# Presentation: 361953

## Comments from EDMVS onrestricted feeding study

- Difficult to evaluate the study without time to examine the data in advance, but generally no major problems with study.
- Effects at 12% reduction in BW gain (RBWG) make a conclusion of no effect at up to 10% questionable.
- Another unpublished study seems to confirm no RBWG effects on PPS, but there were other effects on reproductive organs (epididymal and SV weights).  (VO not tested.) Study to be distributed to EDMVS.

**Notes:**
These are being regarded as Individual comments unless noted.  Some of the statements may be contradictory.

## Restricted feeding study (2)

- Much discussion on need for pair-feeding:
- Not needed if RBWG < 10%
- Not clear how to interpret results if MTD is defined as 10% RBWG
- Not a problem if EPA requires only one dose in the pubertal because controls can be pair-fed.

**Notes:**
Many more nuances not summarized here.

## Dose-setting issues

- MTD should not be based solely on RBWG.  Clinical signs can be the basis.  Stay below signs of pain and distress.
- Use minimum number of animals commensurate with need, usually in the range of 2 to 5.
- Flexibility in dose-setting methods is needed.  Up/down procedure is fine for longer-term assays.  Acute data may suffice for short-term assays.

## Dose-setting issues (2)

- Use all possible sources of data when guessing at initial dose (e.g., HPV data), and consider using in vitro data if no other info is available.
- Be aware of the PK and other differences that can arise by mandating gavage as route of exposure.
- “There is no substitute for good judgment.”

## Dose-setting issues (3)

- Dose-setting must be part of protocol validation, but there is also a need to know what happens when many labs use identical doses.
- For an unknown chemical in validation, it might be reasonable to have all labs do dose-setting independently, with a committee to consider all the results and decide on doses to test in the assay per se.

## Thyroid endpoints inmammalian 2-generation assay

- T3 not useful; thyroid weight useful
- T4 very useful vs. T4 not helpful beyond weight and histopath
- Consider other endpoints (e.g., auditory, ocular) for thyroid effects on neurodevelopment
- Look at what expert review for perchlorate RfD said were useful endpoints for thyroid.
- Follow up on the germinal epithelium effects

## Criteria for Screens

- In response to concerns expressed by the EDMVS it should be noted that:
- The composition of the Tier 1 battery and weight-of-evidence guidelines will be updated based on experience with the assays in validation.
- The desire for redundancy of endpoints in separate assays in the battery will be weighed against cost and animal welfare concerns.

## ER & AR Binding Assays

- EPA will proceed with the validation of the ER uterine cytosol and AR prostate cytosol binding assays developed by EPA taking note of the recommendations for challenge chemicals by the NIEHS expert panel.

## ER & AR Binding Assays

- Although EPA regards the ERTA and ARTA as assays in the research stage (the BRDs could not recommend a protocol or assay system for prevalidation studies) EPA is making substantial progress in the development of stably transfected assays for both ER and AR.  EPA will standardize these assays and investigate options for their validation including partnerships with industry and other countries.