This is just one of the group of protocols approved for John Sheridan & David Padgett. This page and subsequent pages will highlight the numbers of animals used and how they used. The period covered begins in 1995 to the present. Recently published human studies do not even reference the animal experiments being conducted at OSU.

I welcome any input to help make these pages less cumbersome and clear. These experiments should be STOPPED today!

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01A0039 - Stress, Aging and Neuroendocrine-Immune Changes - John F. Sheridan, and David A. Padgett

This protocol was initially approved in January 1995, as protocol 95A0001, and requested the use of 4,320 mice.

Over the years this protocol has added thousands of mice, expanded to a numerous protocols, and continues to inflict pain and distress on the animals used in the experiment. Following these experiments you can begin to understand how animal research continues in spite of the work demonstrating no benefit to human health.

The information is presented in the words of the researcher, animal care personnel, and ILACUC members. The information taken from amendments, e-mails, and protocols.

95A0001 - During the initial approval process there is a question about the prolonged restraint of the mice to be used in the experiment. A memo to Sheridan gives the final approval in April 1995. The pain code for the protocol is 2D.

The committee reviewed the detailed description of your protocol for the prolonged restraint of mice in conical tubes, discussed your references and justification for the use of restraint, and determined that your work could proceed provided that the procedures were first observed by veterinarians of the ULAR staff. ...Upon observation of your procedures, the veterinary staff determined that the 50cc conical tubes with 80-100 air holes (2-3mm) provided room for the mice to move back and forth and had an adequate air supply. After the procedures, the mice were also observed to detect any adverse effects of the restraint. Upon release, the mice displayed normal behavior, and normal eating and drinking activities. The procedure did not appear to cause undue stress or hardship.

Consequently, your protocol for the prolonged restraint of mice in 50ml conical tube for up to twelve hours has been to the best of our knowledge thoroughly reviewed, observed, and therefore approved.

The mice will also be given an influenza virus which will lead to an infection in the mice. Later in 1995, Sheridan submits another protocol, 95A0185 - Neuroendocrine Influences on Wound Healing, which was approved to use 1,100 C57BL/6 and 1,100 SKH-1 mice. On the protocol form restraint is marked yes with the following words:

Prolonged restraint (up to 16 hours) to stimulate stress-induced changes in immune function (Approved protocols 93A0018 & 95A0001)

The mice will also be inoculated with herpesvirus type 1" which induces disease (distress)." Some of the animals will also receive a 3.5mm punch biopsy, a small superficial "wound" will be placed on the dorsal side of the mouse, just below the shoulder blades. All animals will be killed at the end of the experimentation cycle.

Published in Lancet, November 4, 1996, JK Kiecolt-Glaser, another researcher at The Ohio State University, publishes a paper, Slowing Wound Healing By Psychological Stress.

Twenty-six women were used in the study. All subjects underwent a 3.5mm punch biopsy wound. Healing was assessed and it was found that the women under stress did not heal as quickly.

I would imagine the women were allowed to live and went home. Making a worthwhile contribution toward helping to understand stress and wound healing in humans.

December 11,1996, Sheridan submits an amendment to add pain and distress to the mice being used in 95A0001. It reads:

I wish to amend animal use protocol 95A0001 in which mice are infected with influenza virus. I need to alter the section on Criteria for Early Removal....I would like it to read "The endpoint of these studies is mortality. Therefore, animals will not be euthanized prior to completion of a study."

Let me make a couple of points. First, when we have an infection experiment going, the animals are checked twice daily by graduate students, fellows, or technicians. Dead animals are removed in a timely fashion. However, the nature of both the influenza and herpes virus infections is such that an ill animal may look fine at 8am but will be dead by noon. Second, when we use our old animals (generally 20 to 27 months of age) these animals may not be healthy looking to begin with, and they are more susceptible to infection then the young mice. Old mice may lose more than 25% of the body weight and yet survive the infection. Further, these old mice represent an incredible expense costing from $65 to $100 a piece. It is in our best research interests to do all we can to keep them healthy.

The amendment was approved Dec. 17, 1996 and the pain code becomes an E category where pain and/or distress is NOT relieved. ILACUC clarifies that, "Animals that die overnight must be removed by 9:00am. Cages should be checked at 4 hour intervals (9:00am-1:00pm-5:00pm) and dead animals removed."

In January 1997, the number of deaths associated with this project comes to light and Sheridans response is to think up new experimental procedures. He writes up an amendment and e-mails it to the ILACUC chair.

Following a conversation with Dr. Doug Stone (the attending veterinarian for this protocol) last week concerning the high number of moribund, wounded and dead mice in an experiment that represents a joint project with Drs Glaser and Stoner, I want to amend my protocol numbers 95A0001 and 95A0185 to include a behavioral manipulation that has been called social reorganization. The degree of fighting and wounding was totally unexpected in the current experiment and I therefore want to add this experiment procedure to my existing protocols.

General Description of the Model - All studies will involve the use of 4-8 week old, C57BL/6 male mice, maintained on a 12-hr light/dark cycle (lights ON at 0600). Mice will be infected intranasally with one-half of a lethal dose of influenza A/Puerto Rico/8/34 (PR8) virus.

Dr. Stone comments here: "Note that this is already an LD-50 and the stress of Social Reorganization is yet to be added. I suggest you let the full committee review this one to protect you and OSU."

Sheridan cont. - ... We will use social organization (SRO) as a stressor. Upon arrival, male C57BL/6 mice will be divided into cages of 5 animals and allowed to acclimate to the new surroundings for 2 weeks and to allow formation of definable social hierarchies. Subsequently, for SRO, to disturb the social environmental settings, mice will be switched between cages at the beginning of the 12 hour dark cycle (6:00PM). SRO will be performed every second day for 4 cycles. We will use pharmacological interventions to specifically identify neuroendocrine products mediating the effects of SRO on viral pathogenesis and immunity.

Dr. Stone - 'Pharmacological interventions' This is a generic term for 'I will be giving lots of drugs.' This could include urethane or literally anything. I suggest we at least have an idea of what class of drugs so we at least know the biohazard of each. Then there is the volume/route of administration issue.

Sr. Stone when discussing SRO writes - There will be mortality's from this social reorganization. At what point is the study terminated? eg ld50?LD75? From my perspective, ULAR will need some guidance or we could have an LD100 and not report it to the ILACUC. An LD100 does not bother me, as long as the ILACUC is aware of it and does not get angry with ULAR for not reporting it. ...Perhaps a subcommittee review or a written vet report is a solution.

January 27, 1997 ILACUC responses to Sheridan concerning the amendments to 95A0001 and 95A0185.

First with regard to the use of urethane, it is not mentioned specifically in these two protocols. Therefore, we need the details of the proposed administration of urethane including the dose, route, frequency and volume of injections.

As far as the new stress protocol, which you refer to as Social Reorganization, this request cannot be considered simply as an amendment. You will need to present this as part of a complete new protocol. ... This is a particularly troublesome procedure as the large mortality rates, which you have already witnessed, will attest and one which the ILACUC will scrutinize very carefully. ...Be advised, however, that as of now, you are not approved to conduct any social reorganization until you obtain the approval of the ILACUC.

February 4, 1997 Sheridan submits a new animal protocol, 97A0015 - Neuroendocrine Influences on Microbial Pathogenesis and Innate Immunity - which requests approval for 5,500 mice. Stressors are restraint, infection and Social Reorganization. Veterinarian observations will take place for the first several experiments. The protocol is approved.

In two years, an experiment that was originally approved to use 4,000 plus mice is now three protocols, involving NO relief from pain and/or distress, and will use an additional 7,700 mice. Other researchers continue related work using humans.

PART TWO - The Experimental Design - Work Continues and Expands

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