X-Message-Number: 6370 From: Date: Fri, 21 Jun 1996 16:31:32 -0400 Subject: SCI. CRYONICS Pichugin research Dr. Yuri Pichugin, as previously reported, has been doing cryonics-related research with sheep heads and rabbit brains, funded by the Cryonics Institute and the Immortalist Society. He and his colleagues in the Ukraine have repeated CI sheep head work and verified lack of cracking after cooling to liquid nitrogen temperature, and have obtained spontaneous and evoked bioelectric activity (BEA), coordinated in networks of neurons, in rabbit brain pieces rewarmed from liquid nitrogen temperature. (Control specimens, without glycerol solution perfusion, did not show BEA.) Recently Dr. Pichugin visited the Institute of Cell Biophysics in Pushchino, Russia, where there are the most advanced and skilled workers in brain culture or brain 'incubation." Although only 30% of the best researvchers remain--the rest having left the country--Dr. Pichugin conferred at length with Pavel Pakhotin, Ph.D., who has published extensively in western journals such as NEUROSCIENCE, PROGRESS IN NEUROBIOLOGY, AND BRAIN RESEARCH BULLETIN. Dr. Pichugin wants to use relatively long term culture or incubation techniques on brain slices of adult mammals to allow quantitative assessment of viability after cryopreservation. Dr. Pakhotin's work has been mainly with hypothermia of hibernating mammals, studying viability by extracellular investigation of stability of structure-specific electical activity. (Dr. Pakhotin's methods allow survival for THREE DAYS of isolated brains of adult hibernating mammals! Clearly this has possible cryonics implications, early-phase procedures--if the conditions and skills are not too demanding.) Concerning Dr. Pichugin's work with rabbit brain pieces, Dr. Pakhotin expressed high regard for it, but suggested an additional functional test such as vital staining to evaluate the percentage of neurons with damaged membranes, and to measure membrane potentials--work which would be possible, but difficult, to do in the conditions in the Ukraine where Dr. Pichugin works. Dr. Pichugin's achievement with the rabbit BEA is so ground-breaking that major journals might be reluctant to accept his report without the additional verification. Dr. Pakhotin said that, for Dr. Pichugin's purposes, the neocortex is too complicated; the hippocampus zones are more homogeneous and better understood; normal functioning and viability of hippocampal neurons can be determined quantitatively by measuring BEA. Such work requires very pure reagents, water, gases, and high technical equipment and skill. If Dr. Pichugin cannot work effectively in the Ukraine, he would like to do the work in Pushchino. The conference which Dr. Pichugin attended was on "Preservation of Genetic Resources." Among those attending there was very little interest in cryonics, and no willingness to put any burden whatsoever on the future for revival of cryonics patients. Sounds familiar. Meanwhile, Dr. Pichugin is doing more research with sheep brains. Among other things, he is using hematoxylin and eosin for staining, in order better to observe possible cracks and alterations in the neuropil of cryopreserved brain tissue. These changes are hard to observe with Nissl staining, as Mike Darwin also remarked. This work will be reported probably near the end of June. No cracking was found, even with freeze-thaw rates of 60 deg C per hour at 15% glycerol. But there are apparent alterations of the neuropil and white matter, not yet understood. Footnote: Dr. Pichugin visited the Fyodorov Society in Moscow. They gave him a warm reception, but apparently have not done much to establish or advance cryonics in Russia since he and Ben Best visited three years ago. They seem to believe anyone could be revived from DNA in their bones, and they seem to subscribe to the information paradigm of the person (uploaders?) Dr. Khalyavkin was also there, but did not want to talk much about his ideas on gerontology. Robert Ettinger Cryonics Institute Immortalist Society Rate This Message: http://www.cryonet.org/cgi-bin/rate.cgi?msg=6370