups. 

19) 7.14.6.4 Water Consumption: 

This section is replaced with the following: 

Cage water consumption (by weight) will be recorded on PND 21, 24 and 28 (the 

days of body weight collection). Individual water consumption (by weight) will be 

recorded on a twice-weekly basis on the days of body weight collection, beginning on 

PND 28 (when the animals are individually housed) until the day prior to euthanasia. 

20) 7.20.1 Motility/Viability Assessment: 

The fourth and fifth sentences of this section are revised as follows: 

A small sample will be loaded into a 100-um chamber for determination of motility. 

Because sperm motility can be affected by temperature shock, all slides and diluents 

will be warmed prior to use in an incubator, and motility determinations will be 

performed under constant temperature (approximately 37°C) using the HamiltonThorne HTM-IVOS Version 14 computer-assisted sperm analysis (CASA) system. 
p. 8 

WIL-417005 

Page 6 of 14 Protocol Attachment 2 

21) 8.4 Fi Offspring Scheduled for Neuropathology/Brain Weight Measurements 

(Cohorts 2A and 2B) 

The following is added to this section: 

Due to the large number of animals to be perfused for neuropathological assessment, 

perfusions for Cohorts 2A and 2B will be performed over several days. The 

perfusions will be performed such that both sexes and all treatment groups will be 

approximately equally represented across each day. The order of perfusions each day 

will also be counterbalanced by sex and treatment group (i.e., 1 male from Group 1, 1 

male from Group 2, 1 male from Group 3, 1 male from Group 4, 1 female from 

Group 1, etc.). 

22) 8.4.2.2 Neuropathology - Microscopic Examination: 

The last paragraph of this section is replaced with the following: 

Based on the results of the qualitative examination and the professional judgment of 

the pathologist, selected sites in the nervous system and cellular components may be 

further evaluated by the use of additional histopatho logical methods (at additional 

cost). If no neuropathological alterations are observed, subsequent analysis will not 

be performed. 

23) The following section is added to the protocol: 

8.4.2.3 Morphometric Analysis: 

Two coronal sections (homologous between animals) and the midsagittal section of 

pons/medulla will be used for morphometry. Homology will be achieved by stepsectioning tissue blocks at 60-micron intervals as needed and choosing the most 

representative section for each animal. Specific levels to be analyzed are Levels 1, 3 

and 5 and are defined as follows (Paxinos and Watson, 1998). 

Level 1 is a coronal section obtained just rostral to the genu of the corpus callosum 

and contains the following major structures: cerebral cortex, corpus callosum, basal 

ganglia (caudate putamen, nucleus accumbens), and lateral ventricles. Two bilateral 

microscopic measurements are obtained at this level: the vertical height (of the 

hemisphere just medial to the lateral ventricle, and the vertical height of the cerebral 

cortex from the apex of the corpus callosum/cingulum to the dorsal surface of the 

hemisphere. 

Level 3 is a coronal section obtained approximately at the level of the optic chiasm, 

just rostral to the pituitary, and containing the following major structures: cerebral 
Page 7 of 14 

WIL-417005 

Protocol Attachment 2 

cortex, corpus callosum, basal ganglia, hippocampus/dentate hilus, thalamus, 

hypothalamus, and lateral and third ventricles. Four bilateral microscopic 

measurements are taken at this level: the radial thickness of the cortex, the vertical 

height between the layers of the hippocampal pyramidal neurons along a line that 

passes through the termination of the dorsal limb of the dentate hilus, the vertical 

height of the dentate hilus measured at the termination of the ventral limb, and the 

length of the ventral limb of the dentate hilus taken along a straight line from the crest 

to the termination of the ventral limb. 

Level 5 is a sagittal section of the cerebellum and medulla oblongata, taken slightly 

lateral to the midline, and containing the following major structures: cerebellum, 

medulla oblongata (pyramidal tract, trapezoid body, medial longitudinal fasciculus), 

and fourth ventricle. Single microscopic measurements of the height of the 

cerebellum and of the thickness of the base of cerebellar lobule #9 are obtained at this 

level. 

Means of paired measurements for Levels 1 and 3 will be reported and used for 

statistical computations. Measurements on Level 5 will not be paired. A total of 8 

microscopic measurements will be included in the analysis. 

24) 10 Statistical Methods: 

The first sentence in this section is replaced with the following: 

All analyses will be two tailed. 

In addition, the following is added to this section: 

Repeated measures analyses, two-way analysis of variance (ANOVA) models, and 

brain morphometry data analysis will be conducted by BioSTAT Consultants, Inc. 

AH other statistical analyses will be conducted by WIL Research. 

The repeated measures analyses and brain morphometric data analyses performed by 

BioSTAT will be conducted using SAS version 9.2 (SAS Institute, Inc., 2002-2008), 

or higher, software. 

25) 10.1 In-Life Data: 

The first paragraph of this section is replaced with the following: 

Pre-coital intervals, mean estrous cycle length, gestation length, former implantation 

sites, unaccounted-for sites, mean days of attainment of developmental landmarks 

(balanopreputial separation and vaginal patency) and the body weight on the day of 
p. 10 

WIL-417005 

Page 8 of 14 Protocol Attachment 2 

attainment, anogenitai distance (absolute and relative to the cube root of body 

weight), numbers of nipples/areola, and time from vaginal patency to first estrus, will 

be subjected to a parametric one-way analysis of variance (ANOVA) (Snedecor and 

Cochran, 1980). Dunnett's test (Dunnett, 1964) will be applied to the data to compare 

the treated groups to the control group at the 0.05 significance level regardless of the 

outcome of the ANOVA. 

Food and water consumption will be reported but will not be statistically analyzed. 

In addition, the following is added to this section: 

Body weight for Study Days 0 through 91 (males) or 27 (females), Gestation Days 0 

through 20 and Lactation Days 1 to 21 will be analyzed separately with a repeated 

measures analysis of variance (RANOVA). Factors in the model will include: fixed 

effects treatment group (TRT) and day (TIME); and interaction term TRT*TIME. 

The covariance structure for the RANOVA model will be selected by comparing the 

corrected Akaike's Information Criteria (AICC). The Kenward-Rogers adjustment 

for denominator degrees of freedom will be applied. Dunnett's test will be used to 

make pairwise comparisons of each individual treated group with the control. If the 

TRT*TIME interaction is significant at the 0.05 level, Dunnett's test will be 

conducted for each individual day. If TRT*TIME is not significant, Dunnett's test 

will be conducted overall across the pooled days only. 

Body weight gains will be reported, but will not be statistically analyzed. 

26)10.2 Litter Data: 

The last sentence of this section is removed. 

In addition, the following is added to this section: 

Pre-culling (PND 1 and 4) and post-culling (PND 4, 7, 14 and 21) pup body weights 

will be analyzed separately with a repeated measures analysis of variance 

(RANOVA). Factors in the model will include: random effects LITTER; fixed 

effects treatment group (TRT), SEX and day (TIME); and interaction terms 

TRT*SEX, TRT*TIME and TRT*SEX*TIME. 

The first examination will be whether the TRT*SEX and/or TRT*SEX*TIME 

interaction is significant. If either interaction is significant at the 0.05 level, the 

analysis will be conducted, by sex, with a RANOVA. Factors in the model will 

include: random effect LITTER; fixed effects TRT and TIME; and interaction term 

TRTTIME. The covariance structure for the RANOVA model will be selected by 

comparing the corrected Akaike's Information Criteria (AICC). The Kenward-
p. 11 

WIL-417005 

Page 9 of 14 Protocol Attachment 2 

Rogers adjustment for denominator degrees of freedom will be applied. 

In the final RANOVA model, Dunnett's test will be used to make pairwise 

comparisons of each individual treated group with the control. If the TRT*TIME 

interaction is significant at the 0.05 level, Dunnett's test will be conducted for each 

individual day. If TRT*TIME is not significant, Dunnett's test will be conducted 

overall across the pooled days only. 

Pup body weight gains will be reported, but will not be statistically analyzed. 

27) 10.5 Functional Observational Battery, Locomotor Activity, and Auditory 

Startle Data: 

The title of this section is revised as indicated above. 

The first (only) sentence in this section is removed (information moved to 

Section 10). 

28) 10.5.1 Functional Observational Battery Data: 

The title of this section is revised as indicated above, and this section is replaced with 

the following: 

Continuous Functional Observational Battery (FOB) data (including the average of 

3 grip strength trials and 3 landing foot splay trials) will be subjected to a two-way 

ANOVA (Snedecor and Cochran, 1980) including terms in the model for treatment 

group (TRT), SEX, and TRT*SEX interaction. Dunnett's test (1964) will be used to 

compare the control and treated groups. If TRT*SEX is significant at the 0.05 level, 

Dunnett's test will be conducted, by sex, with a one-way ANOVA. Otherwise, 

Dunnett's test will be conducted across the pooled sexes. 

FOB parameters which yield scalar and descriptive data will be analyzed, by sex, 

with Fisher's Exact Test (Steel and Torrie, 1980). 

29) 10.5.2 Locomotor Activity Data: 

This section is replaced with the following: 

Ambulatory activity counts will be analyzed, by session, with a repeated measure 

analysis of variance (RANOVA). Factors in the model will include: fixed effects 

treatment group (TRT), SEX, and time interval (TIME); interaction terms TRT*SEX, 

(TRT*TIME), and TRT*SEX*TIME. The SAS® procedure PROC MIXED will be 

used, and the covariance structure across time will be selected by comparing the 
p. 12 

WIL-417005 

Page 10 of 14 Protocol Attachment 2 

corrected Akaike's Information Criterion (AICC). The Kenward-Rogers adjustment 

for denominator degrees of freedom will be applied. 

The first examination will be whether the TRT*SEX and/or TRT*SEX*TIME 

interaction is significant. If either interaction is significant at the 0.05 level, the 

analysis will be conducted, by sex, with a RANOVA. Factors in the model will 

include: fixed effects TRT and TIME, and the interaction term TRT*TIME. 

In the final RANOVA model, the monotonic dose-response relationship will be 

evaluated using sequential linear trend tests based on ordinal spacing of dose levels. 

The linear dose by time interaction (LinTrt*Time) will be evaluated and, if significant 

at the 0.05 level, trend tests on treatment means will be performed at the 0.05 level 

for each time interval. If the linear dose by time interaction is not significant, the 

trend test will be conducted across the pooled time intervals of the entire session only. 

Nonmonotonic dose responses will be evaluated whenever no significant linear trends 

are detected but TRT and/or TRT*TIME interaction is significant at the 0.01 level. 

Within the framework of the RMANOVA, pairwise comparisons will be made for 

each individual treated group with the control group through linear contrasts. If 

TRT*TIME is significant, the comparisons will be conducted for each time interval. 

If only the TRT effect is significant, the comparisons will be conducted across the 

pooled time intervals of the entire session. These nonmonotonic dose response 

comparisons will be conducted at the 0.01 significance level. 

30) 10.5.3 Auditory Startle Response Data: 

This section is replaced with the following: 

Mean response amplitude (on each block of 10 trials) will be analyzed, by session 

with a RANOVA. Factors in the model will include fixed effects: treatment group 

(TRT), SEX and TRIAL block; interaction terms TRT*SEX, TRT*TRIAL, and 

TRT*SEX*TRIAL. The SAS® procedure PROC MIXED will be selected by 

comparing the corrected Akaike's Information Criterion (AICC). The KenwardRogers adjustment for denominator degrees of freedom will be applied. 

The first examination will be whether the TRT*SEX and/or TRT* SEX*TRIAL 

interaction is significant. If either interaction is significant at the 0.05 level, the 

analysis will be conducted, by sex, with a RANOVA. Factors in the model will 

include: fixed effects TRT and TRIAL; and the interaction term TRT*TRIAL. In the 

final RANOVA, the monotonic dose-response relationship will be evaluated using 

sequential linear trend tests based on ordinal spacing of dose levels. The linear dose 

by trial interaction (LinTrt*Trial) will be evaluated and, if significant at the 0.05 

level, trend tests on treatment means will be performed at the 0.05 level for each trial 
p. 13 

WIL-417005 

Page 11 of 14 Protocol Attachment 2 

block. If the linear dose by trial interaction is not significant, the trend test will be 

conducted across the pooled trial blocks of the entire session only. 

Nonmonotonic dose responses will be evaluated whenever no significant linear trends 

are detected but TRT and/or TRT*TRIAL interaction is significant at the 0.01 level. 

Within the framework of the RANOVA, pairwise comparisons will be made for each 

individual treated group with the control group through linear contrasts. If 

TRT*TRIAL is significant, the comparisons will be conducted for each trial block. If 

only the TRT effect is significant, the comparisons will be conducted across the 

pooled trial blocks of the entire session. These nonmonotonic dose response 

comparisons will be conducted at the 0.01 significance level. 

Latency to maximum response (TMAX) will be reported but not statistically analyzed. 

31) 10.6 Brain Morphometric Data (Cohort 2A): 

This section will be revised in a following protocol attachment. 

32) 11 Quality Assurance: 

The last paragraph of this section is replaced with the following: 

Data analysis performed by BioSTAT Consultants, Inc. will follow the current 

procedural guidelines of BioSTAT Consultants, Inc. BioSTAT Consultants, Inc. will 

provide a statistical analysis report, which will be included as an appendix to the final 

report. Quality Assurance auditing of the statistical report (for internal consistency 

and consistency with the study report) will be conducted under the direction of the 

WIL Research Quality Assurance Department. 

33) 17 References: 

The following is added to this section: 

Paxinos, G. and Watson, C. The Rat Brain in Stereotaxic Coordinates 4th edition. 

Academic Press, San Diego, CA, 1998. 

Stanton, A. An Analytical Method Validation and Stability Study of Ethylene 

Dichloride in Aqueous Formulations. WIL Research, Ashland, OH, DRAFT. 
p. 14 

WIL-417005 

Page 12 of 14 Protocol Attachment 2 

34) Appendix A Study Design: 

References to weekly Detailed Clinical Observations for the Fo and Fi generations are 

removed from this section. 

Reasons for Protocol Modification: 

1) To clarify that animals will not be paired more than once, and that a second 

generation may be warranted based on the results obtained from the FQ animals. 

2) To include the Contributing Scientist (Statistical Analysis). 

3) To update the study schedule. 

4-5) The test substance interacts with plastic. Therefore, these modifications update the 

type of bottles to be used for administration of the test substance. 

6) To allow for more frequent preparation of the test substance formulations. 

7) To indicate that room temperature storage stability was included in a concurrent 

study. 

8) To remove the requirement for homogeneity assessment. 

9) To clarify that same-sex littermates will not be assigned to the same Cohort. 

10-17) Given that formulations may need to be prepared more frequently than once per 

week, these modifications update the schedule for body weight, food consumption 

and water consumption to twice-weekly to accommodate the more frequent test 

substance formulation. 

18) To extend the counterbalancing procedures for behavioral testing to include cage, sex, 

and treatment group in addition to testing times 

19) Given that formulations may need to be prepared more frequently than once per 

week, these modifications update the schedule for body weight, food consumption 

and water consumption to twice-weekly to accommodate the more frequent test 

substance formulation. 

20) To change 100-p.m cannula to 100-um chamber slide, and to update the version of the 

Hamilton-Thorne HTM-IVOS system. 

21) Counterbalancing of perfusions across day, sex, and treatment group is added. 
p. 15 

WIL-417005 

Page 13 of 14 Protocol Attachment 2 

22) To clarify what may occur if the pathologist recommends further analysis of specific 

sites within the nervous system. 

23) To include details for the morphometric analysis. 

24-31) To revise the statistical analyses to be performed. 

32) To include Quality Assurance responsibilities for all statistical analyses performed by 

BioSTAT Consultants, Inc. 

33) Additional references included. 

34) Weekly detailed clinical observations were removed from the protocol via Protocol 

Amendment 1. 
Page 14 of 14 

WIL-417005 

Protocol Attachment 2 

Approval: 

Sponsors approval was obtained via f^mrv^l on J^Jtfl&u 2tiJ3--

Dal* 

WIL Research 

Study Director 

HAP Task Force 

Melissa J. Beck, PhD DaitJ 

"PcterE. Voyt^k, PhD Date / 

Sponsor Representative 
p. 17 

CURRICULUM VITAE 

October 1,2012 

Formal Education: 

2008 

2000 

1995 

1993 

Professional Experience: 

04/24/10 - Present 

PRAGATISAWHNEY CODER 

Developmental and Reproductive Toxicology 

Diplomate, American Board of Toxicology 

Indian Institute of Technology-Bombay (IITB) 

Biotechnology Centre 

Powai, Bombay, India 

PhD Molecular Biology 

Jamia Millia University 

Centre for Bio Sciences 

New Delhi, India 

MS Bio Sciences 

Jamia Millia University 

Centre for Bio Sciences 

BS Bio Sciences 

WIL Research 

Ashland, OH 

Position: Senior Toxicologist 

Developmental and Reproductive Toxicology 

Study Director for developmental, reproductive and 

neurotoxicity studies. Responsible for the planning, conduct 

and successful reporting of assigned studies. Review 

departmental toxicology reports for interpretation and 

presentation of study findings. Provide senior level 

interpretation of data generated by toxicology studies. Consult 

with Study Directors and others to provide advice for 

appropriate analysis, presentation and description of study 

findings. Mentor and advise junior level Study Directors 

regarding scientific aspects of study design and data analysis 

and interpretation. Confer with clients and/or other external 

scientists to provide advice as appropriate. Prepare technical 

papers, publications and reports as directed. Maintain 

competency as a senior toxicologist by researching scientific 
p. 18 

Pragati Sawhney Coder CURRICULUM VITAE 

Page 2 

Professional Experience (Continued) 

literature, attending seminars, scientific meetings and other 

venues for continuing education. Serve as a primary scientific 

contact for key clients and provide on-site support for their 

programs. Develop new capabilities for WIL Research. 

01/16/06 - 04/23/10 Position: Staff Toxicologist 

Developmental and Reproductive Toxicology 

Study Director for developmental, reproductive and 

neurotoxicity studies. Responsible for the planning, conduct and 

successful reporting of assigned studies. Participate in report 

review and generation process. Develop new capabilities for 

WIL Research. Communicate with Sponsors as required. 

The University of Texas at Austin 

College of Pharmacy, Division of Pharmacology and Toxicology 

Austin, TX 

2002 - 2005 Position: Postdoctoral Research Associate 

Laboratory of Dr. John H. Richburg 

Conducted and supervised toxicological research in the lab. 

Performed overall maintenance of the laboratory including 

compliance with environmental health and radiation safety. 

Solely responsible for the development of mouse exposure 

model for the evaluation of cisplatin-induced testicular toxicity. 

2000 - 2002 Position: Postdoctoral Fellow 

Laboratory of Dr. John H. Richburg 

Scientific Trainine: 

2000-2005 Developmental Staging of Rodent Spermatogenesis 

Worked under the direct tutelage of leading experts in the field 

of rodent testicular histopathology and spermatogenesis staging, 

Prof. Rex Hess (Veterinary Biosciences, University of Illinois, 

University of Illinois at Urbana-Champaign) and Prof. Marvin 

Meistrich, (Experimental Radiation Oncology, University of 

Texas MD Anderson Cancer Center). Trained graduate students 

and new appointees in the laboratory of Dr. John Richburg, as 

needed during this period. 
Pragati Sawhney Coder 

Page 3 

Awards: 

WIL Research Company Scientific Publication Award. 2011. 

Continuing Education: 

Stem Cells in Toxicology. 51st Annual Meeting, Society of Toxicology, San Francisco, CA, 

March 11, 2012. 

MicroRNAs in Biology and Toxicology. 51st Annual Meeting, Society of Toxicology, San 

Francisco, CA, March 11, 2012. 

Preclinical Developmental Toxicity Studies. 51st Annual Meeting of the Teratology Society, 

San Diego, CA, June 25,2011. 

Postmarketing Sirveillance Studies and Communication of Risk. 51s t Annual Meeting of the 

Teratology Society, San Diego, CA, June 25,2011. 

The Biology and Toxicology of Peri- and Post-Natal Development. 50th Annual Meeting, 

Society of Toxicology, Washington, DC, March 6, 2011. 

Reproductive and Developmental Toxicology for Industrial and Regulatory Scientists, Noblis, 

Inc. Sponsored by Teratology Society, Falls Church, VA. October 25-28, 2010. 

CNS Functional Teratology. 48th Annual Meeting of the Teratology Society, Monterey, CA, 

June 28, 2008. 

Nanoparticles. 48th Annual Meeting of the Teratology Society, Monterey, CA, June 28, 2008. 

The Biology and Toxicology of Peri- and Post-Natal Development. 46th Annual Meeting of the 

Society of Toxicology, Charlotte, NC, March 25, 2007. 

Vertebrate Embryology. 46th Annual Meeting of the Teratology Society, Tucson, AZ, June 25-

26, 2006. 

Articles: 

Hovey, R.C.; Coder, P.S.; Wolf, J.C.; Sielken, R.L. Jr.; Tisdel, M.O.; Breckenridge, C.B. 

Quantitative Assessment of Mammary Gland Development in Female Long Evans Rat 

Following in utero Exposure to Atrazine. Toxicological Sciences 2011,119(2), 380-390. 

Yao, P.; Lin, Y.; Sawhney, P.; Richburg, J.H. Transcriptional regulation of FasL expression 

and participation of sTNF-a in response to Sertoli cell injury. Journal of Biological 

Chemistry 2007, 282(8), 5420-5431. 

p. 19 

CURRICULUM VITAE 
p. 20 

Pragati Sawhney Coder CURRICULUM VITAE 

Page 4 

Articles (Continued): 

Sawhney, P.; Giammona, C.J.; Meistrich M.L.; Richburg, J.H. Cisplatin-Induced long-term 

failure of spermatogenesis in adult C57/B1/6J mice. Journal ofAndrology 2005, 26(1), 139-

145. 

Seaman, F.C.; Sawhney, P.; Giammona, C.J.; Richburg, J.H. Cisplatin-induced pulse of 

germ cell apoptosis precedes long term elevated apoptotic rates in C57/BL/6 mouse testis. 

Apoptosis 2003, 8, 101-108. 

Giammona, C.J.; Sawhney, P.; Chandrasekaran Y.; Richburg, J.H. Death receptor signaling 

in rodent testis after MEHP exposure. Toxicology and Applied Pharmacology 2002, 185, 

119-127. 

Book Chapters: 

Stump, D.G.; Nemec, M.D.; Parker, G.A.; Sawhney Coder, P.; Sloter, E.D.; Varsho, B.J. 

Significance, Reliability, and Interpretation of Developmental and Reproductive Toxicity 

Study Findings. In Developmental and Reproductive Toxicology: A Practical Approach, 3rd 

Edition; Informa Healthcare Books, 2011. 

Sawhney Coder, P.; Sloter, E.D.; Bowman, C.J.; Stump, D.G.; Nemec, M.D. Evaluation of a 

Male Reproductive Toxicant. In Comprehensive Toxicology, 2 n d Edition; Elsevier Ltd.: 

Oxford, UK, 2010. 

Abstracts: 

Lewis, K.M.; Ferguson, J.; Gleason, T.; Sawhney Coder, P. Effects of intraoperatively applied 

coseal on embryo/fetal development in rats. The Toxicologist 2010,114 (l),9l (Supplement). 

Sawhney Coder,P.; Carpenter, T.M.; Kopp, C.A.; Clevidence, K.J.; Fleeman, T.L.; Stump, 

D.G.;Nemec, M.D. Evaluation Of A Shortened Skeletal Staining (Alizarin Red S) Method 

For Rodent And Rabbit Fetuses. Birth Defects Research, Part A 2009, 85, 341. 

Sawhney Coder, P.; Barrett, R.M.; Varsho, B.J.; Stump, D.G.; Nemec, M.D.; Holson, J.F. 

Comparison of fetal malformation and developmental variation rates between 

Hra:(NZW)SPF rabbits obtained from the Denver or Kalamazoo facilities of Covance 

Research Products. Birth Defects Research Part A 2008, 82, 353. 

Sawhney Coder, P.; Stump, D.G.; Nemec, M.D.; Lampert, B.M.; Trost, L.C. Developmental 

toxicity studies in rats and rabbits with CMX001: a broad spectrum oral anti-viral agent. 

Birth Defects Research Part A 2008, 82, 379. 
p. 21 

Pragati Sawhney Coder CURRICULUM VLTAE 

Page 5 

Abstracts (Continued): 

Sawhney, P.; Birchmeier, P.A.; Varsho, B.J.; Stump, D.G.; Nemec, M.D.; Holson, J.F. A 

comparison of maternal growth curves and reproductive parameters between 

HRA:(NZW)SPF Rabbits obtained from the Denver or Kalamazoo facilities of Covance 

Research Products. Birth Defects Research Part A 2007, 79, 415. 

Nemec, M.D.; Kirkpatrick, D.T.; Sawhney Coder, P.; Knapp, J.F.; Sherman, J.; Tanchevski, 

E.; Donahue, D.A.; Petruzzi, E.E.; Farr, C.H. A Two-Generation Reproductive Toxicity and 

Lactational Inhalation-Phase Exposure Study of Dimethyl Disulfide in Rats. The Toxicologist 

2007, 96(1), 312. 

Sawhney, P.; Richburg, J.H. The XVIIIth testis workshop: Testicular cell dynamics and 

endocrine signaling. Journal of Andrology 2005, 26(6). 

Sawhney, P.; Richburg, J.H. Testicular Sertoli cells survive despite Cisplatin-induced injury 

due to the expression of inhibitor of Apoptosis Proteins that disrupt Mitochondrial-mediated 

apoptotic signaling, The Toxicologist 2004, 78(S1), 401. 

Sawhney, P.; Giammona, C.J.; Seaman, F.C.; Ye, Y.; Richburg, J.H. Trail and death receptor 

response in rodent testis after cisplatin exposure. The Toxicologist 2003, 72(S1), 360. 

Ye, Y.; Giammona, C.J.; Sawhney, P.; Richburg, J.H. The expression of TNF-Related 

Apoptosis inducing Ligand (TRAIL) in rodent Sertoli cells. The Toxicologist 2003, 72(81), 

357. 

Sawhney, P.; Seaman, F.C.; Giammona, C.J.; Castro, C; Richburg, J.H. Establishment of a 

mouse model for assessing the mechanisms of sustained testicular damage after Cisplatin 

exposure. The XVIIt h Testis Workshop: Functional Genomics of Male Reproduction 2003, 

Abstract 69. 

Sawhney, P.; Giammona, C.J.; Seaman, F.C.; Richburg, J.H. Correllation of germ cell death 

and Apoptotoc signalling in testes of C57 mice exposed to Cisplatin. Proceedings of the 93r d 

Annual Meeting, Vol 42. American Association for Cancer Research 2002, Abstract 2625. 

Professional Affiliations: 

Society of Teratology, USA, 2006 - present 

Society of Toxicology, USA, 2002 - present 

American Society of Andrology, 2002 - 2011 

American Association for Cancer Research, USA, 2001 - 2005 

Gulf Coast Chapter, Society of Toxicology, USA, 2000 - 2006 

Symbiotek, Indian Institute of Technology, India, 1995 - present 

Indian Society of Cell Biology, Student Member 1995 - 2000 
Pragati Sawhney Coder 

Page 6 

p. 22 

CURRICULUM VITAE 

Seminars: 

Cisplatin, Copper and Spermatogenesis. Department of Urology, Weill Medical College of 

Cornell University and New York Presbyterian Hospital, 2005. 

The Altruistic Sertoli cell. Center for Research in Reproductive Physiology, The University 

of Pittsburgh School of Medicine, 2005. 

Cisplatin - Therapy for this generation and Apoptosis of the next. Center for Pediatric 

Research, Alfred I. Dupont Hospital for Children, 2005. 

Cisplatin Therapy vs. Spermatogenic Injury. School of Biosciences and BioEngineering, 

Indian Institute of Technology, 2004. 

Cisplatin - Therapy for this generation and Apoptosis of the next. Pharmacology Seminar 

Series at College of Pharmacy at The University of Texas, 2004. 

Cell Death - Apoptosis. Vlth Annual Summer Carcinogenesis Lecture Series at College of 

Pharmacy, The University of Texas at Austin, 2003. 

The Chemistry of Genetics. Department of Chemistry and Biochemistry, Southwestern 

University, 2002. 

Apoptosis - Vth Annual Summer Carcinogenesis Lecture Series at College of Pharmacy, The 

University of Texas at Austin, 2002. 

Primed to Die: Germ cells in the Mammalian Testis. Biotechnology Centre, Indian Institute 

of Technology, 2002. 

Pragati Sawhney Coder Date 
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