X-Message-Number: 7671
From:
Date: Sun, 9 Feb 1997 12:46:12 -0500 (EST)
Subject: Altech handout
The following is a copy of the handout at the Alcor Technology Festival, Feb.
1, minus the color photos. We also showed brief excerpts of the video of the
second sheep heart.
---------------
CRYONICS INSTITUTE
24355 Sorrentino Court
Clinton Township MI 48035
Phone (810) 791-5961, Phone/Fax (810) 792-7062, E-Mail <>
R.C.W. ETTINGER
3326 N. 81 St.
Scottsdale AZ 85251
Phone (602) 941-5591; Fax (602) 947-7759; e-mail <>
CRYONICS INSTITUTE/IMMORTALIST SOCIETY RESEARCH
By R.C.W. Ettinger Jan. 29, 1997
For the Alcor Technology Festival, Feb. 1, 1997
PRELIMINARY: For many years the Immortalist Society and the Cryonics
Institute have
conducted or/and supported research and development, and disseminated
information, in
cryobiology, anti-senescence, general health sciences, cryogenic storage
systems, and related
areas. (The first Immortalist Society disbursement was a small cash prize to
Prof. Isamu Suda for
his cat brain work in the Sixties.) In recent years this work has slowly
grown with our resources
and contacts. This last year has seen further growth, with impetus from the
Visser work and from
cooperation with the Vissers and Alcor and others including Dr. Yuri Pichugin
in the Ukraine.
Any applications of results to CI patient practice is decided among Sally
Bazan, Andy Zawacki,
consultants, and myself, from time to time as appropriate.
Following is a brief summary of past work, with a bit more detail on recent
and current work.
NOTE: Because of time pressures and related uncertainties, as I write this,
it is not clear how
extensive or abbreviated these various segments will be. Some may appear only
in outline as
subject headings. There may be addenda, or some of the material may not be
available in time
for the Alcor Tech Festival. Those interested may find further and related
material in forthcoming
(and past) issues of The Immortalist.
1. STORAGE RESEARCH AND DEVELOPMENT
a) Fiberglass Cryostats
b) New Types of Insulation
c) Intermediate Temperature Storage
[including for economy only]
2. SHEEP BRAINS AND RABBIT BRAINS
a) CI work
b) Dr. Pichugin's work
3. SHEEP HEARTS
Work has begun on applying Visser-related methods to cryogenic storage and
revival of sheep
hearts. This is being done primarily by Andy Zawacki--assisted by a
veterinary and by David
Fulcher--at the CI facility and at a farm his family owns, under my general
direction and with
consultation by Mrs. Visser, Hugh Hixon at Alcor, Dr. Pichugin, and other
collaborators. (We
do not work on live animals; the sheep or organs are obtained from a
slaughterhouse, and if
necessary the sheep are anaesthetized and euthanized by the veterinarian.)
A word may be in order about the rationale of research planned and performed
by people with
little in the way of credentials or experience. First, I note that CI people
had no credentials or
experience to do research and development in cryogenic storage either, but
nevertheless we
planned, fabricated, tested and are successfully using cryostats different
from any other, and in
some respects better than any other.
Cryobiological research of course is much more complex and difficult than
cryogenic storage
research and development. Nevertheless, Mrs. Visser--despite relatively
unimpressive credentials
and experience--succeeded in reviving rat hearts from liquid nitrogen, a feat
that had eluded all
others for decades.
One of the things to be understood is that cryobiology, despite a seemingly
voluminous literature,
has always been a starveling discipline, with relatively few practitioners
and resources. Several
years ago, when told about our proposed approach to brain cryopreservation,
Greg Fahy noted
that it had not been tried before and might be worth a shot. A great many
even of the simple
approaches or variations have not yet been tested.
CI/IS have certain advantages over professional cryobiologists. In
particular, we have less red
tape to contend with. Because we do not keep live animals on our premises, we
have less of a
regulatory burden. Because we are small organizations, we do not require a
committee to consider
every problem, or another committee to review the first committee; we can be
quick and decisive.
Because we are only interested in results--not in credit or profit--we can
minimize paperwork,
in general. ("In general" because we do have certain obligations in
connection with our agreement
with the Vissers and Alcor concerning the Visser technology.)
Of course these advantages may not be enough to assure that we will succeed
where others have
failed, or will succeed in any major way. But the one thing we know for sure
is that our work
will teach us something, and may teach others as well.
We will usually not do work that others can do better or cheaper, if their
results are available to
us--although we must always pre-test possible applications to patients. In
summary, work is
justified if potential benefit, in our best judgment, outweighs the costs.
Following is Andy Zawacki's report of the sheep heart of 1/21/97, slightly
edited, deleting
information that is still confidential under our agreement with the Vissers
and with Alcor.
--------------------
The weight of the sheep was was about 150 lb. It was anaesthetized by the
veterinarian with 20
mg Rompum and 50 mg Ketaset. The wool was clipped from the chest to keep the
area clean.
40,000 units of heparin was injected and given five minutes to circulate.
A bolus of 150 mM KCl was given to stop the heart. After about 20 or 30
seconds the vet said
he could not hear the heart with his stethoscope, and concluded it had
stopped.
The chest was then cut open, and in the process the vet's knife accidentally
cut the heart, about
a one inch cut on the right ventricle. It was not very deep, but at the time
we thought the
experiment was ruined and we stopped recording times and being careful.
When we looked at the heart it was still beating a little. Maybe it was too
weak for the vet to
hear. The heart stopped shortly after that, and we considered different ways
of approaching it
next time. I then decided to remove and wash out the heart just for practice.
I estimate about 8
minutes passed from the time the heart stopped until I started to run the
warm (37oC) Tyrode's
solution through it. Within 30 seconds of starting that, the heart started to
beat and continued to
beat (ventricular and atrial) until the liter of solution was gone, about a
minute or two.
The life support machine was set up in the basement, so we quickly went down
there to continue
the procedure. When we got in the basement the heart was stopped and we
connected it to the
life support machine. It was maybe less than a minute and the heart started
to beat again. The
heartbeat was not quite as strong as it was in the garage when it first
started to beat.
Our video started from the time we got into the basement and prepared to
connect the heart to
the life support machine. The heartbeat was maintained for about 10 minutes.
The heartbeat now
was only atrial, but under the circumstances I was surprised and encouraged.
After about 10
minutes of the heart beating the cold Tyrode's solution was started and the
heart stopped. We
ran about two liters of cold Tyrode's at 1oC through the heart to cool it.
We could observe air
in the coronary artery and the cold Tyrode's did not want to flow very well.
The cannula was then removed and the heart was massaged to try to remove the
air. It must have
worked to some degree because after that the cold Tyrode's went through much
better. After the
cold solution we perfused the heart according to our planned protocol. The
perfusates went
through easily and it appears that the longer the perfusion went on the
better the perfusate flowed
through the heart. As the perfusion went on the color of the heart became
darker.
The cold Tyrode's solution and the cold perfusate were stored in a cooler
with ice until used and
when the temperature was checked it showed it at one degree Celsius. After
the heart was
perfused it was wrapped in cotton gauze appropriately soaked, and put into a
plastic container
which was then closed. The container with the heart was put in a cooler and
covered with dry
ice and we left for the lab. Because we started to get freezing rain it was
about 3.5 hours before
wc got back to the lab.
When we got to the lab the container with the heart was placed in liquid
nitrogen. The container
floated on the liquid nitrogen so the cooling rate would not be as fast as if
the heart was just
plunged into it. The heart is still [Jan. 23] in the liquid nitrogen and we
plan to warm it on
Sunday the 26th. [This has been delayed until the week of Feb. 2, to allow
time for dummy runs
of alternative methods.] I have set up to do another experiment on Tuesday
the 28th and we will
have an extra person just to run the camera so that Dave will be free to help
me and we can get
better video footage.
The weight of the heart was about 250-300 gm. We will weigh it more carefully
after thawing.
--Andrew F.
Zawacki
[color photo]
Heart on life support. Cut visible.
[color photo]
Starting cold Tyrode's to stop the heart.
[color photo]
Heart during perfusion with CPA. The opening shows the pulmonary vein; right
of that is the
inferior vena cava.
[color photo]
Part of the circulation setup. Left box controls temperature and recirculates
filtered solutions.
Container at top provides adjustable pressure head. Container on shelf is for
the heart.
[Another heart was done after this, with better results. At the meeting, we
showed a short excerpt
from the video of the second sheep heart preparation and freezing.]
4. TRANSPLANT-RELATED WORK: One of the Visser projects is to use the Visser
rat heart
technology, or variations thereof, to develop cryobanking of human organs for
transplant, first
proving principle with animals such as pigs. Mrs. Visser is optimistic about
this potential,
although, judging from information I have seen, there are many possible
reasons why the rat heart
procedure, or any simple extensions of it, may not work with larger organs or
different species.
But those who emphasize the possible obstacles, or even known obstacles, have
the wrong focus.
When Rostand revived frog sperm, frozen with glycerol, in 1948, the emphasis
was correctly on
the break-through and not on the difficulty of the long road ahead.
Cryonics Institute directors do not believe it is presently appropriate for
CI to invest substantial
sums in transplant-related work, because it has no direct relevance to
cryonics and financial
returns are far from assured. But if our own people, or collaborators
available at modest rates,
could cryopreserve mammalian organs of approximately human size, this would
be a worth-while
accomplishment, for several obvious reasons. It would be another major
break-through, and the
morale boost alone would justify the relatively modest expense. I omit here
any discussion of
legal and financial ramifications of potentially profitable discoveries we
might make.
Again: To the best of my knowledge, no one anywhere has recovered a beat with
human-sized
hearts warmed from liquid nitrogen tempeature, and no one anywhere is
currently attempting to
do so, other than ourselves. We are not expecting success, if it comes, to be
quick or easy, but
we think the effort is justified.
5. ANTI-SENESCENCE
a) Odens' Work. The late Max Odens, M.D., in 1973 published a paper in the
Journal of the
American Geriatrics Society, Vol. XXI, No. 10, October 1973, titled
"Prolongation of the Life
Span in Rats." He claimed that weekly injections of "DNA solution in water
saturated with
chloroform (3 mg per ml) plus ordinary RNA" resulted in rats in the control
group showing a
doubling of life span (not expectancy) on average for 4 rats, and a trebling
for the fifth rat.
The paper was frustratingly lacking in detail and specifics, so we were (and
as far as I know still
mostly are) faced with the problem of trying to determine whether the work
was valid. One of
the obvious steps was to read all the papers referenced in Odens' paper,
looking for clues to his
procedure. We have found and copied 10 of the 23 referenced papers. A quick,
preliminary look
at these did not suggest anything very promising to me, but I will look at
them more carefully
when I have time. We will also make copies for any other people who need them
and who might
find something useful.
I had also suggested that someone (preferably in England?) might want to
volunteer to hunt down
people previously associated with Odens or with the journal at that time, and
question them for
possible hints as to his procedures, assistants, associates, etc. We repeat
our willingness to pay
any reasonable expenses for such work.
There also remains the possibility of searching literature since the date of
his publication, to find
his paper referenced, on the chance that someone tried to repeat his work, or
did some related
work. Again, if someone will volunteer to do this, we will pay any reasonable
expenses.
b) Combinations of Supplements. Many substances have shown possible
effectiveness in
extending life expectancy or/and life span. In most cases, adequate
experimental studies are
lacking. In even shorter supply are studies of the effects of combinations of
these; are they
independent, or mutually reinforcing, or antagonistic, or are there complex
quantitative relations?
Some organizations, individuals, and institutions are working on these
problems, but as far as I
know there has been little or no coordination. I believe Doug Skrecky is
trying to do some of this
work with Drosophila.
6. MISCELLANY
a) Freeze-Drying
My impression has always been that the results of freeze-drying have been so
poor, relative even
to the crudest freezing, that this approach deserves only a very low
priority, despite the economy
of potential room-temperature storage. However, Doug Skrecky believes the use
of certain sugars
and other variations may change this. We will give it more attention when we
are able.
b) Fixation
Chemical fixation (essentially variations of embalming, e.g. with
glutaraldehyde) is another old
idea for cheap treatment and storage--again with much more damage than
freezing inflicts, as far
as we know, and therefore only a low priority. But we will get to it when we
can.
7. INTER-ORGANIZATIONAL COOPERATION
There are obvious possible advantages to sharing of information and even
coordinated planning
to avoid duplication of effort and expense and to obtain the broadest input
before commitment
to particular efforts. On the other hand--even if adequate good will and
trust were in place--there
could also be losses through discard of approaches that might by committee be
deemed low
priority. Further, energy and enthusiasm can be important ingredients of
success, and these tend
to get lost in committee.
Additionally, even given any amount of good will, there are inherent problems
in sharing among
those with a strictly life-saving, non-profit focus and those with a major
interest in financial
returns. (This does not mean that for-profit companies are concerned only
with money, or that
non-profit organizations would not find more revenues agreeable; it just
means a degree of
difference in emphasis, possibly sometimes a sufficient difference to put
sand in the gears.)
CI and IS are proceeding with our own initiatives, and according to
agreements with Alcor, the
Vissers, and collaborators. Of course we will always listen to new or
additional proposals.
8. CAUTIONARY NOTE
I have generally been a long term optimist but a short term skeptic in many
areas of technology,
and cryobiology is no exception. One of my favorite examples is the "flying
flivver" predicted
in the '30s as foregone within 50 years; more than 60 years later, flying
flivvers are not even on
the horizon--and that is surely relatively simple technology, mere
engineering, not requiring
scientific break-throughs. It is true that we have also accomplished many
things not discussed or
foreseen in the '30s; but it seems prudent not to count on marvels for the
near term.
Brains have much redundancy and (at least in some respects) surprising
freeze-hardiness. Yet the
brain is so complex, and essential aspects conceivably so delicate, that a
full-fledged nanotech
(or equivalent) may be essential not just for revival, but even for inducing
an easily reversible
suspended animation.
After all, for several decades all the cryobiologists in the world failed to
revive a rat heart from
liquid nitrogen, until Mrs. Visser came along. And a heart is scarcely more
than muscle, with
some innervation and vasculature. So--brains? A high-priced and highly
talented team at the
NMRI has been struggling for several years to cryopreserve rabbit kidneys
through vitrification.
They are still predicting imminent success, and I hope they achieve it. But
how anyone could
reasonably extrapolate from that kind of success to realistic optimism about
early success with
brains is a mystery to me. It's like stepping up on a mole-hill, then
proclaiming that the mountain
is just another step away. We don't yet know the physiology of memory, and
still less of
subjectivity, our most crucial feature....We also know of (still
confidential) reported cases where
electron microscope examination looked perfect, but function didn't
follow....My guess is that
reversal of senescence will prove easier and quicker than suspended
animation.
9. THE CRYONICS GUARANTEES
If I may be forgiven for rambling on, it may not be inappropriate to remind
ourselves that--
contrary to what we sometimes hear or read--cryonics organizations can indeed
make certain
guarantees, or assertions solidly rooted in reality.
We cannot, of course, guarantee that we will ever revive our patients, or
even that they will like
it if we do--although I am optimistic on both counts. But we can offer
several forward-looking
statements based firmly on preponderance of evidence.
1. We guarantee our best efforts to preserve and eventually revive,
rejuvenate, and rehabilitate
the patients. (Prospective members should of course check out individual
organizations to satisfy
themselves as to how good those "best efforts" are likely to be.)
2. We guarantee that liquid nitrogen preservation will preserve structure and
potential function
much better than disposal in a grave, or at sea, or in a crematory. (In Fred
Chamberlain's words,
we can guarantee that ice preserves better than earth or water or fire.)
3. We guarantee that, if a person is revived from your remains, that person
will be more like you
than an identical twin would be. The reason is that your DNA (genome) will be
abundantly
available; and future technology will certainly be able to use some of the
internal and external
information to impose or restore at least some of your psyche. This suggests
that such revival,
even if it does not benefit "you," may benefit your friends and relatives (as
theirs may benefit
you). And with a little bit of luck the revived people will actually be you
and those you love.
4. We guarantee that, in the future, tools and viewpoints and reasons for
optimism will be
developed that are now unimagined. This does not rule out the possibility of
calamities, or the
possibility that in the end we will find the universe distinctly
user-unfriendly; but it reminds us
of the fundamental arrogance of pessimism. The convinced pessimist
essentially assumes the
world is circumscribed by his own limitations.
5. We guarantee that, if you absently-mindedly die and stubbornly remain
dead, you will find it
hard to enjoy life.
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