X-Message-Number: 33149 From: Date: Tue, 28 Dec 2010 03:05:51 EST Subject: Scoring Cases Content-Language: en From: Gerald Monroe _ (mailto:) Date: Mon, 27 Dec 2010 12:42:20 -0600 Subject: Re: CryoNet #33139 - #33144 This is a pretty good summary, Gerald. I've interlaced some points that I think are important. >>Simplifying it: the edema damages neurons enough that they won't restart. (nor will a flooded engine, or a jammed machine). But since biological enzymes aren't functional near 0 degrees C, neurons have probably not been able to catalyze the self destructive processes that neurons perform when they think they are damaged beyond repair.>> Edema is certainly a major factor in compromising cellular viability in ischemic and nonischemic hypothermia, but it isn't the only factor. Substrates necessary for the cells to restart function get used up, free radical damage occurs due to multiple factors (including uncoupled electron transport in the mitochondria) and there is undoubtedly biochemical injury of the same kind, if not to the same degree, as occurs in warm ischemia. Many enzymes have rather abrupt inactivation temperatures; in other words they stop working altogether if cooled or heated above a given temperature. However, others behave "erratically' or even continue to function as predicted by the Arrhenius equation. To the extent that these enzymes are mobile, and capable of driving chemical reactions, they will continue to do so: near 0 deg C or even far below it. Catalase, the enzyme that decomposes hydrogen peroxide, is active at dry temperature. I found this out a kid by preparing a liver homogenate in 70% v/v glycerol and adding that to a mixture of hydrogen peroxide in 70% v/v glycerol - with both solutions cooled to dry ice temperature. Within a few seconds, fine white foam begins to appear in the interface between the two solutions. That foam is oxygen from the decomposing H2O2. This only happens when a liver homogenate is added to the peroxide.So, some enzymes, for good or ill, will continue to do their duty as long as they can diffuse to the substrates they are configured to act upon. Another problem is "uncoupling' of linked enzyme systems. Enzymes often function as part of a complex chain of catalyzed reactions, and if one enzyme drops out, or becomes more or less active than it should be, that can result in the failure of the "system,' or worse, in the production of toxic or metabolically problematic compounds. These compounds can inhibit other important reactions, cause direct injury (such as free radicals species do by damaging cellular molecules), or result in excessive consumption of an essential substrate for energy production... Most of the enzymes that degrade cell structure, proteases and lipases, are typically sequestered inside cells; many, but not all of them, in organelles called lysosomes. One positive effect of ultraprofound hypothermia (0 to +5 deg C) is that it greatly slows rupture of the lysosomes. For one thing, the lysosomal membranes are frozen solid at these temperatures - as are most other cellular membranes (including the organelle and plasma membranes). Indeed, it may well be the phase transition of the lipids in the membranous structures of cells that provide a great deal of the protection that is "unexpected' on the basis of the Arrhenius equation (or the Q10 rule) in conditions of hypothermia above the freezing point of water. Nonhibernating mammals, such as humans, are made up mostly of saturated fats which have a freezing point at or above room temperature. When our cells are cooled to a few deg C above freezing, the saturated fats in our cell membranes crystallize, and this not only renders these lipids unavailable for metabolism and more resistant to catabolism, it also has profound effects on the many cellular proteins (including enzymes) that are complexed with those lipids *and with the complexing molecules that attach them to the lipids*, as well. As far back as the mid-1960s, it was understood that cooling can cause destabilization of both the protein-lipid complexes and of their complexing molecules, resulting not only in inactivation of enzymes, but in some instances, possibly "inactivation' of the substrate molecules (upon which enzymes act), as well (Ushakov, B., J. Gem PhysioL 44, 518-560 (1964). >>Hence, if a patient were stored at near 0 C for 24 hours, there's still a reasonable chance that their brain contains the information that cryonics tries to preserve. But it isn't ideal : ideal cryonics preservation freezes the brain before an hour of cold ischemia has occurred (before that point CPR and bypass machines are used to actively prevent ischemia). The reason this is ideal is that the maximum chance of survival according to current science would be to put the brain in state at low temperature that we know is revivable, and then to freeze it preventing any further biochemical change at all from that start. Since the brain is composed of complex molecular structures, even though we cannot prove for certain that revival is possible, we can show that it is virtually certain that the information needed to perform a revival has been preserved.>> It is here that I must disagree with you - principally for the *certainty *with which you make the statement: "Since the brain is composed of complex molecular structures, even though we cannot prove for certain that revival is possible, we can show that it is virtually certain that the information needed to perform a revival has been preserved." I wish that this were so; that it was proven. But that is not the case. We do not yet know what the "information necessary to perform a revival is.' We are increasingly in a position to make educated guesses, but that is not the same as knowing with proven certainty. And it is critically important to define what you mean by "revival.' For instance, it is not too terribly uncommon for people to suffer severe head injury that results in prolonged cerebral edema - brain swelling - who go on to "recover,' but with one teensy little problem: their declarative memory is completely wiped. They have no memory of their name, of growing up, of being married or having children (if such was the case) and most of these people never recover their declarative memories. Generally, their procedural memories are intact: they know how to walk, drive a car, read and so on. Often people so injured will undergo dramatic changes in personality, almost always becoming more, rather than less disinhibited. They may take up profanity, and drink or drugs, become sexually promiscuous, or just generally be more garrulous and obnoxious. This is believed to be due to heavy losses of neurons in the prefrontal cortex. The relevance of such individuals to this discussion is that they raise the question of just WHO exactly survived the accident that caused their brain injury. At what point of memory loss, or personality change, do you stop being you, or become someone else? Or are you just the neuronal circuits that generate your consciousness - and if so, then why is YOUR death such a big deal - since those same circuits will continue to exist in other humans, presumably long after you are dead? In fact, they may well exist in all vertebrates! Cryonics is likely to force the issue of personal identity and personal survival in ways that have previously not been much of an issue, either for individuals, or for society. Cloning of humans is now illegal, but it is patently clear that almost all cryonics patients have sufficiently intact nuclear DNA to allow them to be cloned. Given that medicine and society currently treat amnestic head injured patients who also have profound changes in personality as the SAME person they were before the injury, how is that different from a cryopatient who is " revived' from a single intact nucleus *and who is not afflicted with the frontal neuron loss and accompanying personality alterations that some contemporary brain injury patients experience?* Indeed, by cloning criteria, it is possible to begin reviving almost all of today's cryopatients! The experiment has been a success! While admittedly taking a long way to get here, my point is that we do not yet understand the neurobiology of memory, learning, or personality. We have some ideas, but they are not proven, and there is plenty of room for the possibility that we are doing something with contemporary cryopreservation procedures that wipes memory, or degrades it. While this seems unlikely to me under conditions of "good' cryopreservation (and maybe unlikely to you, too), as a scientist, I've learned to be skeptical; and the older I get the more conservatively skeptical I get - and this despite the loss of so many frontal cortex neurons from aging. Mike Darwin Content-Type: text/html; charset="UTF-8" [ AUTOMATICALLY SKIPPING HTML ENCODING! ] Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=33149