April 2015 • Volume 36:4 How Foundation Research Benefits

Suspended Animation Advanced Neural page 6 Biosciences page 17 The Stasis Foundation page 9 Critical Care Research 21st Century Medicine page 22 page 13 ISSN 1054-4305

$9.95 Improve Your Odds of a Good You have your cryonics funding and contracts in place but have you considered other steps you can take to prevent problems down the road? ü Keep Alcor up-to-date about personal and medical changes. ü Update your Alcor paperwork to reflect your current wishes. ü Execute a cryonics-friendly Living Will and Durable Power of Attorney for Health Care. ü Wear your bracelet and talk to your friends and family about your desire to be cryopreserved. ü Ask your relatives to sign Affidavits stating that they will not interfere with your cryopreservation. ü Attend local cryonics meetings or start a local group yourself. ü Contribute to Alcor’s operations and research.

Contact Alcor (1-877-462-5267) and let us know how we can assist you. Visit the ALCOR FORUMS www.alcor.org/forums/ Discuss Alcor and cryonics topics with other members and Alcor officials. • The Alcor Foundation • Financial • Cell Repair Technologies • Rejuvenation • • Stabilization • Events and Meetings Other features include pseudonyms (pending verification of membership status) and a private forum. Visit the ALCOR BLOG www.alcor.org/blog/ Your source for news about: • Cryonics technology • Speaking events and meetings • Cryopreservation cases • Employment opportunities • Television programs about cryonics Alcor is on Facebook Connect with Alcor members and supporters on our official Facebook page: www.facebook.com/alcor.life.extension.foundation Become a fan and encourage interested friends, family members, and colleagues to support us too. CONTENTS

lume 36:4 April 2015 • Vo 17 Advanced Neural Biosciences Advanced Neural Biosciences is a small neural cryobiology company in How Life Extension Foundation Portland, Oregon, that aims to perfect Research Benefits Cryonics cryopreservation of the brain. The company also studies the effects of ischemia on cryopreservation with the aim of improving outcomes for cryonics patients.

22 Critical Care Research AdvancedBiosciences Neural page 17 Critical Care Medicine specializes in Critical Care Research page 22 ischemia research, and its mitigation page 6 ISSN 1054-4305 by pharmacological and hypothermic The Stasis Foundation page 9 strategies in particular. Their current 21st Century Medicine page 13 $9.95 emphasis is on the development of “liquid ventilation,” a technology that can be deployed in cryonics to rapidly Cover Photo-Image Credit: induce hypothermia without the need Stephen Valentine, Architect for complex surgery.

6 Suspended Animation 28 Non-Cryonics Research Recently Suspended Animation is a standby and stabilization company located Funded by the Life Extension in Florida and California which provides its services to major cryonics Foundation organizations, including Alcor, which uses its services for all non-local The Life Extension Foundation does cases. Suspended Animation pioneers new stabilization equipment and not only fund multiple labs that aim conducts research to evaluate the quality of casework. to perfect the science of human cryopreservation, they also support 9 The Stasis Foundation many other researchers aimed at rejuvenation, slowing the aging process, The Stasis Foundation is located in Comfort, Texas, and is part of an and curing age-related illnesses. ambitious plan to build the most advanced patient care and research facility in the history of cryonics. The Stasis Foundation has conducted 34 Membership Statistics detailed studies into the next generation of patient storage options and How many members, associate is pursuing a number of other relevant research projects. members, and patients does Alcor have and where do they live? 13 21st Century Medicine 21st Century Medicine is the largest cryobiology company in the world 36 Resuscitation Update and engages in research to perfect the cryopreservation of complex Mike Perry surveys the news mammalian organs. Their pioneering research in hypothermic organ and research to report on new preservation, low-toxicity , brain vitrification, and whole developments that bring us closer to body cryopreservation is of profound importance to cryonics and the the resuscitation of cryonics patients. prospect of human suspended animation.

www.alcor.org Cryonics / April 2015 3 2015 Annual Giving Program lcor provides a wide array of services for you the member, and the general Editorial Board public. We inform and educate, we protect and preserve, and we strive to Aremain at the forefront of cryonics technology. Ralph C. Merkle, Ph.D. Since its founding, Alcor has relied on member support to maintain its mission R. Michael Perry, Ph.D. and attract new members. Your support, regardless of size, can provide a better future for all cryonicists. Please act now. Editor Aschwin de Wolf SUGGESTED GIVING LEVELS

Contributing Writers $20 FRIEND Catherine Baldwin Ben Best $60 JUNIOR SUPPORTER Aschwin de Wolf $120 SUSTAINING SUPPORTER Steven B. Harris, M.D. R. Michael Perry, Ph.D. $500 ADVOCATE SUPPORTER Chana Phaedra Stephen Valentine, RA (NCARB) $1,000 LEADING SUPPORTER

Copyright 2015 $2,500 VISIONARY SUPPORTER by Alcor Life Extension Foundation All rights reserved. $5,000 SILVER SUPPORTER Reproduction, in whole or part, without $10,000 GOLD SUPPORTER permission is prohibited. $25,000 TITANIUM SUPPORTER Cryonics magazine is published monthly. $50,000 VANGUARD SUPPORTER To subscribe to the printed edition and/or change your address, please call We encourage every member to donate. Even if you can only afford $5 right now, 480.905.1906 x101or visit the magazine you will make a significant contribution to Alcor’s future. website: Donations may be made via the Donations button on the Alcor website or by www.alcor.org/magazine contacting Alcor’s Finance Director, Bonnie Magee, at [email protected]. Your donation may be made as a lump sum or divided into easy monthly payments.  Please note: If you change your address less than a month before the magazine is mailed, it may be sent to your old address.

Address correspondence to: Cryonics Magazine 7895 East Acoma Drive, Suite 110 The Society Scottsdale, Arizona 85260 Phone: 480.905.1906 ifts have played a fundamental role in the cryonics Toll free: 877.462.5267 movement since its earliest days. Dr. James Bedford, a Fax: 480.922.9027 Gman whose extraordinary vision led him to become the first person to be cryopreserved, and the first to make a bequest to Letters to the Editor welcome: a cryonics organization, exemplified the determination of the early [email protected] pioneers of cryonics. We invite you to follow in his footsteps, and join the James Bedford Society. Advertising inquiries: The James Bedford Society recognizes those who make a 480.905.1906 x113 bequest of any size to the Alcor Life Extension [email protected] ISSN: 1054-4305 Foundation. If you have already provided a gift for Alcor in your estate, please send a copy of your relevant documents Visit us on the web at www.alcor.org to Alcor’s Finance Director, Bonnie Magee. If you’d like to learn more about setting up a bequest, send an email Alcor News Blog to [email protected] or call 480-905-1906 x114 to discuss your gift.  http://www.alcor.org/blog/

4 Cryonics / April 2015 www.alcor.org EDITORIAL

By Aschwin de Wolf

sometimes take for granted that cryonics members are aware ahead of the medical establishment in identifying methods to of ongoing research projects aimed at protecting against protect against disease, slow certain aging processes, and provide I ischemia, improving urgent response capability, and perfecting treatment protocols that statistically improve survival of those whole brain cryopreservation. stricken with serious illnesses. Perfection of whole brain preservation would occur if we could What motivates so many Alcor members to support the Life induce cryogenic temperatures in lab animal brains, rewarm the vitrified brains, and have them regain full function. At cryonics conferences, scientists working at the four full “Perhaps the strongest reason to patronize Life time labs dedicated to improving cryopreservation technologies Extension is the fact that it is operated by long- often make presentations about their recent work. Up until now, however, no one amalgamated all of the various research projects time cryonics members who don’t want themselves, into one descriptive publication. or any of their members to ever die.” This issue of Cryonics magazine is dedicated to enlightening Alcor supporters about the most aggressive research projects in human history whose goal is to improve the quality of cryopreservation in Extension Foundation is that this non-profit group fully funds all a way that will enhance odds of successful resuscitation. four of these research labs and they have set up several organizations None of the research that will be described in this issue is and trusts dedicated to funding reanimation research in perpetuity. conducted by Alcor, but Alcor members derive full benefits from The total value of these various reanimation organizations is today it because the four independent labs conducting this research in the hundreds of millions of dollars. routinely disseminate their findings to Alcor personnel. Alcor The monies to fund these four research labs plus the reanimation conducts its own in-house research and development to improve organizations come solely from membership dues ($75/year) its cryopreservation capabilities. and royalties received on sales of advanced dietary supplements, The most important step cryonics members can take now is to comprehensive blood testing, and small donations. extend their healthy life spans in order to avail themselves of these Long term cryonics members support Life Extension Foundation’s rapidly evolving technologies. efforts by patronizing companies that pay the Foundation a One way of doing this is to become a member of the Life substantial royalty on every product or service. Life Extension Extension Foundation of Ft. Lauderdale, Florida (lef.org). This Foundation then uses these funds to support these research labs. group has a track record dating back to 1980 of being light years I know every vitamin company likes to pretend they’re the best, but Life Extension can prove it with an unparalleled track record of innovation. Perhaps the strongest reason to patronize “This issue of Cryonics magazine is dedicated Life Extension is the fact that it is operated by long-time cryonics members who don’t want themselves, or any of their members, to to enlightening Alcor supporters about the most ever die. No other company on earth can make this claim. aggressive research projects in human history whose After you read the brief articles in this issue about the research programs funded by the Life Extension Foundation, I hope you’ll goal is to improve the quality of cryopreservation in consider joining this group and using their novel technologies a way that will enhance odds of successful revival.” to extend your life years, while simultaneously funding the many research projects they directly support. 

www.alcor.org Cryonics / April 2015 5 By Catherine Baldwin

have the coolest job, ever. Literally. I experience to provide quality care to these work for Suspended Animation, Inc. patients. I (aka “SA”) whose mission is, in the Nearly every year brings us new and simplest terms, all about cooling. Funded improved technologies in the form of by the Life Extension Foundation and smaller computers and batteries, fast contracted by Alcor,Alcor, SASA providesprovides nationwide nationwide fabrication with lighter and stronger standby and stabilization services for Alcor materials, and new laboratory and medical members. And, protecting cells and tissues devices and techniques. With these by rapidly cooling the body is SA’s primary developments come opportunities to aim for stabilizing Alcor members who are incorporate some of these improvements far from Alcor at the time of legal death. into our existing protocols and equipment. There are various methods for bringing For example, there are few measures Suspended Animation the body’s temperature down to near 0 of quality and effectiveness of treatment laboratory in California degrees Celsius to protect the brain and and procedures performed on cryonics other tissues from damage during transport patients. Traditional laboratory methods SA’s laboratory into commercially available to Alcor. The methods haven’t changed all generally involve biopsies and destructive assay plates and analyzed in a real-time that much over the years but there have tissue testing. There may now be some quantitative polymerase chain reaction been improvements, particularly in how alternatives to this type of destructive machine (Applied Biosystems 7500 Fast). they are delivered to cryonics patients, and testing. This instrument is like a copy machine for there are new techniques on the horizon. Newer functional genomic studies and genetic material present in our samples in These are the areas of SA’s research and gene expression analysis technologies have tiny, and unknown quantities. Each plate development. We spend a lot of time trying allowed scientists to identify many of the contains 12 controls and 84 assays of the to figure out how to cool patients faster, genes and signaling molecules expressed key genes involved in regulating cell stress more safely and more effectively. during cell stress, cell injury and cell death. and cell death responses. Over time, by Since opening in 2002, SA has developed There are genes involved in activating examining and quantifying which genes new equipment and adapted state-of-the- pathways that tell the cell to die and others appearing in our samples are consistently art medical equipment and techniques involved in blocking these pathways. While upregulated and downregulated during our for cryonics patient stabilization. SA also these genes are expressed in the cells that procedures, we hope to better assess the created a whole new model for training make up the brain and other tissues, the quality and effectiveness of stabilization and contracting medical professionals molecules also wind up in the blood and treatments. This may also allow us to like thoracic surgeons and perfusionists cerebrospinal fluid (CSF). test the effectiveness of proposed new to perform standbys and stabilizations. SA is experimenting with using treatments. These medical professionals are trained by biomarker and gene expression analysis SA’s present and future cooling devices SA to deal with the special problems of of the blood and CSF to try to assess the are also getting an overhaul and benefitting treating cryonics patients. They keep sharp effectiveness of stabilization treatments, from new research and technologies. Using by working regularly on medical patients both generally and for individual cryonics advanced design modeling, computer in hospitals and other medical facilities, patients. simulations and 3-D printing, SA is but are available on quick notice to travel During animal and human stabilization developing high-powered cooling devices to standbys of cryonics patients, where procedures, serial blood and fluid samples that offer real advantages over previous they use their considerable training and are collected. The samples are processed at systems.

6 Cryonics / April 2015 www.alcor.org adjustable voltage that could power the to a machine that circulates blood and high flow pump for almost a whole day at cools it (called hypothermic perfusion). its maximum output. Cooled blood circulates to the brain and Next, SA prototyped some mask designs other tissues, cooling them. This technique using fiberglass and tubing, testing each for can cool the body by a couple of degrees optimized flow. Each of these designs took in a few minutes or to near 5 Celsius in 30 several days to cut, form and cure—a very minutes to an hour. slow process. To speed things up, SA’s staff SA acquired the same machines and designer recruited a group of industrial modified them to be air-transportable. design students at the Art Institute of We then recruited on-call professional Pittsburgh who were eager to apply their surgeons to perform the surgical procedure skills and create a unique final class project. and perfusionists to run the machines Using the latest computer aided design to perfuse and cool cryonics patients. (CAD) and 3-D modeling software, SA This is part of our standard protocol Gene expression assay using the and the students were able to experiment for stabilizing cryonics patients prior to Applied Biosystems 7500 Fast with more complex, multi-layered designs. transport to Alcor. A caveat is, whether A 3-D printer could produce a mask for a trauma victim or a cryonics patient, Cooling a cryonics patient on the outside, prototype from a design in a few hours. performing this surgical fast cooling particularly their head and neck, is the One mask design quickly emerged from procedure requires a number of highly most basic step in the initial stabilization testing as by far the best. trained people, specialized equipment and protocol. Simply packing a person in ice a suitable facility. This can take time when offers some cooling, but cooling power what we need is immediate, fast cooling. increases by flowing iced water over the Researchers at Critical Care Research, Inc. skin and the more the better. The ice-cold (CCR) and SA are developing a device that water comes in contact with more surface may solve this problem and act as a kind of area and can remove more heat. It’s a bit immediate, fast cooling “bridge” until the like the difference between placing a can or surgical procedure can be performed. Like the surgical fast cooling procedure, bottle in a bucket of unmelted ice versus the new device cools the blood and immersing it in a cooler of ice and water circulating this cooled blood cools the (which cools it faster). Other procedures brain and other tissues. However, the new being administered make it impractical device does not require any surgery to to immerse a cryonics patient so, over access blood vessels. It uses the already the years, a variety of systems of pipes Suspended Animation’s cooling mask open passages through the mouth and and tubes and large electric pumps have throat to access the millions of tiny blood evolved to recirculate ice water within a SA now has this mask fabricated overseas, vessels in the tissue of the lungs. shallow portable ice bath for the patient. blow molding it from just a few ounces of All of the body’s blood passes through Some worked better than others. We felt lightweight, disposable plastic. The mask the lungs to pick up oxygen that we inhale it would be worth the effort to figure out delivers almost 11 gallons of ice water per and release carbon dioxide waste we then how to maximize flow over the head and minute directly to the critical areas of the exhale. The lungs are ideally suited for neck to eke out some additional cooling head and neck of the patient without risk this gas exchange because they contain a benefits while eliminating some weight and of spraying team members nearby. The huge surface area that is dense with blood safety issues associated with other systems. power supply is small and rugged and can vessels at a thin interface. What researchers SA did initial research and testing to quickly charge phones, cameras and other at Critical Care Research found was that identify the best off-the-shelf ultra-light devices in the stabilization kit as well as run the lungs could work very well for heat submersible pump that was inexpensive the high flow pump for hours. exchange as well as gas exchange. enough to be disposed after use and still While fast-flowing ice water over the Similar to passing ice cold fluid over the delivered high flow when converted to run outside of the body can be performed outside of the body to remove heat, passing from a battery instead of being plugged almost immediately and offers some ice cold fluid over the inside surface of the into a wall outlet. We found a 12-volt pump cooling, it’s fairly slow with different parts lungs removes heat also, only much faster. that weighs about a pound and pumps up cooling at different rates, around a degree Cycling cold fluid in and the warmed fluid to 1100 gallons per hour. or two of Celsius per hour. Slower cooling out repeatedly cools the blood moving High flow pumps running for hours means more damage to tissues. So, how can through the blood vessels in the lungs, require significant power. Fortunately, high we cool faster? which cools the patient. Connecting a capacity lithium ion batteries continue to In hospitals treating victims of patient to the device that pumps and cools get smaller and more affordable to support cardiac arrest or trauma, fast cooling is the fluid requires only an endotracheal tube computers and cell phones. We were able accomplished by surgically accessing large in the patient’s airway to guide the fluid in to find a small laptop battery pack with blood vessels in the heart, connecting them and out of the lungs.

www.alcor.org Cryonics / April 2015 7 In 2013, CCR and SA were granted the patent for the “Portable Apparatus and Method for the Administration of Heat Exchange in the Lungs of a Mammal.” In laboratory experiments, this “liquid ventilation” device for cooling cooled as fast as 1.5 degrees Celsius per minute in a live dog. Research by SA continues to further develop this device for use outside the lab. At SA we have been taking advantage of the most advanced microelectronics and Computational fluid dynamics software to analyze heat exchange efficiencies

electromechanical systems to automate the integrate with other medical devices. SA manual functions of the machine. Using engineers are researching new materials and the same technology that allows computers new hardware configurations to maximize and cell phones to grow smaller and more heat exchange with computational fluid powerful, multi-layer circuit boards and dynamics (CFD) software and finite element low power chipsets have made the controls analysis (FEA). Our goal is to make the and interface of the new liquid ventilation device’s components rugged, lightweight, Patient connected to the portable machine lighter, simpler and able to inexpensive and largely disposable.  liquid ventilation device for cooling.

Become An Alcor Associate Member! Supporters of Alcor who are not yet ready to make cryopreservation arrangements can become an Associate Member for $10/month (or $30/quarter or $120 annually). Associate Members are members of the Alcor Life Extension Foundation who have not made cryonics arrangements but financially support the organization. Associate Members will receive:

• Cryonics magazine by mail • Discounts on Alcor conferences • Access to post in the Alcor Member Forums • A dollar-for-dollar credit toward full membership sign-up fees for any dues paid for Associate Membership

To become an Associate Member send a check or money order ($10/month or $30/quarter or $120 annually) to Alcor Life Extension Foundation, 7895 E. Acoma Dr., Suite 110, Scottsdale, Arizona 85260, or call Marji Klima at (480) 905-1906 ext. 101 with your credit card information. Or you can pay online via PayPal using the following link: http://www.alcor.org/BecomeMember/associate.html (quarterly option is not available this way). Associate Members can improve their chances of being cryopreserved in an emergency if they complete and provide us with a Declaration of Intent to be Cryopreserved (http://www.alcor.org/Library/html/declarationofintent.html). Financial provisions would still have to be made by you or someone acting for you, but the combination of Associate Membership and Declaration of Intent meets the informed consent requirement and makes it much more likely that we could move ahead in a critical situation.

8 Cryonics / April 2015 www.alcor.org The Stasis Foundation Research

By Stephen Valentine, RA (NCARB)

INTRODUCTION for cryostorage. While most buildings are or her Pod, he or she will not be removed Timeship, the primary project at the Stasis for well understood activities, just about from that Pod until reanimation. Foundation Research Park, will be a state- everything that will take place at Timeship of-the-art next generation cryonics facility. will be unique and technologically PATIENT & ORGAN PODS Timeship will provide secure long-term demanding, including determining the site There are two types of Pods, “whole- cryostorage for the DNA of endangered of the building in terms of climate, safety, body” Pods and “organ” Pods that are species, organs for transplantation, and infrastructural and social issues. There used for both organs and neuropatients. both neuropatients and whole-body will also be the need for architectural and Pods must provide both impact and patients for future reanimation. It will mechanical systems of high reliability thermal protection for the fragile vitrified also house laboratories for advanced life requiring minimal maintenance for patients. Toward this objective, a complex extension research. extended time periods. These requirements design was developed with multiple layers, necessitate that Timeship be not just an each layer having a protective or thermal architectural and construction project, role in bringing the patient safely to but also a major ongoing research project. eventual reanimation. From the outside- Years of research have been committed in, the layers of the whole-body Pod are as to the project, often involving the world’s follows: first is a protective enclosure tube leading engineers. with removable top and bottom. Within Timeship has been designed to be a this tube goes a stainless steel vacuum- “building of immortality” both in its insulated volume and an aluminum heat- program and its construction. The products conductive liner to assure that the patient of this research to date are contained in a is kept at an even temperature. The patient Model of Timeship viewing the massive multi-volume Program Report capsule is placed inside a liner, and inside lower cryostorage area divided into and summarized in the book, Timeship: The that the patient cradle is inserted. twelve communities. The hierarchy of Architecture of Immortality. Neighborhoods and Communities allows for In this article we will focus on the security and redundancy. The inner square cryostorage systems at Timeship, very is for LN2 storage and cryogenic equipment. briefly list some of the Foundation’s studies of the literature of cryopreservation and The site is approximately 800 acres life extension, and briefly describe the in Comfort, Texas, less than one hour library being established by the Stasis northwest of San Antonio. Plans are Foundation. currently being drawn up for the first phase There are four components to the Stasis of a fabrication and testing laboratory to cryostorage systems: Pods, Transport further develop an advanced intermediate Containers, Temperature Control Volumes temperature storage system referred (TCVs), and Cold Volumes. Until Cold to here as the Temperature Controlled Volumes at Timeship are operational, Volume (TCV), a cryostorage container cryonics patients would have to be shipped with Timeship Cold Volume features. Once in Transport Containers (which are not The multiple components of we have a proven working prototype of addressed in this article) to a secure location the Full Patient Pod this container, the next phase will be the where they would be placed in TCVs. Once development of the Cold Volumes to be Timeship is operational, the patients will The Pod is the basic module of used in Timeship. be removed from the TCVs and placed in Timeship, and the entire building is While there have been decades of Cold Volumes where they will remain until designed outward from the Pod. The Pod research in cryopreservation, there is also reanimation. One of the features of this design must accommodate the shape of the the issue of the contextual requirements system is that once a patient is placed in his patient on the inside, and the way Pods are

www.alcor.org Cryonics / April 2015 9 packed (fit together) on the outside. The Heat conductors are positioned at the COLD VOLUME STORAGE packing geometry of the Pods is crucial to bottom of the TCV so that they can be The objective of Cold Volume storage the efficient cryostorage of a large number immersed in or removed from an LN2 bath. rooms at Timeship is to provide a large- of patients. Adjusting these conductors via control scale economical method for long-term cables changes the thermal resistance storage for human organs, neuropatients,

TCV (TEMPERATURE COLD between the TCV bottom surface and LN2 and whole-body human patients in a secure VOLUME) pool, and changes the heat transfer rate to cryogenic environment with particular

For the most part, since its inception the LN2, thereby allowing control of the attention to equipment simplicity and cryonics has depended on storing patients temperature of the TCV. Small electric redundancy to optimize reliability and in Dewars—vacuum double-walled resistance heaters are located within the safety. This approach was first proposed by vessels like giant Thermos bottles—filled pod’s surface to “trim” and more finely Dr. Steven B. Harris in 1986. with liquid nitrogen (LN2) that provide a control the temperature as required. Each Cold Volume will contain many reliable constant temperature of -196°C. The heat flow of the TCV is modeled Pods. New patients in their Pods will be However, the advancement of methods analytically using the electrical resistance brought into the upper level of the Cold of cryostorage using vitrification requires analogy approach. Volumes through airlocks to protect a higher temperature of approximately against cold escaping and moisture getting -130°C. In addition, future developments in. Working in this upper level, Timeship in vitrification may require different staff will lower the Pod through access temperatures for different patients, and hatches into the appropriate slot in a current techniques may require varying manner that minimizes perturbations to the temperature during cooling to produce the temperature of the Pods and facilitates annealing to minimize fracturing. While safety for staff involved in transfers. commercially available refrigeration may Individual Cold Volumes are referred to be able to meet these requirements, such as “Neighborhoods,” which are 7m × 7m × refrigeration involves compressors, which 5m high and are configured in a 3 × 3 matrix are not reliable over the long time periods of nine each to form “Communities.” required for cryostorage. The Stasis Assuming whole-body storage, a Foundation and its engineers have done Neighborhood could hold 100 patients, a extensive investigations to address these Community could hold 900 patients, and issues. the entire Timeship complex could hold The solution is the TCV, or Temperature Section Drawing: Proposed TCV Version 10,800 patients. If storage included some Control Volume, which was developed [4] Dewar with Full Patient Pods. percentage of neuropatients, the numbers by cryogenics designer Michael Iarocci, would be higher. physicist Dr. , and Stephen Valentine, architect of Timeship. The TCV allows storage at temperatures compatible with vitrification, and will be used as stand- alone units containing the Pods described above until Timeship is operational, at which time the Pods will be removed from the TCVs and placed in Timeship’s Cold Volumes. The TCV must be physically secure, provide cooling and precise and variable temperatures, and have sufficient insulation, structural integrity, and shock resistance to permit some limited transport. In the TCV, a vertical open mouth Dewar houses a second open mouth Dewar, all View looking into a neighborhood with constructed of stainless steel. Both vessels Schematic for modeling the heat flow about half of the pods in place. have vacuum insulation and superinsulation in a Pod with a highly conductive liner contained within a highly insulated Dewar. to minimize heat transfer, and generally the Surrounding each Community in vessel wall vacuum spaces are common to corridor spaces are local LN2 supply Dewars facilitate re-evacuation from the outside. for the cooling of each Neighborhood.

10 Cryonics / April 2015 www.alcor.org The corridors will also provide the space separated from the areas in which there for the process systems that will maintain is LN2 by airlocks similar to those on a the required temperature for each Cold space station. Workers in areas with LN2

Volume. or subject to LN2 leaks will wear breathing

Evaporation of LN2 provides all of the apparatus for their protection and to refrigeration requirements for the Cold prevent their exhalants from introducing Volumes through the heat absorption moisture and other gases into the sealed property known as heat of vaporization. environment.

The LN2 is injected upstream of the cold Timeship must operate without circulating pump into a diffuser section, interruption for up to hundreds of years passes through additional diffusers at and do so economically. The cost of the pump outlet, then passes heaters to operation is strongly influenced by the achieve precise temperature control before Cold Volume demands, type of circulation entering the cold volume. The nitrogen gas equipment selected, the geometry of is then circulated by fans compatible with the packing of the Pods, and the type of Timeship will keep a backup supply of LN should deliveries be interrupted, and a cryogenic environment. The volumetric insulation used. Close packing of the pods 2 will also be able to make its own LN . Neighborhoods and Communities 2 flow is expected to be minimized so cold in the neighborhoods and communities at This diagram shows the distribution of pump work is minimized. With flow selected Timeship is necessary for maximum energy LN2 within Timeship and the extensive and the number of pods identified, the efficiency. The final choice of packing redundancy of supply to each area. total volumetric flow can be determined. geometry must consider the size of the The quantity with consideration for pods in order to accommodate the greatest Then the next step in the development of hydraulic losses in the ducting, Pods, heats range of patients; the ratio of whole-body cryogenic hardware will be the creation of exchangers, etc., will enable the selection of patients to neuropatients and organs; a scaled version of the Cold Volume. the cold circulating pumps. With pumps and the interior volume to surface area ratio, associated hardware selected for circulation which affects heat transfer; the packing of nitrogen gas, the needed quantities of properties of various geometries; and the cooling LN2 injection can be defined. configuration of the Neighborhoods and The Cold Volumes are held at a constant Communities in relation to the design of pressure slightly above atmospheric the building as a whole. pressure to minimize the infiltration of air As mentioned above, the cooling and moisture which would cause a buildup equipment must be redundant at every of frost. level. In addition, each individual Cold The Cold Volumes are required to Volume cooling apparatus shall interface provide precise temporal and spatial with the adjacent Cold Volumes so as to A community of nine neighborhoods with temperature control so that temperatures add another layer of redundancy and access tube. The small pyramids at the top represent the structural truss system of the in each Pod can vary over time in order flexibility. The 2LN in Dewars on-site to facilitate annealing during cooling. shall provide adequate reserves to cover blast-resistant upper enclosure that provides a high degree of security. The open area Therefore it is necessary that Pods be anticipated LN2 delivery interruptions thermally isolated from each other so that and equipment failures as determined by a under the upper enclosure is the patient transfer volume for maneuvering patients to heat does not interfere with annealing. failure mode and effect analysis (FMEA). be lowered into the suspension matrix grid. During the annealing process normal A careful FMEA early in the design phase convection cooling to the particular Pod of the project will assure elimination of INTERNSHIP RESEARCH is not disrupted, so only local Pod heating fundamental design errors and can be a One of Timeship’s mandates is to become a is required to maintain the increased powerful tool in revealing any systemic major repository of information regarding temperature. After annealing the Pods weaknesses. cryostorage, life extension, and related slowly reach the base temperature over The storage area as a whole is isolated subjects. The material will be stored both time. The base or nominal Cold Volume by thermal, structural, blast, and security online and on site in the Stasis Foundation temperature is approximately -130°C. The barriers, as well as by the air locks Research Library. temporal and spatial temperature control mentioned above. The information is being gathered by requirements are +/- 2.5°C and +/- 0.5°C Once the Pods and TCV described teams of recent graduates working as respectively. The cooling scheme proposed above are fully developed and prototypes interns-in-residence at the Stasis Foundation will allow changes in the base temperature are built and tested, the internal Cold properties to survey the literature in various as required. Volume space frame, transfer hatches, and fields and compile reports. The interns are The areas in which the staff works are transfer equipment can be initially designed.

www.alcor.org Cryonics / April 2015 11 generally from Texas universities and are RESEARCH LIBRARY with interlibrary loan access through a coordinated by Stephen Valentine and the The Stasis Foundation established the system that will provide material directly Stasis Foundation Director of Education Stasis Foundation Research Library (SFRL) from patrons worldwide. and Research. The reports done to date, to be a leading resource center for life With ongoing attention to detail the starting in late 2011, include: extension research. SFRL is a unique SFRL will become an indispensable • Preserving Human Cells and Tissues: organization with an ambitious mission: to resource for everyone from researchers Techniques of Cryopreservation provide resources to advance life extension to individuals new to the concepts of life research and to facilitate the innovation extension. As SFRL emerges as a leading • Cryovessels and the History of Liquid and discovery of new technologies that will research institution, the Library will Nitrogen transform lives and the future of humanity. pioneer dynamic, user-focused methods • The Engram of Memory Encoded The collection will include, but will not for retrieval of information for continuing Through Synaptic Plasticity be limited to, methods of cryopreservation, research in life extension. long-term organ storage, nanotechnology • Memory, Consciousness, and Death and brain science. The collection CONCLUSION • Nanomedicine: Applications to development process will be defined As mentioned at the beginning of this Cryopreservation to include a plan for the future growth article, Timeship is as much a research • Organ Bioengineering of the collection, a tool for delineating project as an architectural project. The collecting priorities, and a guide for those demands for up to hundreds of years • Cryopreservation of Gametes and charged with the responsibility of selecting of uninterrupted, precise conditions for Embryos (A Historical Overview) materials. Timeship’s precious cargo are comparable • Human Reproduction: Process of It is a goal of the Foundation to provide only to the demands of the International Collection of Gametes and Embryos for free access to the Library’s collection to Space Station. Timeship’s design and Cryopreservation (IVF) the widest possible audience. This will be engineering must meet this challenge.  accomplished with on-site access along

Reduce Your Alcor Dues With The CMS Waiver

Alcor members pay general dues to cover Alcor’s new minimum for neurocryopreservation members under operating expenses and also make annual contributions this election would be $110,000). Once this election is to the Comprehensive Member Standby fund pool made, the member cannot change back to the original to cover the costs of readiness and standby. Benefits minimums in the future. of Comprehensive Member Standby include no out- To have the CMS fee waived, these are the minimums: of-pocket expense for standby services at the time of need, and up to $10,000 for relocation assistance to the • $220,000 Whole Body Cryopreservation Scottsdale, Arizona area. ($115,000 to the Patient Care Trust, $60,000 for Instead of paying $180 per year in CMS dues, Alcor also cryopreservation, $45,000 to the CMS Fund). provides members the option to cover all CMS-associated • $100,000 Neurocryopreservation ($25,000 costs through life insurance or pre-payment. Members to the Patient Care Trust, $30,000 for who provide an additional $20,000 in minimum funding cryopreservation, $45,000 to the CMS Fund). will no longer have to pay the $180 CMS (Comprehensive Member Standby fund) fee. This increase in minimums If you have adequate funding and would like to take is permanent (for example, if in the future Alcor were to advantage of the CMS waiver, contact Diane Cremeens raise the cost of a neurocryopreservation to $90,000, the at [email protected].

12 Cryonics / April 2015 www.alcor.org 21st Century Medicine

1st Century Medicine (21CM) is a runaway, and projections indicate non-invasively when rabbit kidneys biotechnology company specializing that similar warming should be prepared for vitrification are ready 2in complex system cryopreservation, possible even on human sized to be vitrified; and particularly the cryopreservation of organs, so that rabbit kidney whole mammalian organs. Research on technology should be scalable to the • using similar techniques, 21CM has organ cryopreservation is of great interest size of human organs, potentially been able to show that the limiting to those interested in cryonics, such as those allowing demonstration of large factor for kidney vitrification is who have signed up with Alcor. The full organ survival after vitrification; small variations in gamut of 21CM research is not described concentration from microregion here, but recent research at 21CM that is • fracturing can be prevented in to microregion of the renal inner particularly relevant to cryonics includes human organ sized solution medulla (see YouTube video of the following. samples even after cooling to liquid Dr. Fahy’s symposium presentation nitrogen temperature, and organs at the 50th anniversary meeting • As little as 1-2% ice formation in themselves should be even less of the Society for Cryobiology one part (the inner medulla) of a liable to fracturing than these liquid at https://www.youtube.com/ previously vitrified rabbit kidney samples; watch?v=weeOjfQZtX0 ); during warming (devitrification) • published 21CM data indicate that was damaging but was compatible • extending this observation, the formation of new ice crystals with life support of the kidney 21CM has shown that the same (ice nucleation) in M22 should after transplantation (providing phenomenon of incomplete be almost totally suppressed at the first proof of principle that uniformity of cryoprotectant temperatures of about -140ºC; whole organs can be vitrified and distribution applies to all 40 recovered in a viable state); • published 21CM calculations different tissue types tested in both indicate that temperatures of about rabbits and pigs; • using a new perfusion process and -140ºC should permit safe storage a variety of other new technologies, • highly preliminary evidence of living systems (no biological kidneys can now be perfused with suggests that new 21CM methods deterioration) for extremely long might be able to overcome or at a powerful vitrification solution periods of time (tens of thousands least blunt this limiting factor for (M22) well enough to become of years, or longer); immune to devitrification at vitrification of the kidney and warming rates above 40ºC/min and • the vast majority of vitrified other organs, even when the organs yet maintain life support after this organs of 70 kg (human-sized) pigs are damaged by warm ischemia or new perfusion process; (including the kidney, heart, liver, prolonged cold storage (or even the and brain) can be reliably cooled to combination of the two) prior to • using technology developed at -140ºC without fracturing; cryoprotectant perfusion; 21CM, uniform electromagnetic warming of volumes the size of • using both refractometry and • diagnostic methods for imaging rabbit kidneys can be done at up differential scanning calorimetry, and quantifying ice formation to 160ºC/min without thermal 21CM has been able to predict throughout large, mostly vitrified

www.alcor.org Cryonics / April 2015 13 3-dimensional objects are possible chilling injury, but also provides • 21CM collaborative studies using physical sectioning below evidence that reversible brain have helped to identify new the glass transition temperature to cryopreservation by vitrification is mechanisms of chilling injury and expose structural details and allow not precluded by chilling injury); cryoprotectant toxicity, both in sampling of tissue sub-regions for blood vessels specifically (which detailed investigation, including • brain viability can be maintained are found in all organs, including determination of how much damage for 15 hours or more at 3ºC (which the brain; results are currently being is done by incomplete vitrification; may help rescue trauma victims written up for publication) and in and soldiers, but may also protect systems of potential commercial • amide toxicity can be neutralized in brains during preparation for value (particularly liver slices; kidney, liver, and brain slices (which cryopreservation); results already published), and shows that 21CM technology is 21CM analysis of these studies has • 21CM protection methods allow applicable to all of these systems, suggested for the first time what the brain histology and ultrastructure to again to the potential commercial overarching mechanisms of both be very well preserved after one hour benefit of 21CM, but which also chilling injury and cryoprotectant of warm ischemia in both rabbits implies that the benefits of solutions toxicity may be; like M22 might be applicable to and pigs (which may help trauma and whole brains); heart attack victims as well as soldiers • very recent unpublished major survive longer periods of cardiac breakthrough studies have identified • using sophisticated HPLC methods arrest, but also provides evidence multiple independent interventions developed at 21CM, 21CM consistent with the possibility that have already resulted in radical research has shown that all of that brains can be successfully improvements in cell survival or the cryoprotective agents in M22 cryopreserved even after significant function after M22 exposure in penetrate into multiple organs, preceding warm ischemia); model systems; these interventions including the brain, about equally • brain pre-fixation prior to perfusion appear likely to be helpful when rapidly, even though, in the brain, applied to rabbit kidneys in in vivo in shrinkage is observed; with M22 appears to preserve normal-looking brain ultrastructure studies that are now being planned. • brain (hippocampal) slices from in the cryoprotected state (which both adult rats and adult rabbits can not only may allow 21CM to win In addition to moving ahead with be vitrified and rewarmed without the Brain Cryopreservation Prize rabbit kidney vitrification using new any losses of structural integrity, but also provides an alternative for interventions to negate toxicity, 21CM is viability, or functionality (including some who may prefer this method also currently hoping to extend its work responsiveness to electrical of cryopreservation); and on rabbit kidneys to porcine kidneys. With stimulation, action potential luck, porcine kidneys will be found to be amplitude and conduction velocity, • recent close examination of whether less variable in their response to M22 than etc.); this is critical for 21CM’s 21CM might be able to win the Brain rabbit kidneys, and they and their hosts goal of marketing vitrified organ Cryopreservation Prize via ordinary will be more tolerant of M22 perfusion slices to pharmaceutical companies (potentially viability-preserving) and transplantation. If this works as and might be able to provide a methods of M22 perfusion has hoped, not only will it show that organs method for obtaining cellular indicated that, by employing can be reliably cryopreserved, but it will resolution of the 2-deoxyglucose very high-magnification imaging also show that human-sized organs can be method for probing basic features methods, extensive synapses and reliably cryopreserved. Obviously, if this of brain physiology, as discussed intact membranes are readily visible is achieved, cryonics will become more elsewhere (but it also provides the in the cryoprotected state after all, credible as a byproduct. beginnings of a proof of principle without any particular interventions On another track, 21CM was approached for the possibility of whole brain (which is obviously good news for a few years ago by NASA for advice cryopreservation); the possible feasibility of cryonics). about how to freeze dead astronauts in space. And 21CM has been involved in • complete survival of vitrified- In addition to work done directly discussions with the X-Prize administrators rewarmed brain slices implies that at 21CM, 21CM research has inspired about the prospects for cryopreserving brain slices are not susceptible collaborative and even independent whole mice. While 21CM scientists believe to chilling injury (which not only research in other laboratories. The most the possibility of cryopreserving a whole provides information critical for important examples of this type of inspired mammal and reviving it is remote at this deciphering the mechanism(s) of extramural research include the following: time, even the late Harold Meryman, a

14 Cryonics / April 2015 www.alcor.org foundational figure in the science and history of cryobiology, and no friend to cryonics, acknowledged that, when all other problems had been solved, the ultimate goal of cryobiology would have to be human suspended animation. And Audrey U. Smith, another pillar of the development of cryobiology as a science, undertook very extensive experiments on the revival of frozen hamsters, with illuminating and widely published results. Figure 1. Pig cerebral cortex on the left and hippocampus on the Consequently, when 21CM was right after 1 hr of warm ischemia following 21CM mitigation. approached, several years ago, by a wealthy No cryoprotectant. General brain structure is preserved. family interested in providing grant support for whole mammal cryopreservation, it was decided that the project deserved some consideration. Although the task of reversible mammalian cryopreservation might require 20 years or more to come to fruition, it is also true that it will never happen if no research is ever done to at least explore the boundary conditions that may exist with respect to this problem. The long-term problems of cryopreservation for space migration seemed to deserve Figure 2. Pig cerebral cortex on the left and (in higher magnification) some preliminary study. Plus, there was cerebellum on the right after 24 hours of continuous cold perfusion an additional consideration of some followed by perfusion with 105% of full concentration M22. weight, which was simply that, as much as General brain structure is preserved. The white zones in both cryobiologists and cryonicists have debated images are dilated capillaries, which is a typical osmotic effect. one another and tried to convince each other of the errors of their ways, what has always been missing from the debate has is obtainable with rabbits, pigs, and sheep, Static storage, which is the current norm, been adequately relevant facts. While strong brains being perhaps the most easily vitrified resulted in significant thalamic damage, opinions have always abounded on both organ in the entire body, with preservation whereas continuous perfusion enabled sides of this argument, the cryobiologists of both histology and ultrastructure. After remarkably good preservation of brain have not had sufficiently convincing hard 1 hour of warm ischemia, adequate results structure. In addition, ischemia plus 24 data to dash the hopes of the cryonicists, can still be obtained in both rabbits and hours of continuous perfusion resulted in and the cryonicists have been equally pigs as well (sheep not studied). Similarly, a mean maximized (by devitrification) mass lacking in data sufficient to refute the cold storage or cold perfusion for 24 hours percent ice in the brain of about 0.1%, entirely reasonable negative inferences of adequately preserves brain structure with whereas ischemia plus conventional static the cryobiologists. Finally, the prospect of and without subsequent perfusion with storage resulted in almost 4.5% w/w brain obtaining simultaneous information about M22. In addition, the mean percent of brain ice. the equilibration rates of all organs in the mass that became ice under conditions Thus, these experiments do not resolve body was an interesting side benefit of the that maximized ice formation after M22 the debate between the pro- and the anti- idea of doing research on whole mammals. perfusion (i.e., with devitrification; averaged cryonics camps. They do, however, seem Therefore, although this was not a project over 21 independent samples per brain) to shed considerable new light on this very 21CM would have ever undertaken on its was about 0.13% after 24 hours of static under-researched subject. They also suggest own, the value of receiving supplemental storage and about 0.75% after 24 hours that additional research is justified, and that funding to enable this “luxury” project to of continuous perfusion prior to M22 improvements in cryonics methods might be undertaken was deemed to be persuasive. perfusion, which is perhaps acceptable in allow significant progress in the future.  The results of this “non-partisan” both cases. investigation so far have given both sides However, 1 hour of warm ischemia of the cryonics debate some points. Under plus 24 hours of cold storage gives results good conditions, good brain preservation that depend on the techniques employed.

www.alcor.org Cryonics / April 2015 15 When it Comes to the Health Benefits of a Mediterranean Diet Can Your Omega- Compare?

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To order Super Omega-, call --- or visit www.LifeExtension.com 16 Cryonics / April 2015 www.alcor.org

These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease. When it Comes to the Health Benefits of a Mediterranean Diet Can Your Omega- Compare? By Aschwin de Wolf and Chana Phaedra From supporting heart health and brain function to balancing the inflammatory response, there is no debating the broad- spectrum benefits of omega-3 fatty acids.- Introduction of the effects of cryoprotectants on the phenomenon, especially if the blood is Advanced Neural Biosciences was founded rat brain (blood brain barrier permeability, replaced by an organ preservation solution. There are hundreds of fish oil supplements on the market, but in late 2008 by Aschwin de Wolf and the effects of ischemia, etc.) appear to This finding still stands today and validates only one incorporates lifesaving findings to provide optimal Chana Phaedra as a result of a number of confirm what we are observing in human contemporary cryonics procedures such as omega-3 and olive fruit extracts, along with sesame lignans, in unexpected events favoring its creation; the cryopreservation cases. rapid ice bath cooling and remote blood one molecularly distilled formula—Super Omega- from Life departure of Yuri Pichigan at the Cryonics substitution. Extension®! Institute, whose research equipment and Prior Research supplies were made available to us; a We started our research using the red Fish Oil Olive Fruit Extract generous private donation from Cryonics blood cell (RBC) to investigate the effects “If cardiac arrest is followed + Institute member Alan Mole; free lab space of different cryoprotectants and loading Research confirms that a combination of both fish oil and olive made available in Salem, Oregon, by Jordan protocols on viability. While this model is by rapid cooling and blood oil support a healthy inflammatory response than fish oil alone. Sparks (the founder of Oregon Cryonics), too “crude” to distinguish subtle differences washout with MHP-2, ice-free And only one omega-3 product incorporates the benefits of both and the strong support by Ben Best to between cryoprotectants, or the effects of fish oil and olive fruit extract in one bottle—Life Extension®’s engage in practical cryonics research. cryoprotectants on specific cell types such cryopreservation of the brain is Despite our innocuous name, our as endothelial cells or neurons, it provided Super Omega-. Each two softgel servings supplies the equiva- Super Omega- still feasible for at least 48 hours lent amount of  to  ounces of polyphenol content found in research was motivated from the beginning us a basic method to understand the effects extra virgin olive oil. Item #01482 • 120 softgels • Non-GMO by the aim of validating and improving the of cryoprotectants, as mono-agents or in of cold “bloodless” ischemia.” science of human cryopreservation. From combination. While we quickly moved on Retail Member 2009 to 2014 our company conducted to more advanced research models, we Sesame Lignans Price Price + contract research in Salem, Oregon, initially occasionally have used the RBC model After researching the no-reflow Studies show that when added to fish oil, sesame lignans  bottle $ $ on an uncompensated basis on weekends to look for acute toxicity issues in new phenomenon using ink perfusion we safeguard against oxidation and direct fatty acids toward and later as a part-time endeavor. In late cryoprotectants we are testing. We also used developed a model in which we investigated  bottles $ each $ each pathways that help with inflammatory reactions. 2013, we relocated to a bigger lab space in the model to shed light on a debate about the effects of ischemia on cryoprotectant the cryopreservation of James Bedford, perfusion and vitrification. We have used  bottles $. each $. each Portland, Oregon, and secured sufficient funding to employ the two founders full in whose case it had been (erroneously) this model extensively over the years to = Health Benefits of time. In 2014, we expanded the company claimed that 100% pure concentrations of understand the effects of ischemia on a Mediterranean Diet with a part-time employee and entered DMSO were used, which would have been, cryopreservation of the brain, initially 2015 seeking to grow further. as we observed, highly destructive, at least spurred and supported by the Cryonics No other commercially available fish oil supplement contains Our animal model of choice is the rat. to this type of cell. Institute’s desire to improve the quality of References this level of essential fatty acids, sesame lignans, and olive fruit 1. Public Health Nutr. 2006 Dec;9(8A):1136-40. While the use of small animal models has Our first investigations into the effects cryopreservation after periods of prolonged polyphenols. 2. Am J Prev Med. 2005 Nov;29(4):335-46. often been challenged in aging research of ischemia on the perfusability of the warm and cold ischemia and/or extended 3. J Am Diet Assoc. 2005 Mar;105(3):428-40. 4. Nutrition. 2005 Feb;21(2):131-6. because of the limited relevance to larger brain involved a transcardial perfusion patient transport times. We described our Super Omega- uses a proprietary process to produce a pure, 5. Biochem Biophys Acta. 2004 Jun 1;1682(1-3):80-91. mammals (including humans) because of model in which the brain was perfused findings in more detail in our 2011Long Life stable, and easy-to-tolerate fish oil that exceeds the standards genetic differences, we have found that with India Ink after various periods of magazine article “Human Cryopreservation set by international rating agencies, ensuring any pollutants are Note: While the health benefits of omega-3s from fish oil are universally recognized, the critical importance of olive oil in maintaining healthy in the case of ischemia and cryobiology cold or warm ischemia. Our most robust Research at Advanced Neural Biosciences.” reduced to a virtually undetectable level. vascular function remains largely overlooked. research our results track findings in finding in that research was that rapid (http://www.advancedneuralbio.com/ larger animal models and observations in cooling after cardiac arrest protects the publications/) In short, we validated humans well. In particular, our findings brain against the so-called “no reflow” prior research in the field and observed To order Super Omega-, call --- or visit www.LifeExtension.com www.alcor.org Cryonics / April 2015 17

These statements have not been evaluated by the Food and Drug Administration. This product is not intended to diagnose, treat, cure, or prevent any disease. that as the duration of warm and cold preservation solution” that was made combination of chemical fixation PLUS ischemia increases, the blood-brain barrier available to us by 21st Century Medicine. cryoprotective perfusion and found that it is is progressively compromised, edema of Both solutions, however, were not effective possible to completely inhibit ice formation the brain and the whole body increases, in preventing (severe) whole body edema in a chemically fixed brain, provided there and delivery of the cryoprotectant to the and weight gain during subsequent are no ischemic delays between cardiac brain is poorer, producing ice formation cryoprotectant perfusion, an issue that, to arrest and the start of chemical fixation. upon cryopreservation (despite the use our knowledge, has not been successfully Interestingly, prompt chemical fixation of of a vitrification agent). Most of our solved in the design of hypothermic organ the rat prevents vascular leakage secondary research efforts were aimed at identifying preservation solutions. Preventing this to ischemia, which permits cryoprotective interventions and protocols to reverse this type of vascular leakage (and breakdown perfusion and ice free cryopreservation ischemia-induced perfusion impairment of the blood brain barrier) may require a without edema (!) after up to 2 weeks of at the time of cryoprotective perfusion. transition from “static” organ preservation storage at refrigerator temperatures (~ 4° While ice formation worsened when solutions to intermittent perfusion of perfusion pressure was increased during oxygenated solutions, continued perfusion cryoprotective perfusion in ischemic en-route to a cryonics facility, or conducting “After a thorough examination brains, we did observe benefits when we cryoprotective perfusion in the field. introduced the target concentration of the We also received a grant from LongeCity of the literature and designing a cryoprotectant without gradually increasing that allowed us to investigate whether lot of experiments we discovered the concentration and we observed blood substitution is still beneficial after noticeable improvement when we increased various durations of normothermic a protocol that allowed us to the viscosity of the cryoprotectant. While (i.e., body temperature) ischemia—an successfully recover the rat from the former approach is expected to important practical topic for cryonics improve protection against ice formation at organizations. This research was motivated no warmer than 0° Celsius the cost of (extreme) osmotic damage, the by former President (without blood washout) by approach of increasing the viscosity of the Ben Best’s concern that attempting cryoprotectant appears more appealing and blood washout after warm ischemia could providing adequate metabolic we identified a number of potential carrier actually worsen subsequent cryoprotective support during cooldown and solutions and cryoprotectants that were perfusion by introducing free radical- successful in reversing ischemia-induced mediated reperfusion injury. What we rewarming.” perfusion impairment and ice formation, found, however, is that up to 1 hour of especially in the case of cold ischemia. normothermic ischemia, subsequent blood To our knowledge, this was the first time washout is still beneficial in decreasing Celsius). ischemia-induced perfusion impairment perfusion impairment and ice formation. Advanced Neural Biosciences further during cryoprotective perfusion was At 1 hour of normothermic ischemia we collaborated with Dr. Perry at Alcor to mitigated through modification of the no longer observed any benefits from produce a series of electron micrographs carrier solution. conducting blood substitution (aside from of cortical samples of the rat brain after Our most successful results in a cold accelerating cooling). various periods of normothermic ischemia ischemia model were observed when In collaboration with Dr. R. Michael to complement the warm ischemia (room we replaced the blood of the rat with an Perry at Alcor we investigated how temperature) research that Alcor published organ preservation solution. An important ischemia affects the ability to chemically in 2008. (http://www.ijcep.com/707005A. discovery was that the composition of the fix the brain. As in our cryopreservation html) In 2013 and 2014 we added a series solution is important and the best results experiments, we observed that perfusion of cold ischemia images which includes were seen when we replaced the blood with impairment as a result of ischemia cortical images from rats stored at water Alcor’s MHP-2, a high-potassium, HEPES- produced incomplete fixation of the ice temperature for up to 6 months. In buffered “extracellular” whole body organ brain, resulting in subsequent autolysis of addition to the conventional electron preservation solution that includes the the poorly fixed areas over time. These micrographs we also sent a number of oncotic agent hydroxyethyl starch (HES). results provide a meaningful context to the cortical brain samples that were subjected If cardiac arrest is followed by rapid current debate about the relative merits of to a variety of interventions (e.g., straight cooling and blood washout with MHP-2, cryopreservation or chemical fixation as freezing, ischemia) to 3Scan to create ice-free cryopreservation is still feasible pathways to long-term identity preservation 3-dimensional reconstructions. To our for at least 48 hours of cold “bloodless” because it favors cryopreservation over knowledge, this is the first time that ischemia. This result was not observed chemical fixation in circumstances where these technologies have been used for with saline or other organ preservation start of the procedure is delayed. the characterization of permanent global solutions, with the exception of a “brain In addition, we also looked at the cerebral ischemia and freezing of the brain.

18 Cryonics / April 2015 www.alcor.org This comprehensive series of images will be perfusion. These findings are consistent protocol that allowed us to successfully analyzed by experts in the field of electron with similar studies at 21st Century recover the rat from down to 0° Celsius microscopy and presented together in Medicine and the Cryonics Institute, (without blood washout) by providing a formal paper. Dr. Perry also hopes to including investigations of ice formation adequate metabolic support during use these images to further develop his tendencies in large solutions of VM-1 and cooldown and rewarming. Our next step algorithm to characterize, infer, or predict M22 at dry ice temperature. will be to add extracorporeal perfusion to the ultrastructural signatures of various this model, introduce a low concentration periods of normothermic or cold ischemia. Current Research cryoprotectant, and recover the rat from The images will also be of practical use for Our relocation to Portland, Oregon, and high subzero temperatures. cryonics organizations to employ more our full-time employment, has allowed Another important reason to pursue this empirical evidence to decide the maximum us to pursue a number of new, ambitious model is the ability to recover organized delays beyond which it would no longer be research directions. electrical activity after cooling to ultra- sensible to accept new (third party) patients One of our main interests in the last few profound hypothermic temperatures— after clinical death. Finally, the images can years, and a prerequisite for other research the temperatures at which cryoprotective also be used as a starting point to conduct projects, is whole body resuscitation from perfusion is initiated. A functional (isolated) “reconstructive connectomics,” i.e. the ultra-profound hypothermic circulatory whole brain model should be successful in inferring of the original state of the brain arrest temperatures. We decided to pursue lowering the temperature to 0°C and permit from the damaged state and do “in silico” this model for two reasons. First, because successful recovery of the EEG after repair, which can serve as a template for we believe it is possible to resuscitate rats rewarming before it can be used to study biological repair. from temperatures lower than the lowest the effects of cryoprotective perfusion and We have also produced electron temperatures reached by Andjus, Smith, et cryopreservation. In the model we use in micrographs for the Cryonics Institute to al. While the rat and hamster resuscitation our lab this requires the re-introduction of characterize the ultrastructural effects of experiments from the 1950s and 1960s blood or a blood substitute after the blood VM-1 and alternative modifications of this established the recovery of mammals from in the brain has been replaced with an organ agent on the brain. In 2014 we produced circulatory arrest at low temperatures, Smith preservation solution or cryoprotectant. electron micrographs for Oregon recognized that efforts to move beyond the We have screened a large number of Cryonics, which uses a gentler loading high subzero temperatures that were used isolated head perfusion solutions and protocol for VM-1 that includes more low in these experiments (down to -5° Celsius) protocols to identify a suitable protocol. concentration ethylene glycol steps. These would require extracorporeal perfusion At atmospheric pressures purely aqueous images were produced using a protocol and cryoprotectants. The increasing solutions lack the oxygen-carrying capacity with or without an agent in the carrier interest in small animal extracorporeal for this job but we have been successful solution to modify the blood brain barrier models, and the development of low- in maintaining organized electrical activity to improve penetration of the vitrification toxicity vitrification agents that eliminate in the brain for up to 1 hour with washed solution and reduce dehydration. freezing altogether, now make it possible, red blood cells in a physiological carrier Concerns about the effects of cold in principle, to go below the temperatures solution. Our preference, however, is to use ischemia sustained during transport to of those researchers and pursue real perfluorocarbon-based emulsions which the facility prior to cryopreservation suspended animation research. have a similar oxygen carrying capacity but has renewed interest in two alternative One crucial difference between lower viscosity, are easier to modify, and approaches: (1) continuous perfusion hibernating and non-hibernating animals more practical. We currently have acquired of oxygenated organ preservation is that in the former the temperature can the equipment and supplies to prepare and solutions during transport and (2) “field be lowered and heart rate and respiration validate such blood substitutes. If we can cryoprotection.” Alcor has authorized will gradually decrease and eventually arrest successfully use these perfluorocarbon- field cryoprotection for overseas cases without affecting viability of the animal. based blood substitutes to recover electrical on the reasonable assumption that such a Upon rewarming cardio-respiratory activity in the brain after cooling, we can procedure is superior to a straight freeze. activity will resume again and the animal combine this model with our cryobiological In Alcor’s field cryoprotection protocol the will recover. In non-hibernating animals perfusion model and investigate the effects vitrification agent is introduced in a series (such as the rat), however, lowering the of cryoprotectants and cryopreservation of discrete steps instead of a ramp, after temperature until circulatory arrest occurs on the whole brain with viability as an which the patient is shipped on dry ice does not produce such a synchronized endpoint. to the Alcor facility for further cooldown condition and simply rewarming the animal In addition to this methodologically to cryogenic temperatures. In our lab we is not sufficient for successful recovery advanced whole brain model we also use a simulated this protocol and did not observe from circulatory arrest. After a thorough (hippocampal) brain slice model to measure any ice formation after 48 hours storage of examination of the literature and designing viability after cryopreservation. This model the brain on dry ice after cryoprotective a lot of experiments we discovered a can be used to do a preliminary screen of

www.alcor.org Cryonics / April 2015 19 suitable, low toxicity cryoprotectants to Future Research to the additional damage from cold be tested in the whole brain model but is Our whole brain cryobiology and whole ischemia associated with such an also valuable to understand the effects of body recovery models are still in early immersion protocol? How is the cryoprotectants on neural tissue without stages of development and we expect to brain prepared and loaded with the the presence of a circulatory system or remain quite occupied with this for years cryoprotectant? How will the brain be blood-brain barrier. The model has been to come. We are, however, interested in a isolated and stored? etc. used successfully in conjunction with the number of other topics: 4. “Reconstructive connect- potassium/sodium ratio viability assay to 1. Cryoprotectant toxicity. At this omics.” Our lab created the identify low-toxicity vitrification agents point there is only one lab in the first 3-dimensional ultrastructural (such as M22 and VM-1) in the brain. In world (21st Century Medicine) that reconstructions of the damage addition to this assay, we have identified a can devote considerable efforts and associated with ischemia and ice number of other slice viability assays that resources toward understanding formation. These images can give can be correlated with each other to create the nature and biochemistry of us some idea of the nature of the a compound measure of viability. Our cryoprotectant toxicity. Since damage associated with older or brain slice set-up is also able to directly eliminating cryoprotectant toxicity suboptimal cryonics protocols. But measure spontaneous and evoked electrical is the most fundamental obstacle to answer the question whether the activity, which is the “gold standard” for to reversible cryopreservation of original state can be inferred from brain slice viability. complex organs and true human the damaged state, more advanced suspended animation, a greater methods are required, including, understanding of this phenomenon but not limited to, mathematical “We also are collaborating with is highly desirable, in particular as it modeling of the natural and Alcor to screen its comprehensive pertains to the brain. damaged state. 2. Depressed metabolism. The Our general aim is to engage in what multi-modal stabilization pharmaceutical agent Propofol is we would call “integrated cryonics.” currently the only drug in Alcor’s medications protocol.” Instead of chopping cryonics up into medications protocol aimed at distinct challenges such as “ischemia,” reducing cerebral metabolism after “cryopreservation” and “repair” it is One of our projects in 2015 is a cardiac arrest. We have no real often valuable to look at these issues in a collaboration with the understanding of how effective “holistic” fashion.  to bring together these viability assays and this approach is, how it compares electron microscopy to validate and publish to other anesthetics, and whether a paper about the low-cost vitrification novel approaches could improve agent VM-1 (or a lower toxicity variant). upon the (potential) benefits of We also are collaborating with Alcor Propofol. to screen its comprehensive multi-modal 3. “Universal ice-free cryo- stabilization medications protocol. Instead preservation.” The current of using viability as an endpoint we look situation in cryonics is that only at the efficacy of these medications and patients with little ischemic injury solutions in improving perfusion and can take full benefit of vitrification inhibition of ice formation after various technologies. Our electron durations of cold and normothermic micrographs of cold ischemic brains ischemia. At this point we may have have prompted some observers to identified a novel approach to substantially speculate whether patients in which reduce perfusion impairment and ice cryoprotective perfusion is no longer formation that could be a powerful tool feasible (or is suboptimal) could to increase the number of cryonics cases benefit from immersion of the naked amenable to ice-free preservation. A brain in a vitrification agent to ensure secondary objective of this research is complete distribution of the agent. to have a better understanding of which This possibility raises a number of medications are most important and to practical and research questions. What establish a more compact and practical are the most suitable cryoprotectants administration protocol. for such a protocol? How does the absence of ice formation compare

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Why We are Cryonicists Notes on the First Human Freezing Dear Dr. Bedford How Cryoprotectants Work How Cold is Cold Enough? The Death of Death in Cryonics The Society for The Recovery of Persons Apparently Dead Frozen Souls: Can A Religious Person Choose Cryonics? But What Will the Neighbors Think?! Systems for Intermediate Temperature Storage for Fracture Reduction and Avoidance

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www.alcor.org Cryonics / April 2015 21 Critical Care Research ......

By Steven B. Harris, M.D

ritical Care Research (CCR) arrests are unwitnessed, as they happen in of the brain is badly compromised. In such conducts a number of projects, people who are alone or asleep. In others, cases, the only solution is to quickly freeze Cmostly using the dog model and there is no time to get to the hospital and the patient’s brain, and hope that repair mostly involving cooling, which have it requires time for the paramedics to get techniques in the future are far, far better many potential medical applications. to the victim—time which may not be than are needed for the “average” cryonics Among these are treatment of cardiac available. A person suffering cardiac arrest patient who can have cryoprotection of the arrest brain damage, treatment of shock- in the Emergency Department (ED) still brain. Perhaps strangely, there are times that trauma, and treatment of several types only has a 66% chance of being resuscitated initially “successful” resuscitation is worse of lung injury. Since cryonics involves to normal circulation and finally discharge for cryonics than no resuscitation at all. cooling from normal body temperature to home. Outside the ED this chance drops down to the deep cryogenic temperature by 6% for every minute that resuscitation TREATMENT OF POST of storage (-140°C to -196°C), both CCR (return of heartbeat and blood pressure) is RESUSCITATION BRAIN DAMAGE and its partner, 21CM have input into the delayed. By 10 minutes without resuscitation CardiopulonaryCardiopulmonary support support (CPS(CPS oror CPR)CPR) design of the modern cryonics procedure, outside the hospital, the chances of an adult such as chest compressions while waiting although the procedure itself is handled by being resuscitated and having so little brain for paramedics to deliver an electrical other institutions. damage as to be able to (eventually) return defibrillation shock that restarts the heart, Critical Care Research receives almost home, are close to zero. can increase the brain-damage-free cardiac all of its support from The Life Extension Thus, while it is not true that after four arrest time. But CPS delivers only half of Foundation. to six minutes without blood flow the brain the blood the brain is used to getting, so it “dies” (a popular belief), it is true that after 10 cannot extend the time before heart restart SUDDEN (UNEXPECTED) CARDIAC minutes without blood flow, the brain of an and brain recovery, for more than a few ARREST adult has been damaged to the point that the tens of minutes. Sudden cardiac arrest (SCA) is often brain will almost certainly die in a few days. There are still no drug treatments for defined as having the heart stop beating It often happens, then, that heartbeat and the brain damage that happens in cardiac within an hour of any warning signs (such blood pressure are restored after a cardiac arrest during the time there is no, or greatly as chest pain) in a person who is not dying arrest, yet the brain slowly deteriorates from reduced, blood pressure during CPR. There of another disease (such as cancer) and who the damage it suffered from no oxygen, are many drugs that extend the time of would want to be subjected to resuscitation, until brain capillary breakdown completely damage-free arrest in animals, but all have if possible. SCA is quite common, killing at stops blood from flowing to the brain, failed in human trials, probably because least 200,000 Americans a year, or at least even while the heart continues to beat their effect is too small to show up through 8% of the total of people who die each and the rest of the body recovers. This is the large “noise” of a trial. (There are too year in the US. Most cardiac arrest is due to the “brain death” diagnosis at which it is many variables in a trial from one person a “heart attack” (M.I.) where a clot blocks a pointless to continue, and when the patient to another, including great difficulty in major heart artery and the damage induces is pronounced legally dead, and removed deciding exactly how long they have been a heart dysrhythmia. from the ventilator (“life support”). At in arrest.) Sudden arrest is surprisingly resistant to this point, cryonics perfusion of the brain The only successful treatment for this public health approaches. Almost half of does not work, either, so cryonics rescue damage discovered to date, is to cool the

22 Cryonics / April 2015 www.alcor.org brain after the heart is restarted. This effect become clinically available, which is able to For example, a study in 1993 by Kuboyama was discovered by accident in dogs in the induce the needed 7ºC temperature drop et al. [4] reported that a delay of only 15 early 1990s. Analysis showed that dogs that in less than two hours. It was suggested minutes from resuscitation to beginning of happened to cool a bit (by chance) after that in-hospital cooling might be useful application of hypothermia, negated the experimental resuscitation did better. By for other rhythms and types of in-hospital beneficial effects of cooling. Later studies 2000, experiments had shown the effect to cardiac arrest. [3] extended this time, but generally found that be certain, and large. Just cooling the brain the sooner cooling was applied, the better by 4ºC (7ºF) for 24 hours was enough to for the brain. increase the damage-free arrest time by at “In 2001, Critical Care Animal models of cooling (some least 50%, and sometimes 100%. The effect of the most advanced done at CCR was something like putting a cold pack on Research, Inc. (CCR) published using lung lavage with ice cold Critical Care Research ...... a twisted ankle, after an injury. During a paper [6] showing that FLL perfluorocarbon liquids) have explored the arrest these dogs did not need to be methods of cooling which cool in hypothermic. (Hypothermia can be applied lavage was able to reduce brain 1/25th of the 2 hour time needed to before arrest, and works even better, but temperature in dogs by a rate as induce mildmild hypothermia (7°C(7ºC belowbelow for obvious reasons, medical application is body temperature, oror down to 30°C) inin extremely limited.) fast as 0.5ºC per minute.” humans. In animals, this amount of Human studies soon followed. In 2002, cooling of the brain may be induced in two prospective randomized trials in as little as 5 minutes after insertion of comatose resuscitated survivors of out- ANIMAL MODELS OF POST an endotracheal tube. of-hospital cardiac arrest victims were RESUSCITATION HYPOTHERMIA reported. These trials used patients with Animal models of cardiac arrest, such LIQUID BREATHING AS EXAMPLE return of spontaneous circulation (ROSC) as rodents and canines, have made it OF AN EXPERIMENTAL COOLING but persistent coma after a witnessed possible to investigate the mechanisms METHOD episode of documented ventricular of modulation of post ischemic events Inhalation of water damages the lungs fibrillation (thus insuring that all patients in the cerebrum by use of direct brain osmotically when the fluid is either hypo- had truly experienced no significant brain biopsies and chemical analyses during the osmotic (fresh water drowning) or hyper- blood flow). Groups of untreated patients post resuscitation period. Such studies osmotic (salt water drowning). However, were compared with patients deliberately have shown that the mechanism of mild even iso-osmotic saline causes temporary treated with post-arrest cooling by cold hypothermia protection of the brain in the hypoxia in the lungs after the fluid is packs, or air cooling. The target was about post-ischemia period is nearly as complex removed, via the mechanism of removal of 4ºC of cooling (7ºF) within 4 hours of as the mechanisms of damage of ischemia the water soluble surfactant which allows ROSC, maintained for 24 hours. The itself. A notable feature which stands out the alveoli to remain open in the presence outcomes of these trials were significant, from these studies is that the mild degree of great surface-tension forces. Also, with 55% vs. 39% of patients achieving an of cooling in this type of hypothermia saline does not carry enough dissolved independent life in one study, and 49% vs. does not produce a degree of decrease in oxygen to allow the medical definition of 26% of patients reaching this goal in the brain metabolism sufficient to explain the “respiration” (i.e., movement of enough other. [1] [2] results, so this is not the mechanism by oxygen and carbon dioxide exchange to Since these initial studies, research which the intervention proceeds. However, support metabolism). has focused on new and more rapid the mild degree of cooling does seem to Perfluorocarbons (PFCs) are molecules mechanisms of causing and maintaining substantially decrease all of the known in which all of the hydrogen atoms which hypothermia. Animal studies have post-resuscitation damage-cascades, occupy the non-linking surface positions suggested that the more rapidly the including free radical release, calcium of a “hydrocarbon” molecule (such as the hypothermia is induced, the better its effect, leakage, toxic neurotransmitter release, octane molecule used in gasoline, and many but the exact parameters remain unknown. other types of inflammation reactions, and others) are replaced by fluorine atoms. Extra-corporeal methods are very effective, programmed cell death (apoptosis). Which After this modification the PFC molecules but are thought to remain too invasive for of these are more important is unknown, become chemically inert. Such molecules resuscitation. Peritoneal and pleural lavage just as the order of their importance are liquids at body temperatures, if they with ice-water have been considered. remains unknown in normothermic are heavy and complex enough. These PFC Administration of 30 ml/kg ice-cold saline ischemic damage to the brain. liquids do not dissolve inin eithereither waterwater oror oils.oils solution intravenously brings about half of Animal models have also allowed ey(they form aa new layer under them, when the desired temperature decrease, without investigation of various methods of combined, like the drain cleaning product causing pulmonary edema. A device which experimental cooling which would not “Liquid Plumr,” and poured through employs intravenous heat exchange has also presently be usable in clinical situations. standing water. But PFCs are capable of

www.alcor.org Cryonics / April 2015 23 carrying oxygen and carbon dioxide (CO2), of the lung and assist with compromised It also requires a skilled vascular surgeon. which do dissolve in them. ventilation. It underwent clinical trials PFC lung lavage, by contrast, could be Liquid breathing with perfluorocarbon in premature infants with a particular initiated as soon as an endotracheal tube (PFC) liquids has been investigated since PFC called perfluorobromooctane was in place in the patient’s “windpipe”, 1965, as a means of allowing gas exchange (Perflubron™ developed by the Alliance and in theory could be done in the field, by within the lung by means of a liquid, Pharmaceutical Corporation). However, paramedics. without removal of the critical surfactant. this technique was evaluated but not In the case of fluorocarbon, surfactant is approved by the FDA. In these trials it did DEVELOPMENT OF LUNG LAVAGE not removed because it is not soluble in the not harm the infants but didn’t assist them Unlike other experimental labs, CCR PFC. The lungs can be completely filled enough to be approved. normally allowed many of its experimental with PFC, if it is oxygenated, in a technique Critical Care Research, Inc., beginning in animals (canines) to survive long term, called “Total Liquid Ventilation” (TLV). If a 1997 and first patented [5] and published after the procedure. It was found by this liquid ventilator machine adds and removes [6] in 2001, developed a cooling technique protocol that an asthma-like syndrome the PFC from the lungs, and (while outside which was functionally a hybrid between was produced by high-rate lavage with the lungs) removes the CO2 from it and TLV and PLV. In this technique, which many PFCs, but that this did not become adds oxygen to it, animals can be ventilated we now simply call fluorocarbon lung appreciable until 24 hours after lavage- with liquid alone, without bubbles of gas in lavage (FLL), the lungs are not completely cooling. It was never fatal if lavage was done their lungs. This is the technique seen in the filled with PFC, but instead filled with carefully enough under low pressure, but film The Abyss. Despite that film (where a volume of PFC liquid similar to only a did cause the animals difficulty breathing the liquid breathing scenes in humans were fraction of the lung volume. This liquid for as long as week after the procedure. simulated), it has only been tried on only is then periodically infused and removed, Eventually, as with all PFC procedures, the one human, and then only temporarily. The passing through a heat exchanger to cool it PFC evaporates and disappears from the technique is difficult, the pressures needed between cycles. lungs, and the animals recover completely. close to those which cause lung damage, Initially, the PFC was also passed through Over the course of the last 14 years, CCR and the ability to dissolve CO2 and thus a gas-exchanger in this technique, to remove made an exhaustive study of the necessary remove it from the body is just at the edge CO2 and add oxygen to it. However, as the parameters of lung lavage to minimize of what is needed for dogs and humans. volume of PFC infused was reduced in the post-lavage syndrome. These include In 1984 T.H. Shaffer and colleagues experimentation, it was found that this gas investigation of 10 different candidate PFC investigated PFCs introduced into the replacement was no longer necessary, and fluids (in some cases this was forced by the lungs as a method of cooling animals. that the remainder of the lung was capable manufacturers discontinuing them, as none

The goal was not mild hypothermia, for of removing CO2 from the animal (and of them are made for medical use). CCR this was not then known as a technique. the liquid) and supplying oxygen also, if research has also explored the optimal However, there were other reasons to cool ventilated with pure oxygen gas. Thus, the lavage volume, timing, temperature, and and warm animals, and Shaffer was able gas exchanger for the PFC was not needed, method of delivery and mode of removal. to show that the TLV cooling technique and was eventually discarded. Because PFC is heavy and resists suctioning, worked on anesthetized cats. The animals In 2001, Critical Care Research, Inc. the optimal methods to introduce and were not allowed to survive the experiment, (CCR) published a paper [6] showing remove it turned out to require a dedicated however, so the long term effects of having that FLL lavage was able to reduce brain apparatus and a very finely tuned method the lungs totally full of cooled PFC liquid, temperature in dogs by a rate as fast as of use. This proved quite formidable, and were not investigated. 0.5ºC per minute. Thus, the needed state of the CCR machine required complete re- Another related use of PFCs developed mild hypothermia (4ºC cooling) could be design four times. later was a technique called Partial Liquid induced in less than 10 minutes (allowing At the same time this development was Ventilation (PLV) in which the lungs were for heat transfer delays). This was a factor taking place, Lance Becker at Argonne filled to 1/3rd of the volume of capacity of 12 faster than the 2 hours required by National Laboratories had been working (about the amount of a normal tidal the intravenous techniques, the fastest with tiny ice spheres which could be breath) with PFC, and this was allowed to cooling technique which had been reported suspended in PFC as a slurry, and delivered remain in place while gas ventilation was by any method. to the lungs. The phase change as the ice then carried out “on top” of it. Ventilation Cardiac bypass techniques were known melts provides an extra degree of cooling was accomplished in the rest of the lung to be able to cool in excess of 1°C per power per volume of lavage which PFC by normal gas ventilation methods. This minute, but circulatory bypass involves alone does not, but on the other hand, technique could not be used for heating gaining access to major blood vessels the saline generated in this way also and cooling (since the amount of PFC was (usually the femoral arteries and veins in presumably dissolves some lung surfactant. too small to affect body temperature), but the groin) and there is a necessary delay Becker reported initial tests of this system it was used to open the dependent parts time to initiate bypass after an emergency. in euthanized animals, but did not end up

24 Cryonics / April 2015 www.alcor.org developing it further. As in many of these temperature and the maximum volume heat exchangers already available on the systems, since the animal was not allowed of coolant that can be introduced into commercial market for other uses (these to survive, we do not know the long term the body are significant engineering are capable of nearly 600 watts/degree effects of the procedure. obstacles.” gradient of heat transfer between ice-water Challenge of Becker, et al. to and PFC). CCR then discarded the bulky CHALLENGEChallenge OFof BECKER,Becker, ETet AL al. to and PFC). CCR then discarded the bulky produce ultra-rapid brain cooling in a In reading Lampe and Becker’s article, peristaltic pumps and replaced them with produceTO PRODUCE ultra-rapid ULTRA-RAPID brain cooling BRAIN in a peristaltic pumps and replaced them with large animal. CCR realized that they had seriously largeCOOLING animal. IN A LARGE ANIMAL CCR realized that they had seriously inexpensive commercial fluid diaphragm miscalculated the amount of heat which In 2007, Dr. Becker and Joshua W. miscalculated the amount of heat which pumps, which proved far superior, and able In 2007, Dr. Becker and Joshua W. Lampe would need to be removed during rapid Lampe published a paper [7] in the journal would need to be removed during rapid to run on low voltage D.C. published a paper [7] in the journal Expert cooling, due to their inexperience with Expert Review of Medical Devices, in cooling, due to their inexperience with With the superior suction capabilities Review of Medical Devices, in which they the decrease in effective heat capacity of which they challenged experimenters to the decrease in effective heat capacity of of this device, combined with its challenged experimenters to design a the body, which occurs during very rapid design a cooling system which would lower the body, which occurs during very rapid ability to provide a constant supply of cooling system which would lower brain cooling of the blood. CCR’s previous brain temperature by 4°C in the first 5 cooling of the blood. CCR’s previous perfluorocarbon cooled to 2ºC (from a temperature by 4°C in the rst 5 minutes of experiments had found that only a 60% minutes of application. They wrote: experiments had found that only a 60% reservoir containing only 7 liters of fluid) application. ey wrote: to 70% “core” mass (which includes the to 70% “core” mass (which includes the CCR performed a series of 30 lung- “The induction of mild hypothermia, brain, fortunately) is cooled in the first few “The induction of mild hypothermia, brain, fortunately) is cooled in the first few lavage dog experiments in 2007 and mid lowering body temperature by 4°C, is minutes of cooling, and that if cooling lowering body temperature by 4°C, is minutes of cooling, and that if cooling 2008. The first results were encouraging gaining acceptance as an acute therapy continues past 5 minutes, the extra heat gaining acceptance as an acute therapy continues past 5 minutes, the extra heat enough to apply for a preliminary patent for the treatment of hypoxia and to be extracted in the 6th and 7th minute for the treatment of hypoxia and to be extracted in the 6th and 7th minute on the device in September 2007, and a full ischemia following cardiac arrest and will make up for the later “re-equilibration” patent application was filed (Platt, Battiano, many life-threatening injuries. When patent application was filed (Platt, Battiano, many life-threatening injuries. When which takes place within the animal or Harris) for the device in September 2008 hypothermia is used following ischemia Harris) for the device in September 2008 hypothermia is used following ischemia human as the cold core and brain are re- [8]. (as opposed to before ischemia), it needs [8]. (as opposed to before ischemia), it needs warmed by the periphery (which does A graph from one of the early to be performed rapidly for the greatest A graph from one of the early to be performed rapidly for the greatest not initially cool as fast as they do). The experiments with this device, after design benefit, preferably within 5 min. A experiments with this device, after design benefit, preferably within 5 min. A differential cooling of the thermal core parameters had been partly adjusted, simple model reveals that this poses parameters had been partly adjusted, simple model reveals that this poses during very fast cooling allows a temporary illustrates performance: a significant bioengineering challenge illustrates performance: a significant bioengineering challenge window in which only part of the body This graph shows that Lampe and as the rate of heat transfer is severely This graph shows that Lampe and as the rate of heat transfer is severely (including the brain) needs to be cooled, Becker’s stated goal (which they thought limited, owing to a relatively confined Becker’s stated goal (which they thought limited, owing to a relatively confined with the catch-up period of re-equilibration impossible without a direct application of fundamental parameter space. Current impossible without a direct application of fundamental parameter space. Current later, used for compensatory cooling. a phase-change substance such as ice to the methods of cooling include external a phase-change substance such as ice to the methods of cooling include external Since Lampe and Becker had also body) of 4ºC drop in 5 minutes has been cooling devices, such as cooling blankets body) of 4ºC drop in 5 minutes has been cooling devices, such as cooling blankets assumed that the heat capacity of the body exceeded with the device, which provided or ice bags, which are simple to use, and exceeded with the device, which provided or ice bags, which are simple to use, and is that of an equivalent amount of water (it 5.3ºC drop in the first 5 minutes of relatively inexpensive but slow. Internal 5.3ºC drop in the first 5 minutes of relatively inexpensive but slow. Internal is actually only 70% as large) their estimates application . In addition, in this experiment, cooling has the best ability to cool more application . In addition, in this experiment, cooling has the best ability to cool more of needed heat extraction were off by a CCR was able to cool the animal’s brain and rapidly but current devices are more CCR was able to cool the animal’s brain and rapidly but current devices are more factor of 70% times 70% (a reciprocal heart to 25ºC (77ºF), in just 16 minutes. invasive, costly and most are still not heart to 25ºC (77ºF), in just 16 minutes. invasive, costly and most are still not factor of 2). Also, their estimations of the able to provide cooling within the rapid able to provide cooling within the rapid amount of cold fluorocarbon which could 5-min interval.” 5-min interval.” be delivered to the lungs, was incorrect. At CCR it was realized that the challenge set Lampe and Becker go on to a discussion CCR it was realized that the challenge set of theoretical amounts of heat which can by Lampe and Becker had already nearly of theoretical amounts of heat which can been met. be removed by a number of methods, been met. including cardiac bypass, but note that the surgery for this takes at least 1 hour to WORK ON PORTABLE HIGH surgery for this takes at least 1 hour to CAPACITY LUNG LAVAGE DEVICE implement. They conclude: CAPACITY LUNG LAVAGE DEVICE Beginning in 2007 CCR contracted with “Hypothermia of 3-4ºC must be outside engineering consultants to provide achieved within 5 min for maximum a suitable high capacity (high suction) protection when required after a period machine, which would be completely of ischemia. The human body is well portable (able to run from batteries) and designed to prevent heat loss, creating also able to cool the body more than Later experiments with the device a significant bioengineering challenge 4ºC, should this be required. In designing show that 6-7 minutes of application of in a relatively confined fundamental this machine, CCR made use of the 60 ml/kg/min lung-lavage with ice-cold parameter space. The minimum high-capacity commercial “plate-type” perfluorcarbon (2ºC) results in at least 4ºC

www.alcor.org Cryonics / April 2015 25 drop in brain temperature in the first 5 temperatures begin to track in parallel). into a permanent “whole body” drop minutes, and that application of lavage for Note that blood temperature still exceeds of 4ºC after equilibration of the cold more than 6 minutes results in a permanent brain/core temperature by about 0.4ºC thermal core with the warmer (but less body core temperature drop of 4ºC. All at 900 seconds (15 minutes) because at well perfused) periphery. Thus, the of these results are novel, and should be this time, blood is still carrying heat from brain must be rapidly cooled by 5.5ºC of great importance to the resuscitation the warm periphery of the animal to the to retain a 4ºC drop. community. cooled core tissues, which include the brain Note that all these problems (and (that is, full body tissue and compartment THE FUTURE OF POST benefits!) of having the core of the animal heat equilibration is not complete). RESUSCITATION HYPOTHERMIA substantially cooler than the periphery, only The technique of post-resuscitation happen in extreme rates of cooling, such as cooling, even with simple ice-bags in the are not seen in any other cooling techniques Emergency Department, has been slow applicable to the field in emergencies. to catch on. Problems include logistical However, they now serve as part of the difficulties and even lack of advertising basic technique at CCR. to move information (there are no large The post-lavage asthma syndrome which pharmaceutical companies involved). is exacerbated by the wrong choice of A 2005 survey of 256 emergency room PFC, and by use of the wrong techniques physicians found that despite official during lavage (which result in needless ILCOR endorsement of this technique for overpressures), has also been overcome Advanced Life Support (ALS) in 2003, only by CCR. This syndrome is not known by a quarter of these physicians were using the other groups, because CCR presently has technique on their resuscitated patients.[9] the only large-animal model of lung lavage, Figure: Animal Lavaged with The reasons given were that the technique where the animals are allowed to survive Device in Patent, 2008. In the Figure was difficult or slow, as well as a technically long enough to develop later lung reactions. above temperatures in the venous correct but misguided assertion that the (CCR maintains its own dog colony, with system, arterial system, and brain are ALS guidelines did not incorporate it (these all animals bred “in house” since 2003). recorded. Lung lavage is given for 5.3 guidelines had not been updated since 2000, The present extreme cooling technique minutes. Brain temperature does not at that time, so they had not been able to causes little or no asthmatic reaction in begin to drop rapidly until 45 seconds, incorporate cooling recommendations). dogs, and they are clinically comfortable, which reflects a circulation delay, and survive. as the cooling of the arterial blood The following graph shows the is seen to be immediate. However, “The post-lavage asthma performance of this machine in detail, the overshoot effect occurs after the when used for 5.3 minutes to cool a dog’s procedure is stopped, and the minimal syndrome which is exacerbated by brain by 4.8ºC in 6.75 minutes, slightly brain temperature does not occur until the wrong choice of PFC, and less than the best performance seen in almost 6.75 minutes (a minute and a the graphs above. This dog has been fully half after the procedure has stopped), by use of the wrong techniques instrumented, so that the instantaneous and it is 32.2ºC, which is a 4.8º drop during lavage...has also been change in central venous and arterial blood from a beginning temperature of 37ºC temperatures (in the aorta and inferior (98.6ºF). Over the next hour the brain overcome by CCR.” vena cava) can also be seen. Note the will warm to 33.7 ºC (3.3ºC drop), as excursions in both venous and arterial the cold core of the body equilibrates blood with each of the 40 lavages (one with the warmer exterior (about 20% of Moreover, simple ice-bag techniques as lavage every 8 seconds) in this experiment. the total thermal “mass”). In general, used in emergency departments are often Blood temperatures fall as low as 29.2ºC continuous lavage of about 6 minutes ineffective at cooling. A recent (2009) study (84.6ºF) in this animal which begins at a is required to guarantee a 4ºC drop reported in Journal Watch Emergency Medicine temperature of 37ºC (98.6ºF). At the end at 5 minutes, as well as a permanent reported that post-resuscitative cooling of of the experiment, equilibration of venous brain temperature drop of 4ºC even 287 patients with ventricular fibrillation and arterial blood can be seen, followed by after body and brain equilibration, one arrest, using ice packs, cooling blankets equilibration of the entire blood pool as hour after liquid ventilation cooling has and cooling pads, reached the target a whole within the core of the animal, as been stopped. To cool the body and temperature of 32ºC-34ºC in only 65% represented by the brain temperature (this brain permanently by 4ºC requires an of patients. [10] Despite this, the group happens after about 700 seconds or a bit early and instantaneous brain cooling found that treatment increased survival to less than 12 minutes, when blood and brain of about 5.5ºC, which later develops hospital discharge from 39% to 54% and

26 Cryonics / April 2015 www.alcor.org favorable neurological outcome rate from patients since 2006 and says 52% have after equilibration, after 6 minutes of 15% to 35%. They wrote: “…it is time for survived, compared with single digits lavage), with minimal post procedure lung [Emergency Departments] to implement historically; of those, about 75% have reaction in the following days. Remaining, hypothermia protocols for comatose had a “favorable neurologic recovery,” however, are the difficult and expensive survivors of cardiac arrest.” including many who report a full return clinical trials which must occur in humans. These and other results have continued to normal. Dogs, with more delicate and slightly to amaze physicians who had believed that larger lungs (for their body size) than the traditional 5 to 10 minutes of warm The difference between 10 minutes humans, are a good model for lung lavage. circulatory arrest time (without chest and 20 minutes is particularly important, Although it may be possible to cool a dog compression) was the limit, after which for this is the average paramedic response slightly more quickly than a human by this brain death was certain. A recent Wall Street time for a large fraction of urban areas in method, at the same time, if the technique Journal article quoted one “early-adopter” of the country. Nor are physicians sure that does not damage dog lungs at a given the post-resuscitation cooling protocol [11]: 20 minutes is the limit for resuscitation, pressure, it probably will not damage the as these studies are still being done with tougher human lungs. Even so, for safety’s “We’ve had patients who have mostly conventional methods of cooling sake, initial clinical trials in humans will been stone-cold out for least 20 with ice packs and esophageal ice water need to be carried out at a fraction of the minutes—we know that for sure—and lavage, which takes at least several hours. maximal rates and pressures which have they’ve come back normal or nearly been achieved in canines. normal,” says Michael Mooney, a THE FUTURE OF LUNG LAVAGE Lung lavage remains to be tested after cardiologist who heads the therapeutic Critical Care Research, Inc. has already resuscitation, in an actual large-animal hypothermia program at Minneapolis solved many of the key problems in model of brain ischemia and circulatory Heart Institute. An early adopter of inducing very rapid brain mild hypothermia arrest.  the cooling technique, the cardiology (4ºC drop by 5 minutes of lavage, sufficient practice has treated more than 140 heat removal for a permanent 4ºC drop

REFERENCES:

1. Bernard, S. A., Gray, T. W., et al. (2002). Treatment of cold peruorochemical lung lavage in dogs. (2001) comatose survivors of out-of-hospital cardiac arrest with RESUSCITATION 50: 189-204. (PMID: 11719148) induced hypothermia. N Engl J Med, 346(8), 557. 7. Becker L.B. and Lampe J.W. (2007) Rapid cooling for 2. e Hypothermia After Cardiac Arrest Study Group (2002). saving lives: a bioengineering opportunity. Expert Rev Med Mild therapeutic hypothermia to improve the neurologic Devices. Jul;4(4):441-6. (PMID: 17605679) outcome after cardiac arrest. N Engl J Med, 346(8), 549. 8. (WO/2009/042220) PORTABLE APPARATUS AND 3. Nolan, J. P., Morley, P. T., et al. (2003). erapeutic METHOD FOR THE ADMINISTRATION OF HEAT hypothermia after cardiac arrest: An advisory statement by EXCHANGE IN THE LUNGS OF A MAMMAL (PLATT, the Advanced Life Support Task Force of the International Charles, HARRIS, Steven, B, BATTIATO, Gary. http:// Liaison Committee on Resuscitation. Circulation, 108(1), www.wipo.int/pctdb/en/wo.jsp?WO=2009042220 118. 9. Abella, B. S., Rhee, J. W., et al. (2005). Induced hypothermia 4. Kuboyama, K, Safar P., et al. Delay in cooling negates the is underused after resuscitation from cardiac arrest: A current benecial eect of mild resuscitative cerebral hypothermia practice survey. Resuscitation, 64(2), 181. As quoted in “ e after cardiac arrest in dogs: a prospective randomized study big chill: Improving the odds after cardiac arrest” (2005) Patty [see comments] (1993) Crit Care Med 21:1348-1358. Calver, RN, BSN, eresa Braungardt, BSN, et al. RN volume 68, no 5. 5. Federowicz, M.G., Russell, S.R., and Harris, S.B. Mixed- mode liquid ventilation gas and heat exchange. U.S. Patent 10. Koenig, K. L. Induced hypothermia after VF cardiac arrest 6,694,977, 2004. improves outcomes. (2009) Journal Watch Emergency Medicine. Nov. 6, 2009. 6. Harris, S.B. Darwin, M.G., et al. Rapid (0.5°C/min) minimally invasive induction of hypothermia using

www.alcor.org Cryonics / April 2015 27 Non-Cryonics Research Recently Funded by the Life Extension Foundation

By Ben Best, Director of Research Oversight, Life Extension Foundation

lthough the Life Extension naked mole rat had already been sequenced protein synthesis fidelity discovery caused Foundation (LEF) contributes to discover the basis of naked mole rat the prestigious journal SCIENCE to name substantially to cryonics and cancer-resistance and longevity. Without the naked mole rat “Vertebrate of the A 13 cryobiology-related research, much LEF funding from LEF, Dr. Seluanov would Year” for 2013. The August 2014 issue funding is devoted to anti-aging research have been in danger of losing his naked of NATURE REVIEWS: GENETICS (including anti-cancer research). LEF mole rat colony. Although Gorbunova and featured a review as a cover story on that supports dozens of scientists seeking to Seluanov contributed to analysis of the subject that was primarily authored by the meaningfully extend the healthy human naked mole rat genome,7 this information two scientists.14 NATURE REVIEWS: lifespan through their unique research was insufficient to explain the cancer- GENETICS is the foremost scientific initiatives. resistance and longevity of the naked mole journal (highest impact factor) on the Drs. Gorbunova and Seluanov are a rat. subject of heredity and genetics. LEF was husband-and-wife team at the University OnOn July July 18, 18, 2013 Seluanov and Gorbunova acknowledged as a funding source for the of Rochester in Rochester, New York. made the cover of the prestigious journal review. Both scientists are devoted to finding NATURE with their discovery that high means to reduce aging and cancer so as to molecular weight hyaluronan (hyaluronic extend human lifespan.1 Dr. Gorbunova acid) in naked mole rats protects them “Joao Pedro de Magalhaes, discovered that the sirtuin SIRT6 could from cancer.8 Naked mole rat hyaluronan more than triple the repair of DNA has five times the molecular weight PhD...used funds from LEF damage.2 Increased levels of SIRT6 has of the hyaluronan in humans or mice. to sequence the genome of been shown to extend the lifespan of male Hyaluronan is found in skin products. In mice 10-15%.3 Because DNA damage other mammals, hyaluronan contributes the bowhead whale, the longest- can lead to both aging and cancer, Dr. to wound healing.9 But in the naked mole living mammal, which lives more Gorbunova has been looking for molecules rat, the high molecular weight hyaluronan which will stimulate SIRT6 activity. Dr. causes cancer cells to stop growing.8 A few than 200 years.” Gorbunova was denied government years earlier, Dr. Seluanov had discovered funding on the grounds that there are other that naked mole rat tissue causes cancer to mechanisms of DNA repair besides what stop growing,10 but he had not understood Joao Pedro de Magalhaes, PhD, SIRT6 stimulates. Grants from the LEF the reason. (Senior Lecturer/Associate Professor, have enabled Dr. Gorbunova to continue With LEF funding, the couple Institute of Integrative Biology, Liverpool her work on SIRT6 activation of DNA subsequently discovered that protein University, Liverpool, United Kingdom) repair. synthesis is four times more accurate in used funds from LEF to sequence the Dr. Seluanov has the second-largest naked mole rats than in mice.11 Many genome of the bowhead whale, the naked mole rat colony in the world. neurodegenerative diseases such as longest-living mammal, which lives more Although mice usually die of cancer, Alzheimer’s Disease and Parkinson’s than 200 years.15 With more cells in their cancer has never been reported in a naked Disease are associated with protein mis- bodies, larger animals would be expected mole rat.4,5 Naked mole rats live about ten folding, which is likely in part due to errors to have higher rates of cancer.16 Even tall times longer than mice without evidence of in protein synthesis.12 Precision synthesis humans have a higher cancer risk than aging or age-related diseases.6 Dr. Seluanov of proteins by naked mole rats contributes short humans, independent of all other was denied funding from the government to their cancer-resistance and longevity. risk factors.17 But a variety of anti-cancer on the grounds that the genome of the Both the hyaluronan discovery and the mechanisms allows larger animals to

28 Cryonics / April 2015 www.alcor.org avoid the greater cancer risk that would of old blood. With this information, he will calorie restriction slows aging in rodents30 otherwise accompany their greater size.18 develop strategies to enhance the positive and that lifespan is largely under genetic Dr. Magalhaes’s laboratory is uniquely components as well as to block or remove control in many or all species,31,32 it is only equipped to analyze the genome of the the negative components. He is hopeful in the last two decades that the genes and bowhead whale to determine the means that his results will be ready for clinical pathways regulating lifespan have been by which it achieves such longevity while application in only a few years. discovered. In 1988 mutation in the age- avoiding cancer.19 With funding from LEF, 1 gene was shown to increase the average Dr. de Magalhaes was able to complete lifespan of nematode worms by 40–65%33 genome sequencing of the bowhead whale, “Justin Rebo, MD... is using In 1993 Cynthia Kenyon showed that daf- and publish an analysis of the genome.20 2 mutations could double the lifespan of John Furber, MSc, (CEO, Legendary Life Extension Foundation nematode worms.34 These genes were later Pharmaceuticals, Gainesville, Florida) funding to investigate in detail found to lie in the same genetic pathway, attends many conferences dealing with which when manipulated in mice can the biology of aging every year, and he why shared blood circulation stretch their lifespan by half.35 Two decades is therefore very well-known among between genetically identical mice after the first long-lived mutant in age-1 was biomedical gerontologists. He has designed characterized,33 Dr. Reis found that more a rejuvenation experiment which LEF of different ages rejuvenates the thorough elimination of this gene’s PI3K is funding. Lysosomes are the garbage old mouse, and makes the young gene product can actually extend nematode disposal/recycling centers of biological life span tenfold.36 Dr. Reis believes that cells. Lysosomes contain enzymes that mouse biologically older.” this benefit can extend far beyond worms. digest cellular waste products into reusable Suppression of PI3K in mouse heart components.21 But with time, lysosomes muscle slows many measures of heart accumulate enzyme-resistant age-pigment LEF has been funding the South Florida aging and improves their overall survival.37 molecules known as lipofuscin.22 Some Bone Marrow/Stem Cell Institute (Principal Crippling just one of the normal two copies neurons in the brain can contain up to Investigator Dipnarine Maharaj, MD, of PI3K in all tissues of the mouse is bad 75% lipofuscin.23 Lipofuscin emits toxic FACP) for clinical trials treating cancer for juvenile mice but improves fitness, free radicals. Cells loaded with lipofuscin victims with white blood cells from young metabolism, and survival after maturity.38 cannot be expected to function very well, so donors. A future grant is planned for an Humans who live past age 100 show an lipofuscin may contribute to the maladies experiment to use blood components from inherent genetic bias that produces the of old age. Furber would like to rejuvenate young donors to relieve an elderly person same effects.39 Dr. Reis seeks to identify the cells by removing the lipofuscin. By doing from generalized symptoms of age-related molecules that are directly affected by the an extensive search of scientific literature, frailty and ill health. most beneficial genetic modification, and he identified some chemicals which induce University of California at San Francisco to find drugs that can knock out PI3K and cells to export lysosomal lipofuscin out (Principal Investigator, Cynthia Kenyon, mimic the life-extending benefits observed of the cell.24 With LEF funding he is PhD) has received funding from LEF in previous studies. Nematode worms are experimenting with these chemicals in to screen for drugs that increase stress an ideal biochemical laboratory for life the hope removing lipofuscin from cells resistance in nematode worms. There is span studies of this nature, but Dr. Reis without causing harmful effects when the good evidence that greater stress resistance also experiments with human cells and lipofuscin enters the bloodstream. will increase lifespan, healthspan, and mice. Justin Rebo, MD, (Research Scientist, resistance to aging-related diseases (with In the Fall of 2014 the LEF increased SENS Foundation, Mountain View, application to humans).29 Dr. Kenyon its funding for Androcyte, Inc. for analysis California) is using Life Extension gained fame for her discovery that the of the genomes of centenarians and Foundation funding to investigate in detail lifespan of nematode worms could be supercentenarians (persons over age 110). why shared blood circulation between doubled by genetic manipulation. This Such long-lived persons possess rare genetically identical mice of different ages led to subsequent discovery that genetic protective genes that could benefit everyone rejuvenates the old mouse, and makes manipulation could also extend the life and from the debilities of aging and aging- the young mouse biologically older.25,26 health of rodents. associated diseases. The chief scientific Blood from a young mouse contains more Robert Shmookler Reis, PhD, officer of Androcyte is Dr. George functional white blood cells and stem cells27 Professor at the University of Arkansas for Church, the world-famous Harvard as well as fewer inflammatory proteins than Medical Sciences, Little Rock, Arkansas. geneticist who is at the forefront of the new blood from an old mouse.28 Dr. Rebo’s The research of Dr. Reis has been focused CRISPR gene-editing technology. To date, objective is to determine with precision on the influence of genetics on longevity only one human (Timothy Ray Brown) all of the positive components of young and the diseases of aging. Although we have has been documented to be cured of the blood and all of the negative components known for the better part of a century that HIV virus. Brown was suffering from

www.alcor.org Cryonics / April 2015 29 both leukemia and AIDS. Brown received of their shortening telomeres. Cancer cells CT can give rapid and accurate information a hematopoietic stem cell transplantation usually use telomerase enzyme to multiply, about tumor size, location and rate of that fortunately gave him two copies of the which is why so many researchers have been growth. CCR5 gene that blocks HIV.40 If CRISPR attempting to stop cancer by telomerase As useful as PET imaging can be, technology can be perfected and made safe, interference. But cancer cells have another statistical errors can at times result in “false not only could AIDS be cured, but genes trick to multiply, known as ALT (Alternate negative” and “false positive” reporting. from supercentenarians could protect Lengthening of Telomeres).47 Very few Other issues that may trigger errors include everyone from aging-associated diseases, researchers have been working on ALT, improper PET scanner calibration with like Alzheimer’s Disease. but if researchers find a way of blocking errors which can at times result in false LEF is funding Howard Chang, telomerase in cancer cells, cancer cells will patient body weight, and the variability in MD, PhD, at the Stanford School of increasingly use ALT to multiply. Cancer FDG uptake depending on the elapsed time Medicine. Dr. Chang has been studying cells utilizing ALT to lengthen telomeres from when the radiotracer was injected gene expression (epigenetic) changes in can be identified by ALT-associated into the patient. But the most egregious stem cells that occur with aging. He has bodies in the cell nucleus,48 but ALT errors are perhaps due to incomplete or identified a number of pathways that affect researchers have been hampered by the inconsistent scan interpretations caused by stem cell epigenetics, and is seeking drugs time-consuming and expensive methods inadequate training and a lack of overall that will alter those pathways. Rejuvenation required to study ALT. LEF funded Dr. standards for the quantified reporting of of stem cells will improve wound healing Haroldo Silva of the SENS Foundation to results. Incorrect PET scans are common as well as recovery from a variety of disease develop an improved method of screening today and can result in improper treatments conditions. The elderly often die of immune for ALT, which should boost the entire for cancer patients. system failure,41 which could be prevented field of ALT research. by stem cell rejuvenation.42 Dr. Chang Dr. Victoria Belancio of the Tulane was an Ellison Foundation Scholar until School of Medicine has been researching “In the Fall of 2014 the his grant expired. Without LEF funding retrotransposons (“jumping genes”) that he could not have continued his research cause genome instability. Retrotransposon LEF increased its funding for aimed at intervening in the aging process. activity increases with age, resulting in Androcyte, Inc. for analysis of Aside from the Life Extension Foundation, increased cancer and other aging-associated it is difficult to find sources of funding diseases.49 With funding from LEF, Dr. the genomes of centenarians for aging research. The National Institutes Belancio has established that shift workers and supercentenarians of Health only funds research on specific are at greater risk for retrotransposon- diseases and the Ellison Foundation has induced cancer because of light at night (persons over age 110).” ceased funding anti-aging research. reducing melatonin secretion.50 Another former Ellison Foundation Orn Adalsteinsson, PhD, who is Scholar being funded by LEF is James chairman of the Life Extension Foundation Working with radiologist Richard Black, Shorter, PhD, Associate Professor Scientific Advisory Board, supervises a MD, the International Strategic Cancer of Biochemistry and Biophysics at the number of anti-cancer projects and clinical Alliance adopted invaluable PET reporting University of Pennsylvania. Dr. Shorter trials funded by LEF, many of which are practices in its LEF-supported laser- has discovered that heat shock proteins can conducted in the Bahamas. Only a couple assisted immunotherapy breast cancer trial. reverse the aggregation of toxic proteins, of these will be described. Dr. Black has interpreted more than 80,000 including those causing Alzheimer’s The rapid expansion in the use of Positron PET/CT studies, and his methodology Disease, Huntington’s Disease, Parkinson’s Emission Tomography, or PET scans, to for an across-the-board upgrade in PET Disease, ALS (“Lou Gehrig’s disease”), obtain metabolic information about cancer scan reporting should be incorporated at among others. With funding from the lesions can provide oncologists and their the national level to provide oncologists Life Extension Foundation he has made patients with extremely valuable diagnostic and their patients with the full potential progress in developing heat shock proteins and treatment management information. PET technology has to offer. The five key that reverse the amyloid-beta protein PET scans use an injected radioactive tracer features of Dr. Black’s approach will assure aggregation that is believed to initiate material like fluorodeoxyglucose (FDG) to that oncologists receive the same kind and Alzheimer’s Disease. He is now focusing produce functional imaging that can help quality of information on each and every his attention on the aggregation of the tau differentiate benign from malignant masses, scan, regardless of who interpreted the protein that kills neurons in the later stages evaluate tumor stage, monitor response to scan, or where it was taken. of Alzheimer’s Disease.43.44.45.46 therapy and detect tumor recurrence in a Cancer cells have the ability to multiply variety of malignancies.51 Coupled with 1. Quantitative Reporting: Stan- without restraint, unlike other cells which the precise anatomical imagery produced dardized uptake values, or SUV are limited in their ability to multiply because by computerized tomography, FDG PET/ readings are collected for every

30 Cryonics / April 2015 www.alcor.org object of concern in the scan, not Dr. Black presented his initial findings small iron crystals (25-50 nanometers in just narrative descriptions. at one of LEF’s Scientific Advisory Board diameter), become powerfully magnetized Meetings in 2012; his presentation can be when exposed to the magnetic field of 2. Reproducible Reporting: SUV viewed on the Life Extension website at the an MRI scanner. The injected Combidex readings are standardized to an following URL: www.LEF.org/PET-CT contrast fluid is taken up selectively by area of normal homogenous tissue The critical need to develop superior the macrophages (scavenger cells) that are in the liver to generate a corrected cancer imaging tools cleared a major primarily found in lymph nodes and other SUV for every area of concern. The hurdle in December 2012, when a U.S. inflammatory tissue.53,54 correction factor allows different pharmaceutical giant agreed to sell the Dr. Barentsz is one of the few physicians experts using different equipment shelved research and development rights in the world to have worked extensively to obtain similar results. to Combidex, a revolutionary magnetic with Combidex technology, predominantly resonance imaging (MRI) contrast agent. in prostate cancer cases. In one study, Dr. 3. Index Lesion Focus: “Hotspots” Combidex-enhanced scans can detect Barentsz and his team compared traditional indicating tumor activity must be metastatic cancer lesions too small to be CT scans and Combidex-enhanced MRI monitored from one study to the seen by traditional PET/CT imaging.52 lymphangiography (MRL) for 375 prostate next to enable rapid and accurate Life Extension Foundation continues cancer patients, 16% of whom had lymph measurements of changes over time to be a strong advocate of Combidex node metastases. CT imaging detected or in response to therapy. since helping with the negotiation of the only 34% of the positive nodes, while 4. Comparative Readings Mandate: sale of the Combidex technology package Combidex MRL identified a remarkable PET scan reporting must make to Radboud University Medical Center in 82%. The diagnoses were microscopically reference to the size, SUV, and the Netherlands in 2012. In 2013, world- confirmed by either a lymph-node other features of an index lesion(s) renowned radiologist Jelle Barentsz, MD, dissection or a needle biopsy. The study from previous scans, obligating with the assistance of LEF through Orn group concluded that Combidex-enhanced the current radiologist to request Adalsteinsson, has begun the process of MRL is 96% accurate, and can eliminate those studies for a side-by-side preparing the launching of multi-country the need for highly invasive surgical lymph 55 comparison. research trials, which will ultimately lead to node dissections. new license applications, a commercialized Combidex scans have also been used to 5. Image Snapshots of Index product and widespread patient access. successfully evaluate patients with cancers Lesions: Allows the ordering Combidex (ferumoxtran-10) is of the uterus,56 head and neck,57 kidney,58 physician to visualize the areas of composed of a simple sugar compound, breast,59 and liver.60  abnormality, rather than relying dextran, and superparamagnetic iron solely on a written report. oxide, or USPIO.53 These extremely

REFERENCES:

1. Van Meter M, Seluanov A, Gorbunova V. Forever young? 6. Buenstein R. e naked mole-rat: a new long-living model Exploring the link between rapamycin, longevity and cancer. for human aging research. J Gerontol A Biol Sci Med Sci. 2005 Cell Cycle. 2012 Dec 1;11(23):4296-7. Nov;60(11):1369-77.

2. Mao Z, Hine C, Tian X, Van Meter M, Au M, Vaidya A, Seluanov 7. Keane M, Craig T, Alföldi J, Berlin AM, Johnson J, Seluanov A, Gorbunova V. SIRT6 promotes DNA repair under stress by A, Gorbunova V, Di Palma F, Lindblad-Toh K, Church GM, activating PARP1. Science. 2011 Jun 17;332(6036):1443-6. de Magalhães JP. e Naked Mole Rat Genome Resource: facilitating analyses of cancer and longevity-related adaptations. 3. Kan Y, Naiman S, Amir G, Peshti V, Zinman G, Nahum L, Bioinformatics. 2014 Dec 15;30(24):3558-60. Bar-Joseph Z, Cohen HY. e sirtuin SIRT6 regulates lifespan in male mice. Nature. 2012 Feb 22;483(7388):218-21. 8. Tian X, Azpurua J, Hine C, Vaidya A, Myakishev-Rempel M, Ablaeva J, Mao Z, Nevo E, Gorbunova V, Seluanov A. High- 4. Delaney MA, Nagy L, Kinsel MJ, Treuting PM. Spontaneous molecular-mass hyaluronan mediates the cancer resistance of histologic lesions of the adult naked mole rat (Heterocephalus the naked mole rat. Nature. 2013 Jul 18;499(7458):346-9. glaber): a retrospective survey of lesions in a zoo population. Vet Pathol. 2013 Jul;50(4):607-21. 9. Chen WY, Abatangelo G. Functions of hyaluronan in wound repair. Wound Repair Regen. 1999 Mar-Apr;7(2):79-89 5. Azpurua J, Seluanov A. Long-lived cancer-resistant rodents as new model species for cancer research. Front Genet. 2013 Jan 10. Seluanov A, Hine C, Azpurua J, Feigenson M, Bozzella M, 9;3:319. Mao Z, Catania KC, Gorbunova V. Hypersensitivity to contact

www.alcor.org Cryonics / April 2015 31 inhibition provides a clue to cancer resistance of naked mole-rat. 2012 Oct;33(10):2390-7. Proc Natl Acad Sci U S A. 2009 Nov 17;106(46):19352-7 25. Ludwig FC, Elasho RM. Mortality in syngeneic rat parabionts 11. Azpurua J, Ke Z, Chen IX, Zhang Q, Ermolenko DN, Zhang of dierent chronological age. Trans N Y Acad Sci. 1972 ZD, Gorbunova V, Seluanov A. Naked mole-rat has increased Nov;34(7):582-7. translational delity compared with the mouse, as well as a unique 28S ribosomal RNA cleavage. Proc Natl Acad Sci U S A. 26. Conboy IM, Conboy MJ, Wagers AJ, Girma ER, Weissman 2013 Oct 22;110(43):17350-5. IL, Rando TA. Rejuvenation of aged progenitor cells by exposure to a young systemic environment. Nature. 2005 Feb 12. Lee JW, Beebe K, Nangle LA, Jang J, Longo-Guess CM, Cook 17;433(7027):760-4. SA, Davisson MT, Sundberg JP, Schimmel P, Ackerman SL. Editing-defective tRNA synthetase causes protein misfolding 27. Dykstra B, Olthof S, Schreuder J, Ritsema M, de Haan G. and neurodegeneration. Nature. 2006 Sep 7;443(7107):50-5. Clonal analysis reveals multiple functional defects of aged murine hematopoietic stem cells. J Exp Med. 2011 Dec 13. Breakthrough of the year 2013. Notable developments. Science. 19;208(13):2691-703. 2013 Dec 20;342(6165):1435-41. 28. Villeda SA, Luo J, Mosher KI, Zou B, Britschgi M, Bieri G, 14. Gorbunova V, Seluanov A, Zhang Z, Gladyshev VN, Vijg J. Stan TM, Fainberg N, Ding Z, Eggel A, Lucin KM, Czirr E, Comparative genetics of longevity and cancer: insights from Park JS, Couillard-Després S, Aigner L, Li G, Peskind ER, Kaye long-lived rodents. Nat Rev Genet. 2014 Aug;15(8):531-40. JA, Quinn JF, Galasko DR, Xie XS, Rando TA, Wyss-Coray T. e ageing systemic milieu negatively regulates neurogenesis 15. Austad SN. Methusaleh’s Zoo: how nature provides us with and cognitive function. Nature. 2011 Aug 31;477(7362):90-4. clues for extending human health span. J Comp Pathol. 2010 Jan;142 Suppl 1:S10-21. 29. Epel ES, Lithgow GJ. Stress biology and aging mechanisms: toward understanding the deep connection between adaptation 16. Nagy JD, Victor EM, Cropper JH. Why don’t all whales have to stress and longevity. J Gerontol A Biol Sci Med Sci. 2014 cancer? A novel hypothesis resolving Peto’s paradox. Integr Jun;69 Suppl 1:S10-6. Comp Biol. 2007 Aug;47(2):317-28. 30. Weindruch R. e retardation of aging by caloric restriction: 17. Albanes D. Height, early energy intake, and cancer. Evidence studies in rodents and primates. Toxicol Pathol. 1996 Nov- mounts for the relation of energy intake to adult malignancies. Dec;24(6):742-5. BMJ. 1998 Nov 14;317(7169):1331-2. 31. Curtsinger JW, Fukui HH, Resler AS, Kelly K, Khazaeli 18. Caulin AF, Maley CC. Peto’s Paradox: evolution’s prescription for AA. Genetic analysis of extended life span in Drosophila cancer prevention. Trends Ecol Evol. 2011 Apr;26(4):175-82. melanogaster. I. RAPD screen for genetic divergence between selected and control lines. Genetica. 1998;104(1):21-32. 19. Li Y, de Magalhães JP. Accelerated protein evolution analysis reveals genes and pathways associated with the evolution of 32. Ebert RH 2nd, Cherkasova VA, Dennis RA, Wu JH, Ruggles mammalian longevity. Age (Dordr). 2013 Apr;35(2):301-14. S, Perrin TE, Shmookler Reis RJ. Longevity-determining genes in Caenorhabditis elegans: chromosomal mapping of multiple 20. Keane M, Semeiks J, Webb AE, Li YI, Quesada V, Craig T, oninteractive loci. Genetics. 1993 Dec;135(4):1003-10. Madsen LB, van Dam S, Brawand D, Marques PI, Michalak P, Kang L, Bhak J, Yim HS, Grishin NV, Nielsen NH, Heide- 33. Friedman DB, Johnson TE. A mutation in the age-1 gene in Jørgensen MP, Oziolor EM, Matson CW, Church GM, Stuart Caenorhabditis elegans lengthens life and reduces hermaphrodite GW, Patton JC, George JC, Suydam R, Larsen K, López-Otín fertility. Genetics. 1988 Jan;118(1):75-86. C, O’Connell MJ, Bickham JW, omsen B, de Magalhães JP. Insights into the evolution of longevity from the bowhead whale 34. Kenyon C, Chang J, Gensch E, Rudner A, Tabtiang R. A C. genome. Cell Rep. 2015 Jan 6;10(1):112-22. elegans mutant that lives twice as long as wild type. Nature. 1993 Dec 2;366(6454):461-4. 21. Chen HY, White E. Role of autophagy in cancer prevention. Cancer Prev Res (Phila). 2011 Jul;4(7):973-83 35. Bartke A, Wright JC, Mattison JA, Ingram DK, Miller RA, Roth GS. Extending the lifespan of long-lived mice. Nature. 22. Terman A, Kurz T, Navratil M, Arriaga EA, Brunk UT. 2001 Nov 22;414(6862):412 Mitochondrial turnover and aging of long-lived postmitotic cells: the mitochondrial-lysosomal axis theory of aging. Antioxid 36. Ayyadevara S, Alla R, aden JJ, Shmookler Reis RJ. Remarkable Redox Signal. 2010 Apr;12(4):503-35. longevity and stress resistance of nematode PI3K-null mutants. Aging Cell. 2008 Jan;7(1):13-22. 23. Brunk UT, Terman A. Lipofuscin: mechanisms of age-related accumulation and inuence on cell function. Free Radic Biol 37. Inuzuka Y, Okuda J, Kawashima T, Kato T, Niizuma S, Tamaki Med. 2002 Sep 1;33(5):611-9. Y, Iwanaga Y, Yoshida Y, Kosugi R, Watanabe-Maeda K, Machida Y, Tsuji S, Aburatani H, Izumi T, Kita T, Shioi T. Suppression 24. Julien S, Schraermeyer U. Lipofuscin can be eliminated from of phosphoinositide 3-kinase prevents cardiac aging in mice. the retinal pigment epithelium of monkeys. Neurobiol Aging. Circulation. 2009 Oct 27;120(17):1695-703.

32 Cryonics / April 2015 www.alcor.org 38. Foukas LC, Bilanges B, Bettedi L, Pearce W, Ali K, Sancho 51. Yoon KT, Kim JK, Kim do Y, Ahn SH, Lee JD, Yun M, Rha SY, S, Withers DJ, Vanhaesebroeck B. Long-term p110α PI3K Chon CY, Han KH.Role of 18F-uorodeoxyglucose positron inactivation exerts a bene cial eect on metabolism. EMBO emission tomography in detecting extrahepatic metastasis in Mol Med. 2013 Apr;5(4):563-71. pretreatment staging of hepatocellular carcinoma. Oncology. 2007;72 Suppl 1:104-10. 39. Tazearslan C, Huang J, Barzilai N, Suh Y. Impaired IGF1R signaling in cells expressing longevity-associated human IGF1R 52. Harisinghani MG, Barentsz J, Hahn PF, Deserno WM, alleles. Aging Cell. 2011 Jun;10(3):551-4. Tabatabaei S, van de Kaa CH, de la Rosette J, Weissleder R. Noninvasive detection of clinically occult lymph-node metastases 40. Hütter G, Nowak D, Mossner M, Ganepola S, Müssig A, Allers in prostate cancer. N Engl J Med. 2003 Jun 19;348(25):2491-9. K, Schneider T, Hofmann J, Kücherer C, Blau O, Blau IW, Hofmann WK, iel E. Long-term control of HIV by CCR5 53. Barentsz JO, Fütterer JJ, Takahashi S. Use of ultrasmall Delta32/Delta32 stem-cell transplantation. N Engl J Med. superparamagnetic iron oxide in lymph node MR imaging in 2009 Feb 12;360(7):692-8 prostate cancer patients. Eur J Radiol. 2007 Sep;63(3):369-72

41. Gruver AL, Hudson LL, Sempowski GD. Immunosenescence 54. Corot C, Robert P, Idée JM, Port M. Recent advances in iron of ageing. J Pathol. 2007 Jan;211(2):144-56. oxide nanocrystal technology for medical imaging. Adv Drug 42. Melamed D, Scott DW. Aging and neoteny in the B lineage. Deliv Rev. 2006 Dec 1;58(14):1471-504. Blood. 2012 Nov 15;120(20):4143-9. 55. Heesakkers RA, Hövels AM, Jager GJ, van den Bosch HC, 43. Jackrel ME, DeSantis ME, Martinez BA, Castellano LM, Witjes JA, Raat HP, Severens JL, Adang EM, van der Kaa CH, Stewart RM, Caldwell KA, Caldwell GA, Shorter J. Potentiated Fütterer JJ, Barentsz J. MRI with a lymph-node-speci c contrast Hsp104 variants antagonize diverse proteotoxic misfolding agent as an alternative to CT scan and lymph-node dissection in events. Cell. 2014 Jan 16;156(1-2):170-82. patients with prostate cancer: a prospective multicohort study. Lancet Oncol. 2008 Sep;9(9):850-6 44. Jackrel ME, Shorter J. Reversing deleterious protein aggregation with re-engineered protein disaggregases. Cell Cycle. 2014 May 56. Laghi A, Paolantonio P, Panebianco V, Miglio C, Iafrate 1;13(9):1379-83. F, Di Tondo U, Passariello R. Decrease of signal intensity of myometrium and cervical stroma after ultrasmall 45. Jackrel ME, Shorter J. Potentiated Hsp104 variants suppress superparamagnetic iron oxide (USPIO) particles administration: toxicity of diverse neurodegenerative disease-linked proteins. an MR nding with potential bene ts in T staging of uterine Dis Model Mech. 2014 Oct;7(10):1175-84 neoplasms. Invest Radiol. 2004 Nov;39(11):666-70.

46. Jackrel ME, Tariq A, Yee K, Weitzman R, Shorter J. Isolating 57. Curvo-Semedo L, Diniz M, Miguéis J, Julião MJ, Martins P, potentiated Hsp104 variants using yeast proteinopathy models. Pinto A, Caseiro-Alves F. USPIO-enhanced magnetic resonance J Vis Exp. 2014 Nov 11;(93):e52089. imaging for nodal staging in patients with head and neck cancer. J Magn Reson Imaging. 2006 Jul;24(1):123-31. 47. Shay JW, Reddel RR, Wright WE. Cancer. Cancer and telomeres--an ALTernative to telomerase. Science. 2012 Jun 58. Guimaraes AR, Tabatabei S, Dahl D, McDougal WS, 15;336(6087):1388-90. Weissleder R, Harisinghani MG. Pilot study evaluating use 48. Osterwald S, Wörz S, Reymann J, Sieckmann F, Rohr K, Ere H, of lymphotrophic nanoparticle-enhanced magnetic resonance Rippe K. A three-dimensional colocalization RNA interference imaging for assessing lymph nodes in renal cell cancer. Urology. screening platform to elucidate the alternative lengthening of 2008 Apr;71(4):708-12. telomeres pathway. Biotechnol J. 2012 Jan;7(1):103-16. 59. Daldrup-Link HE, Rydland J, Helbich TH, Bjørnerud A, 49. Kines KJ, Sokolowski M, deHaro DL, Christian CM, Turetschek K, Kvistad KA, Kaindl E, Link TM, Staudacher Belancio VP. Potential for genomic instability associated with K, Shames D, Brasch RC, Haraldseth O, Rummeny EJ. retrotranspositionally-incompetent L1 loci. Nucleic Acids Res. Quanti cation of breast tumor microvascular permeability with 2014;42(16):10488-502. feruglose-enhanced MR imaging: initial phase II multicenter trial. Radiology. 2003 Dec;229(3):885-92. 50. deHaro D, Kines KJ, Sokolowski M, Dauchy RT, Streva VA, Hill SM, Hani n JP, Brainard GC, Blask DE, Belancio VP. 60. Yoo HJ, Lee JM, Lee MW, Kim SJ, Lee JY, Han JK, Choi BI. Regulation of L1 expression and retrotransposition by melatonin Hepatocellular carcinoma in cirrhotic liver: double-contrast- and its receptor: implications for cancer risk associated with light enhanced, high-resolution 3.0T-MR imaging with pathologic exposure at night. Nucleic Acids Res. 2014 Jul;42(12):7694-707. correlation. Invest Radiol. 2008 Jul;43(7):538-46.

www.alcor.org Cryonics / April 2015 33 Membership Statistics

2015 JAN FEB MAR APR MAY JUN JUL AUG SEP OCT NOV DEC Members 1016 Patients 134 Associate 151 Total 1301

International MembersPatients Country

Australia 10 3 Canada 43 2 Germany 5 0 Hong Kong 1 0 Israel 1 1 Italy 3 0 Japan 3 0 Number of Alcor members Mexico 4 0 Monaco 1 0 Netherlands 2 0 New Zealand 2 0 Norway 1 0 Portugal 4 0 Singapore 1 0 Spain 3 1 Thailand 3 1 United Arab Emirates 1 0 United Kingdom 23 2

Number of Alcor patients TOTAL 111 10 ARE YOU GETTING Curcumins ’ How Much Curcumin Are You Absorbing?

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These statements have not been evaluated by the Food and Drug Administration. ThisItem product #  is not intended to diagnose, treat, cure, or prevent any disease. Resuscitation Update Reported by R. Michael Perry

CRISPR Gene Edits research and development in a number the loss of brain cells, we can now work Human Stem Cells of sectors, ranging from electronics and toward finding a means to develop drugs energy to biomedicine and consumer that might mimic the protective effects of Precisely and Efficiently products. Nanotechnology has led to cold on the brain.” Scientists already know new materials—such as graphene—and that lowering body temperature can protect A powerful “genome editing” technology microscopic devices that include new the brain. known as CRISPR has been used by surgical tools and medicines. Up until researchers since 2012 to trim, disrupt, now however, nanotech R&D has been Kate Kelland, Alison Williams / Reuters replace or add to sequences of an hampered by the fact that it has not 14 Jan. 2015 organism’s DNA. Now, scientists at Johns been possible to achieve simultaneous http://www.reuters.com/ Hopkins Medicine have shown that the information on 3D structure, chemical article/2015/01/14/us-health-brain- system also precisely and efficiently alters composition and surface properties. This is cooling-idUSKBN0KN25J20150114 human stem cells. Previous research has what makes the UNIVSEM project, due for shown that CRISPR (third generation gene completion in March 2015, so innovative. editing) can generate genomic changes or By integrating different sensors capable of $4 Million in Funding to mutations through these interventions far measuring these different aspects of nano- develop Artificial General more efficiently than other gene editing sized materials, EU scientists have created techniques, such as TALEN (2nd generation a single instrument that enables researchers Intelligence gene editing). The researchers compared to work much more efficiently. the ability of both genome editing systems AGI Innovations Inc, (www.AGi-3.com) to either cut out pieces of known genes in Community Research and Development an R and D company focused on advancing iPSCs or cut out a piece of these genes and Information Service (CORDIS) Artificial General Intelligence (AGI) has replace it with another. As model genes, the 12 Jan. 2015 secured $4 million in funding to support researchers used JAK2, a gene that when http://cordis.europa.eu/news/ research on long-term AGI development. mutated causes a bone marrow disorder; rcn/122251_en.html AGI is the AI discipline concerned with SERPINA1, a gene that when mutated developing systems with human-like causes an inherited disorder that may cause cognitive abilities such as general learning, reasoning, and problem solving. AGI lung and liver disease; and AAVS1, a gene Cooling Brain Protein Could that’s been recently discovered to be a “safe Innovations Inc, also known as AGi3, was Aid Search for Alzheimer’s harbor” in the human genome for inserting formed early in 2014 to continue research foreign genes. Treatment originally spearheaded by Adaptive A.I. Inc (a2i2), an AGI R and D company formed NextBigFuture Scientists have found a mechanism that in 2001. On completing its first generation 12 Jan. 2015 kicks in when the body is cooled and AGI engine in 2008, a2i2 launched. Peter http://nextbigfuture.com/2015/01/ prevents the loss of brain cells, and say their Voss, founder of these companies, explains crispr-gene-edits-human-stem-cells.html find could one day lead to treatments for that while commercialization helped to brain-wasting diseases such as Alzheimer’s. validate the AGI approach taken by the Studying mice, the researchers were able company, it also totally shifted focus away Providing a Clearer Picture to simulate the effects of body cooling from general AI to providing specific and pick apart the workings of a so-called solutions to its customers. “Our AGI of Nanotechnology’s “cold-shock” protein in the brain, RBM3, research was essentially halted. Fortunately, Full Potential which has previously been linked with we now have a new dedicated and funded preventing brain cell death. “We’ve known company focused entirely on long-term A new tool capable of carrying out for some time that cooling can slow down AGI development” Voss said. simultaneous nano-sized measurements or even prevent damage to brain cells, could soon lead to more innovative but reducing body temperature is rarely NextBigFuture nanotech-based products and help feasible in practice (because) it’s unpleasant 21 Jan. 2015 boost the EU economy. Indeed the tool, and involves risks such as pneumonia http://nextbigfuture.com/2015/01/agi- developed by scientists cooperating and blood clots,” said Giovanna Mallucci innovations-secures-4-million-in.html through the EU-funded UNIVSEM who led the research. “By identifying how project, has the potential to revolutionize cooling activates a process that prevents

36 Cryonics / April 2015 www.alcor.org Scientists Upload a Worm’s Stephen Larson told CNN. “It’s certainly gelatinous liquid inside the egg into a solid. Mind into a Lego Robot exceeded my expectations.” The project Unboiling is probably not something that aims to digitally model the worm entirely most people will ever need to do in the A humble roundworm is leading the race in a virtual environment; creating a robot kitchen. However, in science the same basic in artificial intelligence, showing that it with an elastic body complete with stretchy process has the potential to reduce the cost may be possible one day to upload our muscles. of cancer treatment, food production and brains to a computer. Called the Open other important areas of the biotechnology Worm Project, the research brings together Gant Daily industry, which is worth $160 billion scientists and programmers from around 21 Jan. 2015 globally. “Yes, we have invented a way to the world with the aim of recreating the http://gantdaily.com/2015/01/21/ unboil a hen egg. In our paper, we describe behavior of the common roundworm scientists-upload-a-worms-mind-into-a- a device for pulling apart tangled proteins (Caenorhabditis elegans) in a machine. The lego-robot/ and allowing them to refold. We start with open source project recently had its first egg whites boiled for 20 minutes at 90 major breakthrough when its software— degrees Celsius and return a key protein modeled on the neurons of the worm’s Researchers Unboil Egg Whites in the egg to working order,” said Gregory nervous system—independently controlled Weiss, UCI professor of chemistry and a Lego robot. The machine’s sensors, Researchers at the University of California molecular biology & biochemistry. without any prior programming, made Irvine (UCI) along with colleagues in the robot behave in a similar fashion to Australia have successfully unboiled egg Justin Beach, National Monitor C. elegans, approaching and backing away whites, meaning that they have managed 25 Jan. 2015 from obstacles or stimulated by food. to quickly and inexpensively restore the http://natmonitor.com/2015/01/25/ “We’ve been working on it for four years proteins in the egg to their original state. researchers-uncoil-an-egg-with-profound- and while we have a lot more to achieve When an egg is boiled, the heat causes the implications-for-food-medicine-and- it’s been the most surprising project I’ve proteins in the egg to tangle and clump biotech/ been involved in,” project coordinator together. This is the process that turns the

A Roadmap to Resuscitation uccessful rejuvenation of cryonics patients will Magazine (July-August 1977):80-83. Reprinted in Cryonics Srequire three distinct technologies: (1) A cure for the 29:4 (4th Quarter 2008),14-17. disease that put the patient in a critical condition prior to cryopreservation; (2) biological or mechanical cell Corey Noble, “A ‘Realistic’ Scenario for Nanotechnological repair technologies that can reverse any injury associated Repair of the Frozen Human Brain,” in Brian Wowk, with the cryopreservation process and long-term care at Michael Darwin, eds., Cryonics: Reaching for Tomorrow, low temperatures; (3) rejuvenation biotechnologies that Alcor Life Extension Foundation, 1991. restore the patient to good health prior to resuscitation. OR it will require some entirely new approach such as (1) Ralph C. Merkle, “The Molecular Repair of the Brain,” mapping the ultrastructure of cryopreserved brain tissue Cryonics 15(January 1994):16-31 (Part I) & Cryonics using nanotechnology, and (2) using this information to 15(April 1994):20-32 (Part II). deduce the original structure and repairing, replicating or simulating tissue or structure in some viable form so the Ralph C. Merkle, “Cryonics, Cryptography, and Maximum person “comes back.” Likelihood Estimation,” First Extropy Institute Conference, Sunnyvale CA, 1994. The following list is a list of landmark papers and books that reflect ongoing progress towards the resuscitation of Aubrey de Grey & Michael Rae, “Ending Aging: The cryonics patients: Rejuvenation Breakthroughs That Could Reverse Human Aging in Our Lifetime.” St. Martin’s Press, 2007 Jerome B. White, “Viral-Induced Repair of Damaged Neurons with Preservation of Long-Term Information Robert A. Freitas Jr., “Comprehensive Nanorobotic Content,” Second Annual Conference of the Cryonics Control of Human Morbidity and Aging,” in Gregory M. Societies of America, University of Michigan at Ann Arbor, Fahy, Michael D. West, L. Stephen Coles, and Steven B. April 11-12, 1969, by J. B. White reprinted in Cryonics Harris, eds, The Future of Aging: Pathways to Human Life 35:10 (October 2014), 8-17. Extension, Springer, New York, 2010, pp. 685-805.

Michael G. Darwin, “The Anabolocyte: A Biological Chana de Wolf (now Phaedra), “Reconstructive Approach to Repairing Cryoinjury,” Life Extension Connectomics,” Cryonics 34:7 (July 2013), 26-28.

www.alcor.org Cryonics / April 2015 37 MEETINGS

ABOUT THE ALCOR FOUNDATION BRITISH COLUMBIA (CANADA): The Alcor Life Extension Foundation is a nonprofit tax-exempt scientific and The contact person for meetings in educational organization dedicated to advancing the science of cryopreservation the Vancouver area is Keegan Macintosh: and promoting cryonics as a rational option. Being an Alcor member means [email protected]. knowing that—should the worst happen—Alcor’s Emergency Response Team is ready to respond for you, 24 hours a day, 365 days a year. OREGON: The contact person for meetings in Alcor’s Emergency Response capability includes specially trained technicians and the Portland area is Aschwin de Wolf: customized equipment in Arizona, northern California, southern California, and [email protected] south Florida, as well as many additional certified technicians on-call around the See also: https://www.facebook.com/ United States. Alcor’s Arizona facility includes a full-time staff, and the Patient portland.life.extension Care Bay is personally monitored 24 hours a day. ALCOR PORTUGAL ARIZONA Alcor Portugal is working to have good FLAGSTAFF: SAN FRANCISCO BAY: stabilization and transport capabilities. The Arizona without the inferno. Cryonics Alcor Northern California Meetings group meets every Saturday for two hours. group in beautiful, high-altitude Flagstaff. are held quarterly in January, April, July, For information about meetings, contact Two-hour drive to Alcor. Contact eric@ and October. A CryoFeast is held once Nuno Martins at n-martins@n-martins. flagstaffcryo.com for more information. a year. For information on Northern com. The Alcor Portugal website is: www. California meetings, call Mark Galeck at alcorportugal.com. PHOENIX (650) 969-1671, (650) 534-6409 or email VALLEY OF THE SUN: [email protected]. TEXAS This group meets monthly, usually DALLAS: in the third week of the month. Dates FLORIDA North Texas Cryonauts, please sign up are determined by the activity or event Central Florida Life Extension group for our announcements list for meetings planned. For more information or to meets once a month in the Tampa Bay (http://groups.yahoo.com/group/ RSVP, visit http://cryonics.meetup. area (Tampa and St. Petersburg) for cryonauts-announce) or contact David com/45/ or email Lisa Shock at lisa@ discussion and socializing. The group Wallace Croft at (214) 636-3790 for details alcor.org. has been active since 2007. Email of upcoming meetings. [email protected] for more AT ALCOR: information. AUSTIN/CENTRAL TEXAS: Alcor Board of Directors Meetings and We meet at least quarterly for training, Facility Tours—Alcor business meetings are NEW ENGLAND transport kit updates, and discussion. For generally held on the first Saturday of every CAMBRIDGE: information: Steve Jackson, 512-447-7866, month starting at 11:00 AM MST. Guests The New England regional group [email protected]. are welcome to attend the fully-public board strives to meet monthly in Cambridge, meetings on odd-numbered months. Facility MA—for information or to be added UNITED KINGDOM tours are held every Tuesday and Friday to the Alcor NE mailing list, please There is an Alcor chapter in England. at 2:00 PM. For more information or to contact Bret Kulakovich at 617-824-8982, For information about meetings, contact schedule a tour, call Marji Klima at (877) [email protected], or on Alan Sinclair at [email protected]. 462-5267 x101 or email [email protected]. FACEBOOK via the Cryonics Special See the web site at www.alcor-uk.org. Interest Group. CALIFORNIA LOS ANGELES: PACIFIC NORTHWEST Alcor Southern California Meetings— A Yahoo mailing list is also maintained For information, call Peter Voss at for cryonicists in the Pacific Northwest (310) 822-4533 or e-mail him at peter@ at http://tech.groups.yahoo.com/group/ optimal.org. Although monthly meetings CryonicsNW/. are not held regularly, you can meet Los Angeles Alcor members by contacting Peter.

If you are interested in hosting regular meetings in your area, contact Alcor at 877-462-5267, ext. 113. Meetings are a great way to learn about cryonics, meet others with similar interests, and introduce your friends and family to Alcor members!

38 Cryonics / April 2015 www.alcor.org What is Cryonics?

ryonics is an attempt to preserve and protect human life, not reverse death. It is the practice of using extreme cold to attempt to preserve the life of a person who can no longer be supported by today’s medicine. Will Cfuture medicine, including mature nanotechnology, have the ability to heal at the cellular and molecular levels? Can cryonics successfully carry the cryopreserved person forward through time, for however many decades or centuries might be necessary, until the cryopreservation process can be reversed and the person restored to full health? While cryonics may sound like science fiction, there is a basis for it in real science. The complete scientific story of cryonics is seldom told in media reports, leaving cryonics widely misunderstood. We invite you to reach your own conclusions.

How do I find out more?

he Alcor Life Extension Foundation is the world leader in cryonics research and technology. Alcor is a non- profit organization located in Scottsdale, Arizona, founded in 1972. Our website is one of the best sources of Tdetailed introductory information about Alcor and cryopreservation (www.alcor.org). We also invite you to request our FREE information package on the “Free Information” section of our website. It includes:

• A fully illustrated color brochure • A sample of our magazine • An application for membership and brochure explaining how to join • And more!

Your free package should arrive in 1-2 weeks. (The complete package will be sent free in the U.S., Canada, and the United Kingdom.)

How do I enroll? Signing up for a cryopreservation is easy! Step 1: Fill out an application and submit it with your $90 application fee. Step 2: You will then be sent a set of contracts to review and sign. Step 3: Fund your cryopreservation. While most people use life insurance to fund their cryopreservation, other forms of prepayment are also accepted. Alcor’s Membership Coordinator can provide you with a list of insurance agents familiar with satisfying Alcor’s current funding requirements. Finally: After enrolling, you will wear emergency alert tags or carry a special card in your wallet. This is your confirmation that Alcor will respond immediately to an emergency call on your behalf.

Not ready to make full arrangements for cryopreservation? Then become an Associate Member for $10/month (or $30/quarter or $120 annually). Associate Members will receive: • Cryonics magazine by mail • Discounts on Alcor conferences • Access to post in the Alcor Member Forums • A dollar-for-dollar credit toward full membership sign-up fees for any dues paid for Associate Membership

To become an Associate Member send a check or money order ($10/month or $30/quarter or $120 annually) to Alcor Life Extension Foundation, 7895 E. Acoma Dr., Suite 110, Scottsdale, Arizona 85260, or call Marji Klima at (480) 905-1906 ext. 101 with your credit card information. You can also pay using PayPal (and get the Declaration of Intent to Be Cryopreserved) here: http://www.alcor.org/BecomeMember/associate.html

Call toll-free TODAY to start your application:

877-462-5267 ext. 132 • [email protected] • www.alcor.org You’re going to great lengths to avoid death. Why not do something to prolong life. Join the Life Extension Foundation® now so you can live a longer, healthier life. We’ll give you all the support you need, starting with cutting-edge medical information.

You get it three ways. rough our monthly Life Extension Magazine®… lled with cutting-edge research ndings and global medical breakthroughs even your doctors may not know about. rough our Disease Prevention and Treatment book, lled with breakthrough protocols on over 130 dierent diseases of aging. And with free phone access to our knowledgeable Health Advisors (naturopaths, nurses, nutritionists, even personal trainers). ey’re available every day of the year to address your health concerns and guide you in structuring a personal regimen of diet, exercise and nutritional supplements designed to extend your healthy life span. As a Life Extension® member, you’ll save far more than money with preventive blood screening to head o health problems … and advanced nutritional supplements that are light-years ahead of the commercial marketplace. ese are formulas guaranteed for purity, potency and e„cacy that you simply won’t nd anywhere else. All formulated to keep your body functioning youthfully for more years than you ever thought possible. So while you’re busy planning for the future, try spending a moment to prolong it. Call 1-866-820-4967 toll-free or visit www.LifeExtension.com/PIM501X to join the Life Extension Foundation now.

1-866-820-4967 • www.LifeExtension.com/PIM501X 502.62A4 0115 Your $75 annual membership dues will fund innovative anti-aging research that will ultimately benet you.

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