X-Message-Number: 2909 Date: 18 Jul 94 10:53:53 EDT From: "Steven B. Harris" <> Subject: CRYONICS.Perfusion Tradeoffs Dear Cryonet: In response to Robert Cardwell, the potential problems with doing less than 4 vessel brain perfusions are as I indicated, and the 4-vessel aorta-based cryoprotectant perfusion of the brain should be considered state-of-the-art. Thus, if you are a cryonics organization offering services to the public, there is little excuse for offering anything less in your main lab if you advertise yourself as being state-of-the-art. A good cryonics organization will end up doing at least some fraction of its suspensions this way. But what about the emergencies and one or two day delays between cardiac arrest and cryoprotectant perfusion which will always occur to even the best organizations occasionally (and which will occur regularly to some organizations?). What about clients who are stuck in cardiac arrest out of the country on a weekend, or who have experienced for one reason or another so much warm ischemia that any further warm-time carries the real possibility of not being able to perfuse the brain at all? What is the best choice if you have to choose between a 2-vessel perfusion of the brain with a portable pump followed by a dry ice freeze before shipping, *versus* 3 extra days at water ice temps followed by a fancy 4-vessel perfusion and more careful decent to dry ice temps? How do you judge the tradeoffs? I wish I knew. Research will tell some of the answers, but not all of them. One problem is that human brain perfusion anatomy is not at all like that of any animal, and although we can perhaps draw a few conclusions from basic brain anatomy and animal work (as I have tried to do), still perfusion after clinical death at low temperatures with glycerol is bound to have some major differences from perfusion with blood in life. We can learn some of the answers here for certain by watching many perfusions of the whole cortex of many patients with fiberoptics and dye systems during less than optimal "field" conditions. On the other side of the coin, we can learn something about brain fine structural deterioration (which is likely to be similar in all mammals) by doing animal delay experiments at ice temps. Lastly, animal experiments of the sort CI and the Russians are doing (and also we ourselves at BioPreservation) will ultimately tell us something about how much we gain at the ultrastructural level by perfusion with cryoprotectants at all. At some point we know that besides the extra cracking produced by cryoprotectant, micro-damage from time at zero Centigrade *waiting* for cryoprotectant (and a slow freeze) will outweigh the extra micro-damage that occurs due to an immediate "fast" dry ice freeze with no cryoprotectant. The question is: when does that time come? And how to factor in all the intermediate possibilities involving less-than-optimal cryoprotectant surgical introductions, done at earlier times? And what do we do in the meantime? Real metabolic experiments on real brains suggest that the temperature sensitivity scale for brain ischemia is not the simplistic factor of 2 for 10 C sometimes used as rule of thumb by chemists, but more like a factor of 4 or 5. Near the ice temp we can expect something like an equivalent of 1 minute of warm ischemia for every 1 hour of real-time, or about 24 minutes per day. One day on ice alone puts us at about the limit even optimistic conventional brain resuscitation workers suggest, due to their estimates of irreversibility of cell damage. An additional 1/2 to 1 hour warm-ischemia equivalents is generated even in the best of circumstances while cooling the patient. Thus, even one day of ice preservation after best cooling results in 1 to 1.5 hours total warm-ischemia equivalent time, and insides of brain cells and capillary walls are seriously disordered at much beyond this. Is any of this structure associated with memory? We don't know, but it seems best not to find out. My best guess is that anything beyond a day or two at the ice temp, even with an optimal cooling history and no downtime to speak of, is very unwise. If it can be avoided by more rapid (but cruder) early perfusion followed by crude dry ice freezing, perhaps it should be. Steve Harris Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=2909