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Worked in a monkey lab?

 

Ei Terasawa, Revisited. Again.

Update (9-20-07): The protocols cited below are now available in their entirety minus the university's redactions. There is considerable duplication throughout each individual document. These are large files.

G00146-4-09-96
G00146-4-02-00
G00146-4-01-03
G00452-3-04-04
G00146-4-05-04
__

We recently acquired five extensive documents related to Ei Terasawa's research. These include her past and currently approved protocols associated with experiments she has been performing since at least 1977.

G00146-4-09-96 (3 Projects; Last amendment approved 6-98.)
Title: The Study of the Neuroendocrine Hypothalamus
Hypothalamic Control of Puberty
Hypothalamic Control of Gonadotropin Secretion
Aging of the Neuroendocrine Hypothalamus

G00146-4-02-00 (4 Projects; Last amendment approved 5-02.)
Hypothalamic Control of Puberty
Hypothalamic Control of Gonadotropin Secretion
Aging of the Neuroendocrine Hypothalamus
Estrogen Influences on Neuroendocrine Aging

G00146-4-01-03 approval for Teresawa to continue drawing blood from six monkeys (their serial numbers have been censored, probably to protect their privacy from concerned members of the public!) used previously in the experiments that were suspended.

G00146-4-05-04 (5 Projects; Last amendment approved 1-04.)
Hypothalamic Control of Puberty
Hypothalamic Control of Gonadotropin Secretion
Aging of the Neuroendocrine Hypothalamus
Estrogen Influences on Neuroendocrine Aging
Neuroendocrine Actionas of Prenatal Androgens in Females

G00452-3-04-04 (Last amendment approved 2-06.)
The Study of the Neuroendocrine Hypothalamus

Together, these documents total about 250 pages including many details and much discussion regarding Terasawa's experiments on monkeys at the Wisconsin National Primate Research Center.

In the first document, G00146-4-09-96, she was approved to use 150 monkeys and 60 rats. She explains part of the procedure used in Project 1:

In some animals hypothalamic perfusates will be collected from the stalk-median eminence using a push-pull cannula at a 10-min interval. Artificial cerebrospinal fluid (aCSF) will be perfused through push cannula, while perfusates are collected through a pull cannula. Various neuroactive substances listed below will be tested. The experiment will be performed on monkeys with pedestals on their crania. Because this experiment requires fine adjustment for balance of flow, we need temporary restraint of the animal in the chair. (Unfortunately, we cannot use anesthesia during the collection of hypothalamic perfusates, since anesthesia changes the state of hormone release from the hypothalamus.) At least one month prior to the experiment monkeys are adapted to chairs, starting with a short period and then increasing to a longer period. The chair is padded with sheepskin materials when needed. A 2-3 day waiting period is necessary after placing the cannula due to elevated levels of cortisol and to allow clearance of cellular debris around the cannula, which causes blockage of the perfusion. The monkey is allowed to eat and drink ad libitum and interacts by vocalizing and establishing visual contact with other monkeys present in the same room. During the waiting period animals will be checked at least every 2 hours in the day and every 8 hours in the night by our research staff and/or animal care staff, and by veterinarians when needed. During the push-pull perfusion experiments, an additional 6 h of perfusion will be conducted on the 2 nd day of the perfusion. Therefore, the maximum amount of time in the chair will be 4 ½ days. All records on animals, such as eating and drinking behavior, urination and defecation, during the entire experiment will be kept in a notebook. All procedures are aseptic.

The "median eminence" is a small stalk- or stem-like structure that connects the neurohypophysis with the rest of the brain (it connects to the hypothalamus.). Here's a good drawing - the purple or bluish area is the neurohypophysis. The neurohypophysis is one of two lobes of the pituitary gland. This image gives a good idea of its location. The pituitary is encased in bone.

Terasawa drills a hole through the top of monkey's skulls and then pushes a thin metal pipe down through their brain and aims to pierce the small median eminence that connects the pituitary to the hypothalamus. Then she threads a double-barreled catheter through the pipe. Through one barrel, she pumps various drugs into the delicate tissue. Under pressure from the pump attached to the tube, these chemicals are forced through the tissue, induce the cells to secrete, and, though the other barrel, a pump sucks out the resulting concoction, or perfusate.

In this image, the rich blood supply to this area is apparent.

In the second document, G00146-4-02-00, she was given permission to use 247 monkeys and 60 pregnant rats.

Terasawa lists 27 separate chemicals that she will perfuse through the monkeys' brains. She notes that some of the monkeys will be infused every 2 hours for 2 years through a chronic indwelling cannula in their brain and that these monkeys will be housed alone for the entire period. She says that there will be no side effects.

Terasawa writes: "... some animals will receive more than one drug. Choice of drugs will be based on the the hypothesis to be tested in each project and each animal will receive a maximum of 60 different drugs."

"Despite the many advantages of the push-pull perfusion method, the use of this technique has some drawbacks....When a partial oclussion occurs anywhere in the the system, the balance of the push-pull pressure is not maintained and the volume of perfusate collection becomes reduced. This leads to tissue damage.... Tissue damage is a great concern for this method. Although our approach using short term [repeated] cannula placement does not cause the extensive gliosis [scarring of brain tissue] associated with permanent placement of the cannula, it does cause some gliosis. In addition, infusion must cause tissue damage; ... Nevertheless, it appears that damage is not extensive and that some regeneration of the capillaries occurs, since we can conduct an average of seven or .... [page missing.]

Here, she describes Project 4.

Interestingly, Terasawa makes a clear distinction (above) between push-pull perfusion and microdialysis. Later, in the public relations fiasco that erupted after the USDA inspectors called attention to the problems in her lab, the chair of the UW oversight committee, Eric Sandgren, told her to begin referring to the procedure as microdialysis, and subsequent correspondence and documents began using that term. Maybe it sounds less gruesome.

In the third document, G00146 (Approval for blood draws only), it is more of the same, but worth noting is the fact that following the embarrassing public disclosures of a failed oversight system, the oversight committee began micro-editing her protocols.

“The committee would like to see the following reflected in a rewrite of protocol G146:

*Q#17: Project 4, item 3—In the first sentence clarify that only aged monkeys will be euthanized

*Q9c – Replace “severe physical crippling” with “physical impairment that precludes chairing.”

*Q17b – Paragraph on acclimatization, eight lines down, change “ulcers” to “skin abrasions or bruising.”

She was eventually given approval to proceed. In this version, she had requested 199 female monkeys (24 purchased elsewhere) and 60 rats she planned to purchase form Holtzman Company/SASCO Company.

These two pages may be of interest.

The list of drugs above, from 2003, is nearly identical to the list of drugs she was infusing in 1998. Apparently, she just keeps pushing many of the same chemicals through monkeys' stalk-median eminences year after year and pulling out the resulting chemicals.

Once the USDA inspection had been made public and the university thoroughly embarrassed, the committee tried to close the lab door, but the damaged had been done. In response, the committee asked her questions about a protocol that had been going on for 17 years.

USDA sets a regulatory limit on the number of hours that a monkey can be restrained. The limit is 12 hours of continuous restraint. Then the monkey must be given 1 hour of unrestrained exercise.

Title 9 (the Animal Welfare Act); Part 3; Subpart D; Sec. 3.81; (d) Restraint devices. “Nonhuman primates must not be maintained in restraint devices unless required for health reasons as determined by the attending veterinarian or by a research proposal approved by the Committee at research facilities. Maintenance under such restraint must be for the shortest period possible. In instances where long-term (more than 12 hours) restraint is required, the nonhuman primate must be provided the opportunity daily for unrestrained activity for at least one continuous hour during the period of restraint, unless continuous restraint is required by the research proposal approved by the Committee at research facilities.”

In the case of Terasawa's research, this “unrestrained activity” means simply being able to stand up.

Here we see the committee getting “tough” and Terasawa's response. The restraint chair modifications to allow the “exercise,” her narrative of some monkeys' experiences, and an example of the records she (now) keeps.

Document G00146-4-05-04 amounts to the committee saying, “Ok you got caught at your daily work of hurting animals and embarrassed us, so we're going to make you work hard to get your next protocol amendment approved. But, since you have finally groveled, we'll reward you.” They made her rewrite it 3 times but gave her permission to use 245 more monkeys and 60 more rats.

In this document she claimed that the monkeys would be chaired for only 18 hours at a time. Here's the list of chemicals she forced into their brains.

And here they are again micro-editing her descriptions. Terasawa is so worn out at this point (poor baby) that she doesn't even mention the fact that in her earlier versions (approved by this committee) she had discussed at length that she uses the “push-pull perfusion method” and not “microdialysis.” But at this point she'll call a monkey a rat if the committee will approve her studies and let her get back to cashing those rich NIH checks.

This might explain this letter; she probably knew that it was unlikely that they would allow her to keep restraining monkeys for days on end.

Interestingly, for some reason, her description of the procedures she uses to collect semen is different in this document than in the one below. Here she writes:

Semen collection – Adult male monkeys will be electro-ejaculated. Males will be briefly restrained in a metal restraint apparatus and aluminum foil will be placed on the penis. Certain males will spontaneously ejaculate at this point while other males will require slight pulsatile current to stimulate ejaculation. Depending on the male, some individuals may be lightly anesthetized with ketamine (5-10 mg/kg i.m.) for chemical restraint. One hour prior to initiating ejaculation, monkeys may receive diazepam 0.25 mg/kg i.m. or p.o. to reduce anxiety. Successful use of Viagra (sildenafil citrate) for promoting reproductive function in male rhesus monkeys has been reported. This option may be explored to improve breeding with genetically important but socially deficient males. Viagra will be administered p.o. at a dose 0.92 mg/kg (based on doses used clinically in humans). Receptive females will be placed with males 1 hr. after dosing.

[Recommended ketamine dosage varies between species of macaques. The notion that the dosage of Viagra will be the same between rhesus monkeys and humans is pure gibberish.]

And that brings us to document G00452-3-04-04.

Here's the semen collection part: (scroll down)

In this document she is given permission to use 115 monkeys and 12 pregnant rats.

This is basically a study of the effects of sex hormones. Infant monkeys are variously ovariectomized at one week of age or 12 months of age and then killed between the ages of 2 and 37 months so that their brains can be examined.

Terasawa claims that her research will lend insights into polycystic ovarian syndrome, and anorexia.

She uses the semen to artificially fertilize monkeys to create timed pregnancies because she also performs “fetectomies,” abortions by Caesarean section, because she wants fetal tissues at various stages of development.

And finally, she ovariectomizes monkeys at various ages and then implants (or doesn't) estrogen capsules and then kills them at a later date to study their brain tissue. She claims that her data will aid in treating osteoporosis.

The main scientific problem with Terasawa's work (as opposed to the grotesque cruelty) is that she does not seem to understand the scientific method of limiting and controlling variables.

She is pumping dozens of neuroactive chemicals with complex bioactivity into monkeys' brains and then sucking out the chemical products. This is nuts. And she's been doing it for two decades.

See also:

The Disturbing Case of Ei Terasawa

Wisconsin University Union Chair Files Lawsuit, Claims UW Animal Research Oversight Akin to McCarthyism, Guantanamo

Oversight

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