Document QgeVqMBwMoywEw9jNvLqjJ2xL

Molecular Dosimetry of Vinyl Chloride James A. Swenberg, D.V.M., Ph.D. Research Report and Budget Request July 28, 1997 BFG40943 rfom: *ame9 4, Swnora "o. Wenay Sfiermart Oait: 3/2;.Br nm*: Pag* i of 4 Research Report Year 1 (1996-1997 Budget) The laboratory portion of this project began this spring by sending teams of 3-4 of our laboratory personnel to New Jenev to collect tissues and liver cell types from rats that were exposed to 0. 10. 100. or 1100 ppm vinyl chloride (VC) for 1-20 days. Procedures were rather difficult during the first set of necropsies, but went much better for subsequent procedures after we sent an additional person and shipped up our own centrifuge. We were able to harvest good yields of cells and tissues for the experiment as a whole. It was unfortunate that the [UC,]-VC experiment was the first to come off. as this was the least smooth running of all. Even so. adequate numbers of cells were able to be harvested to meet our objectives. We are currently isolating DNA from the 4-week hepatocytes. as these are the most plentilul cells. A new iinmunoaffinity chromatograph-GC MS method is replacing our previous low pressure strong cation exchange-GC/MS method for the analysis of ethenoguanine. After this shakedown, we will begin working with the [UCJ-VC liver cells. Parallel to this effort will be the analysis of urines for excretion of ethenoadenine using immunoaftinity chromatography and LC-MS. Likewise, we will begin examining the major DNA repair pathway for VC DNA adducts including methyl purine giycosyiase (MPG) and AP-endonuclease (AP). Year 2 (1997-1998 Budget) During Year 2 wc will continue to work up the many samples collected in New Jersey. Priority will be given to establishing exposure response relationships, identifying cell-specific differences in DNA repair, determining effects of exposure on endogenously formed DNA adducts, and doing initial studies related to incorporating tiiese data into a PB/PK risk assessment model. V e are also developing new methods for the1N. 2 -ethenoguanine adduct, which has never been characterized in vivo. Year 3 (1998-1999) We expect to complete this research during die third year. At this time, we will have analyzed all ofthe relevant tissues and cells for DNA adducts, will establish the role of defects in DNA repair and the effect of induction ofDNA repair on exogenous and endogenous DNA adducts, and incorporate these data into a biologically-based risk assessment model. BFG40944 BFG 00443 -ames A. Swenoera To: Wenay Sherman a*e: V2im iime: is Page a of Research Budget Mv understanding ofdie funding available for this project is diat between S50.000-75.000 will be available per year for three years. The cost of this research project will greatly exceed this, with personnel costing -SI40.000 this year. However, since VC lias been and continues to be one of die chemicals that we are investigating under our NIEHS Superfund Basic Research Program Project. I can offset part of the cost providing that the CMA agrees to share acknowledgement widi NIEHS. Should we need additional [UCJ-VC exposures, they can be done at UNC, since the EOIC lias agreed to fund new research that includes funds to set up a small nose-only exposure system for similar studies on ethylene oxide. Thus, we can maximize our future research dollars to fund critical studies. We need to move forward as fast as possible, as the EPA has already developed a new risk assessment on VC dial suggests increased risk compared to their previous estimates. The new assessment incorporates metabolism data, but not information on DNA adducts or DN'A repair. Finally , we incurred major expenses during the four trips to New Jersey that were not included in Year l's budget. These are listed under sample collection in the budget. 1997-1998 Budget request Sample collection Travel expenses Shipping Supplies Research budget Personnel Supplies LC-MS/'MS time (S25/hr) Travel Equipment maintenance agreements S8172 752 11771 35000 24000 2500 3000 2000 S20.695 S66.500 TOTAL BUDGET REQUEST S87.195 LnB Li- i'ILL _mklInli Fax :9i 9--966-6123 Sep 22 '97 11:58 P.01/03 James A. S-^enberg. D.V.M.. Ph.O. Director, Curriculum <o Toxicology Profcjioi, EjjvmminaituJ Science* X Engiaecriac. Nulntnm, 1 Pathology Scttfiolt of Public Health X Medicine THE UNIVERSITY OF NORTH CAROLINA at CHAPEL HILL To: Wendy Sherman Fax: 703-741-6091 From: James Swenberg Phone: 966-6142 Date: September 22,1997 Subject: Calendar of"events'' Pages (including cover): 2 Ubonwy ofMoieeul* Grouofvmu & Muntcoctu CM 7400, KCMHU Hall Qupai Hill, NC2739*7400 (919) 96M142 (Soereaty) <>#) 446-6I39 (Office). (010) MO-6123 iFm; enull: juanjmatmx>0VK.mhi BFG40946 BFG 00445 Sept. 97: Oct. 97: Nov. 97: Dec. 97: Jan. 98: Feb. 98: Mar. 98: Apr. 98: May 98: June 98: Swenberg Laboratories 1997-1998 eG methods development continues DNA from 4 wk whole livers 1" analyses rat whole liver 4 week, all doses start eA analyses in urines - 13C whole liver DNA in eG method DNA from 4 wk heps and NPCs urine analyses urine analyses I3C whole liver DNA in sG method 4 wk heps and NPCs DNA in sG method Urine analyses Abasic site measurement 4 wk whole liver ,3C heps and NPC DNA Urine analyses Abasic site measurement 4 wk whole liver l,C heps and NPC DNA in eG method 32P postlabeling analyses of sA and eC 4 wk whole liver Urine analyses Abasic site measurement 4 wk whole liver 13C heps and NPC DNA in eG method 32P postlabeling analyses of fA and eC 4 wk whole liver l.N^-eG analyses start Abasic site measurement 4 wk whole liver 13C heps and NPC DNA in eG method 32P postlabeling analyses of eA and eC continue l,N1-eG analyses Abasic site measurement 4 wk whole liver 32P postlabeling analyses of eA and eC l^-eG analyses MPG & AP endonuclease assays begin Abasic site measurement 4 wk whole liver 32P postiabeling analyses of eA and sC l,N:-eG analyses BFG40947 .hB CF MCL CfiRCINQ Fax913-966-5123 Sep 22 '97 11:58 P.03/03 July 98: MPG & AP endonuclease assays begin Abasic site measurement 4 wk whole liver 32P postlabeling analyses of eA and cC l,NJ-eG analyses Aug. 98: MPG & AP endonuclease assays begin Abasic site measurement 4 wk whole liver 32P postlabeling analyses of eA and sC l,NJ-eG analyses Year 2:98-99 Begin with brain Year 3:99-00 Lung, Kidney, risk assessment analyses begin BFG40948 BFG 00447 STUDY PLAN (Revised April 22,1997) COMBINED INHALATION TWO-GENERATION REPRODUCTION AND DEVELOPMENTAL TOXICITY TESTING OF VINYL CHLORIDE I. Test Requirements Contained in Study Plan Inhalation Two-Generation Reproduction Study of Vinyl Chloride in Rats Inhalation Developmental Toxicity Study of Vinyl Chloride in CD Rats II. Administrative Official and Project Manager: Wendy K. Sherman Manager, Vinyl Chloride Panel Chemical Manufacturers Associanon 1300 Wilson Boulevard Arlington, VA 22209 Phone: 703/741-5639 Fax: 703/741-6091 III. Testing Facility: Huntingdon Life Sciences, Inc. P.O. Box 2360 Mettlers Road East Millstone, NJ 08875 Study Director: Raymond E. Schrceder M S. Phone: 908/873-2550 'ext 210> IV. Professional Qualifications Curriculum Vitae (C.V.) providing the training and experience of each professional involved in this study were provided as an attachment to the original study plan. Copies of those C.V.s are available upon request from Ms. Sherman. V. Identity of Test Substance An analysis of the material which is being used in the studies was conducted and the results are available from Huntingdon Life Sciences, Inc. BFG40949 00448 bfg VI. Study Protocol The protocol for the combined inhalation two-generation reproduction and developmental toxicity testing was included as an attachment to the original study plan. A copy of the protocol is available upon request from Ms. Sherman. VII. Test Schedule Developmental Toxicity Study: In-life phase starts: In-life phase completed: Progress report: Progress report Final report: Reproduction Study: October, 1996 April, 1997 i \ < In-life phase starts: In-life phase completed: Progress report Progress report: Progress report: Final report: October, 1996 September, 1997 April, 1997 October, 1997 April, 1998 July, 1998 BFG 00449 DEPARTMENT OF HEALTH & HUMAN SERVICES Public Health Service Agency for Toxic Substances and Disease Registry Atlanta GA 30333 M ^ 8 1S9T Ms. Wendy K. Sherman Manager, Vinyl Chloride Panel Chemical Manufacturers Association 1300 Wilson Boulevard Arlington, Virginia 22209 Dear Ms. Sherman: The Agency for Toxic Substances and Disease Registry (ATSDR) commends the efforts of the Chemical Manufacturers Association (CMA) to assist our agency in addressing key gaps in toxicological information through voluntary research. The purpose of this letter is to reaffirm the importance of your efforts. ATSDR is the lead agency within the Public Health Service responsible for implementing the health-related provisions of the Comprehensive Environmental Response, Compensation, and Liability Act (CERCLA) . As such, one of ATSDR's goals is to evaluate relationships between hazardous substances in the environment, exposure, and adverse human health outcomes. In 1991, ATSDR initiated its Substance-Specific Applied Research Program (SSARP) by determining 117 priority data needs for its 38 highest ranking hazardous substances found at Superfund hazardous waste sites. Identifying and addressing these research needs are crucial to ATSDR in determining the types and levels of hazardous substance exposures that may present significant risks of adverse human health effects. ATSDR is pleased that the CMA vinyl chloride study, "Combined inhalation two-generation reproduction and developmental toxicity study in CD rats," has been initiated and that CMA has updated the agency on the study's progress. Vinyl chloride currently ranks number 4 on ATSDR's priority list of hazardous substances, and it has been found in at least 493 National Priorities List (NPL) sites. BFG40951 BFG 00450 Page 2 - Ms. Sherman Research on this substance is not only important to citizens who live near hazardous waste sites, but also will have implications for workers and others who may be exposed to vinyl chloride during remediation of waste sites. We appreciate the opportunity to work with CMA to assure the success of this very important cooperative research effort. Sincerely yours, Assistant Administrator BFG40952 BFG 00451