Male Offspring in Long-Lived Families are Less Fat
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The amount of visceral fat carried by individuals, just like number of calories consumed, has a strong influence on natural variations in health and longevity. More visceral fat is a bad thing, producing chronic inflammation and other less well understood disruptions of metabolism. This study shows that men, but not women, in long-lived families are less fat. That this is the case for only one gender is some defense against the hypothesis that the important factor being passed down here is culture (choosing to eat less) rather than genes, which would put a damper on many claims of genetic associations in longevity.

If the case, this would be an analogous situation to many life span studies in mice in recent decades that failed to control for inadvertent calorie restriction, and thus mistakenly identified various interventions as being life-extending, when in fact it was simply a matter of reduced calorie intake. The consequences of differences in visceral fat tissue, like those of dietary calorie intake, are large in comparison to most other influences on long term health at the present time, and so caution should be the watchword. Read studies carefully.

Familial longevity is marked by an exceptionally healthy metabolic profile and low prevalence of cardiometabolic disease observed already at middle age. We aim to investigate whether regional body fat distribution, which has previously shown to be associated with cardiometabolic risk, is different in offspring of long-lived siblings compared with controls.

Our institutional review board approved the study, and all participants (n = 344, average age in years 65.6) gave written informed consent. Offspring (n = 175) of nonagenarian siblings were included. Their partners (n = 169) were enrolled as controls. For abdominal visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) measurements, a single-slice 8.0 mm computed tomography (CT) acquisition was planned at the level of the 5th lumbar vertebra. In addition, participants underwent prospectively electrocardiography-triggered unenhanced volumetric CT of the heart. Abdominal VAT and SAT areas and epicardial adipose tissue (EAT) volumes were acquired. Linear regression analysis was performed adjusting for cardiovascular risk factors.

Total abdominal fat areas were smaller in male offspring compared with controls (353.0 versus 382.9 cm2). The association between low abdominal VAT areas in male offspring (149.7 versus 167.0 cm2 in controls) attenuated after additional adjustment for diabetes. Differences were not observed for females. EAT volumes were similar between offspring of long-lived siblings and controls. We conclude that males who have genetically determined prospect to become long-lived have less abdominal fat and in particular less abdominal VAT compared with controls.


Brain Preservation Foundation Prize Update
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The Brain Preservation Foundation aims at advancing and validating the state of the art for preserving the fine structure of the human brain, containing the data of the mind. This is a goal of great value for the cryonics industry, and for the possible plastination industry that might arise to be its competitor. All too many people, billions, will die before the advent and widespread availability of working rejuvenation therapies, and it is madness that so little is done to preserve these individuals for a chance at a future life, given the present existence of technologies that can achieve this goal. But that is the world we live in, and one of many things it is worth trying to change.

The Brain Preservation Foundation runs a technology prize to help accelerate and publicize progress in preservation technologies. Here is a recent update on the current batch of competitors:

Brain Preservation Prize competitor Shawn Mikula has just published the first ever paper demonstrating how an entire mouse brain can be preserved at the ultrastructure level for electron microscopic (EM) imaging of its entire connectome. As is well known to all electron microscopists, the traditional protocol for preparing brain tissue for EM imaging only works for small pieces of tissue. The key problem has been that the mix of chemicals used to preserve (and stain) the lipid membrane of cells, is prone to precipitation and barrier formation within the tissue. This has limited high-quality ultrastructure preservation and staining to depths of just a few hundred microns thick. Dr. Mikula's paper shows that this can be overcome by adding a high concentration of formamide to the mix. According to his paper this is sufficient to completely eliminate barrier formation allowing for uniform preservation and staining of an entire mouse brain.

Are these results sufficient for Mikula to win the mouse phase of our Brain Preservation Prize? The short answer is yes - if the claims made in the paper can be verified by our imaging then Mikula will be awarded the mouse phase of our prize. Another key question is whether his 'formamide' technique will be able to be scaled up to a large mammal - like the pig brain required for the final phase of our prize, or a human brain? Dr. Mikula is already working to procure high-quality glutaraldehyde perfused pig brains on which to test his technique. I suspect that to scale up to these large brains his protocol will need to be modified to include vascular perfusion.

I want to also touch on the significant progress that has been made by our other competitor team, 21st Century Medicine (21CM). 21CM's core mission is to develop a cryopreservation protocol sufficiently benign that whole, donated human organs could be vitrified (stored below -130 degrees Celsius without ice formation) and rewarmed when needed for transplantation. They have had great success showing that viability can be restored in vitrified slices of tissue. Unfortunately it is much easier to get cryoprotectant solutions into and out of half millimeter slices than whole brains. The whole rabbit brains that 21CM has perfused with cryoprotectant agents (CPA) for our prize have shown significant amounts of shrinkage due to dehydration from the high concentration and fast ramping of CPA used. Electron micrographs of this tissue are thus difficult to interpret and we have been unable to accurately assess the degree of ultrastructure preservation by this technique. 21CM has ideas on how to overcome this hurdle (which they believe to be one of evaluation rather than preservation) but progress has stalled on those experiments due to the expense involved.

Recently however, 21CM has begun a set of experiments which overcomes this dehydration and shrinkage issue in a very simple and inexpensive, but unorthodox, way. They perfuse the rabbit brain with glutaraldehyde fixative prior to perfusion with CPA and low temperature vitrification! This pre-fixation is of course completely incompatible with recovery of function by simple rewarming, but it has the effect of stabilizing the vascular system and tissue sufficiently to allow long duration room temperature perfusion of CPA. Initial results show that these brains (stored intact briefly at -135 degrees C) are not shrunken by this procedure and electron micrographs of brain ultrastructure appear "textbook-normal".


Neural Stem Cell Transplant Treats Parkinson's in Rats
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The proximate cause of the most visible symptoms of Parkinson's disease is the progressive loss of a small but vital population of dopamine-generating neurons. This loss happens to everyone, but for a variety of underlying reasons, not all of which are clear at this time, people with Parkinson's experience a more rapid loss of these cells. This is the case for many age-related medical conditions: they are a more rapid progression of a process that is in fact happening to all of us, and so the development of therapies is worth keeping an eye on. One approach to the treatment of Parkinson's disease is to attempt to restore the failing population of dopamine-generating neurons via some form of cell therapy, as demonstrated here in rats:

Parkinson's disease (PD) is considered the second most frequent and one of the most severe neurodegenerative diseases, with dysfunctions of the motor system and with nonmotor symptoms such as depression and dementia. Compensation for the progressive loss of dopaminergic (DA) neurons during PD using current pharmacological treatment strategies is limited and remains challenging. Pluripotent stem cell-based regenerative medicine may offer a promising therapeutic alternative, although the medical application of human embryonic tissue and pluripotent stem cells is still a matter of ethical and practical debate.

Addressing these challenges, the present study investigated the potential of adult human neural crest-derived stem cells derived from the inferior turbinate (ITSCs) transplanted into a parkinsonian rat model. Emphasizing their capability to give rise to nervous tissue, ITSCs isolated from the adult human nose efficiently differentiated into functional mature neurons in vitro. Transplantation of predifferentiated or undifferentiated ITSCs led to robust restoration of behavior, accompanied by significant recovery of DA neurons within the substantia nigra. ITSCs were further shown to migrate extensively in loose streams primarily toward the posterior direction as far as to the midbrain region, at which point they were able to differentiate into DA neurons within the locus ceruleus. We demonstrate, for the first time, that adult human ITSCs are capable of functionally recovering a PD rat model.


Linking Mitochondrial DNA Damage and Glaucoma
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Damage to mitochondrial DNA is a consequence of the normal operation of cellular processes, and is one of the contributing causes of degenerative aging. It acts through a convoluted chain of circumstances to generate a population of malfunctioning cells that export harmful reactive molecules into surrounding tissue. Here, researchers provide evidence linking mitochondrial DNA damage, and consequent dysfunction, with the progression of glaucoma, a form of neurodegeneration causing blindness:

Glaucoma is a chronic neurodegenerative disease characterized by the progressive loss of retinal ganglion cells (RGCs). Mitochondrial DNA (mtDNA) alterations have been documented as a key component of many neurodegenerative disorders. However, whether mtDNA alterations contribute to the progressive loss of RGCs and the mechanism whereby this phenomenon could occur are poorly understood. We investigated mtDNA alterations in RGCs using a rat model of chronic intraocular hypertension and explored the mechanisms underlying progressive RGC loss.

We demonstrate that the mtDNA damage and mutations triggered by intraocular pressure (IOP) elevation are initiating, crucial events in a cascade leading to progressive RGC loss. Damage to and mutation of mtDNA, mitochondrial dysfunction, reduced levels of mtDNA repair/replication enzymes, and elevated reactive oxygen species form a positive feedback loop that produces irreversible mtDNA damage and mutation and contributes to progressive RGC loss, which occurs even after a return to normal IOP.

Furthermore, we demonstrate that mtDNA damage and mutations increase the vulnerability of RGCs to elevated IOP and glutamate levels, which are among the most common glaucoma insults. This study suggests that therapeutic approaches that target mtDNA maintenance and repair and that promote energy production may prevent the progressive death of RGCs.


GABA Neuron Transplant Enhances Neural Plasticity in Mice
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Neural plasticity, the ability of the brain to generate new neurons and reshape its pathways, declines with age. Finding ways to temporarily reverse this decline and induce a more youthful state of plasticity may prove helpful in treating many conditions, but research is still at a fairly early stage of exploration:

Researchers have successfully re-created a critical juvenile period in the brains of adult mice. In other words, the researchers have reactivated brain plasticity - the rapid and robust changes in neural pathways and synapses as a result of learning and experience. The scientists achieved this by transplanting a certain type of embryonic neuron into the brains of adult mice. The transplanted neurons express GABA, a chief inhibitory neurotransmitter that aids in motor control, vision and many other cortical functions. Much like older muscles lose their youthful flexibility, older brains lose plasticity. But in the study, the transplanted GABA neurons created a new period of heightened plasticity that allowed for vigorous rewiring of the adult brain. In a sense, old brain processes became young again.

In early life, normal visual experience is crucial to properly wire connections in the visual system. Impaired vision during this time leads to a long-lasting visual deficit called amblyopia. In an attempt to restore normal sight, the researchers transplanted GABA neurons into the visual cortex of adult amblyopic mice. "Several weeks after transplantation, when the donor animal's visual system would be going through its critical period, the amblyopic mice started to see with normal visual acuity." These results raise hopes that GABA neuron transplantation might have future clinical applications. This line of research is also likely to shed light on the basic brain mechanisms that create critical periods.


Trials of Cell Therapies for Heart Regeneration
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Stem cell therapies to repair heart damage were one of the first to become widely available via medical tourism, and have been underway in earnest for more than a decade now. There is evidence from their use that benefits can be attained for patients. Providers and researchers continue to refine their techniques. There are numerous studies. That evidence is not good enough for the conservative end of the scientific community, who require a complete and rigorous understanding of the mechanisms of action before proceeding with enthusiasm, and nor for regulatory bodies in the US and Europe. Thus trials continue, now into their late stages, and treatments remain unavailable in many countries:

Like any new branch of medicine, cardiac cell therapy has progressed in fits and starts. Despite dozens of clinical trials, there's no slam-dunk treatment for improving the cardiac function of heart failure patients, but marginal, statistically significant improvements observed in some of the studies are propelling the cell-based therapies to ever larger, more expensive, and more rigorous trials. Most cardiologists remain underimpressed, says the head of an ongoing Phase 3 trial in Europe that involves injecting bone marrow cells into heart attack patients. "If [the trial] is positive, that's great, but I still think we'll have quite a challenge to convince people. If it's negative, then you get most of the cardiac community saying, 'Yep, we expected that.'?"

Now, it's make or break. Some anticipate that the results of the trial and two other ongoing Phase 3s will finally provide definitive evidence supporting the efficacy of cell therapies for the heart - evidence that has so far been lacking. On the other hand, negative results could spell the end of the approach altogether. "If our Phase 3 doesn't work, I think there's little likelihood any program could succeed in this indication," says the CEO of a Belgium-based firm sponsoring a clinical trial involving bone marrow-derived cells. "In the event they don't work [this time], I think it will be the end."

The idea to pluck cells from a person's bone marrow and shoot them into the heart took root in 2001, when researchers showed that doing so in mice could help regenerate damaged heart tissue. Yet no one knew how the cells worked. At the time, the prevailing thought was that stem cells took up residence in the heart and proliferated to produce new tissue. But this idea has since become a matter of debate. While some researchers claim the cells can form new cardiac muscle, others assert that the cells only very rarely differentiate into cardiomyocytes and instead support cardiac regeneration by other means.

Many scientists now believe that the introduced cells perform a paracrine function, signaling the activation of reparative pathways via growth factors or other secreted messengers. On its own, the heart regenerates about one percent of its tissue per year via the division of cardiomyocytes; perhaps cell therapies simply boost that normal behavior. The absence of a concrete mechanism of action has been one of the main criticisms of the field. On the other hand, most patients don't care how a treatment works, just that it does. "We have more trials than we have meaningful basic science papers. You'd like it to be the other way round. But I understand why there was an explosion [of clinical trials] - because there is such a need." And many researchers disagree that a known mechanism is required for advancing the therapy. "Nothing moves a field forward like actual clinical trials." While mechanisms are important - knowing them can help optimize treatments, for instance - "you can't slow things down because the mechanism of action isn't agreed upon by everybody."


Cryonics is Still in Search of Better Approaches to End of Life Management
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Cryonics is the industry and collection of technologies associated with low-temperature preservation of an individual upon death, necessarily carried out as soon as possible so as to prevent tissue damage in the brain. It is connected to research and development in forms of organ preservation associated with transplant medicine. A good cryopreservation of at least the brain ensures the best chance of future restoration with all the data of the mind intact, encoded in the fine structure of neurons and synapses: a preserved individual has all the time in the world to wait, after all. The odds of success are unknown, but infinitely better than is the case for all of the alternative options for those too old or too ill to wait for the advent of future rejuvenation therapies.

In an ideal world a good preservation would occur because it was scheduled ahead of time: a team and resources must be assembled and on the site, and this is hard and expensive to do at very short notice when there are so few qualified individuals and such a large territory to cover. This is why cryonics is strongly connected to legal issues surrounding self-determination in end of life choices, since in most countries people are forbidden to choose the time and manner of their own death, and doctors are forbidden to assist in enabling that death to be an easy one when the patient is in pain and dying, beyond the capacities of present medical technology. In that ideal world, the cryonics industry would also be large enough to ensure that first responders to medical emergencies, coroners, and other relevant individuals would as a matter of course be trained to understand and respect cryonics arrangements.

The present small size of the cryonics industry and the hostile nature of our legal systems means that we don't live in that world, unfortunately. We are not granted ownership over our own lives and bodies. Cryonics must occur as a last minute emergency effort at short notice in most cases, and the existing services and regulatory bodies must often be fought at the same time. Even people well connected within the cryonics community, who are well aware of the hurdles in the way, can succumb to sheer accident and as a result obtain a poor preservation with an unknown but probably large level of neural damage:

Dr. Laurence Pilgeram, a cryopreservation member of Alcor since 1991, was involved in cryonics early on. He gave a talk at the 1971 Cryonics Conference in San Francisco, California, on "Abnormal in-Vitro Oxidation and Lypogenesis Induced by Plasma in Patients with Thrombosis". Dr. Pilgeram was awarded his PhD. in Biochemistry at the University of California at Berkeley in 1953. In 1954-55 he served as an Instructor in Physiology at the University of Illinois College of Medicine in Chicago. After two years, he accepted an offer to develop and head an Arteriosclerosis Research Laboratory at the University of Minnesota School of Medicine. He later moved to Santa Barbara, California for a time before joining the Baylor College of Medicine in Houston, TX to develop and head the Coagulation Laboratory there.

On April 10, Dr. Pilgeram, collapsed outside of his home of an apparent sudden cardiac arrest. Despite medical and police personnel aware of his Alcor bracelet, he was taken to the medical examiner's office in Santa Barbara, as they did not understand Alcor's process and assumed that the circumstances surrounding his death would pre-empt any possible donation directives. Since this all transpired late on a Friday evening, Alcor was not notified of the incident until the following Monday morning.

Fortunately, no autopsy was performed which at least eliminated any invasive damage but the lengthy delay led to a straight freeze as the only remaining option. The medical examiner released the body to the mortuary that Alcor uses in Buena Park, California and he was immediately covered with dry ice, per our request. Aaron Drake and Steve Graber traveled to California to perform a neuro separation in the mortuary's prep room and then returned to Arizona for continued cool down which began on April 15, 2015.


A Trial of Immunotherapy to Treat Multiple Myeloma
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Any proposed first generation rejuvenation toolkit of future decades must include a robust approach to cancer therapy, at the very least offering reliable detection methods and cures even if not providing outright prevention. An important part of cancer therapies presently under development is the ability to far more accurately target cancer cells, thereby greatly reducing the presently onerous, damaging side-effects of treatment. Of the numerous approaches to targeted therapies, immunotherapy is one of the most advanced towards widespread clinical adoption, as illustrated by the results of this early stage trial of a form of adoptive T cell therapy:

Researchers say they have safely used immune cells grown from patients' own bone marrow to treat multiple myeloma, a cancer of white blood cells. A trial was conducted involving a particular type of tumor-targeting T cell, known as marrow-infiltrating lymphocytes (MILs). "What we learned in this small trial is that large numbers of activated MILs can selectively target and kill myeloma cells." MILs are the foot soldiers of the immune system and attack foreign cells, such as bacteria or viruses. But in their normal state, they are inactive and too few in number to have a measurable effect on cancer.

For the clinical trial, the team enrolled 25 patients with newly diagnosed or relapsed multiple myeloma, although three of the patients relapsed before they could receive the MILs therapy. The scientists retrieved MILs from each patient's bone marrow, grew them in the laboratory to expand their numbers, activated them with microscopic beads coated with immune activating antibodies and intravenously injected each of the 22 patients with their own cells. Three days before the injections of expanded MILs, patients received high doses of chemotherapy and a stem cell transplant, standard treatments for multiple myeloma.

One year after receiving the MILs therapy, 13 of the 22 patients had at least a partial response to the therapy, meaning that their cancers had shrunk by at least 50 percent. Seven patients experienced at least a 90 percent reduction in tumor cell volume and lived, on average, 25.1 months without cancer progression. The remaining 15 patients had an average of 11.8 progression-free months following MILs therapy. None of the participants had serious side effects from the MILs therapy. The overall survival was 31.5 months for those with less than 90 percent disease reduction, but this number has not yet been reached in those with better responses. The average follow-up time is currently more than six years.


Continued Investigations of Very Small Embryonic-Like Stem Cells in Adult Tissues
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It is important in the development of the present and the next generation of stem cell therapies to have cheap, reliable access to patient-specific pluripotent stem cells on demand. These are the basic starting point for generating therapeutic cells of a specific type, and only pluripotent cells have the capacity to create cells of any type. This is why there there is so much interest in developing the technology of induced pluripotency, for example, by which any cell sample from a patient can be used to create pluripotent cells. Some researchers believe that adult tissues contain populations of pluripotent stem cells necessary to continued tissue maintenance over a lifetime. If the case, these would be a useful source of cells for cell therapies. These proposed cell populations are given various names by various different research groups, such as very small embyronic-like stem cells. There is still some debate over whether such pluripotent cell populations actually exist in adult tissues, and whether researchers are accurately characterizing their observations. Research continues, however:

The pancreas is one of three organs (besides lung and liver) with huge regenerative ability. Mouse pancreas has a remarkable ability to regenerate after partial pancreatectomy, and several investigators have studied the underlying mechanisms involved in this regeneration process; however, the field remains contentious. Elegant lineage-tracing studies undertaken over a decade have generated strong evidence against neogenesis from stem cells and in favor of reduplication of pre-existing islets. Ductal epithelium has also been implicated during regeneration. We recently provided direct evidence for the possible involvement of very small embryonic-like stem cells (VSELs) during regeneration after partial pancreatectomy in mice.

VSELs were first reported in pancreas in 2008 and are mobilized in large numbers after treating mice with streptozotocin and in patients with pancreatic cancer. VSELs can be detected in mouse pancreas as small-sized LIN-/CD45-/SCA-1+ cells (3 to 5 μm), present in small numbers (0.6%), which express nuclear Oct-4 (octamer-binding transcription factor 4) and other pluripotent markers along with their immediate descendant 'progenitors', which are slightly bigger and co-express Oct-4 and PDX-1. VSELs and the progenitors get mobilized in large numbers after partial pancreatectomy and regenerate both pancreatic islets and acinar cells.

In this review, we deliberate upon possible reasons why VSELs have eluded scientists so far. Because of their small size, VSELs are probably unknowingly and inadvertently discarded during processing. Several issues raised in the review require urgent confirmation and thus provide scope for further research before arriving at a consensus on the fundamental role played by VSELs in normal pancreas biology and during regeneration, aging, and cancer. In the future, such understanding may allow manipulation of endogenous VSELs to our advantage in patients.


Doubt Cast on GDF-11 Mechanism for Improved Health in Mice
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The path of scientific discovery is never direct, and if something appears simple in an investigation of biochemistry then it is perhaps time to wonder what was missed and when the other shoe will drop. In the past couple of years researchers have demonstrated improved health in aged mice through reactivation of stem cell activity via increased levels of GDF-11. The situation may well be more complex than first thought, however, as a second group is having issues replicating the effect. This work calls into question present hypotheses on the nature of the underlying mechanisms exhibited in previous work on GDF-11, and points to the need for a better and more complete analysis of what is going on under the hood. The observed improvements to health seen in past studies are not yet disputed, but clearly something is missing:

In 2013, a team found that levels of a protein called GDF11 decreased in the blood of mice as they grew older. When the researchers injected the protein into the heart muscle of old mice, it became 'younger' - thinner and better able to pump blood. Two subsequent studies found that GDF11 boosted the growth of new blood vessels and neurons in the brain and spurred stem cells to regenerate skeletal muscle at the sites of injuries. Those results quickly made GDF11 the leading explanation for the rejuvenating effects of transfusing young blood into old animals. But that idea was confusing to many because GDF11 is very similar to the protein myostatin, which prevents muscle stem cells from differentiating into mature muscle - the opposite effect to that seen.

Researchers set out to determine why GDF11 had this apparent effect. First, they tested the antibodies and other reagents used to measure GDF11 levels, and found that these chemicals could not distinguish between myostatin and GDF11. When the team used a more specific reagent to measure GDF11 levels in the blood of both rats and humans, they found that GDF11 levels actually increased with age - just as levels of myostatin do. That contradicts what the former group had found. The researchers next used a combination of chemicals to injure a mouse's skeletal muscles, and then regularly injected the animal with three times as much GDF11 as the former team had used. Rather than regenerating the muscle, GDF11 seemed to make the damage worse by inhibiting the muscles' ability to repair themselves.

Researchers suggest that there could be multiple forms of GDF11 and that perhaps only one decreases with age. Both papers suggest that having either too much or too little GDF11 could be harmful. The more recent research group injured the muscle more extensively and then treated it with more GDF11 than the former group had done, so the results may not be directly comparable. "We look forward to addressing the differences in the studies with additional data very soon."