29. November 2014 · Comments Off · Categories: Health, Science

This article was originally published in Cryonics Magazine, 2011 Issue #1

I am an ardent supporter of Dr. Aubrey de Grey and his work to advance rejuvenation science. The man is priceless and unique in his concepts, brilliance, dedication, organizational abilities, and networking skill. His impact on anti-aging science has been powerful. I have attended all four of the conferences he has organized at Cambridge University in England. For the February 2006 issue of LIFE EXTENSION magazine I interviewed Dr. de Grey, and for the December 2007 issue of LIFE EXTENSION I wrote a review of ENDING AGING, the book he co-authored with Michael Rae.

Dr. de Grey asserts that aging is the result of seven kinds of damage – and that technologies that repair all seven types of damage will result in rejuvenation. His seven-fold program for damage repair is called SENS: “Strategies for Engineered Negligible Senescence”. Dr. de Grey asserts that repairing aging damage is a more effective approach than attempting to slow or prevent aging, and I agree with him. Being an ardent supporter of SENS has not stopped me from simultaneously being a critic of aspects of his program that I think are flawed or deficient. I will attempt to outline some of my criticisms in simple language, assuming that my readers have some knowledge of basic science.

Two SENS strategies cannot justly be described as damage-repair, in my opinion. To protect mitochondrial DNA from free radical damage he wants to make copies of mitochondrial DNA in the nucleus – and import the resulting proteins back into the mitochondria. I would call this an attempt to slow or prevent aging – it cannot be called repair.

Similarly, SENS aims to eliminate cancer by deletion of genes that contribute to cancer, specifically telomerase and ALT (Alternate Lengthening of Telomeres) genes. I am not convinced that this is the best way to eliminate cancer, and I do not believe that deleting cancer-producing genes can properly be called damage-repair.

My criticisms about a procrustean attempt to force two strategies into a model purporting to only be concerned with damage and repair is minor, however, compared to a more fundamental concern that I have that a significant form of aging damage may be being ignored by SENS. I have written a review expressing my concern entitled “Nuclear DNA Damage as a Direct Cause of Aging” that was published in the June 2009 issue of the peer-reviewed journal Rejuvenation Research, [note 1] a journal of which Dr. de Grey is Editor-in-Chief. A PDF of my review is available in the life extension section of my website BENBEST.COM. Those interested in all the citations for claims I will make in this essay are encouraged to read my review. In this essay, I limit my citations to only a few critical articles.

There are many types of DNA damage, but for the purposes of this essay I will focus on breakage of both DNA strands – resulting in a gap in a chromosome. There are two mechanisms for repairing double-strand DNA breaks: Homologous Recombination (HR) and Non-Homologous End-Joining (NHEJ). HR usually results in perfect repair, but HR can only operate when cells are dividing. NHEJ is the more frequent form of double-strand break repair, but it is error-prone. NHEJ is the only DNA repair mechanism available for non-dividing cells. Even in cells that divide, 75% of double-strand breaks are repaired by NHEJ. [note 2]

It is hard to believe that it could be a coincidence that the most notorious “accelerated aging” diseases are due to defective DNA repair. The two most prominent of these diseases are Werner’s syndrome (“adult progeria”) and Hutchinson-Gilford syndrome (“childhood progeria”), both of which are caused by defective nuclear DNA repair, mainly HR. In both diseases the “aging phenotype” is apparently due to high levels of apoptosis and cellular senescence. Apoptosis (“cell suicide”) and cellular senescence (cessation of cell division) are both mechanisms that are induced in cells experiencing nuclear DNA damage that the cell is unable to repair. It is not surprising that victims suffering massive depletion of properly functioning cells should exhibit “accelerated aging”. Mice that are genetically altered to show increased apoptosis and cellular senescence also show an “accelerated aging phenotype”.

Elimination of senescent cells and stem-cell replenishment of cells depleted in tissues by this elimination – as well as depleted by apoptosis – are part of SENS. But these strategies are only applicable to cells that divide – not to non-dividing cells such as neurons. Cryonicists are acutely aware that organs – and even whole bodies – can be replaced, but brains (neurons, axons, dendrites, and synapses, particularly) must be preserved if we are not to lose memory and personal identity. The ability of future medicine to replace all organs and tissues other than the brain would render most of SENS unnecessary – except for the brain.

There is considerable evidence of a significant role for DNA damage in brain aging. There are nearly twice as many double-stand nuclear DNA breaks in the cerebral cortex of adult (180 days) rats as in young rats (4 days) – and old (over 780 days) rats have more than twice the double-strand breaks as adult rats. [note 3] Adult rats show a 28% decrease in NHEJ activity in the cerebral cortex neurons compared to neonatal rats – and old rats show a 40% decrease. [note 4] Declining NHEJ activity with age is at least partially due to ATP decline and cellular damage that SENS is intended to fix. But even if NHEJ activity did not decline with age, nuclear DNA damage in neurons will increase at least in part because NHEJ is so error-prone.

Nuclear DNA damage typically leads to mutation or DNA repair – or apoptosis or cellular senescence when DNA repair fails (a mechanism that is believed to have evolved for protection against cancer). But not all DNA damage is repaired, and NHEJ repair is often defective. Accumulating DNA damage and mutation can lead to increasingly dysfunctional cells.

Cancer is due to nuclear DNA damage, mutations, and epimutations. Dr. de Grey has written that “only cancer matters” for mutation and epimutation to nuclear DNA. His mutation terminology does not even acknowledge DNA damage. He has assumed that damaged DNA either is or becomes a mutation. He has assumed that DNA damage that does not become a mutation is either repaired – or leads to apoptosis or cellular senescence.

Dr. de Grey has made the claim that evolution has required such strong defenses against cancer that residual mutation (and, implicitly, DNA damage) is negligible. But cancer incidence increases exponentially with age up to age 80, so it is likely that the residual increases exponentially at the same time.

As recently as the 1980s it was widely believed that normal aging is associated with extensive neuron loss. Now it is established that functional decline in the aging brain is associated with increased neural dysfunction rather than neurodegeneration. [note 5] This neural dysfunction may or may not be mostly due to cellular damage that SENS is intended to fix – including causes of declining NHEJ activity. How much neuron dysfunction associated with aging is due to accumulating mutations or unrepairable nuclear DNA damage is unknown. SENS assumes without proof that nuclear DNA damage and mutation is negligible as a cause of aging (apart from cancer, apoptosis, and cellular senescence). This may be right or it may be wrong. I believe that without definitive proof, nothing should be assumed, and active investigation to determine the facts should not be neglected.

I believe the situation is not hopeless if nuclear DNA damage proves to be a significant cause of brain aging. Future molecular technologies for detection and repair of nuclear DNA damage could be significantly better than natural DNA repair enzymes. And, to simplify the required effort, the DNA repair technologies could be restricted to genes that are actively transcribed in neurons, rather than needing to repair the whole genome.

Notes

1: Best BP. Nuclear DNA damage as a direct cause of aging. Rejuvenation Res. 2009 Jun;12(3):199-208.

2: Mao Z, Bozzella M, Seluanov A, Gorbunova V. Comparison of nonhomologous end joining and homologous recombination in human cells. DNA Repair (Amst). 2008 Oct 1;7(10):1765-71.

3: Mandaville BS, Rao KS. Neurons in the cerebral cortex are most susceptible to DNA-damage in aging rat brain. Biochem Mol Biol Int 1996 Oct; 40(3):507-14.

4: Vyjayanti VN, Rao KS. DNA double strand break repair in brain: reduced NHEJ activity in aging rat neurons. Neurosci Lett. 2006 Jan 23;393(1):18-22.

5: Morrison JH, Hof PR. Life and death of neurons in the aging brain. Science. 1997 Oct 17;278(5337):412-9.

15. October 2014 · Comments Off · Categories: Cryonics, Science

Cryonics Magazine, July 2013

[The following is a text adaptation of a PowerPoint presentation given on Sunday, May 12, 2013 at the Resuscitation and Reintegration of Cryonics Patients Symposium in Portland, Oregon]

An understanding of probable future repair requirements for cryonics patients could affect current cryostorage temperature practices. I believe that molecular nanotechnology at cryogenic temperatures will probably be required for repair and revival of all cryonics patients in cryo-storage now and in the foreseeable future. Current nanotechnology is far from being adequate for that task. I believe that warming cryonics patients to temperatures where diffusion-based devices could operate would result in dissolution of structure by hydrolysis and similar molecular motion before repair could be achieved. I believe that the technologie for scanning the brain/mind of a cryonics patient, and reconstructing a patient from the scan are much more remote in the future than cryogenic nanotechnology.

Cryonicists face a credibility problem. It is important to show that resuscitation technology is possible (or not impossible) if cryonicists are to convince ourselves or convince others that current cryonics practice is not a waste of money and effort. For some people it is adequate to know that the anatomical basis of the mind is being preserved well enough ― even if in a very fragmented form ― that some unspecified future technology could repair and restore memory and personal identity. Other people want more detailed elaboration.

Books have detailed what nanotechnology robots (nanorobots) will look-like and be capable-of, including (notably) Nanosystems by K. Eric Drexler (1992) and Nanomedicine by Robert A. Freitas, Jr. (Volume I, 1999; Volume IIA, 2003). The online Alcor library contains articles detailing repair of cryonics patients by nanorobots at cryogenic temperature, in particular, “A Cryopreservation Revival Scenario using Molecular Nanotechnology” by Ralph Merkle and Robert Freitas as well as “‘Realistic’ Scenario for Nanotechnological Repair of the Frozen Human Brain.” Despite the detailed descriptions, calculations, and quantitative analyses that have been given, any technology as remote from present capabilities as cryogenic nanotechnology is certain to be very different from whatever anyone may currently imagine. It is difficult to argue against claims that all such descriptions are nothing more than handwaving, blue-sky speculations.

Current medical applications of nanotechnology are mainly limited to the use of nanoparticles for drug delivery.1 Nanomachines are being built, but they are little more than toys ― including a rotor that can propel a molecule2 or microcantilever deflection of DNA by electrostatic force.3 In classical mechanics and kinetic theory of gases, on a molecular level, temperature is defined in terms of the average translational kinetic energy of molecules, which means that the lower the temperature the slower the motion of the molecules. According to the Arrhenius Equation, the rate of a chemical reaction declines exponentially with temperature decline. It would be wrong to conclude that nanomachines would barely be able to move at cryogenic temperatures, however. Nanomachines operate by mechanical movement of constituent atoms, a process that is temperature-independent. In fact, nanomachines would probably operate more effectively at cryogenic temperature because there would be far less jostling of atoms in the molecular structures upon which nanomachines would operate. Nanomachines would also be less vulnerable to reactions with oxygen at cryogenic temperature, although it would nonetheless be preferable for cryogenic nanorepair to occur in an oxygen-free environment.

Although under ideal circumstances ice formation can be prevented in cryonics patients, circumstances too often result in at least some freezing―such as inability to perfuse with vitrification solution, or poor perfusion with vitrification solution because of ischemia due to delayed treatment. Past cryonics patients were perfused with the (anti-freeze) cryoprotectant glycerol, whereas cryonics patients are currently perfused with cryoprotectant solutions that include ethylene glycol and dimethylsulfoxide (DMSO). Unlike water, which forms crystalline ice when solidifying upon cooling, cryoprotectants form an amorphous (non-crystalline, vitreous) solid (a “hardened liquid”) when solidifying upon cooling. The “hardened liquid” is a glass rather than an ice. The temperature at which the solidification (vitrification) occurs is called the glass transition temperature (Tg).

For M22, the cryoprotectant used by Alcor to vitrify cryonics patients, Tg is typically between −123°C and −124°C (depending on the cooling rate). Tg is about the same for the cryoprotectant (VM-1) used for cryonics patients at the Cryonics Institute. Although freezing can be reduced or eliminated by perfusing cryonics patients with vitrification solution before cooling to Tg, eliminating cracking is a more difficult problem. Cryonics patients are cooled to cryogenic temperatures by external cooling. Thermal conductivity is slow in a cryonics patient, which means that the outside gets much colder than the inside. When the outside of a sample cools more quickly than the inside of the sample, thermal stress results. A vitrified patient subjected to such thermal stress can crack or fracture. No efforts have been made to find additives to M22 that would have a similar effect as boron oxide has on allowing Pyrex glass to reduce thermal stress.

If a vitrified sample is small enough, and if cooling is slow enough, the sample can be cooled far below Tg ― down to liquid nitrogen temperature ― without cracking. A rabbit kidney (10 milliliter volume) can be cooled down to liquid nitrogen temperature in two days without cracking/fracturing.6 Cryonics patients are much too large to be cooled to liquid nitrogen temperature over a period of days without cracking. The amount of time required for cooling vitrified cryonics patients to liquid nitrogen temperature without cracking is unknown, and would probably be much too long.

In 1990 cryobiologist Dr. Gregory Fahy published results of cracking experiments that he performed on samples of the cryoprotectant propylene glycol.4 Tg for propylene glycol is −108°C, but in RPS-2 carrier solution the Tg is −107°C. In one experiment he demonstrated that cracking began at lower temperatures for smaller samples, specifically: −143°C for 46 mL, −116°C for 482 mL, and −111°C for 1412 mL. (The last volume is comparable to the volume of an adult human brain.) Dr. Fahy also demonstrated that cracking could be delayed by cooling at slower cooling rates. But when cracking did occur, the cracks formed at the lower temperatures were finer and more numerous.

Based on evidence that large cracks formed at higher temperatures by more rapid cooling results in a relief of thermal stress that prevents the fine and more numerous cracks formed when cracking begins at lower temperature, the Cryonics Institute (CI) altered its cooling protocol for cryonics patients. CI patients are cooled quickly from −118°C to −145°C, and then cooled slowly to −196°C.5 In order to minimize or eliminate cracking in cryonics patients, proposals have been made to store the patients at temperatures lower than Tg (−124°C), but higher than liquid nitrogen temperature (−196°C).6 Such a cryo-storage protocol is described as Intermediate Temperature Storage (ITS). Alcor currently cares for a number of ITS patients at −140°C, but a consensus has not yet been reached about what ITS temperature will be chosen when this service is made available to all Alcor members.

Although Alcor’s vitrification solution M22 can prevent ice formation with some samples and protocols, M22 cannot prevent ice nuclei from forming at cryogenic temperatures. Ice nuclei are local clusters of water molecules that rotate into an orientation that favors later growth of ice crystals when a solution is warmed. Ice nuclei are not damaging, but the fact that ice nuclei can form indicates molecular mobility which could be damaging. Specifically, between the temperatures of −100°C and −135°C, ice nuclei can form in M22, with the maximum ice nucleation rate occurring near Tg. At −140°C the ice nucleation rate for M22 is undetectable. But nuclei will be probably formed in cooling to −140°C.

Although cryostorage at −140°C is an attempt to minimize cracking and minimize nucleation, this ITS neither eliminates cracking nor ice nuclei formation. Cryonics patients slowly cooled from Tg to −140°C will surely experience some ice nucleation. Alcor places a listening device (“crackphone”) under the skull of its cryonics patients for the purpose of monitoring cracking events. My understanding is that for most Alcor patients the crackphone detects cracking at Tg or only slightly below Tg, although there was reportedly one M22-perfused patient for which the first fracturing event occurred at −134°C. The propylene glycol experiments would support the view of cracking occurring slightly below Tg, but vitrified biological samples resist cracking better than pure cryoprotectant solutions.

With ice formation, cracking could occur at temperatures higher than Tg. Although ITS may prevent the formation of cracking that could occur in cooling below −140°C, it does not prevent the cracks that occur in cooling from Tg to −140°C. I have wondered whether there are forms of damage which would occur in a cryonics patient stored at −140°C that would not occur during storage at −196°C. A solid cryogenic state of matter does not prevent molecular motion. Molecular motion in a biological sample held at cryogenic temperature could result in damage to that sample.

Ions generated by radiation are much more mobile than molecules. An ionic species (probably protons) in trimethylammonium dihydrogen phosphate glass is nine orders of magnitude more mobile than the glass molecules—and sodium ions in sodium disilicate glass are twelve orders of magnitude more mobile than the glass molecules.9

Cryobiologist Peter Mazur has stated that below −130°C “…viscosity is so high (>1013 Poise) that diffusion is insignificant over less than geological time spans.” He adds that “…there is no confirmed case of cell death ascribed to storage at −196°C for some 2-15 years and none even when cells are exposed to levels of ionizing radiation some 100 times background for up to 5 yr.”10 Frozen 8-cell mouse embryos subjected to the equivalent of 2,000 years of background gamma rays during 5 to 8 months in liquid nitrogen showed no evident detrimental effect on survival or development.11

In attempting to evaluate damaging effects of temperature and radiation, it could be valuable to analyze chemical alterations, rather than complete cell death or viability. Acetylcholinesterase enzyme subjected to X-ray irradiation shows conformational changes at −118°C, but no conformational changes when irradiated at −173°C.12 X-ray irradiation of insulin and elastase crystals resulted in four times as much damage to disulfide bridges at −173°C compared to −223°C.13 Another study showed a 25% crystal diffraction lifetime extension for D-xylose isomerase crystals X-ray irradiated at less than −253°C compared to those irradiated at −173°C.14

One study showed that lettuce seeds show measurable deterioration when stored at liquid nitrogen temperature for periods of 10 to 20 years. Rotational molecular mobility was quantified. A graphical plot was generated showing increasing times for when 50% of lettuce seeds would fail to germinate as a function of decreasing temperature. Those times were estimated to be about 500 years for −135°C and about 3,400 years for −196°C.15 Translational vibrational motion has been given as an explanation for seed quality deterioration at cryogenic temperatures.16 The mean square vibrational amplitude of a water molecule is not even zero at 0 Kelvins (−273°C), and has been determined to be 0.0082 square Angstroms. The mean square vibrational amplitude is 0.0171 square Angstroms at −173°C and 0.0339 square Angstroms at −73°C.17

Realistically, however, 3,400 years is much longer than cryonics patients are likely to be stored. Storage in liquid helium at −269°C or in a shadowed moon crater at −235°C18 would certainly be more trouble than it is worth. Northern wood frogs spend months in a semi-frozen state at −3°C to −6°C, and are able to revive with full recovery of heartbeat upon re-warming.19 An empirical study of a cryoprotectant very similar to M22 (VS55)
showed viscosity continuing to increase exponentially below Tg, just as viscosity increases exponentially with temperature decrease above Tg.20 The exponential decrease in viscosity (molecular mobility) that makes ice nucleation cease at −135°C indicates that there is probably little molecular mobility at −140°C, despite the possibility of damage from ionic species or vibrational motion. All things considered, however, my personal preference is for storage in liquid nitrogen, rather than some intermediate temperature above −196°C. I would also prefer for cryogenic nanorobot repair to be at liquid nitrogen temperature.

I am by no means a nanotechnology expert, but I can give a brief description of my own views of how cryogenic nanotechnology repair of a cryonics patient would proceed. I must thank Ralph Merkle for his assistance in allowing me to consult with him to formulate and clarify many of my views. I believe that repair of cryonics patients at cryogenic temperature would be a combination of nano-mining and nanoarcheology. Nanorobots (nanometer-sized robots) would first clear blood vessels of water, cryoprotectant, plasma, blood cells, etc. The blood vessels would become mining shafts that would provide access to all body tissues. Nanometer-sized conveyor belts or trucks on rails could remove blood vessel contents. Where freezing or ischemia had destroyed blood vessels, artificial shafts would be created. Unlike the nano-mining that simply removes all blood vessel contents, the creation of artificial shafts would have the character of an archeological dig. Care would be taken in removing material to avoid damaging precious artifacts that might indicate original structure ― which could
be discovered at any unexpected moment.

Section 13.4 of K. Eric Drexler’s book Nanosystems provides diagrams and details of a nanorobot manipulator arm. Such a “diamondoid” component would contain about four million atoms, and could be fitted with a variety of tools at the end of the arm. A variety of tips with varying degrees of chemical reactivity could allow for reversible, temporary chemical bonds that could be used for grabbing and moving molecules. These could range from radicals or carbenes that would form strong covalent bonds, to boron that can form relatively weak and reversible bonds to nitrogen and oxygen, to simple O-H groups that can form even weaker hydrogen bonds. Tools for digging need not be so refined. The manipulator arm is depicted as being 100 nanometers long and 50 nanometers wide, although nanorobots would need to be larger to include capability for locomotion, computation, and power. A complete nanorobot could be as large as a few thousand nanometers in size. A capillary is between 5,000 to 10,000 nanometers in diameter, so there should be plenty of room for many such nanorobots to operate. Ralph Merkle estimates that 3,200 trillion nanorobots weighing a total of 53 grams could repair a cryonics patient in about 3 years.21,22 Like many of the calculations associated with nanotechnology, I take these figures with a pound of salt. It is certainly true, however, that it could take years to repair a patient, and that there should not be a rush to finish the job.

Merkle & Freitas have suggested that nanorobots be powered by electrostatic motors. Stators and rotors would be electric rather than magnetic. Tiny moving charged plates are easier to fabricate than tiny coils and tiny iron cores, but more fundamentally, magnetic properties do not scale well with reduced size (i.e., molecular-scale magnetic motors don’t work), whereas electrostatic properties do scale well with reduced size. Electrostatic actuators are already being used in microelectromechanical systems (MEMS).23 High density batteries could provide power for days, and recharging stations could be located throughout the patient. Alternatively, nanotube cables could bring power to the patient from the outside. Such cables could also be a means of transmitting and receiving computational data. Nanotube cables could also be used to reunite fracture faces
created by cracking. Scanning and image processing capabilities would need to evaluate what needs to be fixed.

As much as possible I would favor replacement rather than repair, which would greatly simplify the process. It would be much easier to replace a kidney than to repair the diseased kidney of an elderly patient who died of kidney disease. Curing disease and rejuvenation would thus become part of the repair of a cryonics patient. Of course, neuro patients would require an entirely new body. The brain would be the major exception to replacement strategy because the brain could not be replaced without loss of memory and personal identity.

Even within the brain, however, it could be feasible to replace many components without loss of memory and personal identity. It could be feasible to replace many organelles such as mitochondria, lysosomes, etc., and many macromolecules such as proteins, carbohydrates, and lipids. DNA could be repaired, and possibly even modified to cure genetic disease, but epigenetic expression in neurons may be critical for reconstruction of synaptic structure. Synaptic connections would not only be restored, but the quantity and quality of neurotransmitter contents should be restored. It is not simply a matter that some neurotransmitters are inhibitory and others are stimulatory. There are more than 40 different neurotransmitters used in the brain, and there must be a good reason why such variety is necessitated.

Part of the repair process could involve removal of ice nuclei, nearly all of which would be extracellular. Re-created blood vessel contents would include fresh cryoprotectant, water, plasma, and blood cells without the original ice nuclei. Although some repair scenarios favor different types of repair above cryogenic temperature, I doubt that this is necessary or desirable. Alternative repair scenarios involve splitting the brain in half, and halving the halves repeatedly at cryogenic temperature—with digitization at each step—until the brain has been totally digitized.21,22 Or digitization could be done by repetitive nano-microtomes at cryogenic temperature. The digital data could be used for full reconstruction. Some people might object that if one individual could be created from digital data, many such individuals could be created—raising questions of which are duplicates and
which is the original. There is detailed discussion of the duplicates problem/ paradox in the philosophy section of my website BENBEST.COM.

Although other repair scenarios could prove to be feasible, I believe that cryogenic nanotechnology will be required for all cryonics patients in the foreseeable future until the problem of cryoprotectant toxicity can be solved. With effective nontoxic cryoprotectants, sufficient cryoprotectant could be used to prevent ice nuclei formation at all temperatures, prevent devitrification (freezing) upon rewarming, and eliminate all toxic damage. In such a case, there could be true reversible cryopreservation (suspended animation).

What is needed to create the nanotechnology required for repair of cryonics patients? Small machines will need to build parts for smaller machines, which would in turn build even smaller machines. Many details of machine
operation must be perfected at each stage. Current modern technological civilization began with cave people pounding on rocks. Ralph Merkle has said that compared to future technology, current technology is pounding on rocks.

References

1. Chi AH, Clayton K, Burrow TJ, Lewis R, Luciano D, Alexis F, D’hers S, Elman NM. Intelligent drug-delivery devices based on micro- and nano-technologies. Ther Deliv. 2013 Jan;4(1):77-94.

2. Kudernac T, Ruangsupapichat N, Parschau M, Maciá B, Katsonis N, Harutyunyan SR, Ernst KH, Feringa BL. Electrically driven directional motion of a four-wheeled molecule on a metal surface. Nature. 2011 Nov 9;479(7372):208-11.

3. Zhang J, Lang HP, Yoshikawa G, Gerber C. Optimization of DNA hybridization efficiency by pH-driven nanomechanical bending. Langmuir. 2012 Apr 17;28(15):6494-501.

4. Fahy GM, Saur J, Williams RJ. Physical problems with the vitrification of large biological systems. Cryobiology. 1990 Oct;27(5):492-510.

5. Best B. The Cryonics Institute’s 95th Patient. Long Life. 2009 Sept-Oct; 41(9- 10):17-21.

6. Wowk B. Systems for Intermediate Temperature Storage for Fracture Reduction and Avoidance. 2011 Third Quarter;32(3):7-12.

7. Okamoto M, Nakagata N, Toyoda Y. Cryopreservation and transport of mouse spermatozoa at -79 degrees C. Exp Anim. 2001 Jan;50(1):83-6.

8. Angell CA. Entropy and Fragility in Supercooling Liquids. Journal of Research of the National Institute of Standards and Technology. 1997 March-April; 102(2):171-185.

9. Mizunoa F, Belieresa J.-P, Kuwatab N, Pradelb A, Ribesb M, Angell CA. Highly decoupled ionic and protonic solid electrolyte systems, in relation to other relaxing systems and their energy landscapes. 2006 Nov;352(42/49):5147- 5155.

10. Mazur P. Freezing of living cells: mechanisms and implications. Am J Physiol. 1984 Sep;247(3 Pt 1):C125-42.

11. Glenister PH, Whittingham DG, Lyon MF. Further studies on the effect of radiation during the storage of frozen 8-cell mouse embryos at -196 degrees C. J Reprod Fertil. 1984 Jan;70(1):229-34.

12. Weik M, Ravelli RB, Silman I, Sussman JL, Gros P, Kroon J. Specific protein dynamics near the solvent glass transition assayed by radiation-induced structural changes. Protein Sci. 2001 Oct;10(10):1953-61.

13. Meents A, Gutmann S, Wagner A, Schulze-Briese C. Origin and temperature dependence of radiation damage in biological samples at cryogenic temperatures. Proc Natl Acad Sci U S A. 2010 Jan 19;107(3):1094-9.

14. Chinte U, Shah B, Chen YS, Pinkerton AA, Schall CA, Hanson BL. Cryogenic (<20 K) helium cooling mitigates radiation damage to protein crystals. Acta Crystallogr D Biol Crystallogr. 2007 Apr;63(Pt 4):486-92.

15. Walters C, Wheeler L, Stanwood PC. Longevity of cryogenically stored seeds. Cryobiology. 2004 Jun;48(3):229-44.

16. Wowk B. Thermodynamic aspects of vitrification. Cryobiology. 2010 Feb;60(1):11-22.

17. Leadbetter AJ; The Thermodynamic and Vibrational Properties of H$_2$O Ice and D$_2$O Ice. 1965 Sep;A287:403-425.

18. Paige DA, Siegler MA, Zhang JA, Hayne PO, Foote EJ, Bennett KA, Vasavada AR, Greenhagen BT, Schofield JT, McCleese DJ, Foote MC, DeJong E, Bills BG, Hartford W, Murray BC, Allen CC, Snook K, Soderblom LA, Calcutt S, Taylor FW, Bowles NE, Bandfield JL, Elphic R, Ghent R, Glotch TD, Wyatt MB, Lucey PG. Diviner Lunar Radiometer observations of cold traps in the Moon’s south polar region. Science. 2010 Oct 22;330(6003):479-82.

19. Costanzo JP, Lee RE Jr, DeVries AL, Wang T, Layne JR Jr. Survival mechanisms of vertebrate ectotherms at subfreezing temperatures: applications in cryomedicine. FASEB J. 1995 Mar;9(5):351-8.

20. Noday DA, Steif PS, Rabin Y. Viscosity of cryoprotective agents near glass transition: a new device, technique, and data on DMSO, DP6, and VS55. Exp Mech. 2009 Oct;49(5):663-672.

21. Merkle, RC. The Molecular Repair of the Brain. Cryonics. 1994 Jan;15(1):16-31.

22. Merkle, RC. The Molecular Repair of the Brain. Cryonics. 1994 Apr;15(2):18-30.

23. Fennimore AM, Yuzvinsky TD, Han WQ, Fuhrer MS, Cumings J, Zettl A. Rotational actuators based on carbon nanotubes. Nature. 2003 Jul
24;424(6947):408-10.

11. October 2014 · Comments Off · Categories: Cryonics, Neuroscience

Cryonics MagazineSeptember-October 2012

On Saturday, July 7, 2012, I attended the Symposium on Cryonics and Brain-Threatening Disorders in Portland, Oregon. The symposium was the “brain child” of Aschwin de Wolf, who also kindly invited me to give a presentation on treatments to mitigate Alzheimer’s Disease (AD). The symposium was organized by the Institute for Evidence-Based Cryonics and Cryonics Northwest.

It has been said that cryonics arrangements are made by people who think about things other people would rather not think about – in this case, one’s personal mortality. Like the sun in the sky, we can be aware of its presence, but prefer not to look at it. Dementia is in the same category. Despite the fact that anyone who lives long enough (cryonicists are usually life-extensionists) is much more likely than not to get dementia, even cryonicists are often reluctant to plan for becoming demented. Aschwin deserves a lot of credit for not only being a cryonicist, but for organizing (with his wife Chana) the world’s first symposium/conference dealing with the subject of cryonics and dementia. It is all the more impressive because Aschwin is a man in his 30s.

The symposium required no registration, registration fee, or notification of attendance. One man attended because another attendee had informed his wife of the event while on an airplane to Portland. There were only about 30 people at the event, but the quality of the attendees and presenters was very high. The event was held at Kaos Softwear, a manufacturing company where Chana is a manager. All the talks were allotted one full hour.

Chana, who has a master’s degree in neuroscience, was the first presenter. Her topic was neurogenesis — the creation of new neurons. Although neurogenesis was discovered in 1965, because neurons are post-mitotic (are non-dividing cells), the discovery was viewed with skepticism until the discovery of neural stem cells in 1992. Neurogenesis only occurs in two discrete areas of the mammalian brain: in the olfactory system and in the hippocampus. The latter is more crucial, although the exclusion of the cerebral cortex is of great concern insofar as that is the probable location of memory, identity, and decision-making. The hippocampus prepares new memories for long-term storage in the cerebral cortex. Chana asked lots of questions for which there are yet no answers. Why does the hippocampus need to create new neurons in the creation of new memories? How is neurogenesis used? How is neurogenesis regulated? Neurogenesis declines with age, and is enhanced with exercise or ischemia. Ultimately, endogenous neurogenesis does not appear to hold much promise as a repair strategy for AD or other forms of dementia. However, it is a worthwhile endeavor to understand neurogenesis in order to guide our own attempts at neuronal repair and/or replacement.

Aubrey de Grey began his talk by acknowledging that none of the work being funded in the 2012 $4.5 million budget of his SENS (Strategies for Engineered Negligible Senescence) Foundation is focused on repairing the brain, although there is a project determining the rate of accumulation of epimutations, that is not focused on repair. He spent the first half-hour reviewing the SENS program, and the next 15 minutes explaining why 3 of the 7 SENS strategies are particularly applicable to dementia: (1) Neurofibrillary tangles and soluble amyloid in Alzheimer’s disease (AD), and their counterparts in other neurodegenerative diseases, are intracellular junk, (2) amyloid plaque in AD is extracellular junk, and (3) late-stage neurodegeneration involves cell loss. Dr. de Grey said that intracellular junk shows signs of failed autophagy. He said that most of the intracellular junk in dementia is protein. It should be easier to dispose of than the cholesterol degradation products which are the focus of SENS lysosome work on atherosclerosis, but which are not properly delivered to the lysosome. He outlined the circumstantial evidence that the main problem may be the same as in atherosclerosis, i.e. oxidized cholesterol poisoning the lysosome. He spoke of the current clinical trials for having microglia eliminate extracellular junk (amyloid plaques). The first human clinical trials had shown great promise, but were halted because 5% of the patients developed brain inflammations. The newer trials have apparently corrected that problem. Aubrey noted the widespread belief that the amyloid would be removed without being of much benefit – expressing his belief that this misses the point, because major postponement or reversal of AD will require fixing all three main problems, hence lack of benefit from fixing one is not evidence that that one need not be fixed. I am one of the skeptics because follow-up autopsies on the first trials showed that even when amyloid plaques had been completely removed, no reduction in degeneration had occurred [THE LANCET; Holmes,C; 372:216 (2008)]. By the time AD is diagnosed, neurodegeneration is too far along to be helped by removing amyloid (though there is rapid progress in improving very early diagnosis). Immunization to remove amyloid would be more effective if begun in the 20s or 30s, much like shots for measles or polio — as prevention rather than cure. Although amyloid may serve a positive function in repair or it would not have evolved. [Aubrey notes: who says it evolved? “Aging is a product of evolutionary neglect, not evolutionary intent” (Hayflick)]. Concerning cell loss, Aubrey was sanguine about Jean Hebert’s work exploiting the fact that certain neural progenitor cells are highly migratory, potentially facilitating widespread distribution of new neurons throughout the neocortex via stem cell therapies. Even if neurons can be replaced in the neocortex, I wonder how that would compensate for the loss of synaptic connections and strength of synaptic connections. Of the three approaches mentioned by Dr. de Grey, I would say that removal of intracellular junk has the best chance of being of benefit on its own, because it is the neurofibrillary tangles that tend to cause cell death rather than the amyloid plaques, which are an upstream event.

My talk was basically a summary of the “Alzheimer’s Disease: Molecular Mechanisms” page in the life extension section of my website BENBEST.COM. I wrote the page in 2003 between leaving my job as bond database support for Scotiabank in Toronto, Canada, and becoming president of the Cryonics Institute in Michigan. For the subsequent 9 years I have become increasingly displeased about how out-dated the webpage was becoming. So I was pleased at the opportunity to do the massive research required to update that webpage for this symposium. Unfortunately, it was all I could do to finish the updating before catching my flight to Portland. Aschwin and Chana allowed me to crash at their condominium. I missed the Friday evening social for those attending the symposium because I spent all evening and a couple of hours the next morning creating my PowerPoint. I was pleased with the result, however, and pleased with the presentation I was able to deliver.

I encourage anyone interested in the content of my talk to consult my Alzheimer’s webpage because that page has detailed linkable references which I could not include in my presentation. I believe that the most promising therapy is the targeting of copper with PBT2, which removes copper from amyloid without chelating essential element metals. Etanercept, which antagonizes the inflammatory cytokine TNF-alpha has also shown promising results. Possibly also, passive immunization with tau antibodies would be of greater benefit in stopping neurodegeneration than immunological approaches against amyloid. Concerning prevention, exercise, curcumin, pomegranate juice, and folic supplementation have shown good results. Seemingly conflicting results would indicate that ginko biloba can slow cognitive decline in Alzheimer’s patients, but is of no benefit in preventing the disease.

Mike Perry’s topic was Early Detection of Alzheimer’s Disease. On that subject he reported that the CerebroSpinal Fluid (CSF) is low in amyloid beta and high in phosphorylated tau protein. I had put much more detail on this subject into the biomarkers section of my webpage on Alzheimer’s Disease – which I showed to Mike later in the day. In his presentation Mike noted even for people who do not get AD, dementia of some kind is still very probable with aging. He commented that AD is not a terminal illness, which is defined as an illness in which two physicians have certified that the patient probably has no more than six months left to live. No AD patient dies of AD — the cause of death is usually infection (pneumonia, bedsores, urinary tract infection, etc.). I expressed concern that suicide by VSED (Voluntary Stopping of Eating and Drinking, as Mike calls it) by an AD victim could lead to autopsy. Mike denied that this was necessarily the case.  I was told that for anyone who had died by refusing food and water the cause of death would be obvious, and no autopsy would be required, though circumstances and policies will vary. Mike Darwin, however, noted that VSED could be harmful to the brain as cardiac arrest draws near, due to low respiration rates. Aschwin responded that this kind of brain damage is still relatively benign in comparison to the alternative (advanced dementia). James Swayze, who is a paraplegic with cryonics arrangements and was in attendance at this event, has expressed concerns that dehydration causes brain damage. Dehydration may reduce brain functionality, but brain dehydration is a key process in removing water from the brain in the vitrification point of view and is probably a benefit rather than a harm for cryonics purposes.  Alzeimer’s patients nearly always die of infection, and because infection may also occur early in the disease,  Mike Darwin recommended that anti-microbial treatment be refused by an Alzheimer’s victim as a way of hastening cryopreservation. If infection does not occur early in the disease, however, refusing antibiotics may not produce the desired result.

Keegan Macintosh, who recently graduated from a Canadian law school, presented on the subject of Thomas Donaldson’s 1988 lawsuit in California to be cryopreserved before his brain cancer destroyed too much of his brain to make cryonics a worthwhile effort. Keegan criticized the attorneys involved in the appeal for arguing that Donaldson’s right to “premortem cryopreservation” stemmed from a constitutionally-protected right to assisted suicide, rather than the right to pursue a risky, but potentially life-saving procedure. By framing the case this way, the Court was able to avoid having to consider Donaldson’s unique and crucially relevant motive, and thus the possibility of cryonics succeeding, for him or anyone else. Acknowledging, however, that options for assisted suicide could be of use to cryonicists with brain-threatening disorders, Keegan examined developments in American law on the issue, and then turned to Canadian jurisprudence. He pointed out a number of potentially significant differences between the U.S. Supreme Court’s substantive due process analysis in the more recent physician-assisted suicide cases, Washington v. Glucksberg and Vacco v. Quill, and Supreme Court of Canada’s approach to section 7 of Canada’s Charter Rights and Freedoms (right to life, liberty and security of the person) in Rodriguez v British Columbia, and cases since. The government’s position is presumably influenced by a desire to avoid a “slippery-slope” that disvalues human life. Keegan noted that although formerly other countries looked to the American Constitution for guidance, Canada’s constitution is now the world’s most popular role-model. Section 2 of Canada’s Charter of Rights and Freedoms emphasizes “freedom of conscience and religion.”

What would be the effect of someone acting on the belief that pre-mortem cremation is the road to salvation? “Freedom of conscience” implies that secular morality is as important as religious belief and there is some emerging jurisprudence to that effect. Would the belief that good-quality cryopreservation is necessary to live again at some future time not then receive equal protection to analogous beliefs and practices of religious origin? Keegan believes that an appeal such as Donaldson’s – and indeed any constitutional challenge against a law impeding access to cryonics – would have a better chance of success in Canada than in the United States.

Max More spoke without slides on the subject of “Survival, Identity, and Extended Mind.” The objective of Max’s talk was to consider how it could be possible to back-up personal identity-relevant information and then reintegrate that information to restore personality if cryopreservation has been imperfect. If cognitive processes and their inputs can be external to the brain, Max would like to take advantage of this to improve the chances of reviving people suffering from brain-threatening disorders. Andy Clark and David Chalmers wrote an authoritative paper entitled “The Extended Mind.” According to Max, for an outside object or process to be considered part of the mind, it has to produce results that are reasonably comparable to the components normally seen internally and biologically/neurologically. Clark and Chalmers propose three conditions for considering externally-located processes to be part of an individual’s cognitive processes: 1) constancy (the external component has to be there reliably); 2) accessibility (a natural ease of use of that component); and 3) automatic endorsement (the person must trust the component as they would trust any comparable part of their natural body). Max noted that a few years before the Clark/Chalmers paper he had considered the related issue of when an external technology could be considered part of the self (in chapter 4 of his dissertation: “Technological Transformation and Assimilation”). Although Max doubted Ray Kurzweil’s claim that an externally-convincing simulation of his father (made out of traces available) would actually have a self, Max did not argue that no well-simulated person could have a self. Max suggested that a notebook could be part of the thinking process, rather than just a tool. Nonetheless, he was dubious about the value of keeping lots of diaries, although it has been suggested that biographical information could assist in reconstruction of a cryonics patient and that cryonics organizations should take a more proactive role in fasciliting storage of identity- and memory relevant information. Max was also dubious that a computer that could convincingly simulate a person would have a self. He raised the question “What is self?” He referred to David Hume’s claim to introspectively only be able to discover thoughts and feelings, but no self. Dennett called self an illusion. This would lead me to believe that neither Hume nor Dennett should have much concern with their own survival (like most people?). Max said that he could lose a few memories without feeling his self was compromised — because he believes that personal identity is more than memory. It includes dispositions, values, and so on.

After the presentations there was a panel of all the presenters, plus Aschwin the host. I requested that each panelist explain what they would do if diagnosed with AD. Aschwin said he would immediately proceed to terminate his life under conditions favorable to cryopreservation provided that the diagnosis was credible and there are no short-term cures on the horizon. Keegan said that he would see first what, if any, time he had before symptoms such as apathy and denial would be expected to set in, and take some conservative portion of that time remaining to spend some quality time with family and friends. Keegan noted that, despite our best efforts, cryonics may not work, and thus it is rational to seek meaningful experiences in the moments one knows they have left, if such can be done without irreparably compromising one’s cryopreservation. I noted that Robert Ettinger also said he would terminate life by hypothermia in a cold bathtub at the end of a party with friends – but delayed such an action to the point where he lost consciousness and lost the ability to do any such thing at the age of 92 when he deanimated. I said that I would probably spend about a year attempting to confirm the diagnosis, and might delay further trying to determine if a cure was possible or forthcoming soon. Max said that he would want a second opinion, but like Keegan wanted to have some joyful time before self-termination. Chana said that she would be very concerned about how the decision to self-terminate would affect others, in particular how to explain to her family why she was ending her life while outwardly being in good health. Chana and Aschwin spoke of being sensitive to each other’s feelings about the matter. Aschwin noted that those who care for AD family members to a natural death often suffer from severe caregiver depression. Chana said that once she had decided to pull the plug that she would “find a way to take a bath in Agent Orange and take advantage of Oregon laws.” By this she meant she would find a way to give herself an aggressive form of cancer that would cause two Oregon physicians to declare that she is a terminal patient. Once this is done, an Oregon physician can write a prescription for phenobarbital which the patient can use for suicide by overdose at the time and place of their choosing. Aubrey said that he would delay the decision without worrying too much about loss of neurons. In addition to delaying because of diagnosis confirmation and evaluating hope for a cure within a short time period, Aubrey added evaluating the likelihood that cryopreservation procedures would be improved by waiting. Mike Perry said that he would try to confirm the diagnosis and if sure about it, “get it [deanimation] over with as soon as possible.”

I mentioned the case of a CI Member dying of cancer who, with her husband, called Suspended Animation, Inc., to be present at their suicide. Her 30-year-old husband was in good health, but did not want to live without his wife and planned to die along with her. CI terminated both their memberships and established a policy of reserving the right to cancel cryonics contracts in cases of suicide. Aschwin strongly disapproved of this CI policy. In his opinion, cryonics organizations should never encourage or condone suicide but should not refuse cryopreservation to those who have taken their own lives. I believe cryonics organizations cannot be seen as encouraging the hastening of death on the ground that cryonics may work, and must ensure that others do not get that impression. Not enough was said about what policies would be most appropriate for cryonics organizations.

I asked Aubrey if he thought that an AD patient would ever be so advanced that SENS could not save the self. Aubrey agreed that could happen, but it would be difficult to say when. The case is similar with straight frozen patients or patients with varying amounts of ischemic damage. The concept of “information theoretic death” is meaningful, but difficult to determine. Even if SENS methods could not recover enough memory and identity to save a person, some future molecular archeology might be able to do so.

There was some discussion about the most promising treatments for Alzheimer’s disease. Aschwin pointed out that both early-onset Alzheimer’s and late onset Alzheimer’s have a strong genetic component, which should favor the use of gene therapy.

Mike described the activities of the Venturists, which is offering to save Venturist Members who are being cryopreserved by a cryonics organization that fails. Another project of the Venturists is that they are seeking $50,000 for Mike Darwin, who lost his cryopreservation arrangements with Alcor due to financial difficulties.

14. January 2013 · Comments Off · Categories: Cryonics

I moved to Florida a number of months ago, but I am only now starting a cryonics group in the Broward County, Florida area. As a first event I have booked a table at the Peking Tokyo Buffet restaurant on 1219 South Federal Hwy, Deerfield Beach, Florida for dinner in the early evening (7 P.M. to 9 P.M.) on Thursday, January 24th. All those interested in cryonics in Broward county and adjoining areas are invited to attend.

This will be the first meeting of this group so the main goal will be to meet other cryonicists or cryonics-interested people in the South Florida area. All-you-can-eat buffet with a wide selection of foods for only $10.95, but no purchase or meal is required for those who simply want to socialize and discuss cryonics while others eat. Drop-in any time between 7 P.M. and 9 P.M., but closer to 7 P.M. would be preferred. Use exit 41 from I95, drive East along SW (becomes SE at Dixie) 10th Street, and turn right on Federal Highway (US Route 1) to reach the Peking Tokyo Buffet in Deerfield Beach. Anyone interested in cryonics is welcome to attend.

I have been phoning cryonicists in the area and quite a few seem certain to attend. I have also started groups on Facebook and Meetup for this purpose, which I invite others to join, but don’t get the wrong impression that the buffet event on Thursday, January 24th is going to be as poorly attended as the Facebook or Meetup groups might make you imagine.

Here is the Facebook group and event:

http://www.facebook.com/groups/cryonics.boward/

http://www.facebook.com/events/522418457791831/

Here is the Meetup group and event:

http://www.meetup.com/Cryonics-Meetup-of-Broward-County-Florida/

http://www.meetup.com/Cryonics-Meetup-of-Broward-County-Florida/events/98480242/

For those wondering about my move to South Florida, here is some background information.

21. June 2012 · Comments Off · Categories: Cryonics, Science

From June 3 to 6, 2012 I attended the annual Society for Cryobiology meeting, which in 2012 was held in Rosario,Argentina.

Attending with Argentine biogerontologist and Cryonics Institute Member Rudy Goya may have reduced the interaction I had with the cryobiologists. There were fewer sessions than usual, and thus more free time. The welcome reception was not held until the evening of the first day of the sessions.

The first session dealt with an aspect of Argentine cultural heritage, the Llullaillaco children — three Inca children who had been mummified by dehydration high on a volcano and preserved for over 500 years. Two of children were selected by the Incas because they were “perfect” (beautiful and pure) at 6 or 7 years of age. It was believe to be an honor to go directly to heaven, not really death or sacrifice. The children were given an intoxicant and buried alive atop the Llullaillaco volcano. Much of the session focused on the conditions that caused the children to be so well-preserved, and the conditions the curators should use to preserve the children for the future — involving careful regulation of temperature, atmosphere, humidity, and an environment inhospitable to most microbes.

If reanimated cryonicists receive anything like the care these children are receiving, there should be no concerns about being welcome in the future. In a sense, the Incas had it right when thinking they were sending the children to heaven. Of course the Inca children were deprived of life and are unable to experience or enjoy their treatment by modern curators — and cryonicists should not encourage hastening death based on reliance on unproven future technologies.

At this conference there were special “How to do it?” sessions overlapping part of the lunch hour that focused on practical techniques unrelated to the experimental results and theoretical considerations covered in the regular sessions. Sunday’s topic was proteomic analysis, which covered removal, isolation, and identification of proteins from cells. The presenter (from the Institute of Molecular Cell Biology in Rosario) claimed that instrumentation allowing high throughput and resolution had given proteomics a maturity comparable to genomics.

The afternoon sessions were concerned with cell and tissue preservation. Elza Cabrita reported on improved cryopreservation of fish sperm through a combination of cryoprotectants and antioxidants. Locksley McGann reported on experiments sequentially exposing human articular cartilage to four CPAs (DMSO, glycerol, propylene glycol, and ethylene glycol) at lowering temperature (0ºC, −10ºC, −15ºC). Vitrified samples were cooled to liquid nitrogen temperature, and demonstrated 75% cell recovery when rewarmed.

Adam Higgins reported on an improved procedure for washing glycerol from red blood cells. Currently about 99% of banked blood is stored at refrigerator temperature (2-4ºC), with a shelf life of 42 days. Only 1% of blood (mostly rare blood types) is cryopreserved with glycerol and stored at −80ºC, with a shelf life of ten years. A major deterrent preventing more blood from being banked at −80ºC is the 30-60 minute glycerol washout procedure. Adam’s group developed a procedure that can wash the glycerol out in 30 seconds, but 5 seconds longer or shorter results in too much hemolysis. A three minute washout procedure is less time sensitive (one minute longer or shorter is tolerable), but the method needs to be scaled-up from the 0.5 milliliter test volumes being used.

On Monday, Peter Mazur reported that in vitrifying mouse oocytes, it is the warming rate and not the cooling rate that is most critical for success. He spoke of microwave warming and the problem of thermal runaway (uneven warming). Ice blockers would not cross cell membranes, and thus would not be of use against intracellular ice formation. Pier Morin reported on miRNA microarray assessment of miRNA expression of the freeze-tolerant insect goldenrod gall fly at control (+5ºC) and freezing (−15ºC) temperatures. mIR-210 was down-regulated and mIR-1 was up-regulated at freezing temperature (the latter is involved in cell cycle regulation).

Ali Eroglu reported on epigenetic perturbation resulting from human oocyte cryopreservation techniques. Both the slow freezing and vitrification methods he used resulted in down-regulated expression of H19 and Ube3a genes. Igf2r was down-regulated by vitrification, but not by slow freezing.

Monday’s “How to do it?” session described a combination of nanotechnology and stem cells for tissue engineering. Specifically, electrospinning can be used to create a nanometer scale web of biodegradable fibers that can be populated with mesenchymal stem cells by electrospraying. The main challenge is vascularization of the tissue. Vascular Endothelial Growth Factor (VEGF) increases cell adhesion, but not necessarily vascularization.

Barry Fuller reported on successful hypothermic perfusion of liver. A kidney hypothermic perfusion machine has been in operation for ten years, but liver has been more challenging, because of its large size and the fact that two vessels supply the organ (hepatic artery and portal vein). The liver hypothermia perfusion machine uses two pumps.

PhD student Na Guan described her study of gene expression changes associated with chilling injury of rat liver slices. Cryoprotectant solutions supplied by 21CM (Greg Fahy) were used to ensure no ice formation interfered with the process. ATP levels indicated that the cryoprotectant solutions used were causing no damage, although the composition of those solutions was not disclosed. 1108 genes were observed, of which 251 were up-regulated and 77 were down-regulated by chilling at −15ºC. Focusing on the top ten up- and down-regulated genes: inflammatory and DNA repair genes were considerably up-regulated, and genes associated with biosynthesis of cholesterol and polyunsaturated fatty acids were down-regulated. The latter seems paradoxical in light of the up-regulation of cell surface-linked signaling pathways, which indicate cell membrane injury.

During the question period, both Andreas Sputtek and Arthur Rowe were sharply critical of the undisclosed composition of the 21CM cryoprotectant solutions being used. Sputtek said that because science is about disclosure of methods and materials, Guan’s work was not science. Guan said she had begged 21CM for disclosure, but said she was told that anyone wanting to replicate the experiments could buy the solutions from 21CM. Tiantian Zhang said that gene analysis only done 30 minutes after chilling injury does not give the whole picture. She said that in her own work doing gene analysis of fish oocytes or embryoes after chilling injury, gene expression changes dramatically with time — that it is a mistake to only analyze the expression 30 minutes after exposure as Guan had done. After the presentation, Arthur Rowe spoke with Guan telling her how much trouble he has had over the years with her collaborator (Dr. Fahy) in connection with the non-disclosure issue. I spoke with Guan myself after her presentation. She told me that the greatest chilling injury occurs at −90ºC. She also said that she would be getting her PhD in July and did not know who would be continuing her work. When I spoke to Dr. Fahy about the presentation, he told me that the composition of the vitrification solution had been disclosed and that Guan was mistaken in believing that she could not disclose the composition.

Tuesday morning had been scheduled to begin with a lecture by Ken Storey. Storey typically has no interest in what other cryobiologists have to say, is fairly ignorant of areas of cryobiology outside of hibernation and effects of low temperature on animals in nature, and only comes for his own presentation before leaving. His ignorance is on display when journalists get him to do cryonics-bashing, which he does with relish, but the general public only sees the comments of a respected cryobiologist, not the ignorant misunderstandings of cryobiology. I would not have expected Storey to come all the way to Rosario, Argentina only for his own presentation, but this is what he attempted to do — and he missed one of his flight connections. Ironically, this year Storey was honored by being made a Fellow in the Society for Cryobiology. Storey does, admittedly, have a fabulous knowledge of molecular biology, and is an outstanding scientist in connection with his own work.

To compensate for Storey’s absence the conference organizers arranged a makeshift follow-up session on the Llullaillaco children. This wasn’t entirely a waste, because many issues had not been addressed in the first round. I was going to question using a 2% oxygen and 98% nitrogen atmosphere for the children rather than pure nitrogen, but Barry Fuller raised this objection before I was called upon. I did, nonetheless, suggest that the goal should be to perfect the preservation environment rather than try to recreate the conditions of the mountain. Even this had not been done because the relative humidity had been raised to 70% on the bad advice of an expert rather than held to the 40% present on the volcano. The children were reportedly gaining 300 grams per year, probably from the humidity. There is a lower humidity limit below which no microorganisms can grow, but 0% relative humidity in the −20ºC preservation chambers would run the risk of freeze-drying.

For the second session on Tuesday, John Crowe had been scheduled to lead a symposium composed of 3 other speakers besides himself, but all of the other 3 speakers cancelled-out. John, nonetheless, did an excellent job of speaking for the whole session on the basis of his own work. John is an expert in dehydration and freeze-drying of organisms as well as on tardigrades and trehalose. Drying DNA with trehalose prevents fragmentation, and drying proteins with trehalose prevents denaturation. John discovered that drying liposomes with trehalose prevents membrane fusion — although he lost most of the patent rights on commercially valuable processes by publishing too soon. Dehydration of samples containing sucrose drives the glass transition temperature (Tg) from 20ºC to 60ºC, but dehydration of samples containing trehalose raises the Tg from 20ºC to 120ºC. More recently, however, it has been found that LEA proteins can be as protective as trehalose, but in a way that is distinctive and complementary to trehalose — stopping liposome fusion, preventing protein aggregation, and changing sample Tg. Yeast cells are protected against dehydration damage not only by trehalose, but by the trehalose transporter protein which exports the trehalose to the exterior membrane surface and imports the trehalose to the internal membranes of organelles such as mitochondria. But although the genome of tardigrades has been sequenced, the tardigrade trehalose transporter has not yet been identified.

Barbara Reed is probably the world’s foremost expert on plant cryopreservation, and she has spoken a lot about the benefits of antioxidants for cryoprotection. But the presentation Barbara gave on Wednesday gave me the strongest indication that oxidative stress could be a significant mechanism of cryoprotectant toxicity. Not only because a variety of cryopreserved plants show improved viability with Vitamin E, Vitamin C (if iron is removed), lipoic acid, glutathione, and melatonin — but because oxidative damage was shown to increase significantly associated with cryoprotectant loading.

Roland Fleck works with the UK Stem Cell Bank. The Bank conducted studies indicating that a 2-step freezing protocol results in better viability than vitrification. But examining the results of 8 technicians showed that in the hands of the most experienced technician vitrification was as effective as the 2-step freezing protocol. Protocols should not be so highly dependent upon technician expertise. After his presentation, Roland told me he was concerned that he was only able to assay viability by the use of trypan blue, which only indicates membrane integrity and does not provide a very fine measure of cell function. He said that the requirement to use the trypan blue viability assay was imposed by bureaucrats or scientists who do not have much knowledge of cryobiology.

Igor Katkov said that he believes any sperm cell can be vitrified simply by choosing the right cooling and warming rate. He said he was advised by his patent attorney to drop seven slides from his PowerPoint presentation.

At the business meeting the Society membership was reported to be down to 186. The journal CRYOBIOLOGY continues to be profitable. CRYOBIOLOGY has a 33% rejection rate, a 1.83 impact factor, and 33 Members on the Editorial Board. The Society has $300,000, which the IRS thinks is too much for a charitable organization, but the IRS is allowing the society to retain tax-exempt status. Increasing travel awards is the preferred use of money, but there is a problem that on the one hand travel awards are a taxable benefit, and on the other hand it is illegal to pay the taxes on travel awards. The 2013 conference (the 50th annual conference) is to be held in Washington, DC, where the first conferences were held. The 2014 conference might be Istanbul, Turkey and the 2015 conference could be in Isreal, but definite decisions have not been made.

Last year’s new Society for Cryobiology Fellows Barbara Reed and John Crowe each gave presentations reviewing their careers. Barbara Reed began as a plant biologist in 1985, but was brought into the field of cryobiology by a need to preserve germ tissue. John Crowe said that after the Sputnik shock of 1957 the US government sought to encourage more young people to go into science, including him. As a teenager, John was sent to a number of different science laboratories on his summer vacations. John considers himself more of a “dryobiologist” than a cryobiologist. He entertained us with photos taken in the many exotic countries he and and his wife have visited since his retirement.

The two new Fellows for 2013 are Ken Storey and Mehmet Toner.

This conference was attended by not more than about 80 people, at least half of whom were South America. There were maybe 30 or so hard-core Society for Cryobiology Members. This was my 9th annual meeting in a row, but for the most part I made little effort to relate to the cryobiologists, although one of my intentions in attending these meetings has been to soften the hostility of cryobiologists to cryonicists. I sat near the front of the meetings with Rudy who told me that he learned a great deal about the cryobiology behind cryonics practices by attending this conference. Very many of the cryobiologists were reporting on using vitrification at this conference, and including articular cartilage and plant tissue as well as single cells. I was fairly active in my questioning and comments — about which a few of the cryobiologists complimented me.

I lost my sense of urgency about talking to Peter Mazur. Peter recently told a journalist that although it is not possible to prove that the chance of cryonics patients being reanimated are zero, “you can, I think demonstrated that the probability of its being done is so extremely low that effectively it is zero”  [CANADIAN MEDICAL ASSOCIATION JOURNAL; Monette, M; The Church of Cryopreservation; 184(7):749 (2012)] I am curious about the demonstration Peter has in mind, but I am also committed to learning from cryobiologists rather than arguing with them about cryonics. Peter walked away a few years ago when I asked him when solution effects rather than mechanical damage cause injury to cells due to slow cooling, so that may be a touchy subject with Peter as well.

I did, however, pepper John Crowe with questions — finding him to be friendly and informative. John confirmed what Peter Mazur had told me about cells being able to tolerate the loss of all osmotic water (freezable water, which constitutes at least 80% of cell water) without injury — a matter of great relevance in the vitrification of cryonics patients (assuming inter-cellular effects are not of great significance).

I sought-out Ali Eroglu, with whom I have had little interaction in the past, calling his attention to an article in the most recent issue of CRYOBIOLOGY about transfection of mammalian ovary cells with trehalose [CRYOBIOLOGY; Chakraborty,N; 64(2):91-96 (2012)]. Ali has microinjected oocytes with trehalose (along with low concentrations of DMSO to protect the mitochondria) [BIOLOGY OF REPRODUCTION; Eroglu,A; 80(1):70-78 (2009)]. Ali had not seen the CRYOBIOLOGY article, but he told me that ovarian tissue is easier to work with than oocytes.

At the final banquet I sat next to one of the conference organizers. He told me that John G Baust had been supposed to conduct a symposium, but had cancelled the whole thing a month before the conference without giving any explanation. He agreed with the comments I had made about the Llullaillaco children, and told me that a committee of cryobiologists was going to supplement the questionable advice that the Argentine government has been getting from a single advisor in New York. He told me that National Geographic had discovered the children and attempted to remove them from Argentina on a midnight flight, but the Argentine government got wind of the plan and intervened. Nonetheless, the children were simply kept in −20ºC freezers for several years while planning and building better preservation chambers.

The return bus trip to BA on Thursday took the entire afternoon — much longer than I would have expected. I sat next to Adam Higgins on the bus, and spoke with him much of the time, mostly about his life and work, as well as about our experiences in Argentina. Adam knew Spanish fairly well because he has spent four months of language immersion living in Equador (and visiting the Galapagos Islands). If he gets a patent for deglycerolizing blood, the University would get half the royalties and he would split his half with his collaborators. The advantages of his method would be the ten year rather than 42-day shelf life for banked blood, and the greatly reduced washout time. The latter is a significant savings in labor costs, but would have to be weighed against greater electrical costs for a −80ºC freezer as opposed to refrigeration. Even if he is successful in perfecting his methods, he thinks that the blood banking industry is too conservative to be captivated by superior storage methods. Adam has attended most of the annual conferences since I began attending in 2004, and told me that he would like to become a Governor of the Society. Not once did Adam ask me what work I do, and he evidently does not know because he was surprised when I told him I am not a Member of the Society for Cryobiology. Whether or not I am formally accepted as a Member, my attendance at these conferences is implanting me into the consciousness of the cryobiologists as being a member of their community.

On October 27-29 I attended CR VII, the 2011 Calorie Restriction Society Conference held in Las Vegas, Nevada.

Members of the Calorie Restriction Society restrict their calories while maintaining adequate nutrition as a means of extending their lifespan (or improving their healthspan), as has been proven to work in lower animals.

Although I was still in a wheelchair as a result of falling from a ladder and hip surgery, I got my airline to give me handicapped-support (wheelchair assistance), and I rented a wheelchair in Las Vegas.

CR VII was the seventh CR Society conference held in the ten years since the first such conference was held in the same city, in the same hotel, and in the same meeting-room ten years earlier in 2001. Thursday, October 27 featured presentations by Calorie Restriction Society Members, whereas Friday and Saturday featured presentations by PhD scientific researchers. I am a CR Society Member, so I was invited to speak on cryonics on Thursday. It was a small conference, so there were not many more than forty people attending on any of the days.

My presentation was preceded by a presentation by Peter Voss, who is both a CR Society Member and a Member of Alcor. Peter and his companion Louise Gold were the only CR Society Members other than me attending  the conference who are cryonicists. Peter spoke of the ultimate goal of indefinite lifespan, sharing his wisdom based on his experience practicing calorie restriction, describing cryonics as a “safety net of unknown fabric”, and mostly speaking of his goal of developing Artificial General Intelligence to accelerate research in life extension technologies. Concerning his CR practice, he noted that CR is not binary, and that people receive the benefits to the degree that they restrict their calories. He said that he does not count calories, but simply weighs himself and adjusts his calories appropriately, which is the practice I have adopted. Peter is not worried about hostile AIs because he believes that rationality is positively associated with morality. (See http://www.adaptiveai.com/ for a sample of Peter’s work.)

Although it was not a large group, I expected that such a group of dedicated life extensionists willing to go to extremes in restricting their calories would be very receptive to the practice of cryonics. On the other hand, Shannon Vyff warned me that although CR Society Members can be enthusiastic to hear about cryonics, they don’t sign-up. I gave considerable thought to the marketing aspect of my presentation. I decided to be very up-front about being a salesman, while nonetheless attempting to side-step salesmanship (and sales resistance) by concentrating on the technical issues and encouraging a technical discussion (although I did mention prices and insurance funding).

Alcor Member (and long-time cryonics promoter) Brenda Peters lives in Las Vegas, so I invited her to be my guest at the CR Society Conference. My thought was simply that Brenda and I could renew our friendship while enjoying the conference together.

I began my presentation by describing my and experience and mistakes in practicing calorie restriction as well as my fall in September which resulted in hip surgery and no prospect of walking again for many weeks — and how this had interrupted by exercise/CRAN program. When I asked who felt familiar with their knowledge of technical issues of cryonics, I was surprised that none of the non-cryonicists raised their hands.

After giving my presentation of the technical issues in cryonics I asked the audience to pair-up to discuss both their understanding of my presentation, and reasons they may have for thinking that cryonics may not work. After the paired discussions I asked for questions and objections. Brenda was more enthusiastic than I expected about raising her hand to comment. I somewhat bluntly said that I would rather hear from anyone but her, which was apparently confusing to people who weren’t aware that we knew each other. I was wanting to hear the unvarnished objections to the idea of cryonics which CR Society Members might have. I did not mean to hurt Brenda’s feelings, and I blame myself for not discussing my expectations with her beforehand. I did, nonetheless, allow Brenda to speak a couple of times.

It proved to be hard work getting CR Society Members to explain whatever objections they might have to cryonics. One fellow expressed his belief that not enough is known about the mind to know that cryonics can preserve it. I replied that the mind is based on the synaptic “connectome” and that minds recover from low-temperature surgery in which there is no electrical activity in the brain. Another fellow wanted to hear the experimental evidence that cryonics patients have been revived, to which I could only reply that cryonics is dependent on technologies which do not yet exist, and that revival seems inevitable to me if technology continues to progress and the anatomical basis of mind is preserved. One man believed that dogs had already been cryopreserved and revived, but I corrected his misconception by stating that the dogs have only been revived from cooling down to just above the melting temperature of water. When someone said that most businesses don’t last long, I replied that it is a mistake to compare the durability of cryonics organizations to efforts to start a diner in a location where the success is uncertain. One woman raised the overpopulation issue, which I noted is no more a plausible threat than the danger that too many people will practice Calorie Restriction. I added that the same logic would ban all medical research, especially research into preventing infectious diseases.

Although there were not many objections, neither did I hear much enthusiasm for cryonics. Perhaps they were stunned by an unfamiliar idea, and it takes time for resistance to be overcome. I had been hoping for some sign-ups. I had placed Membership forms on the literature table. It was as if they had no objections to cryonics, but still weren’t interested. Which left me thinking that I shouldn’t have asked for reasons why they think cryonics won’t work, but instead asked for reasons why they won’t sign-up.

A number of people complimented me on the quality of my presentation. But during subsequent discussions with CR Society Members at the conference, I heard further objections to cryonics. One CR Society Member told me that he hoped my presentation would motivate him to sign-up for cryonics. He said that he had mentioned cryonics to his mother several years ago, but she was freaked-out by the thought of being reanimated in a strange and alien world. Since then she had become demented, and he thought it would be wrong to foist cryonics upon her while she is in that condition.

Another CR Society presenter spoke of his project to develop an eco-friendly farm with local barter and community-building that would be sustainable through the disastrous global warming and prolonged depression he was expecting. His bleak vision of the future of technology left no possibility for cryonics, but at least he corrected himself when he started to say “cryogenics”.

Another fellow I spoke with later was concerned that cryonics organizations could not survive in light of the acrimony he saw between Members. His biggest concern, however, was that people of the future would be vastly superior, and treat him with contempt or worse upon his revival. A female CR Society Member told me that she is restricting calories entirely to increase her health-span, not her lifespan. She does not think that life is very good, and she has the hope and belief that the afterlife will be better.

Over lunch, one fellow suggested promoting cryonics as a means of cutting the astronomical health-care costs that so many people incur in their last year of life. I replied that any association of euthanasia with cryonics or any hastening of death on the expectation that cryonics may work would be disastrous for cryonics — and all the moreso if done as a cost-cutting measure.

I had difficulty moving around in the conference room due to the tables and my wheelchair, which made it difficult to chat with people during breaks. I had a similar problem during meal breaks. Whether I would have gotten a better understanding of why no-one seemed eager to sign-up for cryonics if my mobility had been better remains to be seen. I would think that after years of giving presentations about cryonics I would become blunted to lack of interest, but each such experience remains uniquely poignant and disappointing.

I learned much from the scientific presentations, but I won’t attempt to summarize very much. I was, however, very impressed by the extent to which a linkage was made between the blockage of the insulin/IGF-1 pathways in lower organisms and the practice of calorie restriction by humans. There is evidence that protein restriction may be the essence of calorie restriction, and that low protein diets are associated with reduced levels of IGF-1, but only when protein is less than 12% of macronutrients. Increasing insulin sensitivity seems to be the key to extending lifespan, yet although exercise is the most powerful intervention increasing insulin sensitivity, exercise does not increase lifespan.

Stephen Spindler and Luigi Fontana are scientists who have a long and intimate relationship with the CR Society. Both were speakers at this conference. Luigi in particular has been conducting studies on the physiology of long-time calorie restriction practitioners, and the benefits that are seen in the risk factors for various aging-associated diseases. He has published many studies of this research:

http://www.ncbi.nlm.nih.gov/pubmed/21402069

http://www.ncbi.nlm.nih.gov/pubmed/21841020

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2724865/

http://ajpheart.physiology.org/content/294/3/H1174.long

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2829867/

A DVD of the presentations is being made by the CR Society, and will be available for sale within a few weeks, I expect.

August 31 to September 4, 2011 I attended fifth biannual SENS Conference (SENS5, Strategies for Engineered Negligible Senescence) at Cambridge University in the United Kingdom.

People who attend SENS conferences are the demographic that is the most receptive to cryonics of any identifiable group I have yet found. They are mostly scientists interested in intervening in the aging process. Quite a number of attendees are already cryonicists, including Aubrey de Grey, the originator of SENS and the organizer of the conference. But cryonicists are nonetheless a distinct minority. In previous years I brought a few Cryonics Institute brochures, which were soon taken. This year I brought enough brochures for as many of the 240 attendees as might want one (there were many left over).  I also brought a few copies of my “Scientific Justification of Cryonics Practice” (the published write-up of my SENS3 cryonics presentation) which I gave to a few attendees who seemed most receptive.

In addition to my oral presentation on cryonics I also had a poster. Scientific conferences usually have poster sessions where scientists present research, reviews, or ideas in the form of a poster. Poster presenters stand by their posters at scheduled times to discuss their work on a one-to-one basis with individuals rather than to an audience. My poster dealt with challenging the concept of biological age and denying the possibility of a biomarker of aging that could determine biological age. I contended that biological age and biomarkers of aging assume a singular underlying aging process, which I denied on the grounds that aging is multiple forms of damage. I sought to make maximum use of the one-to-one interaction by preparing Socratic questions to stimulate thinking and discussion with the attendees. The process also gave me another means of meeting and speaking to those attending. One interesting person I met was a Torontonian who is currently studying for his PhD at University of Glasgow. His work involves developing gene vectors that can precisely target and modify genes on chromosomes. I consider gene therapy to be an essential tool for the ultimate implementation of SENS, and a deficiency of SENS that there is so little attention paid to this technology. I don’t see how SENS can be implemented by any means other than genetic re-programming. LysoSENS, for example, would require new genes to create new, more effective enzymes for the lysosomes. MitoSENS would require all mitochondrial proteins be made in the nucleus and imported into the mitochondria.

Partly in this connection, was my aggressive lobbying of Aubrey de Grey to have Argentinian biogerontologist and Cryonics Institute member Rodolfo Goya as an invited speaker at SENS5. I began lobbying in January when Dr. de Grey was at ConFusion 2011. Aubrey was initially reluctant based on the first batch of Dr. Goya’s papers that I sent, but a later batch in which Dr. Goya was principle investigator proved to be effective. In Dr. Goya’s presentation at SENS5 he described his use of viral vectors attached to magnetic nanoparticles to deliver IGF-1 genes to senescent female rats to rejuvenate dopamine-producing cells in the hypothalamus. He injects the particles into the venticles, so the technique is somewhat invasive. Another speaker, Matthew Wood, described exosome nanoparticles which can cross the blood-brain barrier so I am hopeful that Dr. Goya can adopt this technique. Dr. Goya ended his presentation with a short pitch for cryonics (showing CI’s cryostats), which even I found embarrassingly awkward. I introduced Dr. Goya to a number of other cryonicists attending SENS5, including Igor Artyuhov, who is the scientific advisor for KrioRus, and Alcor Member Maria Entraigues, who is the SENS volunteer co-ordinator, and a native of Argentina (now living in Los Angeles).

Russian biogerontologist Alexey Moskalev reported on decreasing the number of single-strand DNA breaks and increasing the maximum lifespan in fruit flies by overexpressing the stress response/DNA repair gene GADD45 in the nervous system. That such a presentation would be included in SENS5 was of special interest to me insofar as I have contended that (and debated with Aubrey de Grey concerning) nuclear DNA damage possibly being a significant cause of aging damage that is missing from SENS:

http://www.benbest.com/lifeext/Nuclear_DNA_in_Aging.pdf

http://www.alcor.org/magazine/2011/02/28/deficiencies-in-the-sens-approach-to-rejuvenation/

http://www.alcor.org/magazine/2011/06/07/sens-a-reply-to-ben-best/

Alexey later told me that he had read my paper in REJUVENATION RESEARCH, and I’d like to think that I helped inspire his work.

Alexey announced that there will be a genetics of aging conference in Moscow in April 2012. I entertained the thought of going, partly because of my desire to see KrioRus, but I would rather go later when KrioRus is established in its new building, and has a research program in full swing.

Alexey’s research was partly funded by the Science for Life Foundation (the organization of the wealthy life-extensionist Russian Mikhail Batin). Maria Konovalenko (who was featured in LONG LIFE magazine) reported on her work at the Science for Life Foundation to build an open web-based database of age-related changes (molecular and phenotypic). Maria has her own blog.

I am not going to attempt to describe the other very excellent SENS5 presentations other than to say that great progress has been made in starting research programs on each of the SENS strategies, and by 2012 research on all the strategies is expected to be in progress.

Alcor President Max More was an invited speaker, which means that he had a half-hour time-slot immediately preceding my 15-minute time-slot near the end of the program. Max gave an overview of cryonics, whereas I concentrated on technical and scientific issues associated with vascular and neuronal injury from ischemia and reperfusion. During the question period I was asked if we are interacting with hospital staff to limit pre-mortem ischemia in cryonics patients. I said that the current legal environment limits such interactions, but that pre-mortem anti-oxidant protocol has been recommended and used.

I arranged to send more information to a few people in the audience, including a man who was interested in hydrogen sulfide to limit ischemic injury in cryonics, and an Italian neuroscientist who is interested in neurophysiology studies of vitrified brain tissue as well as contact information for Italian cryonicists.

At the final banquet I sat with CI Member Dr. Gunther Kletetschka, who is now living in the Czech Republic and is pursuing a number of imaginative cryonics-related research projects. One of these involves carbon nanotubes to deliver non-toxic metals to cells to use magnetocaloric cooling. Such a technique could cool tissues uniformly rather than externally, thereby eliminate the thermal stress that causes cracking when vitrified cryonics patients are cooled at cryogenic temperatures.

The last day was spent punting on the Cam River, with dinner in the evening. This provided an opportunity for more networking and information exchange, although most of this was in connection with biogerontology.

There was much biogerontology to be learned at SENS5. What I learned at SENS5 can potentially extend my life and that of others. To postpone cryopreservation by life extension is to benefit from technical advances, to extend the time in which I can contribute to technical advancement, and to enjoy more present life. In the best case, rejuvenation will become a reality in my lifetime and I won’t need to be cryopreserved at all. I work for this possibility as well as for improved cryopreservation. Moreover, in doing research for my cryonics presentation at SENS5 — and in giving the presentation — I learned many things that can help me make more informed choices in directing the research that Aschwin and Chana de Wolf do for the Cryonics Institute.

A video of my presentation may eventually be placed on the SENS5 YouTube site.

July 24-27 I attended the 2011 annual Society for Cryobiology conference in Corvallis, Oregon.

A number of the first presentations were concerned with means to *avoid* cryopreservation. Room temperature storage is much less expensive and troublesome, and improves ease of transport, especially in remote areas. One such technology “shrink wrapped” DNA in a glass  and another used trehalose to protect lipid membranes in a similar manner. Applied to cells, such technologies are viewed as a form of room-temperature vitrification.

Another researcher had successfully freeze-dried hematopoietic stem cells using trehalose and other additives without losing the ability of the stem cells to differentiate. Stress proteins in combination with trehalose allowed for desiccation of mammalian embryonic kidney cells without loss of viability. Late Embryogenesis Abundant (LEA) proteins also assist trehalose in dehydration tolerance.

Christoph Stoll showed that depleting red blood cell membranes of cholesterol can increase
trehalose uptake, but when I asked him in person about it, he said that the uptake was not enough to make much difference. Depleting cell membranes of cholesterol makes them more vulnerable to chilling injury, so I don’t think cholesterol depletion is a very good idea.

Masakazu Matsumoto spoke about some of the interesting anomalous properties of water.

Andrew Brooks spoke about the largest University cell and DNA repository in the world at Rutgers University.  They store DNA by plunging in liquid nitrogen.  He told me that 10 freezings and thawings does not impair DNA quality. That is encouraging for CI’s tissue/DNA storage program, because we plunge our samples into liquid nitrogen. Brooks gave data  showing that RNA is much less hardy in liquid nitrogen than DNA.

David Denlinger noted that HSP70 RNAi can block cold tolerance in insects. He also mentioned a Czech study which found that insect larva fed proline could survive liquid nitrogen. Perhaps we should be feeding proline to terminal cryonics patients.

In preparation for this conference, I had done a lot of reading on the subject of chilling injury and was hoping to question researchers on the subject. Steve Mullen showed a video of meiotic spindles dissociating at low temperature.

Spindles are a form of microtubules. Microtubules are known to dissociate at low temperature, but can spontaneously re-associate upon rewarming. But that would not be so beneficial when the microtubules are functioning as centrosomes because the reassembly would not be a reconstruction of the original structure. This is probably why cell division often  stops at low temperature.

Tiantian Zhang is one of the two candidates to become the new Society for Cryobiology President. Her field of study is cryopreservation of fish embryos and oocytes, which are especially vulnerable to chilling injury.

Fish are useful scientific models because they have a much simpler genome than mammals. 50% of endangered species are fish, but fish don’t get anywhere near the concern that pandas do. In both her lecture, and when I spoke to her in person, Dr. Zhang had apparently not learned any more than what was in her 2009 paper.

Why does reducing yolk content reduce chilling injury? Why is methanol the most non-toxic cryoprotectant for fish embryos, and so protective? If microtubule dissociation were a mechanism of chilling injury, it is indeed ironic that a 2006 Society of Cryobiology meeting presentation found that methanol causes proteolysis.

Kevin Brockbank spoke on the oxygenated hypothermic machine perfusion that he used to preserve pig livers at 4-6deg C for 12 hours. As a somewhat off-the-wall question, I asked him if he had assayed for chilling injury. This was off-the-wall because I have never heard of anyone assaying chilling injury. He responded that he had not, but that there were plans to use gene arrays to assay for chilling injury. This is like gene arrays to assay for aging — it requires deeper analysis, especially if chilling injury — like aging — is due to multiple mechanisms, the mechanisms are controversial, and no one mechanism is dominant. Northern wood frogs, arctic insects, and polar fish don’t have problems with chilling injury, although their adaptations include heat shock proteins and highly unsaturated cell membranes.

Much to my frustration, I have not had a good conversation with Peter Mazur (the uncrowned guru of cryobiology) since he got me to tell him I am a cryonicist several years ago. I have repeatedly asked him questions, and he has repeatedly been rude and dismissive. This year was different, for some reason. When I asked him about frozen water expansion contributing to mechanical damage he noted that cells could tolerate a 9% expansion without lysis even if freezing was intracellular. When I asked him how much dehydration cells could tolerate without damage, he said cells could lose all of the osmotic water (90% of cell water), and could lose more in freeze-drying with proper protectants (like trehalose). I was somewhat stunned by this answer, which takes no account of intracellular electolyte concentration increasing on dehydration. Next year I will be more optimistic about the possibility of talking with him, and I will prepare questions more carefully.

I spoke to Society for Cryobiology President John Crowe about his negative remarks concerning trehalose, in light of the fact that he is very aware of many of its benefits. John told me that a new method of manufacturing trehalose from starch is making trehalose as inexpensive as sucrose. If trehalose is used on bakery sugar, the sugar will not melt and run after a couple of days, as happens with sucrose. I mentioned to John that Robert Ettinger had just died. I had imagined that he might ask me to say a few words about the matter to the cryobiologists at their business meeting, but John treated the matter as a non-event, and I got the distinct impression that he would have preferred that I had not mentioned it.

At the business meeting it was noted that membership has dropped from close to 300 in 2008 and 2009 to just above 200 in 2011. There is concern that web access to the journal
CRYOBIOLOGY is becoming so easy that the incentives for membership have dropped. Or the global financial crisis is taking its toll on Society for Cryobiology membership. CRYOBIOLOGY journal impact factor has fallen to 1.830 from a high of 2.044 in 2002.

I appreciate being able to attend the business meetings, but one of the vehemently anti-cryonics cryobiologists gives me dirty looks. I have not been kicked-out yet, though, and decreasingly worry that I will be. A similar thought goes through my head as when I attend an Alcor meeting: “Spy in the House of Love.” But I really want the Society to prosper and grow, not be harmed, because I appreciate their good work (as with Alcor), even if they view me as a threat.

I had a brief chat with the cryonics-friendly Treasurer, who asked me when I think a cryonics patient will be reanimated. When I told him not less than 50 years, he said that a lot of surprising things can happen in 20 years. He is a more optimistic cryonicist than I am! At least as remarkable is that he is currently working with biotechnologists who are engineering scaffolds that can be used for growing organs from stem cells. That is a very cryonics-relevant project!

Every year I exchange a few words with Arthur Rowe (the cryobiologist who repeatedly compares cryonics to restoring a cow from hamburger — as he did in “Death in the Deep Freeze” – a comparison which probably originated with Peter Mazur). This year Arthur spent a lot of time hanging out with John G. Baust (the man who compared publishing cryonics science research with publishing Nazi hypothermia experiments). At the end of the conference I lost patience trying to catch Arthur alone, so I approached Arthur to say “hi”. Arthur said that he had seen on TV that Robert Ettinger had just died. He asked me about Robert’s educational credentials, and about my taking Robert’s place as CI President. Then he introduced me to John Baust. John was politely quiet, and said very little.

As with the 2010 Cryobiology Conference, I felt decreasingly paranoid as the meeting proceeded, but my level of paranoia was nonetheless very high near the beginning of this meeting. Overall, the amount by which I “came out” as a cryonicist was modest this year, and my softening of the hostility of cryobiologists to cryonics was modest this year compared to the previous one. The 2012 Society for Cryobiology Conference is scheduled to be held in Argentina.

On the evening of Thursday, May 19 and on Friday, May 20, I attended the 2011 (2nd annual) Teens & Twenties young cryonicists gathering which preceded the Suspended Animation, Inc. conference in Fort Lauderdale, Florida. The Teens & Twenties gathering (for young cryonicists having human cryopreservation contracts in place with some cryonics organization) is an offshoot of the cryonics Asset Preservation Group. Like the Asset Preservation Group, this event was created-by and is run-by Cairn Idun. Bill Faloon funds Teens & Twenties through a Life Extension Foundation education grant. Members of the Asset Preservation Group, such as myself, are permitted to attend despite being more than 30 years old. Of the 52 people who attended, ten were Asset Preservation Group members, and 42 were young cryonicists.

When asked who did not want to be photographed, only one person in the group raised his hand. I will refrain from mentioning any of the young cryonicists by name. Writing about this very people-oriented event without mentioning individual young cryonicists is like writing about lemonade without mentioning lemon. Some of the personalities were particularly colorful and memorable. But I know that many of the individuals do not want the publicity, and in my experience people get very emotional about what is said and not said about them. Even with explicit permission I am concerned that many of the young cryonicists might not fully appreciate the kinds of problems writing about them in connection with cryonics might cause for their future careers.

This year the demographics of the young cryonicists more closely matched what is typical of cryonicists. Last year about one third of those attending were female, and there was a high representation of people from the entertainment industry. This year, the attendees were overwhelmingly male, with most of the females being companions of males (which is not to say they were not cryonicists). Many members of this group were impressively highly educated, mostly in computer technologies, and secondarily in biotechnologies.

EXCELLENT MEALS WERE INCLUDED IN THE SCHOLARSHIPS

There were six Russians: five from KrioRus, and one from CryoFreedom. KrioRus is located near Moscow, whereas CryoFreedom is further south in Russia, closer to Ukraine. Dr. Yuri Pichugin (formerly the Cryonics Institute’s cryobiologist, is associated with CryoFreedom. CryoFreedom advertises neuropreservation for $7,500. Although it currently has no human patients, two pets are in liquid nitrogen. (I also learned that there is a man named Eugen Shumilov who is working to start a new cryonics company in St. Petersburg, Russia, but there was no representation of Shumilov’s organization at this event.

There are two overlapping goals of the Teens & Twenties event. One is the opportunity for members of the Asset Presevation Group to meet the young cryonicists. The other is the opportunity for the widely dispersed young cryonicists to become acquainted with each other, and to build lasting networks (community building). Cairn Idun has designed a number of “getting to Know You” exercises to facilitate the networking.

There are two self-introductions: the first lasting one minute, and the second lasting two minutes. I was the most anal-retentive of any of the participants in these exercises. I wrote-out my self-introductions, and practiced reading them to myself until I was sure I was within a few seconds of the one and two minute allocations. The one-minute self-introductions were on Thursday evening, and the two-minute self-introductions were Friday morning.

The Thursday evening self-introductions were followed by the exercise wherein participants classified themselves by “color”: (Green:Conceptual, Curious, Wise, Versatile), (Red:Adventuresome, Skillful, Competitive,Spontaneous), (Gold:Responsible, Dependable, Helpful, Sensible), and (Blue:Warm, Communicative, Compassionate, Feeling), as described in my write-up of last year’s Teens & Twenties event.

Once again, Greens were most numerous, followed by Reds. Cairn directed the participants to gather into groups by color. No directions were given for these meetings, so it was to foster socialization between “like-colored” individuals.

Last year a number of people had little to say in their second self-introductions, imagining that they had said all that could be said about themselves in their first self-introduction. I concerned myself quite a bit about how to prevent this from happening again. I made a number of suggestions in the Young Cryonicists Facebook Group, as did others. Cairn had participants list wants and “not-wants” of various kinds before the second self-introductions as a means of facilitating self-awareness. I tried to make my second self-introduction very personal in the hope that it would inspire others. There weren’t too many who were at a loss for words in the second self-introductions this year. Many of the participants passed-out business cards or other self-descriptive materials in conjunction with their second self-introduction.

There was a breakout session in which those with special interests had an opportunity to discuss their interests or how they might work together on those interests. The interest areas were entertainment, research, computer sciences, communication networking, and psychology/philosophy of self.

INTEREST GROUPS

Bill Faloon encouraged the participants to share thoughts about types of research that could lead to reanimation — with the thought that many of the young cryonicists would be in charge of large revival trust funds with income that can be used for research on reanimation technologies. I won’t attempt to summarize the thoughts of others, but I can say a few things about what I said.

Some people don’t want cryonics because they are afraid that they will not be restored in their original condition. The mother of one cryonicist is a stroke victim, and she has had a frightening first-hand experience of losing mental & movement capacity. Hollywood plays into this vision by depicting reanimated beings as zombies who are criminally insane.

Few people want to be the first of those reanimated — they would prefer that many others be reanimated first to ensure that the process works perfectly. I suggested that the first people reanimated might be brought back by next-of-kin who are overly eager to see their loved-ones as soon as possible. The idea of reviving pets first would not be popular with many pet owners. Reanimation technologies might be perfected on non-pet animals, although even today there is increasing sentiment against animal research. Animal rights activists seek legislation to protect animals from “unnecessary research”, which would likely include anything cryonics-related. Austria banned research on apes in 2006, and the number of countries with similar legislation continues to grow [SCIENCE; 332:28-31 (April 2011)]. Even if reanimation research was conducted on apes, the extrapolation of restoring ape consciousness/identity to restoring of human consciousness/identity is non-trivial.

I worry that as more wealthy cryonicists are cryopreserved, their only concern will be for reanimation research. Many of them will not appreciate that improved cryopreservation methods will advance cryonics and thereby enhance their chances of reanimation.

The next “getting to know you” exercise was what Cairn calls “speed dating”. Each participant is to spend two minutes with every other participant having a one-to-one conversation. For myself, it gave me an opportunity to talk to many people I would not have spoken with otherwise, and to have personal conversations with many individuals that I cannot imagine happening in any other way. Spontaneous socializing more often results in people talking only to those they already know. This exercise is a good ice-breaker, but it does involve some effort. It can be a strain to be starting conversations again-and-again, and again-and-again having to break them off once they become interesting — but the result was well worth the effort for me. Having a personal connection with individuals enables me to interact with them more productively, and this must also be true of the others. I rate speed-dating as the most valuable of all of the exercises, along with self-introductions.

Participants filled out a sheet indicating their interest level in cryonics — including such things as whether they planned to have a cryonics-related career, do volunteer work for a cryonics organization, or simply be a consumer.

GATHERING FOR THE GROUP PHOTO

he final event was the group photo, after which was a dinner and then reception for the Suspended Animation conference. The photographer who made the group photo was employed to make photographs only intended sor private use of Suspended Animation, Inc., but we did not learn this until later (even the photographer did not know).

All the young cryonicists had the fees, hotel expenses, and meals associated with the Suspended Animation conference paid-for. The opportunity for some of the young cryonicists who have an interest in science to directly interact with current cryonics researchers could eventually lead to large scientific dividends for cryonics research in the future.

There were reportedly many exaggerated rumors about what happened in the evening hot-tub sessions in the 2010 Teens & Twenties gathering. I brought my bathing suit this year, but did not spend a great deal of time in the hot tub. The conversation was a bit more playful than it was in other contexts, and there was more of a party-spirit in the hot tub — which some of the participants relished. I would guess that about half of the Teens & Twenties participants spent at least some time in the hot tub.

Despite all of the intensive social interaction and “getting to know you” exercises, I would have a hard time making a connection between names, faces, and biographies of at least a third of the young cryonicists. I don’t believe that I am unique in that regard. The “speed-dating” exercise was particularly helpful in strengthening and deepening the name/face/biography connections. Memories of the individuals and their personalities are likely to be more easily refreshed in the future thanks to the meetings and exercises of this gathering.

YOUNG CRYONICISTS VISIT WITH SAUL KENT

Robert Ettinger‘s book Youniverse: Toward a Self-Centered Philosophy of Immortalism and Cryonics is a book containing many insights and deep thoughts, yet has such an informal writing style that many readers might not take it seriously. I know of no other work of philosophy in which the author begins a sentence with “Anyway,”. Ettinger writes that the first cryonics-related organization was founded “in 1962 or 1963, I forget which”, then says “Why don’t I look it up?” and justifies himself by reference to a Woody Allen movie. This is not the kind of writing one expects from a philosophy treatise.

Ettinger may not take himself too seriously, but he is even more dismissive of most of the world’s foremost philosophers and religious figures. The writings of Aristotle are called “ramblings”. In describing William James’s statement that James was only able to understand Hegel while under the influence of nitrous oxide, Ettinger notes how appropriate it is that nitrous oxide is also called laughing gas. Ettinger wrote that “Rousseau has been extravagantly praised, and not only by himself”, but dismisses Rousseau as unoriginal, incoherent, not profound, and frequently wrong. Ettinger describes the philosopher G.E. Moore as being “definitely confused as well as confusing, abounding in contradictions and non-sequiturs, sometimes substituting assertions for arguments.” Ettinger often seems himself guilty of the last accusation. He faults Isaac Asimov for the “absurdity” that without the “saving grace of death” the rigid views of the old would prevent further progress — but leaves a critique of Asimov’s argument “as an exercise for the reader”. Ettinger writes that “Paeans of praise have poured from the pens of platoons of panting pundits” concerning Godel’s Incompleteness theorem, which he dismisses as a linguistic trick associated with the failure of physics to correspond identically with formal (mathematical) systems. By finding the quote from Wittgenstein “I don’t know why we are here, but I am pretty sure that it is not in order to enjoy ourselves”, Ettinger has massively deflated my respect for the philosopher Ludwig Wittgenstein. Ettinger describes the modern “self-styled bioethicist” as a “new type of vermin or parasite” whose major accomplishment has been to create “the illusion of looking down on people far above them.”

Ettinger wrote that “fear of God” is generally really fear of parents, neighbors, and a lifetime of conditioning. He says people too readily submit to tradition rather than use reason. To be “normal” is to have the same delusions as the neighbors. He says loyalty “is frequently a worthy habit”, but sometimes nothing more than an unjustified habit. Ettinger says faith is arrogant certainty in the absence of evidence, which ultimately “boils down to sacrificing your integrity for a bit of comfort”. To Ettinger it is obvious that non-human animals have consciousness and feelings, and that a God that disregarded the suffering of animals on the grounds that animals have no soul “would have less compassion than the average human”. Like many physicists, Ettinger seems accepting of the idea that time and the universe began with the Big Bang, but wonders where God would be before He created time and the universe. Ettinger can make no sense of an omniscient, omnipotent God creating people who need to live their lives to prove whether they deserve Heaven or Hell. Ettinger says that a benevolent God would forgive the skeptics, who should therefore have no reason to compromise their integrity and disbelief.

Ettinger’s irreverence extends to the legal system. Frequent use of appeals courts and split decisions in the Supreme Court are given as evidence that laws are unclear or that bias is pervasive. He describes juries as “ignorant, stupid and readily swayed by irrelevancies and by histrionics”. In connection with the adversarial system, Ettinger wrote “All lawyers are frightening, and specialty litigators are terrifying. Some firms are said to keep their lead litigators chained in a tower room and fed raw meat until needed.” I asked Mr. Ettinger what his beloved son (a lead litigator at a prestigious law firm) had to say about the law chapter, but I got no definitive response.

As the book title YOUNIVERSE implies, Ettinger believes that “me-first” and “feel-good” are the only possible basis for conscious motivation. He also states that a person ought to want whatever will maximize future “feel-good”, and that people do not always want what they ought to want. Ettinger believes that “figuring out what we ought to want is the primary problem of philosophy”. He says that a main aim of YOUNIVERSE is to debunk the views that values are arbitrary or externally given.

Ettinger challenges the claim of David Hume that “You can’t derive an ‘ought’ from an ‘is'”, and — like Ayn Rand with her Objectivist Ethics — he does so by reference to values being rooted in biology. Ettinger disparagingly dismisses Rand’s views as narcissism, “me generation”, and “looking out for number one” without explaining how this differs from “me-first”. Rooted in biology, Rand makes survival the basis of her ethics, rather than “feel-good”. Ironically, Ettinger writes more approvingly of Nietzsche’s self-centeredness, although Ettinger faults Nietzsche’s belief in the importance of power over other people as a core value. (Ettinger notes that Nietzsche believed Russians and Jews, rather than Germans, would be the “master races” of Europe.)

I disagree with the arguments of Rand and Ettinger for deriving “ought” from biology. Biology dictates that animals value food and water, but many humans have committed suicide by refusing food and water. To assert that such people are “wrong” and did not do what they ought to have done would be attempting to externally impose values upon them. Ettinger could argue that such people were acting in such a way as to maximize their satisfaction — “me-first” and “feel-good” (he gives the examples of a woman rushing into a burning building to save her baby, or “saints” who gain personal satisfaction from ascetic service to others). But by that argument they were wanting what they ought to want. The point Ettinger seems to be making is that people should not allow others to impose their values upon them — should not be driven by guilt, social pressure, the need to conform. But if people are driven by these motives, they are nonetheless still maximizing their satisfaction. Ettinger might say that such people are acting without integrity by not being true to themselves, but why should people be blamed for valuing the opinions of others and for this being important to them? If it is “impossible to be motivated by anything other than self interest, because motivation means what is important to the self”, then the word “ought” is inappropriate. If “me-first” and “feel-good” are the only possible bases for conscious motivation, then the word “ought” is inappropriate. The only reason that people fail to want what they ought to want is because of matters of fact, not matters of value — people failing to appreciate the consequences of their actions in the context of their values.

The issue of determinism and free will is a subject about which I have thought, read, and written about considerably (see A Case for Free Will AND Determinism ), yet I found Ettinger’s chapter on this subject impressively thoughtful and informative. I mostly agree with Ettinger’s views, about which we are both very much in the minority. I won’t say much about the issues or insights I gained in the determinism chapter, but I will comment on how he applies determinism to cryonics. Ettinger notes that “determinism is very nearly equivalent to” conservation of information, which implies that any human who ever lived could be reconstructed without having been cryonically preserved — except that there may never be adequate computing power.

Although I can conceive of retaining my personal identity in the total absence of any memories that I have, I nonetheless find the idea hard to relate-to. I am even less comfortable about the idea that the essence of my personal identity is feeling. Ettinger has firmer opinions on these subjects than I do, but I sense that his emphasis on feeling as the essence of personal identity contradicts his admonishments about the use of reason against intuition, tradition, and conditioning.

Ettinger skims over the subject of ischemic damage in cryonics, and I think he is wrong to say that “cryothermic damage will in most cases be the most difficult to reverse”. Freezing damage is like broken pieces that are nonetheless intact, whereas ischemic damage is like dissolution or decomposition of structure. Nonetheless, I cannot quantify my argument in terms of “most cases”. I think Ettinger is wrong to cling to the word “immortality” as meaning “indefinitely extended life” when its literal meaning is “eternal life”. His use of the word “immortality” presents cryonics as an alternative to religion rather than an extension of medicine.

Although Ettinger acknowledges that death will mean an end to suffering, he sees a number of disadvantages, including
“…it’s hard to enjoy life when you’re dead.
…daisies are prettier when viewed from above.
…you can only vote in Chicago.
…you need extra strength deodorant.”
But mainly, “Life is better than death because it is more interesting.” (For my own views on the subject, see: Why Life Extension?)

In his lifetime of reading Ettinger has collected numerous notable quotes, and these gems are liberally sprinkled throughout YOUNIVERSE. Some of my favorites include “‘Love thy neighbor as thyself’ presupposes that you love yourself” (Miguel de Unamuno), “The greatest part of our happiness depends on our disposition, not our circumstances” (Martha Washington), and Will Rogers’s WWII suggestion for getting rid of German U-boats: “Boil the Atlantic Ocean. How do we do that? Hey, I’m just an idea man, I leave the details to the engineers.”

Ettinger also has a chapter called “Misunderstandings” which deals with his insights into a wide variety of subjects. Indicative of my “anti-intellectual” bias, is the fact that my favorite is Ettinger’s observation that torque (force X lever arm length) has identical units to work (newton-meters), despite the fact that work and torque are completely different. He offers no solution or explanation, however.

A consequence of Ettinger’s informal writing style is that there is much autobiographical material throughout YOUNIVERSE. But the last formal chapter (I am not counting the Appendix) is explicitly autobiographical. He says “I have perhaps a few thousand admirers, hardly any of whom give me much thought or attention”. Ettinger speaks of his loneliness in having experienced the loss of all his friends and family of his generation, and that there is nobody left whom he wants to impress. Indicative of Ettinger’s world-weariness is his quote of a comment made by his brother that all of life is “killing time and amusing oneself while waiting to die”.

Ettinger’s final comments concern his plan to have a pre-mortem “jolly wake” with music, speakers, toasts, and other festivities prior to a suicide intended to improve the conditions of his cryonic preservation. Ettinger notes earlier in the book that “many people are more afraid of seeming cowardly than of facing danger”, which is why suicide with an audience of friends and family would boost his courage. The last line of the chapter reads “If I never wake up, my last experience will have been better than most — a very brief comfort, to be sure.”

Although there are some cryonicists who believe that Robert Ettinger would be the perfect cryonicist to win sympathy for voluntary self-euthanasia to improve cryopreservation, I am not one of them. How can you justify voluntary euthanasia in a non-terminal person when there is no way of knowing how many years of life that person could be expected to live? How can you justify voluntary euthanasia for ANYONE not suffering from a terminal disease, or expect the public to be sympathetic to voluntary self-euthanasia under these conditions? Even for terminal cryonics patients, I would not be to eager to see a public association of cryonics with self-euthanasia or physician-assisted suicide. Cryonicists would be accused of taking advantage of mentally-compliant sick and elderly people for monetary reasons, which would lead to even more cryonics-unfriendly legislation.

And there are practical problems, not the least of which is the danger of autopsy. Many cryonicists, myself included, cling to life tenaciously — much more tenaciously than the average person. I would find it very difficult to euthanize myself or have myself euthanized. The ideal situation is when death is nearly certain to occur within a week. But this is the condition in which standbys are typically initiated, not the condition in which standbys fail to occur. Heart attack is a common cause of death, and this is most often unexpected. Most cryonicists who receive standby are people dying of cancer, and whose slide toward death is along a more predictable path. The ability of cancer victims to euthanize themselves would make the standby process easier, but that would have no effect on reducing the number of cryonicists who deanimate without standby, despite having arranged for standby. There are no convincing arguments that simplifying self-euthanasia or physician-assisted suicide will lead to the majority of cryonics cases having greatly improved cryopreservation by significantly reducing the number of cryonicists deanimating under unfavorable conditions.