X-Message-Number: 11685 Date: Thu, 6 May 1999 07:47:34 -0400 From: Mike Darwin <> Subject: Cryonics: Yesterday's Technology Tomorrow WARNING: The following is likely to be offensive to many of the persons who subscribe to this list. If you do not wish to read or be exposed to materials which could make you angry, defensive, or otherwise upset you, please stop now. This post is also moderately technical, if you don't enjoy reading about technical matters, this post probably isn't for you either. -------------------------------------------------------------- I have not posted to Cryonet in a long time, and probably will not post to it again for a long time, if ever. I am not a regular "reader" but I skim Cryonet with some frequency, usually reading no further than the subject line, or a line or two from a paragraph here or there. The principal reason I continue to open the file when it is downloaded is that Doug Skrecky occassionally posts things from the scientific literature which I find interesting. I stopped participating in Cryonet and cryonics by and large for the reasons Charles Platt cited when he unsubscribed with such "relish" awhile back. My attitude is more easy going than tha.t. I think people should talk about what they want to in an un-moderated forum. (I will also say that since Cryonet was started, I've come to realize via participation in other un-moderated forums that un-moderated forums almost always decay or fail in various ways.) Anarchy is another word for entropy. When I quit posting to Cryonet and distanced myself from cryonics it was for many comlex reasons, not all of which are accessible to examination by me, let alone others not familiar with the history of cryonics in general, or my 30+ years involvement with it. However, at the time I made the decision to stop my public dialogue with the cryonics world, I made a promise to myself that, when or _if_ a certain event ocurred, I would point it out, on Cryonet, and in other appropriate venues in the cryonics community. The last posting I show a record of to Cryonet was 03/11/97, I feel sure Keith Lynch will correct me if there are later ones. I figured the odds were pretty good I'd have to write this post, but I didn't know how long it would take. Then, on 04 May, 1999 the following post appeared: >CryoNet - Tue 4 May 1999 > #11670: Nitric Oxide confirmed as a regulator for Cell Death [Olaf Henny] >Message #11670 >Date: Mon, 03 May 1999 08:53:25 -0700 >From: Olaf Henny <> >Subject: Nitric Oxide confirmed as a regulator for Cell Death >I am re-posting below an article from Cryonet after obtaining Steve's >permission. I believe it will be of great interest in this group. >Best, >Olaf >Message #11641 >From: "Stephen Bogner, P.Eng." <> >Subject: Nitric Oxide confirmed as a regulator for Cell Death >Date: Thu, 29 Apr 1999 13:25:38 -0600 > >Researchers at the Howard Hughes Medical Institute (HHMI) at Duke University in collaboration with scientists at the Dana-Farber Cancer Institute have >now found that nitric oxide (NO), a well-studied molecule implicated in a >host of communication pathways in and between cells, is also a switch that >controls whether cells live or die. >Stamler and his colleagues found that NO molecules occupy a critical site on >the enzyme caspase, a molecular "executioner" within human cells. When >occupying this site, NO effectively plugs a communication pathway that >activates caspase and triggers cell death. >"We showed that nitric oxide sits on the site and keeps the enzyme >inactive," said Stamler. "Conversely, the nitric oxide is removed in cells >programmed to die. Simply put, if you block nitric oxide production within >the cell, you make the cell more susceptible to cell death. And if you add >it back, you prevent cell death." >Apoptosis can be triggered through a biochemical chain of events known as >the Fas pathway. When activated, the Fas pathway initiates a cascade of >signals within the cell that ultimately turns on caspase. When NO occupies >the site on caspase, however, the death message is blocked. Fas somehow >manages to pop the nitric oxide off the cells that are programmed to die. > "We'd like to think modulation of the nitric oxide system can be used for >therapeutic gain," he said. > >The complete article can be found at: http://www.hhmi.org/news/stamler.htm > >The reason that I think that this might be of interest to cryonet is because >it may have implications for the medication protocol that might be used to >limit damage prior to or during suspension. This is not my area of >expertise - so someone knowledgeable may want to comment. > >Regards (and long life); > >Steve. >Stephen Bogner, P.Eng. The above post from Steven Bogner and Olaf Henry was the trigger I had waited two years for. The substance of the post is that the nitric oxide radical might be a source of damage in ischemia, might be responsible for delayed neuronal death, and "The reason that I think that this might be of interest to cryonet is because it may have implications for the medication protocol that might be used to limit damage prior to or during suspension. This is not my area of expertise - so someone knowledgeable may want to comment." I accept Dr. Bogner's invitation. Since I left Alcor in circa 1992 I have been involved in research into the mechanisms of cerebral and systemic ischemia-reperfusion injury and, in particular, into the development of multimodal therapies aimed at ameliorating this injury. Our endpoint has been _complete_ neurological recovery to control levels following prolonged periods of warm (normothermic) ischemia. Our goal is to reach the point where we can routinely and uniformly (~90% of the time) resuscitate dogs from 30 minutes of normotheric _total body_ cardiac arrest. This is much more difficult than demonstrating recovery, acute or chronic, after isolated brain ischemia where the multiple organ systems of the body continue to be perfused without damage: damage which will greatly complicate and increase the difficulty of achieving cerebral survival following such a long interval of cardiac arrest. We have currently reached the 17-minute mark. We have not done any further work in this area for the past year because we have been involved with development of a novel enabling technology that we believe will be critical reaching the 30 minute mark. That work is now complete, and manuscripts are in preparation. When we began this work in 1992 all dogs subjected to 10 minutes of ischemia were uniformly and catastrophically neuroinjured (persistent vegatative state). Dog and man share a unique and surprisingly tight overlap in both time-course and mechanisms of injury after cardiac arrest. We learned early on that any initial recovery was followed by setbacks and that the only truly valid model was evaluation of performance at 3 months or more _post insult._ We strongly suspected apoptosis (programmed cell death) and delayed necrotic cell death to be a major culprit, and were soon able to prove that this was so. FYI, the particular caspases involved in triggering aopoptic death are caspase 1 and, more importanly, caspase 3. By 1993 I had a very good idea of what the primary mechanisms of injury were in both the CNS and the body as a whole as a result of doing a dipstick check for a urinary tract infection in a post-resuscitation dog who had received the _very best_ in pharmacological, hemodynamic, and hypothermic intervention, with the net result being a persistent vegetative state. The test was positive, but the bacterial cultures were subsequently and definitively negative, as was the evidence from the necropsy. Subsequent evaluations showed the presence of high levels of nitrite in the urine of all ischemic dogs. Nitrite is normally present in urine only as a result of the action of bacteria on nitrogen containing compounds in the urine such as urea, so even increased nitrogen catabolism from the ischemic insult would not account for the nitrite. Furthermore, control dogs had higher nitrite levels than experimentals. I will not bore you with the tortuous trail that I followed to understand WHERE the nitrite was coming from. Suffice it to say that I was well aware that the distribution of blood flow to tissues in health and disease was largely mediated by nitric oxide. Further, I was intensely aware that nitric oxide was produced in the brain by a special enzyme, bNOS, which is distinct from those present in the circulatory system (i.e., inducible nitric oxide synthase = iNOS and endogenous constitutive nitric oxide synthase = ecNOS). In fact, just a few years before I left Alcor, Hugh Hixon had been running around the Alcor faciliuty in Riverside waving a copy of SCIENCE NEWS and expressing wonderment that the brain used nitric oxide as a neurotransmitter (I've news for you Hugh, it uses carbon monoxide too!). In 1993 I had a number of papers from a symposium held in Shizuoka, Japan on nitric oxide's role in neurotoxicity. These papers were in Japanese, but short synopses were available in english. A year later CRC press published the proeedings of that conference NITRIC OXIDE: Roles in Neuronal Communication and Neurotoxicity. I paid the nearly $200 that slim volume cost and have never regretted the expense. As it turns out, the really nasty radical isn't nitric oxide, but rather peroxynitrite (.ONOO-). I (and Jerry Leaf and I earlier) had already tried a number of approaches to inhibiting ischemic injury including the use of two free radical scavengers. All with dismal outcome. I was becoming increasingly convinced that cerbral ischemia (and systemic) ischemia-reperfusion injury was multifactorial and thus would require multimodal intervention. As I saw it, a major "error" in past thinking, just as in the case of trying to block inappropriate immune inflammatory signal molecules in the systemic immune inflammatory response (SIRS) (something all slowly dying cryopatients experience and which ends in multisystem organ failure (MSOF)), has been to single out one species of free radical as the culprit in some injury, and go after it in isolation. Early on, in ischemia and SIRS research, the focus was on the hydroxyl radical (.OH-), then it shifted to the superoxide radical ion (.O2-), and more recently has focused on the interaction of the nitric oxide molecule (.NO), which also happens to be a free radical, with superoxide, which is another radical, which together interact to create the very damaging peroxynitrite radical (.ONOO-). New species of reactive radicals in the body seem to be discovered regularly. I might also add that each of these radical species can generate other damaging radicals, via multiple pathways. When a radical molecule takes an electron from another molecule which already has all of its electrons in pairs, it creates a radical in the other molecule when it stops becoming one itself. Thus, a long game of musical chairs begins, with each molecule stealing from another, until the chain is finally ended by two radicals reacting with each other (often one of these is a sacrificial antioxidant vitamin molecule, like vitamin C, which can take two extra electrons, and wind up with one less double bond). Meanwhile, this game of musical chairs has potentially damaged vast numbers of biomolecules -- some of which are criticalto cell survival. The modern organism, after a long evolution of living in oxygen (a double radical molecule with two unpaired electrons, each in a separate outer orbital), has as a consequence developed many, many different traps and defenses against free radical damage from having to metabolize oxygen. The take-home message here is that it is likely that most, if not all, of these radical species, are significant sources of injury in SIRS and certainly in ischemia. Dealing with one of them in isolation, such as the use of PEG-conjugated superoxide dismutase for the superoxide radical, or mannitol for the hydroxyl radical, is likely to be clinically futile. A good analogy would be locking just one of your car doors to protect against against car-thieves. Most of these radicals are alike, most of them do their damage by stealing electrons from the double bonds of unsaturated fatty acids in cell membranes, and making mutations and breaks in your DNA. But, alike doesn't mean the _same._ The work of many other fine scientists was proceeding in this area during this tme. In particular, Szabo Sabo and his colleagues began to unravel just how damaging peroxynitrite was, and Szabo identified the mechanism by which lipid peroxidation of the mitochondrial membrane resulted in the formation of permanent holes or pores that allowed the leakage of cytochromes into the cytoplasm where they triggered two very nasty processes: poly-ADP-ribosylation synthase activation, and the activation of caspases 1 and 3. The former resulted in rapid and sustained depletion of ATP levels in a futile attempt to repair DNA damaged by all the free radicals generated during ischemia-reperfusion, and the latter triggered programmed cell death, or apoptosis as it properly called. My focus was to develop a multi-modal approach to treating ischemic injury and above all, to find a way to deliver a diverse array of radical scavengers across the blood barrier in a time frame of _seconds_. At the top of my list was a peroxynitrite scavenger. Further down were iNOS and bNOS inhibitors which did not do more harm than they did good. Unselective NOS scavengers were disasters, and caused acute death of the animals. Something sophisticated was needed. In 1994 we did the first experiment using a "sophisticated approach." The dog was walking around 24 hours after 14 minutes and 45 seconds of normothermic ischemia and was christened "Cerberus." She was neurologically normal that day, and she recognized the staff and wagged her tail vigorously when Sandra Russell (her favorite person) approached. People in the cryonis community were kept appraised of this. Certainly the leaders of the cryonics organizations were informed. So too was a broad cross section of the membership. After the Visser fiasco I was asked to present our work on Cerebral Resuscitation at the Alcor Cryonics Conference held in Phoenix, Az in April of 1998. One of our 16 minute dogs was brought to the conference, and the general mechanics of the approach used to achieve successful systemic and cerebral resuscitation were outlined in considerable detail; including the critical importance of selective inhibition of nitric oxide and the criticality of scavenging the peroxynitrite radical. This work, done by Steve Harris, Sandra Russell, Joan O'Farrell, Chris Rasch, Carlotta Pengelley and myself, was apparently understood by the 150 or so people present. The direct structural damage done by free radicals during _both_ ischemia and reperfusion is profound. However, it pales in comparison to the damage done by SIRS and MSOF which many cryonics patients will experience as part of the dying process before their hearts ever stop beating. The technology to block or mitigate this injury has existed since 1994 and has been greatly improvd upon since. Similarly, for those who attended the 21st Century Medicne Conference it should be obvious that even more dramatic advances have been made in mitigating cryoinjury. Few who did not come to the conference ordered the tapes, and fewer still seem to have watched them. It is certainly the case that almost no one cared enough about any of this progress to do anything about it. I opened CRYONICS magazine today and found an article by Linda Chamberlain about the horrors of HMO healthcare and her own sad experience. Linda pointed to a system that is essentially collectivist and concerned not with the doctor-patient relationship, but rather with decision making for the benefit of the "membership" and cost containment as being the root of her own agonizing experience with what can only be termed as in my opinion, a grossly negligent series of medical mis-steps that lead to a pneumothorax, severe pain, and a very real risk of loss of life. Linda was fortunate. Doubly fortunate. She lived _and_ she got the opportunity to complain and warn others about the problem. She is unikely to settle for substandard medical care in the future, and she is very likely to be conscious of both the risks, and the advantages to upgrading (when you can afford it) to a higher level of health care services. Certainly she will be more vigilant and assertive when putting her life on the line. Before Linda's bad experience she was enthusiastic about the quality of medical care her HMO provided (it actually _is_ one of the better ones). I vividly recall several conversations with her and Fred on just this matter; in particular my loathing of the gutting and socialization of medicine and the destruction of the doctor-patient relationship. But what does Linda's plight have to do with this post, or cryonics for that matter? Quite a lot, really. Cryonics organizations, as it turns out, are much like HMOs. They make decisions about the technology their members get -- or don't get "for their own good." They make these decisions with a cross-section or a majority of their "members" in mind. They make these decisions with cost-containment and affordability in mind. They do not make them on a one on one basis with members as physicians used to do, explaining what is available, and what the cost-benefit trade offs are. They determine what is available and you trust them to make the right choices for _you._ You can see how well this is working out in healthcare. In fairness to cryonics organizations, they have little incentive to behave otherwise. They are virtually all non-profit socialist enterprises with their members each expecting to be served steak and lobster when dinner time comes, and not worrying much about the bill or who's picking up the tab. Even more to the point, the members of these organizations are in no position to complain once dinner has been served -- or treatment given. Cryopreservation is, after all, for the forseeable future anyway, a one-way trip with no reports of happy outcomes or adverse reactions. There are no cryopreserved patients struggling for breath, trying to remember who they were, or dealing with a fragmented and disrupted sense of self. Or, for that matter dealing with no self at all. There is no feedback. No normal corrective market mechanisms; no crippled patient, no person in pain, no loss suffered and reported upon. Not yet. And, just like Linda, and millions of others like her, they won't worry about the problem with HMOs or cryonics, until they become personal. The problem is, they may not be as lucky as Linda was on either score, and in any event, they certainly won't be telling any of us NOW about how happy or dissatisfied they are with their cryonics care. So we have a unique situation now. For 30 years I worked and dreamed of a day when the cryonics community would have access to technology that would vitrtually eliminate ultrastructural damage to the brain after cryopreservation. I dreamed of being able to recover animals from 15 minutes of normothermic ischemia, let alone 17. And, if you think those two extra minutes don't count for much, I'll quote the dean of cerebral resuscitation himself, Dr. Peter Safar, as he recently summed up his career to me: "When we strated this work 30 years ago we could, with great effort, recover most dogs from 8 minutes of normothermic ischemia with lasting good neurological outcome. We are now able to do this after 11 minutes about half to three-quarters of the time. That works out to about 1 minute of progress per decade of work." One minute per 10 years. We've added 6 minutes in 5 years. Four years, in reality, since the last year has been spent on perfecting non-invasive rapid induction of hypothermia using liquid ventilation: again with outcome = long term _normal_ survival of the animals as a goal. So, you can imagine my amusement when I read Bogner's and Henry's post reproduced above. WOW! Nitric oxide might be important in ischemia. Maybe we should _DO_ somethig about this. Well, gentlemen, the first cryonics patient who had something done about it was a CryoCare member named James Gallagher who had the advantage of peroxynitrite scavenging, iNOS inhibition, bNOS inhibition and multimodal treatment of ischemia coupled with rapid and effective restoration of good cerebral blood flows and oxygenation. This happened in December of 1995. It could have happened to any other cryopatient frozen with any reasonable warning of impending death essentially any time after early 1994. But it didn't. I suppose that in the next 12 months somewhere between 3 and 12 people will cryopreserved in the United States. If it is a bad year, that number might be as high as 20 (I participated in 10 cases the last 12 months of my tenure at Alcor). The technology to vastly decrease both ischemic damage and cryoinjury now exists and is implementable in a cost-effective fashion. It will NOT be used on these people. In fact, arguably it will not be used on anybody for a long while yet. My dog Cannibal may be the first to get it, and he unarguably deserves it more than the vast majority of the rest of you. With a handful of exceptions, only people such as Saul Kent, Bill Faloon, and the stalwarts that have worked, and supported the work, to make these advances are the only other people remotely deserving of benefit from them. I will be leaving an active role as a service provider in cryonics as of 25 June, 1999. It will have been after 31 years. When I first got involved with cryonics I was 13 years old and I expected a lot of surprises. It is fair to say I was certain only that cryonics represented a true marvel: the technology of tomorrow available today. I leave considerably wiser. And the irony is not lost on me that cryonics as it exist now is not tomorrow;s technology today, but yesterday's technlogy today _and_ tomorrow (and probably the day after, too). If we here at the lab are fortunate enough to re-start our resuscitation work in a month or so as planned, cryonics will soon be the day _before_ yesterday's technology -- tomorrow. I have said my piece. Now, it is time for me to follow Voltaire's advice spoken so wisely by Candide: "It is time to return to work in the garden." To the rest of you cryonicists, men and women of not inconsiderable intelligence or resources, I say with honesty and with no malice, return to your Panglossian debates "in this, the best of all possible worlds," and I promise, for my part, not to interrupt you so rudely again. Mike Darwin Speaking solely on his own behalf and in consultation with no one regarding this post. Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=11685