X-Message-Number: 3117 Date: 12 Sep 94 03:54:35 EDT From: Mike Darwin <> Subject: SCI.CRYONICS brain research Bob Ettinger expresses interest in and reservations (regarding accessability) about BPI brain cryopreservation research: >There seems to be a problem, however, with the proprietary aspects of these >studies. Any available information on results of this work would be >appreciated, and we are willing to pay a reasonable amount for the trouble of >sending it. >The Cryonics Institute sheep head work was published in its essentials about >two years ago; the current Ukrainian work is being published in THE >IMMORTALIST as we receive it (although only with some of the photos; >complete photos available), and sometimes also posted in whole or in part on >CryoNet. >If the profit angle rules out any extensive or timely cooperation, then I >suppose those not privy to particular procedures will just have to wait for >the patents to be available. (It may also turn out that the non-proprietary >work will be better or sooner.) Bob needn't have concerns about "propreitary aspects" of this work. We plan a full and open disclosure, and indeed, most of the work I've done (the overwhelming majority, in fact!) has been publically disclosed. I would urge Bob (and others with interest in this area) to request copies of the Lake Tahoe presentations which I and another cryobiologist made on ischemia and cryoinjury (some of the former data collected from a human brain). Extensive histological and electronmicroscopic data was presented and, believe it or not, even the slides projected on the screen during the talk are still usefully visible on the video. In fact, as I have said before, *I* would really appreciate a copy of these tapes as my only set was left at Alcor (and, like Bob, I would be willing to pay all reasonable duplicating costs). I suggest this as a good starting place because some of what we are finding currently is extending on those results (sad to say). As to our current research I'm not (at the moment) in any position to make disclosures. There are several reasons for this: 1) The work is far from completed. One thing which I can say is that I am finding it incredibly frustrating trying to sort out various kinds of *artifacts* from real injury. Here I can give some examples: a) Glycerolization, even at fairly slow rates, results in considerable dehydration of the tissue. The dehydration makes it almost impossible to see ultrastructure: everything is packed in on top of everything else: sort of like trying to read the print on a piece of onion skin paper which has been stuffed into a thimble. b) Removing the glycerol, however, introduces all kinds of arifacts too. And, as Bob's Ukraine investigators have noted, getting glycerol out isn't easy. Yes, we've mastered that achievement, but the question then becomes what is artifact (i.e., injury) from the dehylcerolization vs. "real" injury from glycerolization and/or freezing. Currently we are experimenting with different ways of fixing the tissue and different conditions of fixation in an effort to arrive at "the truth" of what is really happening. c) Freezing and thawing introduce whole other sets of complications. Because we are constrained to thaw slowly (about 36 hours for a whole dog) the animal is exposed to not only the many hours of multimolar glycerol it was exposed to during the slow freezing, but also during rewarming. So, how much of the injury is from thawing? d) In order to get a better idea of what condition the tissue is actually in in the frozen state we will be doing some freeze substitution studies. Unfortunately, these take a lot of time and we are just getting started. e) Glycerol interacts with the fixative in a negative way. This problem gets worse with high glycerol concentrations. 2) As most people who know me know (and who have followed my work in CRYONICS and here on the net), I am very opposed to less than full disclosures. By this I mean that giving bits and pieces of the results often results in misunderstandings, hostility, accusations of sloppiness, and so on. Thus, at a minimum, I want to have a full disclosure of my materials and methods as well as my results. I also want to have some confidence my results MEAN something. What I am saying here is that right now I'm looking at a bunch of EMs, scratching my head and saying "What the hell does that mean!" I don't think anyone would appreciate seeing results presented under those conditions and with that degree of uncertainty. However, once the data is solid and the work done, I'll be the first to publish. Some work here is proprietary. However, survey work on the effects of current cryopreservation protocols is NOT proprietary. We are a long way from making hurculean strides towards marketable suspended animation. Open sharing of results is thus highly desirable. One small side project related to this work is a paper on the gross, histological and ultrastructural effect of immersion fixation of intact dog brains. A draft of that paper has been completed and circulated for commentary to a few people. I will send the second round (preposting/prepublication) to Bob and CI. If Bob would like copies of my EMs from the mid 80's on the cat brains and other organs, as well as the paper, I will be happy to supply them (some of them, anyway: there are over 1,000 images and the cost of duping the color slides would be astronomical!). Better still, if Bob would like to borrow the EM's (originals) he can do so as long as he looks at 'em quick and gives 'em back (that way he can copy what he wants). I would very much be interested in seeing good quality copies of the CI brain EMs. I mean no offence by this at all, but the pix which have been published in THE IMMORTALIST are virtually useless for evaluation. The text accompanying them *is good* however and gives an idea of what is there. But, in my case this because the text might as well be describing the pictures *I* get from straight frozen brains (so I can almost see them in my mind's eye when I read Dr. Pichugin's descriptions). If Bob would like to enter a private and detailed correspondence (perhaps including Dr. Pichugin as well) about our current work and results, I would be happy to do that -- in which case we would be be exchanging data in a collaborative fashion and working together (however distantly) to solve the same sorts of problems. Mike Darwin Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=3117