X-Message-Number: 17803 From: Date: Mon, 22 Oct 2001 01:19:44 EDT Subject: Quality Control and Cryonics Charles Platt and others have commented about quality control in cryonics and have referenced my earlier proposals for patient sampling, in particular brain biopsies of cryopatients. No one has got it quite right (at least from my point of view) but then it was a long time ago, and, as Charles has noted a simple matter became contentious. I'll number my points for clarity. 1) Animal research can be remarkably predictive of outcomes in humans if you are clever, wise, and lucky. Most cancer research has been historically done with rodents and particularly mice. Countless billions of dollars have been spent on various models of human cancers in murine (mouse) models. It is now possible to cure a large number of cancers in mice. However, this did not proved to be true for humans. Almost all of the money spent since President Nixon's declaration of a war on cancer in the 1970s was misspent. Animal rights activists may now take a bow: this stupidity gave you vast and accurate ammunition against the animal research community. Monkeys would seem better models to evaluate treatments for head injury and prolonged cardiac arrest in humans since they are phylogenetically closer to us than, say, dogs. But the fact is that monkeys are much more resistant to ischemia than humans. Dogs map humans in terms of timeline, degree and distribution of neuronal injury more closely than any other animal I have researched (and I am not alone in this conclusion). They also demonstrate truly awful behavioral deficits with increasing ischemic times which are comparable to those observed in humans similarly injured. Sheep and other grazing animals can suffer devastating neuronal losses with none or very little change in behavior. Choosing and validating the right animal model is critical. Even in the veterinary world there is enormous interspecies difference. The drug you see advertised on TV for arthritic and elderly dogs (Rimadyl made by Pfizer) does not work on cats. Similarly, 4-methylpyrazole, a drug which can rescue dogs from ethylene glycol toxicity is useless in cats. Closer to the home front, glycerol does not penetrate bovine erythrocytes significantly below 10 C and several cryoprotectants are impermeant to the intact rat brain. Rat brains have historically been avoided as models in cryobiology (perhaps not with justice) because of the extremely dense neuronal architecture and the idiosyncratic response to cryoprotectants observed by some scientists in the little work that has been published in this area. Scientists engaged in animal research now call this work comparative medicine and there are staggeringly knowledgeable and prolific Internet lists which provide both ongoing discourse and a vast reservoir of knowledge to draw upon. The take home message is that you pick your model and then validate it as many ways as you can in other relevant systems. This not a criticism of CI or anyone else. The plans laid out by David Pascal that Yuri Pichugin plans to follow seem reasonable. Similarly, Charles' concerns about applying research insights to humans have some merit. You don't really know till you try it, which brings me to point 2) below: 2) Human cadavers obtained with reasonably short postmortem intervals are obtainable for about $2,000 each in the greater Los Angeles area. Timing is the hard part, but if you are a cryonics organization and plan this kind of work in advance that issue should not be onerous since you should be prepared for it anyway. The ethical issues do not trouble me, nor do I think they trouble other researchers in cryonics. Several of us have already had access to human brains post mortem which were cryopreserved and subsequently dissected and examined in some cases. The results were presented at meetings of cryonicists without objection (i.e., the Lake Tahoe Cryonics meeting in the 1980s and 1990s). If people don't want cryonics but do want to make a contribution by donating their bodies and brains for irreversible destructive research this is morally and ethically acceptable. Indeed, acting contrary to this wish would constitute the assault on the person and the ethically unacceptable behavior. To do this kind of work is no different from to work in hospital (as I have) and to follow a patient's instructions to discontinue dialysis or other life sustaining treatment even though the quality of life is still high (by my own standards and those of most of the rest of society). This is done all the time. To do otherwise constitutes false imprisonment and battery. It also bespeaks a deep disrespect for the right of the individual to make an informed choice. Yes, people make bad decisions in this regard. And yes, they are irreversible. But it is their right to make them and this has been acknowledged by both social norms and the legal system. Greater evil has historically come from _imposed_ decisions by governments or individuals who are not party to all the information or who are driven by ideological or political agendas. 3) The spinal cord is a good surrogate for the brain, especially as applied to cryopreservation. Spinal cord samples can be removed from neuropatients for a variety of evaluations. In the cryopreservation of James Gallagher (a CryoCare patient) BioPreservation removed a large section of spinal cord and lumbar nerves. We also removed cryoprotected samples of kidney, liver and heart. These fairly substantial samples were cooled with the patient at the same rate and under the same conditions. These samples (which are CryoCare/BPI property) are, as far as I know, still in the neurocan with the patient. The purpose of this kind of sample taking was to evaluate by both freeze substitution electron microscopy and post-thaw electron microscopy how effective (or damaging) the cryopreservation technique had been. Unfortunately, my research objectives changed and BPI did not have the money to pursue this investigation. However, the opportunity still exists and should be exploited. Had we been better prepared we could have taken a second set of samples and fixed them immediately following glycerol perfusion and then examined them under light and electron microscopy. These would have served as pre-freeze controls for the frozen samples. We were not prepared for this maneuver (which requires that terminal CPA concentration solution be made up containing fixative for immediate immersion of the samples after collection). 4) As to brain biopsies via the burr hole: this was one of the most ludicrous episodes in the history of cryonics which has had many a ludicrous episode. The proposal was to take two needle biopsies of non-critical areas of the cerebral cortex via the burr holes. The amount of biopsied material would have been equal to about the same volume of tissue contained in a 16 g blood collection needle (for those of you who have donated blood). A superficial cortical sample such as this has never caused anyone any detectable neurological deficit. If you've had as much experience as I have (and other medically knowledgeable people in cryonics) you would realize that a) this kind of thing is done every day without problems and b) most patients with brain tumors (and those around them) do not notice any change in mentation until the mass is at least the size of a walnut and usually the size of a golf ball; if not larger. We are not talking about the midbrain here, or the hippocampus. This procedure was simple, safe and a "no-brainer" as the slang of the time went. The tissue would be handled in two ways: One sample would be cooled with the patient at the same rate and under the same conditions. (Today, if one were vitrifying the patient the sample could be inserted in a removable capsule in the esophagus or terminal pharynx to give a worse case cooling rate and then be removed following cooling.) The other sample would be processed by immediate post-removal fixation so that the effect of cryoprotective perfusion could be separated from that of subsequent cryopreservation. While not perfect, this kind of sampling would return an incredible wealth of general information and a similar bonanza of information about the quality of how that particular patient was treated. It is also possible to collect a sample prior to the start of cryoprotective perfusion to see what condition the brain is in following legal death and initial cooling and stabilization. This language is, I believe, already standard in the CI contract. The issue could have easily been resolved by simply having the client say "no" if they did not want this procedure done. And, of course, the procedure would have to be explained in the client's agreement with the cryonics organization. This did not seem a major obstacle. Because this practice has not become routine or even occasional, there is virtually no objectifiable feedback from _any_ cryonics society about what condition their patients are in. Thus, there is endless and unnecessary argument that testing could resolve. I did remove cord biopsy samples from patients frozen by Trans Time in the 1970s and these samples were incredibly valuable and led to major alterations in practice. Mike Darwin Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=17803