One concern about prolonged cardiopulmonary support in cryonics is that its decreasing effectiveness may not be able to meet cerebral oxygen demand, and may even become detrimental. Some investigators have  observed that severely reduced flow (cerebral blood flow less than 10% of control) to the brain may actually be more harmful than no flow at all.  Explanations of why incomplete (“trickle flow”) ischemia may be worse than complete ischemia include aggregation of slow moving blood cells,  glucose-induced excessive lactate production, and oxygen-induced free radical damage to membranes.

In contrast, a study by Steen et al. concluded that some blood flow is better than no flow at all. The authors found that dogs could sustain only 8 to 9 minutes of complete ischemia but 10 to 12 minutes of incomplete ischemia (cerebral blood flow less than 10% of control) without neurological impairment. These results are at odds with the findings of Hossmann et al. who found better electrophysiological recovery in cats and monkeys after complete ischemia, and studies by Nordstrom et al. who observed increased metabolic recovery in rats after complete ischemia.

The authors speculate that these differences may reflect the different durations of (in)complete ischemia. Hossmann et al. studied 60 minutes of ischemia and Nordstrom studied 30 minutes of ischemia. The authors note that the durations they studied (8-14 minutes) are more clinically relevant because neurological recovery with contemporary technologies is not possible after 30 or 60 minutes of cerebral ischemia. Although these findings provide support for restoration of any kind of cerebral circulation after cardiac arrest, it does not offer much guidance in evaluating the practice of prolonged cardiopulmonary support in cryonics.

The authors also draw awareness to the difficulty of correlating electrophysiological and metabolic recovery to neurological recovery. They quote a study by Salford et al. who observed some return of metabolism even though histological abnormalities had already been developed. Such studies warrant caution about using return of electrophysiological activity as an indicator of cerebral viability because it is not likely that such viability can be sustained over the long term, let alone predict functional recovery of the brain.  It is doubtful that viability in the latter, stricter, sense can be maintained during stabilization of most, if any, cryonics patients. At best, the studies that demonstrate recovery of electrophysiological and metabolic activity after prolonged cerebral ischemia offer hope that such periods of circulatory arrest do not produce acute information-theoretic death.

No metabolic or histological evidence was found to support the implication of no-reflow, lactate accumulation, and free radical damage in incomplete ischemia.  Again, the authors speculate that no-reflow may be more pronounced during longer periods of incomplete ischemia, an observation that seems to be indirectly supported by Fisher et al. who observed progressive impairment of perfusion for longer periods of ischemia.

Cryonics patients often experience shock, blood coagulation abnormalities, and decreased cerebral perfusion prior to pronouncement of legal death and cardiopulmonary support.  An additional complicating factor in cryonics is that cardiopulmonary support is often supplemented by induction of hypothermia and administration of vasopressors and neuroprotective agents. Although the paper by Steen et al. addresses a lot of issues that are important to evaluate cryonics procedures, it is clear that for real empirical guidance regarding the wisdom of prolonged cardiopulmonary support specific cryonics research models are required.

16. July 2008 · Comments Off · Categories: Cryonics · Tags: , ,

From: ExtroBritannia

Cryonics: Why it has failed, and possible ways to fix it – with Mike Darwin

The next ExtroBritannia event is scheduled for Saturday August 2, 2008; 2:00pm – 4:00pm.

Location to be announced asap.

Lead Speaker: Mike Darwin, President of Alcor Life Extension 1983-1988, Research Director 1988-1992. Described by Wikipedia as “Second only to Robert Ettinger as one of the most influential figures in the controversial field of cryonics”.

The talk will draw on the speaker’s extensive personal experience with cryonics – the low-temperature preservation of humans and other animals that can no longer be sustained by contemporary medicine, until such time in the future when resuscitation may be possible.

The talk will cover: the audacious ambition and vision of cryonics, practical details of how it works, a whistle-stop history of cryonics, issues with the governance of cryonics organizations, factors influencing public perception of cryonics, and reasons for both fear and hope for the future of cryonics.

Special attention will be given to the decline of cryonics in the UK and the failure of UK cryonics to establish a robust, full-service beachhead in Britain. The talk will also highlight what can be done to re-establish cryonics in the UK as a stable enterprise that will deserve the confidence of both its members and the public as a competent, high quality undertaking offering services which meet the highest ethical, scientific and biomedical standards.

The meeting is sponsored by the UK Transhumanist Association. There is no charge to attend. Join the debate!

Discussion is likely to continue after the event, in a nearby pub, for those who are able to stay.

There’s also the option of joining some of the UKTA regulars for drinks/lunch beforehand, starting c. 12.30, in a pub (to be announced). To find us, look out for a table where there’s a copy of Aubrey de Grey’s book “Ending Aging” displayed.

Venue: To Be Announced – somewhere in Central London

15. July 2008 · Comments Off · Categories: Cryonics, Death · Tags: , , ,

Most life extensionists and transhumanists do not buy into many of the myths about cryonics. But one perspective that is sometimes voiced by futurists is that cryonics is a rational backup plan until aging is cured. This position has some serious shortcomings and potentially lethal implications.

Human cryopreservation is the practice of placing terminally ill patients who have exhausted contemporary medical treatments into long term cryogenic care, allowing them to benefit from future medical treatments. Although aging-associated diseases are an important cause of death, they are not the only cause of death. Even when biological aging becomes optional, a person will still be vulnerable to accidents and violence.

The mindset that cryonics will become redundant as soon as aging is conquered is especially dangerous when it leads (young) people to forgo or postpone making cryonics arrangements because they expect to benefit from  rejuvenation technologies and dietary supplementation during their lifetime. This may not only reflect wishful thinking regarding the rate of progress in overcoming aging, but it will also leave them vulnerable to other causes of death.

As long as humans (or post-humans) are vulnerable to injury that cannot be treated with contemporary medical technologies, human cryopreservation will remain important as a form of critical care. In other words, as long as there can be situations that warrant metabolic arrest to avoid information-theoretic death, there is a need for cryonics or similar technologies to induce metabolic arrest, like molecular warm biostasis.

There are a lot of people who believe in the technical feasibility of cryonics and intend to make cryonics arrangements….when necessary. As cryonics observers know, this is an extremely risky attitude because when people need cryonics the most, they often are unable to communicate their wishes, may meet resistance from relatives who benefit from their not making cryonics arrangements, or lack financial resources because life insurance is no longer an option to fund cryonics.

The best time to make cryonics arrangements is when it seems least likely that you need them soon. This is also evidenced by the fact that young healthy people can get excellent rates on life insurance.

The latest issue of Life Extension Magazine (August 2008) contains an encouraging report about off-label use of etanercept (commercial name: Enbrel) to reverse the cognitive deficits associated with Alzheimer’s disease. Etanercept is a tumor necrosis factor (TNF) blocker that is used to treat diseases such as ankylosing spondylitis, juvenile rheumatoid arthritis, psoriasis, psoriatic arthritis, and rheumatoid arthritis. Pilot studies and case reports not only reported improved cognitive function after weekly perispinal administration (injection  into the back of the neck) of etanercept, but rapid clinical improvements have also been observed within minutes of administration of the drug in at least one person, a result that the Life Extension Foundation (LEF) reportedly was able to reproduce in a pilot study of a 91-year-old female patient with advanced Alzheimer’s disease.

The Life Extension Foundation (LEF) is currently seeking to launch an expanded study to investigate the effects of weekly Enbrel injections plus nutrients that help suppress the production of excess TNF-alpha in people who have early-stage Alzheimer’s disease. Although the normal costs of weekly injections is around $675, LEF will not charge people enrolled in the studies for these treatments. Patients who have, or people who know someone with, early-stage Alzheimer’s disease are encouraged to contact LEF for enrollment. The study will be conducted in the Fort Lauderdale area in south Florida.

Future installments of this blog will review the research on TNF-alpha modulation for the treatment of Alzheimer’s disease in more technical detail. We also recommend a recent article on long-term brain maintenance on the blog Existence is Wonderful, which also discusses cryonics in a favorable context.

11. July 2008 · Comments Off · Categories: Health · Tags: , , , ,

One problem in assessing the merits of taking a specific dietary supplement (ranging from vitamins to  exotic multi-ingredient compounds) is widespread selection bias in the documentation that is supposed to support the use of the supplement in question.  The sheer number of scientific studies combined with variation in research methodologies virtually guarantees that for every supplement a supporting study can be found. For example, the recent issue of Life Extension Magazine (August 2008) has an article on the multiple health benefits of melatonin with 81 references. All these studies discuss either the biochemical properties of melatonin or show beneficial effects. This is what is what is seen. What is not seen are the studies in which melatonin is not effective or has adverse effects.  Or the studies that never got published as a result of “publication bias.” Granted, melatonin seems to be a remarkably effective agent for a diverse number of conditions, including its use as a neuroprotective agent in stroke, but such selective presentation of biomedical research seems to be a mainstay in the marketing of dietary supplements.

Another limitation of such documentation is that the studies that are used to recommend the taking of a supplement often solely address the (short-term) effects of that compound on the medical condition in question. Although it would not be practical to report on all the studies that investigate (chronic)  administration of the compound on other systems in the body, such unrelated adverse effects should not be ruled out when considering prolonged use. It is a major leap from demonstrating beneficial effects of a compound in rodents and preliminarily studies in humans to “recommending” the use of that compound for prolonged use in humans. And it is a giant leap to go from such studies to combining different effective compounds in very high dosages in a single product.

Promoting the use of supplements with a hodgepodge of  encouraging in-vitro studies, small animal studies, and observations in humans is not necessarily wrong, nor constitutes deliberate selection bias. Human biochemistry is extremely complex, and rigorous  research would require enormous resources and longitudinal experiments.  In real life there is a need to make informed decisions based on the evidence at hand. Still, our current state of knowledge and our ignorance about how all that we know adds up for specific individuals should induce modesty and, perhaps, moderation. For those who take supplements as a means to radical life extension, making cryonics arrangements remains the irreplaceable  cornerstone of such a program because it increases the odds to reach a time where truly meaningful (molecular) life extension technologies will be available, aside from the protection cryonics offers against most “lethal” accidents.

Induction of hypothermia can reduce injury to the brain when it is deprived of oxygen. How fast do we need to cool a patient during cardiac arrest or stroke to prevent irreversible injury to the brain?

It is an established fact that induction of hypothermia prior, during, or after circulatory arrest can reduce brain injury. As a general rule, the lower the temperature is dropped, the longer the brain can tolerate circulatory arrest. The neuroprotective effects of hypothermia are often expressed using the Q10 rule which says that for every 10 degrees Celsius drop in temperature metabolic rate decreases by 50%. Or to put it differently, the Q10 rule states that ischemic damage susceptibility is decreased by a factor of 2 for every 10 degrees Celsius temperature drop.  Q10 may vary between species and in different organs and cells. For example, different temperature sensitivities were observed for release of the neurotransmitters glutamate, aspartate, glycine, and GABA during cerebral ischemia by Nakashima et al. Because even very modest reductions of brain temperature can have profound neuroprotective effects, the Q10 rule may not tell the complete story.

Other things being equal, it would be very useful to have a measure of brain injury when hypothermia is induced prior and/or during cardiac arrest. At least two authors have made an attempt to produce such a measure of ischemic exposure. In Cryonics Magazine (2nd Quarter, 1996), Michael Perry started initial work on this in an article called “Toward a Measure of Ischemic Exposure” (PDF).  Perry’s Measure of Ischemic Exposure (MIX) calculates how long the patient has been at a given temperature, with a higher weighting used for higher temperatures. A related measure has been proposed be Steve Harris called the E-HIT. E-HIT stands for Equivalent Homeothermic Ischemic Time. In his (unpublished) manuscript, Harris uses the E-HIT formula to calculate the equivalent normothermic ischemic time for different cryonics case scenarios and real cases. Clearly, the availability of such a measure (and its routine calculation in case reports) would constitute a major contribution to cryonics as evidence based medicine. It could aid in deciding if viability of the brain was maintained during cryonics procedures by estimating the equivalent warm ischemic time.

What makes such a measure complicated during cardiopulmonary resuscitation (CPR), or cardiopulmonary support (CPS) in cryonics stabilization procedures is that hypothermia may only constitute one intervention to mitigate brain injury. In an ideal cryonics case, pronouncement of legal death is followed by rapid restoration of oxygenated blood flow to the brain by (mechanical) cardiopulmonary support, administration of neuroprotective drugs and induction of hypothermia. Such a combination of interventions might avoid any injury to the brain, reducing the equivalent warm ischemic time to zero. A more realistic scenario is that such a combination of interventions may reduce the extent of ischemic injury compared to cooling only. Another complicating factor is that oxygenation in combination with low perfusion pressures might produce more injury than “anoxic cardiopulmonary support” (chest compressions without ventilation). It is clear that calculating a measure of equivalent ischemic time for real cryonics cases can become very complicated.

It would be interesting to know the cooling rate that would be necessary to stay ahead of brain injury, using contemporary medical criteria, during circulatory arrest. For this purpose we use some very simplifying assumptions:

1.The patient is not ischemic prior to pronouncement of legal death.

2. Cooling is initiated immediately after pronouncement of legal death.

3. There is no cardiopulmonary support or administration of neuroprotective agents.

4. Brain injury starts at 5 minutes of warm ischemia.

5. Q10 is 2.0: for every 10 degrees Celsius we decrease the temperature , metabolism is dropped 50% , which doubles the time a patient can tolerate ischemia.

6. No other forms of injury occur other than ischemic injury.

7. Ischemic injury is completely eliminated at the glass transition temperature of the vitrification agent M22 (-123.3°C).

8. A constant cooling rate is assumed.

Using these assumptions, Alcor’s Mike Perry calculates that a cooling rate of 2.89 degrees Celsius per minute is necessary to stay ahead of the equivalent of 5 minutes of warm ischemia.

Let Ehit = total ischemic time limit in hours, 1/12 corresponding to 5 min
Q10 = factor of decrease in metabolism per 10 degrees
Tdrop = desired temperature drop, from 37 degrees (body temp) down to -123.3= 160.3 degrees Celsius
ch=desired cooling rate in deg/hour
cm=desired cooling rate in deg/min = ch/60

Then

ch = 10*(1-exp(-Tdrop*ln(Q10)/10))/(Ehit*ln(Q10))

For Q10=2, Tdrop = 160.3, cm = 2.89 deg/min

If some of the assumptions are slightly changed we find the following for Q10=2.2

For Q10=2.2, Tdrop = 160.3, cm = 2.54 deg/min

If we assume negligible ischemic insult below 0 Celsius and only worry about cooling down to that temperature, so Tdrop is only 37 rather than 160.3, it doesn’t change these amounts drastically:

For Q10=2, Tdrop = 37, cm = 2.66 deg/min
For Q10=2.2, Tdrop = 37, cm = 2.40 deg/min

Clearly, such high cooling rates cannot be achieved during either conventional cardiopulmonary resuscitation or cardiopulmonary support in cryonics. The cooling rates we can hope for during the initial stages of cryonics procedures may exceed 1.0 degrees Celsius per minute at best. It is therefore not realistic to assume that cooling alone may be able to limit brain injury to a degree that allows resuscitation without adverse neurological effects using contemporary medical criteria. This should strengthen the case for the use of other interventions such as administration of neuroprotective agents and oxygenation of the patient. Although the latter intervention may produce adverse effects on the brain itself, the calculations above indicate that anoxic cardiopulmonary support is not compatible with maintaining viability of the brain as the objective of cryonics stabilization procedures. The case for rapid stabilization of cryonics patients remains strong.

08. July 2008 · Comments Off · Categories: Cryonics · Tags: , , , ,

Today’s post on 21st Century Medicine’s vitrification agent M22 completes the series on vitrification agents in cryonics. To date, three different vitrification agents have been used for cryopreservation of humans: B2C (at Alcor from 2001-2005), VM-1 (at the Cryonics Institute since 2005) and M22  (at Alcor since 2005).

Perhaps the most encouraging development in cryonics is that Alcor’s current vitrification agent, M22, is not only the least toxic cryoprotectant in the history of cryonics, it is also the state of the art in mainstream cryobiology research for vitrification of complex organs.

It is doubtful if the state of the art in vitrification in cryobiological research would be where it is today without the incentives provided by cryonics to search for a cryoprotectant that enables reversible vitrification of the brain without ice formation and minimal toxicity.

The first vitrification agent in cryonics: B2C

Vitrification agents in cryonics: VM-1

Vitrification agents in cryonics: M22

M22 represents the culmination of decades of work in applied cryobiology by researchers Gregory Fahy , Brian Wowk, and others to develop a vitrification agent that can recover complex organs (such as the kidney) from cryogenic temperatures without ice formation and minimal toxicity. In 2005, M22 was licensed by the patent holder 21st Century Medicine (21CM) to the Alcor Life Extension Foundation to replace their previous vitrification agent B2C. As a result, the least toxic vitrification agent for complex organs that has been documented in peer review journals is currently being used for cryonics patients at Alcor.

M22 incorporates a number of important discoveries in cryobiology:

1. High concentrations of a cryoprotective agent (or a mixture of different cryoprotective agents) can prevent ice formation during cooldown and warming.

2. The toxicity of some cryoprotectants can be neutralized by combining them with other cryoprotective agents.

3. The general toxicity of a vitrification agent can be predicted by using a measure called qv*, allowing for the rational formulation of less toxic vitrification agents.

4. Within limits, non-penetrating agents can reduce the exposure of cells to toxic amounts of cryoprotectants without reducing vitrification ability.

5. Synthetic “ice blockers” can be included in a vitrification mixture to reduce the concentration of toxic cryoprotective agents necessary to achieve vitrification.

6. Substituting methoxyl (-OCH3) for hydroxyl groups (-OH) in conventional cryoprotective agents can decrease viscosity, increase permeability, and reduce the critical cooling rate necessary to avoid ice formation.

7 Chilling injury can be eliminated by introducing the vitrification agent with a hypertonic concentration of non-penetrating solutes.

8. In cryonics, with a minor proprietary modification, M22 can be used for whole body perfusion without causing severe edema that has been a problem for some other solutions.

Vitrification is the solidification of a liquid without crystallization. When a solution is cooled down to the glass transition point (-123.3°C for M22) the extreme elevation in viscosity will produce a glass in which all translational molecular motions are arrested. Although water vitrifies at cooling rates exceeding a million of degrees Celsius per second, such cooling rates are relaxed when other solutes are substituted for water. In cryobiology solutions with high concentrations of cryoprotective agents can be used to vitrify complex organs such as the kidney or the brain.

Vitrification has a number of clear advantages over conventional cryopreservation. The most important advantage is the elimination of ice formation. Although the adverse effects of ice formation can be mitigated by the use of cryoprotective agents (glycerol, DMSO) and optimization of cooling rates, massive ice formation does not permit recovery of complex organs with full viability. Another advantage is that vitrification eliminates the need to strike a balance between the risk of intracellular freezing induced by fast cooling on the one hand, and cell dehydration and solution concentration induced by slow cooling on the other hand.

The challenge in formulating successful cryoprotective agents is to design vitrification solutions that are non-toxic but allow for vitrification at realistic cooling and warming rates. For more than a decade the least toxic vitrification agent was Greg Fahy’s VS41A, which is an 55% weight/volume equimolar mixture of DMSO and formamide plus propylene glycol. The “1A” in VS41A reflects the solution’s ability to vitrify at normal atmosphere pressure (as opposed to an older, more dilute solution, VS4, which requires 1000 atmospheres of pressures to vitrify). The equimolar concentrations of DMSO and formamide reflect Baxter and Lathe’s research who concluded that amides can neutralize the toxicity of DMSO, a finding that Greg Fahy later revised in favor of the theory that it is actually DMSO that neutralizes the toxicity of formamide. The ability of DMSO to neutralize the toxicity of formamide (up to certain concentrations) allows for the formulation of vitrification agents with reduced toxicity. This finding has been so fundamental that an equimolar concentration of DMSO and formamide remains the core of M22.

Another major step was made when the researchers at 21CM found that high concentration of (penetrating) cryoprotectant agents do not necessarily increase toxicity. Contrary to conventional cryobiology expectations, Fahy et al. found that weaker glass formers favor higher viability. They proposed a new compositional variable called qv* to predict the general toxicity of vitrification solutions. Using qv* they made the “counter-intuitive” decision to substitute a higher concentration of the weaker glass former ethylene glycol for propylene glycol to create a solution called Veg, which produced a substantial improvement in terms of viability as measured by K+/Na+ ratios.

Because cells contain higher concentrations of protein, the intracellular space is more favorable to vitrification than the extracellular space. As a consequence, the concentration of penetrating (toxic) cryoprotectants can be reduced in favor of non-penetrating polymers like polyvinylpyrrolidone (PVP). Variations of Veg in which the concentration of DMSO and formamide was reduced in favor of PVP increased viability without decreasing its ability to suppress ice formation. The concentration of penetrating cryoprotectants can be further reduced by inclusion of non-penetrating “ice-blocking” polymers. These ice-blockers also reduce the critical cooling and warming rates necessary to avoid ice formation, which is an important requirement for solutions that are used to vitrify complex organs such as the human brain.

Because concentrated vitrification solutions depress the homogeneous nucleation temperature (Th) below the glass transition temperature (Tg), a major obstacle to successful vitrification is the presence of heterogenous nucleators. Some organisms have antifreeze proteins (AFPs) and anti-freeze glycoproteins (AFGPs) that mitigate heterogenous nucleation by binding to nucleators. Because adding such anti-nucleating proteins to vitrification solutions would be prohibitively expensive and less effective, Greg Fahy proposed the creation of synthetic ice-nucleation inhibiting polymers. In 2000 Wowk et al. published work that showed the effectiveness of a co-polymer of polyvinyl alcohol (PVA) and vinyl acetate in inhibiting heterogenous ice-nucleation. This co-polymer is now being sold by 21CM under the name “X-1000″. X-1000 is particularly effective in glycerol solutions, presumably because glycerol itself is a poor anti-nucleation agent. Increasing the concentration of X-1000 in vitrification solutions decreases ice formation and relaxes minimum cooling rates. Although X-1000 is presumed to be non-toxic, the maximum concentration in vitrification solutions does not exceed 1% w/v because no further benefits were observed beyond this concentration. In 2002, 21CM announced the discovery of another synthetic “ice-blocker” called Z-1000. Z-1000 is the polymer polyglycerol (PGL), which specifically inhibits ice nucleating activity caused by the bacterium Pseudomonas syringae. Mixtures of PVA and PGL are more effective in inhibiting ice formation than either agent alone, suggesting the PVA and PGL complement each other by inhibiting different sources (bacterial and non-bacterial) of ice nucleation.

A variant of Veg that includes the low molecular weight polymer polyvinylpyrrolidone K12, X-1000, and Z-1000 named VM3 improved viability in renal cortical slices and decreased the critical cooling and warming rates necessary to avoid ice formation and de-vitrification (ice formation during rewarming) while maintaining the same molar concentration as VS41A. The transition from Veg to VM3 reflects the two breakthroughs mentioned above: reduction of cryoprotectant toxicity by inclusion of non-penetrating polymers and ice blocking agents. VM3 also was the least toxic agent in vitrification of rat hippocampal brain slices, which is of particular importance for cryonics. The first vitrification agent ever to be introduced to cryonics was a hyperstable variant of VM3 called B2C. B2C was used until late 2005, when it was replaced by M22.

M22 takes advantage of two other discoveries: the ability to design better glass formers by methoxylation of conventional polyols, and inhibition of chilling injury by delivering the vitrification agent as a hypertonic solution. Because hydroxyl groups can bind either to water or hydroxyl groups on other cryoprotective agents, substituting methoxyl groups for hydroxyl groups should decrease interaction between cryoprotectants and increase interaction between the cryoprotectant and water. As a result, methoxylated compounds have stronger ice inhibiting ability, thus reducing the critical cooling rate for vitrification or reduce the concentration of (toxic) cryoprotective agents in a solution. Methoxylated cryoprotectants also decrease viscosity and increase cell permeability, allowing for shorter perfusion times, and thus reduced cryoprotectant exposure at higher temperatures. For example, the methoxylated glycerol derivative 3-methoxy-1,2-propanediol has a higher glass transition point and vitrifies at ~ 5% lower concentration than the corresponding conventional cryoprotective agent. Complete exploitation of these advantages is limited by the fact that they are more toxic than their non-methoxylated compound, as predicted by qv*. As can be seen in the table, the major difference between VM3 and M22 is the reduction of PVP K12 in favor of the penetrating cryoprotectants 3-methoxy-1,2-propanediol and n-methyl-formamide, and increased concentration of the ice-blocker Z-1000. The final molar concentration of 9.345 M demonstrates that more concentrated vitrification agents do not necessarily have to be more toxic.

VS41A

Veg

VM3

M22

Dimethyl sulfoxide

3.10 M

3.10 M

2.855 M

2.855 M

Formamide

3.10 M

3.10 M

2.855 M

2.855 M

Propylene glycol

2.21 M

-

-

-

Ethylene glycol

-

2.71 M

2.713 M

2.713 M

N-methylformamide

-

-

-

0.508 M

3-methoxy-1,2-propanediol

-

-

-

0.377 M

Polyvinyl pyrrolidone K12*

-

-

7% w/v

2.8% w/v

X-1000 ice blocker*

-

-

1% w/v

1% w/v

Z-1000 ice blocker*

-

-

1% w/v

2% w/v

Total Molarity

8.41 M

8.91 M

8.41 M

9.345 M

* Non-penetrating polymers are in w/v

M22, so called because it was intended to introduced at -22 degrees Celsius, constitutes a major landmark in vitrification of complex organs. In 2005 Fahy, Wowk et al. announced routine recovery of rabbit kidney slices from temperatures around -45 degrees Celsius. Although consistent recovery of vitrified organs is not yet feasible, continued progress in solution composition and perfusion techniques inspire optimism that this may be possible in the future. In 2007, Greg Fahy of 21CM reported recovery of electrical activity in vitrified brain slices and induction of long-term potentiation (LTP), which indicates that the structures for processing memory are maintained after vitrification, storage and rewarming of brain tissue. Visual evidence that M22 can preserve the ultrastructure of the brain better than B2C was published on the Alcor website in 2005.

M22 also needs to be used in a suitable carrier solution to support cell metabolism at low temperatures and decrease oxidative injury and edema. The carrier solution for M22 is called LM5 to reflect the 50% reduction of glucose (as compared to the older carrier solution RPS-2) in favor of equimolar concentrations of mannitol and lactose, to address compatibility problems with the ice blockers. The combination of the isotonic LM5 plus the non-penetrating polymers in M22 creates a hypertonic solution, which has been shown to eliminate chilling injury, which is the injury that is caused by exposure to low temperatures as such. For cryonics, the composition of M22 is further enhanced by including a proprietary components that allows perfusion of whole body patients without edema.

The research breakthroughs discussed above allow for a global reconstruction of the composition of M22 using the table. Maintained is the equimolar combination of DMSO and formamide from Fahy’s older vitrification solutions to reconcile strong glass formation ability and minimal toxicity. The discovery of the  compositional variable qv* allows for substitution of higher concentrations of the weaker glass former ethylene glycol for propylene glycol. Substitution of a non-penetrating polymer, PVP K12, and the ice-blockers X-1000 and Z-100 allow for further reduction of DMSO and formamide, reduction of critical cooling rates, and increased stability against ice formation. In M22, PVP K12 is reduced to optimize hypertonicity of the non-penetrating agents for suppression of chilling injury. Added are the methoxylated cryoprotectant 3-methoxy-1,2-propanediol and the highly permeable amide n-methyl-formamide, producing the least toxic but most concentrated vitrification solution to date.

The most striking differences between Alcor’s old perfusate and the newer vitrification agents licensed from 21CM are complexity and cost. Until 2002, Alcor patients were perfused with high molar glycerol in an MHP-2 based carrier solution. M22 itself consists of 8 (!) different components, putting the total number of components of M22 in carrier solution above 15. Such perfusates makes great demands on preparation skills and quality controls. Components such as the ice blockers and 3-methoxy-1,2-propanediol have put the cost of Alcor’s whole body perfusate alone close to the cost of complete cryopreservation arrangements at the Cryonics Institute (CI). This raises obvious questions about costs and benefits. As evidenced by CI’s VM-1, potent protection against ice formation can be achieved with a vitrification agent that solely consists of DMSO and ethylene glycol. It is plausible to assume that vitrification lessens demand on future repair technologies, but it speculative to assume that minor differences in toxicity between different vitrification agents will translate in earlier resuscitation and less expensive repair protocols. However, more toxic vitrification solutions, such as CI’s VM-1, may cause acute injury to endothelial cells. As Brian Wowk notes, “good cryoprotection depends on good perfusion, which depends on preservation of vascular integrity during perfusion. The ability to perfuse M22 into whole bodies with tolerable edema is likely to be intimately related to its low toxicity to vascular endothelium.” And of course, there are also PR advantages to the fact that a cryonics organization uses a vitrification agent that is also the state of the art in conventional cryopreservation of organs.

M22 produces substantial brain shrinking during perfusion of (non-ischemic) patients. As a matter of fact, cerebral dehydration may be a major contributing factor to vitrification of the brain and even allow for reduced concentrations of M22 for brain preservation. This does not mean that the (expensive) non-penetrating polymers could be replaced for any high molecular weight polymer because the ice blockers and non-penetrating cryoprotective agents also protect the extracellular space against ice formation and are effective in ischemic patients with a compromised blood brain barrier (BBB). The limited ability of some components of M22 to cross the BBB and, and differences in permeability of the various components of M22, does raise questions about the exact composition of M22 beyond the BBB and within brain cells after completion of cryoprotective perfusion.

Patients outside of the US may not fully benefit from cryopreservation with M22 because of the of long cold ischemic times during transport. This raises the question if cryonics patients can be perfused outside of the US and shipped in dry ice. Experiments with VM-1 in bulk solution indicate that this solution is very stable against ice formation, even during long storage periods. M22 in bulk solution seems to form ice crystals overnight if stored in dry ice. This does not necessarily mean that M22 cannot be used in combination with dry ice for overseas patients because human tissue perfused with M22 (or any cryoprotective agent) is not the same as M22 in pure solution. But regardless of M22′s compatibility with dry ice shipping, cryonics organizations may benefit from formulating a highly concentrated inexpensive vitrification solution that is extremely robust against formation of ice, which can be used for simple perfusion of non-US patients in combination with dry ice shipping. The decreased cold ischemic times of such a solution may outweigh the increased toxicity of such solutions.

How do you teach a child about something that is so far “unproven”?  How do you bring up the subject of cryonics and how it may allow someone to be reanimated in the future?

I am a cryonicist, I’ve been a signed member for years, I’m also a mother, social activist, environmentalist and author.  I teach religious education at my church, and I volunteer in my children’s schools.  My book “21st Century Kids”, set in the year 2008, is about two children who ‘die’ now but are cryonically preserved and then reanimated 200 years in the future.  The book deals with how they view the society then, and how that society views them.  The book is of course science fiction, but it is based on things that scientists see as possible now.  When I talk to an eager classroom of 9-& 10-year-olds at a school about my book, I read passages out of it that are exciting and imaginative like the nano-tech and simulated artificial reality parts, but I also make sure the subject of cryonics comes up.  I’ve talked with dozens of classrooms, and hundreds of children at my own church about cryonics.  I know how hard it can be, death is a reality—it is a fear for children, or it is a sadness when someone they loved died—they may think that person is in heaven, and they will see them again–when cryonics comes up, the children become animated sharing stories, and what they think.

I love children for their open-mindedness.   Of all the children I’ve talked to, many have said cryonics sounded neat or cool—I even had a few say they were going to tell their parents they wanted to sign up for cryonics.  I’ve amazingly heard back from several parents over the past few years, asking me for information.  I give them cryonics magazines, and talk with them about life insurance and how easy it is to set up—the importance of being signed if something unexpected happens so you don’t have to be a ‘last minute case’ and I go over the basics of just what cryonics is with them.  This blog piece would turn into a book if I listed all the things I say and children ask—but I’ll go over a few of my ‘sound bytes’.

I tell children that some people choose cryonics because they’ve seen studies that showed cat brains have been preserved at colder than ice temperatures for several years and had normal looking electrical activity when re-warmed and given new blood, but we can not yet re-animate the whole body and all of the organs.  I say that there are scientists right now looking at how to better preserve organs for humans in hospitals, like when a person in a hospital in Texas needs a new kidney, and a person in California who has a kidney that would match, dies—how to get that kidney to the person in Texas fast enough, through ultra cold transport and planes.  I say cryonicists also like that some children have been born and grown into healthy adults after having been preserved cryonically as embryos. This makes them think that the procedure might work on a whole human someday.   I tell children that I, and my own children (usually the kids I’m talking to know one of my three kids) are signed up for cryonics, and when we die–like if a car accident happened tomorrow—we will be preserved, and most importantly our brains will be preserved in case scientists in the future figure out how to get it running again.  I say that even if they don’t, scientists from now are very interested in the mummified Egyptian bodies from over 3 thousand years ago—and have even been able to better understand some diseases now by looking at the diseases the mummified bodies had, and how those diseases have evolved since then.  I say that my body will be donated to science, and that if my brain is not made to have full awareness after several hundred years then I hope that some things can be learned by the future society about the preserved bodies from now.  Kids want to know how long I think it would take to work, I say I think over 500 years—and say that it would be likely to work if society, technology and medicine keep advancing as they have over the last 500 years.

It is hard to predict when talking with a group of children, where the talk will go—I ask them questions, like if they know something their great grandparents didn’t have a hundred years ago, we end up talking history and then talking about what could be.  Children sometimes bring up very sad stories about someone dying, and I say yes even with cryonics when someone is dead—they are gone from now, to us and their family is sad, they don’t know if they will ever see them again.  I’ve had the heart wrenching experience of an 11 year old talking about how his dad died of cancer, and I’ve had a few children in the 5-7 range who share about a grandparent who died and how much they miss them.  I empathize with their loss.  I say to children that  I believe their loved one is in a better place, that many cryonicists too want to go to a better place they believe in, like heaven—and they think they some day will, but if cryonics works they’ll have more time here on Earth to do good deeds—to try to help with some of humanities problems, before they go on.  It is hard to talk about death, but children will share deeply as they have their own fears about death and that gives me solace, the section about cryonics is deep, is profound but we always move on.

Children like the idea of cryonics, but they also like to talk about “future weapons” they see on T.V., and each group I talk to always brings up space travel.  Cryonics is a short part of our conversation, and wherever our conversation goes I try to keep it exciting and to stimulate their imaginations.   Having a group of children to speak with about futurist issues at a school or a church and covering cryonics is a lot of fun, and I always try to stay sensitive to what other parents might say when their children tell them what they ‘learned that day’.  People wonder if I talk differently with my own children, and the answer is not really.  Sure we make more jokes about “if cryonics works then….”Or if doing something dangerous joking “make sure I’m preserved if…” but in the end, I say the same things to them that I would to a group of children that are not my own.  Cryonics is not proven, it is just a chance, it would be fun if it worked and some of the research into it can help people now and I’m proud to be a cryonicist.  I tell my children that I’m happy that they are too. I also tell my children that I hope that after they are adults and choose partners in life to start their own familes with, that their families will also be cryonicists.

In the end, I’d encourage other cryonicists to share their views with children they know—they could even present “21st Century Kids” to a group of children, or simply give it as a gift, or read it to kids they know.  Teaching kids about cryonics is simply sharing what could be, it is not giving false hope—but hope that is based on science and studies that show it could work someday. Even though I share about cryonics with many adults, I have the most fun teaching children about it, and I hope you too get to engage in fascinating conversations about the future and how it could be, with a wide-eyed eager child who is in awe with life.

Feedback on this article is encouraged at the Immortality Institute forum.

“The only thing that retards aging is calorie restriction. As genetic studies go forward, we’ll find out why.” Roy Walford

Our society in America currently as of 2008 has more overweight people than average-weight people.  ‘Healthy weight’ Americans consist of only around 40% of the population, according to the US Center for Disease Control and Prevention, the ‘CDC’.  The amount of children considered overweight has grown alarmingly in the past decade, to the point now where 1 out of every 3 children in the US is considered overweight the Academy of Pediatrics and Adolescent Health said in 2006.  One Gallup Poll survey from 2003 showed that out of 215 million US adults 20% felt they had been discriminated against in their workplace for their weight.  This figure was only slightly less than the 25% percent of smokers that felt discriminated against in their workplace for their smoking and due to anti-smoking laws being passed in society.  Interestingly only 25% of the US population at the time of the poll were smokers, and 43% were considered overweight—demonstrating that smokers are generally scorned by the American public more than overweight people.  Today in ’08, smokers are still treated with more open hostility than are overweight people.  A smoker in a public place is harming other people’s lungs with their smoke, an overweight person is only harming himself or herself and this is generally seen, as it should be, as free will.

I agree that overweight people have the right to be overweight, if they are happy with it—but what about the children?  Many doctors pressure parents to help their children lose weight, for the sake of the child’s health and since overweight children have a much higher chance of being overweight adults. Excessive weight has been shown in countless studies to cause many health problems from diabetes, gallbladder disease, high blood pressure, gout, cancer, heart disease and stroke to name just a few.  I myself spent 6 years being 185, to 245 pounds on my five foot ten inch frame, very overweight.  Six months after the last of my three pregnancies, when I was 26 years old—I was two hundred and five pounds and sick of being so big, my eyesight was worse than when I was in my teens, my left knee constantly sore, I was tired and felt unattractive so I started to look at diets.  I’d signed up for cryonics when I was 21 years old and in a high risk pregnancy so I’d read some of the life extension sites and had seen calorie restriction mentioned as the only known thing to extend life, as compared to hormones, supplements and exercise.  At 25, I was looking at all the fad diets, and remembered what I had read about calorie restriction—I ordered the book ‘Beyond the 120 Year Diet’ by Roy Walford.  I was intrigued when it came, my 205 pound self read through the book three times, ear marking and highlighting certain passages—looking up more information on-line. I then got the software so I could track my calories and most importantly what vitamins and minerals I was taking in each day so I would not be malnourished.  Over six months I kept a food diary, used the software to plan my meals and I lost 85 pounds.  At 120, I felt like a new person—to my friends and family I looked like a new person, my knee pain went away, I had more energy and I even passed my drivers license test without my glasses—my eyesight had improved to near 20/20.  I experienced first hand the health benefits of losing weight, and most importantly I did this while my children were young.  If I had raised them, eating the way I had been while I was overweight—they too may have had problems.  Instead, I showed how to work to accomplish a goal and as I learned about nutrition, they learned about nutrition as well.

It would be unethical for a child to be on calorie restriction, as it would stunt their growth. My children would eat mega-muffins (like ‘lab chow’ for humans) for snacks (with chocolate chips so they’d eat them yet still get the protein, vitamin and minerals of a complete meal) but my children have never been calorie restricted.  I taught them that being hungry is ok, and actually good for you but they know about what different vitamins do, and how our bodies use them—they know how to evaluate the nutritional qualities of a meal. We have meal times together at our dining table, and along with our normal conversations of the day, we will comment on the nutritional qualities of the food—in this way over time they are learning.

I feel that I set an example to them of how they could be if they want when they grow up.  Each of my three children right now at ages 11, 9 and 6 have different views on food, one of them eats more healthy than the other two currently.  I do not chastise children for their food choices, or praise one in front of the others, I will just say what I would and would not eat.  We do not have hydrogenated oils, artificial dyes and preservatives, fried foods, or highly processed foods in our home.  We do have healthy deserts for the children such as ‘Healthy Choice’ brand ice cream bars, carrot cake (for the vitamin A), raw walnuts and dark chocolate – mom just does not eat dessert, dad will however and the children do as well.  I can practice CR easily while living with my family, and not impose it upon them—as a parent I hope that as they grow they will always make healthy choices.  I don’t know if any of them will choose CR, but I’m happy that I modeled losing weight and then healthy eating over the years.

One does not have to be practicing calorie restriction to teach children about nutrition.  All children need to be taught about proper caloric intake, the importance of healthy fats and complex carbohydrates.  They should know what vitamins and minerals do in the body, there are sites on-line such as Nutrition Mission, Monster Nutrition or the USDA My Pyramid Blast Off Game that can help you teach them.  There are great nutrition books for kids like The Edible Pyramid, The Race Against Junk Food, or Dr. Seuss’s ‘The Things You Can Do That Are Good For You’.  Those books are all for the early elementary aged child, and are good to read with a picture book of the inside of a human body. Reading a book to your child enables you to engage them in conversation and find out what they think, you can fill in gaps in their knowledge.  The best thing you can do though for your children, is simply to practice healthy eating and to talk about it from time to time.

Feedback on this article is encouraged at the Immortality Institute forum.