The above diagram shows the locations of the Sirtuin family of proteins as they are found in the cell. Sirtuins are a family of seven proteins that regulate cellular health. The sirtuins are present in the mitochondria (Sirt 3,4,5) and in the cell nucleus (Sirt 1,6,7). The sirtuins represent one of the most important pathways in the body. Remember that the cell and its organelles are analogous to computer hardware while the pathways represent the computer software. As I have said it is much easier to modify the software rather than the computer. When we modify the “software” we potentially modify upstream causes of aging. The Sirtuins are responsible for a host of functions in the body which can make the difference between health and disease. The following diagram shows just some of the functions. Let us briefly talk about each aspect of this illustration. SIRTUINS AND CHROMATIN REGULATION Chromatin is the material that makes up a chromosome that consists of DNA and protein. The major proteins in chromatin are proteins called histones. Their primary function is packaging long DNA molecules into more compact, denser structures. This prevents the strands from becoming tangled and also plays important roles in reinforcing the DNA during cell division, preventing DNA damage, and regulating gene expression and DNA replication. We see the names facultative and constitutive heterochromatin. All we need to know about these is that constitutive heterochromatin can affect the genes near itself; while facultative heterochromatin results in genes that are silenced but under specific conditions of developmental or environmental signaling cues, it can lose its condensed structure and become transcriptionally active. Thus, we can see that the facultative heterochromatin might be able to be manipulated. This leads to the field of Epigenetics. We will see down the road this can have profound implications in Anti-Aging medicine and disease control. CELL CYCLE CONTROL The cell cycle represents the essence of life. It deals with how cells reproduce. Typically, problems in the cell cycle result in many degenerative diseases and cancers. As cells move through the cell cycle, do they breeze through from one phase to the next? If they're cancer cells, the answer might be yes. Normal cells, however, move through the cell cycle in a regulated way. They use information about their own internal state and cues from the environment around them to decide whether to proceed with cell division. This regulation makes sure that cells don't divide under unfavorable conditions (for instance, when their DNA is damaged, or when there isn't room for more cells in a tissue or organ). Again, we can see the importance of the Sirtuins and the cell cycle. Our health is very dependent upon the proper functioning of the cell cycle. SIRTUINS AND CELL FATE DECISIONS The cell fate control is one of the Holy Grails of Regenerative Medicine. Sirtuins have a direct effect on what type of cell a stem cell may become. Sirtuins seem to have importance in the process of creating an IPS (induced pluripotential stem) cell. iPS cells are derived from skin or blood cells that have been reprogrammed back into an embryonic-like pluripotent state that enables the development of an unlimited source of any type of human cell needed for therapeutic purposes. Again, the relationship between sirtuins and stem cells is extremely important. It can have profound implications on Regenerative Medicine procedures. We can see how the sirtuins can control stem cell fate by taking notice of various environmental clues. Sirtuins give clues to the stem cells as to what to do and when to do it. We are leaning more and more about the relationship between the Sirtuin proteins and their effects on stem cells. They maintain stemness which is an essential characteristic of a stem cell that distinguishes it from ordinary cells namely self-renewal. SIRTUINS AND DNA REPAIR The sirtuin pathways are very important in the repair of DNA damage. The following diagram give us some insight into this process. The ssDNA is damage to a single strand of DNA while DBS repair represents a double strand break in the DNA. The following illustration shows the difference between single (ssDNA) and double Strand (DSB) DNA breaks. DNA safekeeping is one of the most important functions of the cell, allowing both the transfer of unchanged genetic material to the next generation and proper cellular functioning. Therefore, cells have evolved a sophisticated array of mechanisms to counteract daily endogenous and environmental assaults on the genome. These mechanisms rely on the recognition of the damaged DNA and its subsequent signaling. If DNA repair is not accomplished, this can lead to a variety of medical problems from cancer to a variety of degenerative diseases. SIRT1, SIRT3 and SIRT6 are involved in the signaling of different DNA repair pathways through key signaling factors. They help accomplish this with a variety of enzymes. Actually, DNA repair is intimately tied into chromatin regulation. Again, we go back to the histones and their importance. The Sirtuin histone targets are essentially gene targets. A major enzyme the sirtuins work with is PARP. The main role of PARP (found in the cell nucleus) is to detect and initiate an immediate cellular response to metabolic, chemical, or radiation-induced single-strand DNA breaks (SSB) by signaling the enzymatic machinery involved in the SSB repair. One very significant fact is that the PARP enzyme is a very big consumer of NAD+. This is one of a number of reasons that as we age we need to increase the amount of NAD+ or its precursors that is available to our body. More about NAD+ in a bit. SIRTUINS AND MITOCHONDRIA Mitochondria play a critical role in energy production, cell signaling and cell survival. Defects in mitochondrial function contribute to the aging process and aging-related disorders such as metabolic disease, cancer, and neurodegeneration. As time goes on we are realizing the mitochondrial malfunctions leave their footprints on most diseases. Alterations in the expression/activity of SIRT3, SIRT4, SIRT5 are linked with many different diseases. Overall, mitochondrial sirtuins regulate mitochondrial protein networks, orchestrate mitochondrial function, and allow cells to adapt to metabolic stresses. In addition, emerging evidence indicates that sirtuins regulate yet another important cellular process, autophagy. Autophagy is extremely important in the body’s recycling process. It allows the orderly degradation and recycling of cellular components. The sirtuins and mitochondria are intimately involved in this process. If you encourage autophagy you encourage anti-aging. SIRT3, SIRT4 and SIRT5 (mitochondrial sirtuins) belong to the sirtuin family proteins and are located in the mitochondria. They catalyze NAD+ into a variety of different compounds. They modulate the function of various targets to regulate the metabolic status in cells. Emerging evidence has revealed that mitochondrial sirtuins coordinate the regulation of gene expression and activities of a wide spectrum of enzymes to orchestrate oxidative metabolism and stress responses. Mitochondrial sirtuins act in synergistic or antagonistic manners to promote respiratory function, antioxidant defense, insulin response and adipogenesis to protect individuals from aging and aging-related metabolic abnormalities. The next diagram shows how mitochondria produce energy, namely ATP, the body’s energy currency. ATP keeps the cells alive and it is instrumental in many of the body’s repairs. This is the Krebs cycle and other energy producing cycles. HOW DO THE SIRTUINS ACCOMPLISH THEIR VARIOUS TASKS? The basic role of sirtuins is that they remove acetyl groups from other proteins. An acetyl group is a small molecule made of two carbon, three hydrogen, and one oxygen atoms. When Acetyl groups are added to or removed from other molecules that may affect how the molecules act in the body. In the above diagram the Acetyl group is represented by Ac in the green hexagons. Acetyl groups control specific reactions. Sirtuins work with acetyl groups by doing what’s called deacetylation. This means they recognize there’s an acetyl group on a molecule then remove the acetyl group, which tees up the molecule for its job. One way that sirtuins work is by removing acetyl groups (deacetylating) biological proteins such as histones. For example, sirtuins deacetylate histones, proteins that are part of a condensed form of DNA called chromatin. Here we see the DNA strands wound about the histones. The histone is a large bulky protein that the DNA wraps itself around. Think of it as a Christmas tree, and the DNA strand is the strand of lights. When the histones have an acetyl group, the chromatin is open, or unwound. This unwound chromatin means the DNA is being transcribed, an essential process. But it doesn’t need to remain unwound, as it’s vulnerable to damage in this position, almost like the Christmas lights could get tangled or the bulbs can get damaged when they’re unwieldy or up for too long. When the histones are deacetylated by sirtuins, the chromatin is closed, or tightly and neatly wound, meaning gene expression is stopped, or silenced. Sirtuins deacetylate a multitude of targets including histones, transcription factors, and metabolic enzymes. Taken together, sirtuins have been implicated in numerous cellular processes including stress response, DNA repair, energy metabolism. Sirtuins are a family of proteins that act as metabolic sensors. They deacetylase a coenzyme NAD+ into free nicotinamide. Basically, they break down acetyl from proteins to maintain their functioning for longer. The ratios of NAD+ and NADH determine the nutritional status of the cell and sirtuins are there to respond to it. NAD+ is an essential currency for energy metabolism and DNA repair. Sirtuins are proteins that evolved to respond to the availability of NAD in the body. The preceding diagram spells out the relationship between the Sirtuins and NAD+. We see that the sirtuins in response to NAD+ levels influence a number of other pathways which are responsible to a variety of different diseases. The bottom line is the Sirtuins are a family of proteins that act as metabolic sensors. They deacetylase a coenzyme NAD+ into free nicotinamide. The ratios of NAD+ and NADH determine the nutritional status of the cell and sirtuins are there to respond to it. NAD+ is an essential currency for energy metabolism and DNA repair. Sirtuins are proteins that evolved to respond to the availability of NAD+ in the body. SIRTUINS AND OXIDATIVE STRESS The sirtuins help regulate oxidative stress and inflammation. The above diagram shows this very well. In this diagram we see that the sirtuins have a direct effect on a pathway called the NRf2 pathway. This is the major pathway in the body that helps reduce inflammation. Consider it a thermostat of inflammation. When this pathway is stimulated it produces very powerful antioxidant enzymes which will significantly lower inflammation. The antioxidant properties also tie into the NAD/NADH ratio which ideally, we like to be 700/1. HOW TO INCREASE SIRTUINS There’s a lot of evidence pointing to the longevity benefits of increased sirtuin activity. If not in over-expression, then increasing sirtuins can still be good for your health in most cases. Glucose restriction extends the lifespan of human fibroblasts because of increased NAD+ and sirtuin activity. Inhibiting insulin shuttles SIRT1 out of the cell’s nucleus into the cytoplasm. Cancers use primarily glucose and glutamine for fuel with the exception of some ketones in rare cases. Caloric restriction and fasting increase SIRT3 and deacetylate many mitochondrial proteins. Reduction of calorie intake without causing malnutrition is the only known intervention that increases the lifespan of many species including primates. It’s thought that these effects in longevity require SIRT1. The following illustration demonstrates a variety of methods to help increase sirtuin activity. The chart is broken down into a few different categories. The input and effector categories are the most important to us. We see the sirtuins are very important sensors. Activating AMPK elevates NAD+ levels, leading to increased Ketosis. Ketone bodies like beta-hydroxybutyrate promote sirtuin activity. This is essentially a ketogenic diet. Thus, one simple way of increasing the sirtuins is by following a somewhat ketogenic diet. Natural ways of caloric restriction and fasting are still the best ways of signaling energy deprivation which promotes longevity. In an everyday context, a low carb diet is also pro-sirtuin to a certain extent because of the low levels of insulin and glucose. Exercise has anti-inflammatory effects and it increases SIRT1. The long-term benefits of exercise are even thought to be regulated by SIRT1. Cyclic-AMP (CAMP) pathway activates SIRT1 very rapidly to promote fatty acid oxidation independent of NAD+. CAMP is linked with AMPK (another very important pathway in the body) which gets activated under high energy demands while being energy deprived. This can be accomplished with cold exposure and high-intensity exercise training. Heat exposure and saunas increase NAD+ levels which promote SIRT1 as well. Sweating, cardio, yoga, or infrared saunas will probably have a similar effect on activating heat shock proteins which can increase sirtuins. Chronic oxidative stress and DNA damage depletes NAD+ levels and decreases sirtuin activity. This will then disrupt DNA repair and impair mitochondrial functioning. That’s why you want to keep stressors acute and followed by recovery. Melatonin can activate sirtuins and has anti-aging effects. It’s also the main sleep hormone and a powerful antioxidant that helps the brain get more recovery from deeper stages of sleep. Sirtuins also affect the circadian clocks so keeping a consistent circadian rhythm is incredibly important for longevity. NAD+ is under circadian control and when you you are misaligned you’ll have less energy and lower SIRT1 and SIRT3 activity. The enzyme SIRT1 increases FOXO DNA binding by deacetylating FOXO in response to oxidative stress. FOXO is another very important pathway. FOXO proteins get increased in response to cellular stress and increased energy depletion. PERHAPS NAD IS THE BEST WAY TO INCREASE SIRTUIN FUNCTION Maybe the best way to consider NAD+ is to consider it as the energy currency to purchase increased Sirtuin activity. Of all the methods to increase Sirtuin activity NAD+, especially when given intravenously, may be one of the most effective sirtuin stimulators. In the above diagram, STACs are sirtuin activating compounds. In addition to Resveratrol, there is also Pterostilbene which is a better effector than Resveratrol. THE BOTTOM LINE IS EAT THE ABOVE FOODS, EXERCISE, ESPECIALLY INTERMITTENT HIGH INTENSITY EXECISE, INTERMITENT FASTING, AND CALORIE RESTRICTION WILL INCREASE YOUR SIRTUINS. TAKE PLENTY OF NAD OR ITS SUBSTRATES. WHEN YOUR SIRTUINS ARE INCREASED IT WILL HAVE A WATERSHED EFFECT ON THE OTHER PATHWAYS IN THE BODY. THE DIAGRAM BELOW SAYS IT ALL. Thanks, Dr. P

In 1972, Easter Island, called Rapa Nui, famous for its Moai statues, offered a new wonder: the discovery of the drug rapamycin named after the island's Polynesian name, Rapa Nui. Over the past three decades, rapamycin, which was isolated from soil bacteria, has been applied as an immuno-suppressor in a multitude of ways, including to coat coronary stents and to reduce the immune responses in people who receive organ transplants. Currently, it’s garnering attention because of its potentials in anti-cancer and neuroprotection as well as anti-aging therapies. Rapamycin has extended the expected lifespan of middle-aged mice by 28 percent to 38 percent. In human terms, this would be greater than the predicted increase in extra years of life if cancer and heart disease were both cured and prevented. Aging researchers currently acknowledge only two life-extending interventions in mammals: calorie restriction and genetic manipulation. Rapamycin appears to partially shut down the same molecular pathway as restricting food intake or reducing growth factors. We can see in this illustration of this concept. What we do notice is that caloric restriction intimately tied in with longevity. Rapamycin works by targeting a master regulator of cell growth in our cells called mTOR pathway. mTOR is a kinase which is a protein that phosphorylates (adds a phosphate group) to an amino acid. This is important because the phosphate group basically acts as a flag telling other proteins to bind on top of the phosphate. A bit more about kinases. As one of the most abundant gene families in humans, protein kinases tightly control cell signaling via pathways and cell function through protein phosphorylation. What is a pathway and how do we put it into the proper prospective? I saw an excellent article that gave a great analogy on how pathways function in the body. Think of the cell and all its different organelles (such as mitochondria) as the computer hardware and the biological pathways act as the software. We might not be able to change our hardware but we can modify the software. This is the essence of Anti-Aging medicine. When we want to truly alter the course of aging we typically need to influence the pathways or the “software”. All too often aging problems are attacked by altering the downstream causes of aging. This is typically not an effective method of treating aging. When rapamycin targets mTOR, it inhibits cell growth. Many of the problems that come with aging arise from uncontrolled growth or aging cells that have accumulated so much internal cellular debris that they lose healthy functionality. This makes rapamycin a potent anti-cancer drug, since cancer is marked by uncontrolled cell growth. The inhibition of mTOR also triggers autophagy, a process by which lysosomes, the so-called recycling centers of cells, clean up misfolded proteins and damaged organelles. During stress conditions, autophagy can lead to the cell survival by degrading dysfunctional components and providing the building blocks of cells, such as amino acids and lipids for reuse by the cell. mTOR is involved in every aspect of cellular life and existence. The following diagram shows some aspects of the mTOR pathway. In the case of mTOR we are actually trying to apply the brakes to cell growth and proliferation. In addition, the mTOR pathway is a direct target of the IGF-1 signaling pathway, which is a major driver of aging. Despite talking about mTOR as if it is just one molecule, it functions, in fact, as two separate complexes called mTORC1 and mTORC2 as we see from the following diagram: mTORC1 and mTORC2 which are composed of discrete protein binding partners to regulate cell growth, motility, and metabolism. These complexes are sensitive to distinct stimuli, as mTORC1 is sensitive to nutrients while mTORC2 is regulated via growth factor signaling. mTOR can affect cell division processes, the response to stress, and general cell and protein tasks. These signaling pathways are very complicated and depend on many feedback loops, energy supply, and a great variety of other molecules or signals. During certain diseases and aging, the function of mTOR can become deregulated. What does all of this have to do with aging? Many processes that lead to better longevity outcomes, like calorie restriction, fasting, and protein restriction, stimulate autophagy – a process within cells that cleans up broken proteins, bacteria, and viruses. Autophagy stimulates longevity. Aside from regulating cell growth and metabolism, mTORC1 also controls autophagy, an intracellular process that allows orderly degradation and recycling of cellular components. mTORC1 negatively regulates autophagy. The following illustration demonstrates Autophagy. The lysosomes are essentially recycling centers. mTOR SUPPRESSES AUTOPHAGY, WHICH SUPPRESSES LONGEVITY, THUS SUPPRESS mTOR INCREASE AUTOPHAGY Activating mTOR prevents autophagy from occurring. Autophagy seems to be a crucial component of longevity. Therefore, suppressing mTOR which rapamycin does should have longevity benefits. It is this effect, plus others like its suppression of cell division and growth, that immediately raised interest for its study in the field of aging sciences. But herein lies a cautionary tale for all potential aging therapeutics. Remember, mTOR affects many processes important for biological functioning. We cannot just eliminate mTOR without subsequent problems. We need to walk a fine balance line. mTORC1 and mTORC2 generally promote an anabolic response by stimulating protein synthesis, glycolysis, mitochondrial functions, and lipid synthesis to influence proliferation and survival, effector and memory responses, innate training and tolerance as well as hematopoietic stem cell maintenance and differentiation. Deactivation of mTOR restores cell homeostasis after immune activation and optimizes antigen presentation and memory T‐cell generation. Antigen presentation and memory T-cell generation are the basis of the Covid-19 vaccines. These findings show that the mTOR pathway integrates information of the environmental and cellular energy status by regulating cellular metabolic responses to guide immune cell activation. Elucidation of the metabolic control mechanisms of immune responses will help to generate a systemic understanding of the immune system and may eventually help us in dealing with the Covid-19 virus. Should we be concerned when we utilize rapamycin to block the mTOR pathways? Remember, the mTOR pathway affects many processes to keep our cells functioning. Suppressing it too intensely is clearly not good; we need a lighter touch. Like many things in life we need a fine balance. Not only do we need to establish the right dose for us, but we also need to identify the right treatment regimen. In some clinical studies, it took about two weeks to demonstrate significant suppression of mTOR1C. Sensible usage strategies (if there are any) for rapamycin could, for example, be for those aged 60 years and older to either use a low daily dose or to use it for 30 days at a time every 6 months or so. Low-dose, short-term rapamycin in healthy humans has been found to be safe, with few or no side effects. It’s possible that rapamycin can be given to healthy humans with fewer side effects than occur in transplant recipients. Healthy men given a single dose of rapamycin of roughly 16 mg/kg had no significant difference in the incidence of adverse effects between treatment and placebo. Healthy elderly volunteers given everolimus (a rapalog of rapamycin) at 0.5 mg daily, 5 mg weekly, or 20 mg weekly, for six weeks, had no serious adverse effects associated with the drug. The most common associated side effect was mouths ulcers. The treated groups had higher antibody titers in response to flu vaccines compared to controls, implying better immune function. Note that this is a much lower dose than the doses given in mouse studies of rapamycin. A rapalog is a class of molecules that has rapamycin-like activity but is not confined to chemical derivatives of rapamycin. A rapalog is also known as an analog. Unfortunately, treatment with rapamycin and its analogs/ rapalogs is associated with negative side effects that limit their potential utility as anti-aging therapies. These side effects include immunosuppression, glucose intolerance, an increased risk of type 2 diabetes, and disruption of lipid homeostasis. Although short term, low dose, and/or intermittent rapalog-based regimens may partially limit side effects, and trials of rapamycin in the elderly have begun, the efficacy of such treatment regimens in treating age-related diseases remains to be determined, particularly as the effects of rapamycin on longevity are dose-dependent. All three FDA-approved rapalogs sirolimus, everolimus, and temsirolimus have similar effects on the glucose metabolism and immune cell profile of mice. It is clear that there is an urgent need for new molecules that inhibit mTOR signaling with reduced side effects. There are some studies that claim the Metformin may also target mTOR. Taken together, rapamycin and metformin are promising candidates for life and healthspan extension; however, concerns of adverse side effects have hampered their widescale adoption for this purpose. It appears that the two may cancel out the unwanted side effects of each. Recent studies have demonstrated that a number of natural products, nutraceuticals, isolated from plants (e.g. fruits, vegetables, spices, nuts, legumes, herbs, etc.) also inhibit PI3K/Akt/mTOR pathway, and exhibit potent anticancer activities and probable antiaging activities. As most of the natural products occur in our diet every day, and are very safe, the results suggest that those natural products may be explored for cancer prevention and treatment. This special issue selects apigenin (present in many fruits, vegetables), curcumin, cryptotanshinone (Asian herbal compound), fisetin (compound from various berries), indoles (Cruciferous vegetables), isoflavones (genistein and deguelin), quercetin, resveratrol, and tocotrienol (a form of Vitamin E). Immunologically, mTOR has a fundamental part in controlling and shaping diverse functions of innate and adaptive immune cells, in particular, T-cell subsets differentiation, survival, and metabolic reprogramming to ultimately regulate the fate of diverse immune cell types. We must remember m TOR is a kinase. As one of the most abundant gene families in human, protein kinases tightly control cell signaling and cell function through protein phosphorylation. There are 634 protein kinases encoded in human genome to date, and 85% kinases are observed dysregulated in human disorders. However, only 49 small molecule kinase inhibitors have been approved for treating human diseases, urging for more in-depth investigations on the rest 77% possibly druggable kinome. Among all protein kinases, mTOR (mechanistic target of rapamycin, previously known as mammalian target of rapamycin) has drawn extensive attention. This is due to its indispensable roles in regulating key cell function such as proliferation, metabolism, autophagy, ageing, and others, and its ability to sense environmental changes including nutrients and growth factors to adjust cell physiology. Dysregulation of mTOR signaling is observed in solid tumors, epilepsy, obesity, and diabetes. The bottom line for the mTOR pathway is intermittent fasting, calorie restriction, a number of different supplements, and some intermittent rapamycin in the prescribed dosages and regimens which include Metformin. I suspect, as we refine things, we will see much more of rapamycin use. I suspect mTOR inhibition is just the tip of one of the icebergs. Thanks, Dr. P

The above diagram depicts the various pillars of aging. Unfortunately, these causes of aging seem to gang-up on an individual. When we look at aging we need to look at “Hallmarks of Aging”. The next diagram is an excellent synopsis of Aging. It demonstrates what I call the upstream causes of aging. We can see the primary hallmarks of aging. The first four niches represent the true upstream causes of aging. The other five hallmarks are responses to aging. Any of the above are capable to cause us to age more rapidly. Intervention in any of these hallmarks can have significant effects on the aging process especially the first four, the true upstream causes of aging. The real question becomes what biomarkers (a measurable substance in an organism whose presence is indicative of some phenomenon such as disease, infection, or environmental exposure) can be used to predict and measure aging? The following diagram is a take-off of the first diagram showing a variety of biomarkers. The question at hand is which of these biomarkers can be of benefit in predicting and slowing down aging and showing disease risk in a practical sense. More importantly, can it demonstrate if in fact that certain modalities are slowing down aging? Aging is the biggest risk factor for all chronic diseases including arthritis, cancer, heart disease, diabetes, Alzheimer’s, Parkinson’s and many more. The problem is that there is no good consensus on what biomarkers to use to measure aging. The preceding diagram shows some of the biomarkers utilized yet there is much controversy as to which are the most important and practical. There are two kinds of age: chronological age, which is the number of years one has lived, and biological age, which is influenced by our genes, lifestyle, behavior, the environment and other factors. Biological age is the superior measure of true age and is the most biologically relevant feature, as it closely correlates with mortality and health status. The search for reliable predictors of biological age has been ongoing for several decades, and until recently, largely without success. They may be part of the of the Holy Grail of Anti-Aging. The above diagram is an attempt to give some ideas of what can be utilized as biomarkers of aging. The biomarkers are very complicated and may not actually be all that reliable. Some are very well known while others are esoteric. Let us look at the more well-known markers of aging. TELOMERES Telomeres are DNA timers that limit the lifespan of a single cell. On the individual cell level, telomeres are the best marker of aging. However, we are composed of trillions of cells and each of them has different age and expected lifespan. Some studies show that there may be some importance on Telomere aging of the immune system. A recent large study concluded that telomere tests are not a good predictor of age-related health status. At one time Telomere testing was considered the gold standard of age assessment. I still consider it an important test but now there are other tests that may give us a better picture of aging. A recent large study concluded that telomere tests are not a good predictor of age-related health status. It is an important test but now there are other tests that may give us a better picture of aging. Telomere aging can directly affect the immune system. The younger your immune system the better equipped you are to fight off diseases. There are studies demonstrating V-cells can make the immune system younger. PROTEOSTASIS Protein homeostasis or 'proteostasis' is the process that regulates proteins within the cell in order to maintain the health of both the cellular proteome and the organism itself. Proteostasis involves a highly complex interconnection of pathways that influence the fate of a protein from synthesis to degradation. THE ABOVE PROCESSES ARE IMPORTANT IN THE DEVELOPMENT AND MAINTANCE OF PROTEINS. The tree diagram of Proteostasis is a good summation of what happens when our proteins run into problems. When discussing Proteostasis, we still do not have a good handle on the appropriate biomarkers of aging. Proteins and their problems are important for assessing aging but they still need leave much to be desired. The repair, recycling, and elimination of damaged macromolecules/organelles have emerged as key processes in maintaining cell integrity and function. At this time there are not really good biomarkers for Proteostasis. MITOCHODRIAL AGING Mitochondria influence or regulate a number of key aspects of aging and suggest that strategies directed at improving. Mitochondrial quality and function might have far-reaching beneficial effects. Mitochondria are organelles found in all human cells, and their primary role is energy production through oxidative phosphorylation. They are also involved in signaling by producing ROS, as well as by regulating cellular metabolism, apoptosis‐programmed cell death, and other functions that are biologically important but cannot be reliably measured in real life. The mitochondria seem to have a direct effect on most other aspects of aging. Their effects are far ranging since they influence almost all other aspects of aging from Telomere attrition to Stem Cell Exhaustion. This is easily seen on the following diagram. There still does not seem to be a consensus of opinion as what mitochondrial biomarkers are of value in assessing aging. However,we know that mitochondria are extremely important when dealing with aging. The mitochondrial theory of aging proposes that accumulation of damage to mitochondria and mitochondrial DNA (mtDNA) induces aging by reducing energy availability and increasing production of ROS that damage macromolecules. Unfortunately, these are difficult to measure. Luckily, there are a number of measures that can help keep mitochondria healthy. Many of these treatments are available in our clinic. CELLULAR SENESCENCE Cells continually experience stress and damage from exogenous and endogenous sources, and their responses range from complete recovery to cell death. Proliferating cells can initiate an additional response by adopting a state of permanent cell-cycle arrest that is termed cellular senescence. Aging is a complex biological process involving the continuous accumulation of changes in our bodies. Except for wrinkles and grey hair, among other changes in physical appearance, getting older also entails gradual deterioration of various bodily functions, leading to numerous age-related diseases and conditions. Although human aging is believed to go hand-in-hand with cellular senescence, what we currently know about this relationship is just the tip of the iceberg. Senescence is a key hallmark of chronological aging in humans, senescence levels may vary significantly between organs within the same person. Therein is where the problem lies. It is difficult to measure Senescence in humans. We can treat Senescent cells with Senolytic agents but we are still not sure if we are actually slowing down the clock of aging. Cellular senescence is a natural and irreversible process resulting from cells having the potential to multiply for only a limited number of times (also known as Hayflick limit). Senescent cells are old cells that can no longer divide. Even though the limited potential for division acts as a brake on old damaged cells from becoming cancerous, over time, senescent cells can cause damage to other cells and trigger aging and age-related illnesses. The problem with Senescence is that we do not have a good methodology of how to measure Senescence with biomarkers. Luckily, as was the case with mitochondrial aging we do have effective methods in our clinic to handle senescent cells. Nutrient Sensing and Cell Communications These are certainly important aspects of aging and manipulating these pathways can indeed have significant impacts on aging. Although these pathways produce measurable entities in the blood we cannot necessarily gain much information from their measurements. The reason for this is that these values are constantly changing depending on diet and exercise, even the time of the day. Thus, there are not dependable biomarkers. STEM CELL EXHAUSTION Stem cell exhaustion is influenced by the other pathways of aging. One of the most obvious characteristics of the aging process is the progressive decline in the regenerative potential of tissues. Adult stem cells are critical for rejuvenating tissues and persist throughout our lifespan. However, stem cell function declines during the aging process in tissues such as the brain, blood, skin, intestinal epithelium, bone, and skeletal muscle. This demise may contribute to tissue degeneration, organismal aging, and age-related diseases. Again, the problem is that there is no good biomarker test to assess stem cell exhaustion. Luckily, we have devised methods of maintain stem cell numbers by combinations of supplements and growth factor patches. SAVING THE BEST FOR LAST ... EPIGENETICS The term, "epigenetics," encompasses the ensemble of mechanisms that modulate gene expression programs that adapt to environmental cues and define stable characteristics from differentiated cell types. Epigenetics seems like a concept pulled from a futuristic science-fiction movie where a drop of blood is fed into a machine, in which an algorithm churns through an accumulation of chemical groups coating a strand of DNA and spits out an individual's realistic age reflecting a lifetime of experiences and exposures. In simplistic terms, epigenetics involves putting a methyl group on a strand on DNA. A methyl group is a biochemical compound made from hydrogen and carbon. What we are really dealing with is called DNA methylation. DNA methylation occurs when a methyl group is added to a DNA strand. DNA methylation is a tool to typically lock genes into their off position. This is seen in the following illustration. Aberrant DNA methylation, which is a nearly universal finding in cancer results in disturbed gene expression. DNA methylation is modified by environmental factors such as diet and exercise that may modify cancer risk and tumor behavior. Abnormal DNA methylation has been observed in several cancers. These alterations in DNA methylation may play a critical role in cancer development and progression. Dietary nutrient intake and bioactive food components are essential environmental factors that may influence DNA methylation. In recent decades, researchers have learned a great deal about DNA methylation, including how it occurs, where it occurs, and they have also discovered that methylation is an important component in numerous cellular processes. In the following illustration we see some aspects on how to influence epigenetic modulation. Actually, these will influence many of the pillars of aging. Epigenetic biological clocks have recently taken on a good bit of interest and importance. The three major influences on epigenetic clocks are DNA methylation, histone modification, and noncoding RNA. Among these three, a growing body of science emphasizes the role of DNA methylation in aging and age‐related chronic diseases in humans. In part, this is because DNA methylation is easily assessed in circulating cells and is relatively stable over time. Thus, it appears that as far as Epigenetics is concerned DNA methylation seems to be a realistic biomarker. It has everything we are looking for. The next diagram is a very realistic view of the Methylation Epigenetic Clock. We can see the protective factors and the risk factors. What we are able to do in our clinic is enhance the protective factors by modalities such as EBO2, V cells, and a host of intravenous products such as NAD. We now have at our disposal some real-time methods of measuring the DNA methylation clock. When talking about Methylation clocks there are a few different types. The main ones are called the Horvath and Hannan clocks. The science community has known since the 1960s that DNA methylation has strong effects on aging. The first demonstration that DNA methylation levels can generate age predictors was published by a group of researchers at UCLA. They developed an “epigenetic clock” that analyzes the effects of age on tissues. A study conducted by Dr. Horvath found that women’s breast tissue ages faster than the rest of their bodies. They also found that cancerous tissue, on average, was 36 years older than other tissues. Overtime we have seen the development of several age estimators that use different sets of information from different tissues and age spectrums. The one we use looks at methylation markers which happen through epigenetics. This test was created by Dr. Steve Horvath and Dr. Greg Hannum in 2013. Dr. Horvath’s work has stood out in particular because of its ability to accurately predict age across all age types. Multi-tissue age estimator is first-ever accurate age predictor that works across most tissues and cell types. The epigenetic clock can officially predict one’s biological age with more accurately than chronological age and has shown that epigenetic age in humans can be reversed. Estimated DNA methylation age is associated with age-related conditions and predicts lifespan. The following is a diagram of different DNA methylation clocks. As can be seen there are three main clocks here. Our clinic is now offering DNA methylation tests which look at certain biomarkers. Biomarkers are able to capture aspects of biological aging. There is now a test to accurately measure DNA methylation. This is called the TruAgeTM test. TruAge uses blood to perform its analysis. It uses a powerful algorithm and computer learning software by analyzing almost a million data points from over 10,000 patients. By looking at how it has shown very tight correlations to chronological age and how the body methylates its DNA, TruDiagnosticTM is able to use their precise and reliable algorithm to predict one’s biological age. With the constant development of statistical technology and acknowledgment of aging and disease, we are now able to estimate biological age like never before. How is this different from genetic testing, like "23andMe?" Genetics is the sequence of the DNA that you are born with and you die with. This means that our DNA sequencing is not influenceable, meaning you are stuck with it. But 60% of your DNA is controllable thanks to epigenetics. Due to epigenetics, we are able to have control over the way our genes are expressed, due to certain influences we put on the body. Lifestyle changes, such as diet and exercise, are one way we can influence gene expression. Other companies look at the genetic sequencing of an individual, which we have no control over. It is like knowing how a car engine works and finding out the motor stopped running but you aren’t given the tools to fix it. By addressing the modifiable parts of our DNA, we are able to take control of things like disease risk and rate of aging, rather than sitting by and waiting for problems to happen. DNA methylation is an excellent test but has its limits. For example, Horvat’s clock is based on the analysis of over 300 pieces of epigenetic information. One of the questions in the science of Anti-Aging medicine is the DNA methylation test may have too many components. Although it is comprehensive can we actually see if we are responsible for improvements that they patient might have with the various modalities that they may receive. DNA methylation is a very important weapon in our battle to control aging. There is already an excellent example of a clinical trial to reverse DNA methylation. This clinical trial is called the TRIIM TRIAL. TRIIM TRIAL A COCKTAIL TO REVERSE METHYLATION GROWTH HORMONE METFORMIN DHEA Based on an epigenetic age estimator, this is the first-ever and only report of an increase in human lifespan by means of anti-aging interventions. What medications have been shown to help reverse the epigenetic aging rate? Dr. Fahy and his team ran a small (9 patients) human clinical trial known as the Thymus Regeneration Immunorestoration, and Insulin Mitigation (TRIIM). The purpose of the TRIIM trial was to investigate if the immune system of older people could be rejuvenated to make them biologically younger and work better, as previously shown in mice. For one year, the 9 subjects took DHEA, metformin, and growth hormone, and on average, shed 2.5 years of their biological age. Based on an epigenetic age estimator, this is the first-ever and only report of an increase in human lifespan by means of aging interventions. It certainly will not be the last. Before you go out and use the medications used in the TRIIM trial a word of caution. Growth hormone may not always be anti-aging. GLYCANAGE It has been stated on many different occasions, sugar is the enemy of healthy living. This is certainly the case when we are assessing the GlycanAge of someone. Almost all proteins in our body are covered by sugar molecules, called glycans. If you study a protein without its glycan coat is like studying a bird without its feathers. These glucose molecules attach to proteins, resembling branches of a tree via highly complex process, called glycosylation. Glycosylation is the enzymatic process by which carbohydrates called glycans are attached to proteins or lipids, typically on the cell surface or in the bloodstream. This is an illustration of this: Our DNA dictates the protein structures in our bodies, but the proteins get further modified by the thick layer of glycans which are very much dependent on the lifestyle we live. Glycans play a vital role in keeping us healthy, but are also involved in most major diseases that affect mankind. As we age, the balance of pro-inflammatory and anti-inflammatory glycans in our body changes. Various inflammatory effectors provoke an imbalance between the two and lead to low-grade systematic inflammation, which can speed up the process of aging and lead to many diseases. The following chart is an excellent one when studying a patient’s bio markers. Glycans are impacted by a variety of genetic, biological and environmental factors, glycans are subject to change and the decisions you make can influence them! The following chart shows the Glycan aging biomarkers and how they seem to stand out. Glycan age markers are the only ones which seems to measure responsiveness to the interventions that beneficially affect the biology of aging. The bottom line is unless you are able to see how your intervention is working you are working as if you had blindfolds on. GlycanAge is different because it measures your IgG glycosylation, which directly correlates with the level of inflammation in your body. IgGs are a type of antibody representing approximately 75% of the antibodies found in the serum. In humans, IgG is the most common type of antibody found in blood circulation. It is instrumental in fighting most infections. It will give you information about the immune balance of your body that changes with age, health and life circumstances. GlycanAge is based on a single molecule and its functional regulation. While the methylation clock measures information, Glycanage measures direct effectors. In other words, how different treatments one receives affects their aging. An individual's repertoire of glycans changes with age and environmental factors such as such as smoking and poor diet. The type of glycans attached to IgGs affects their pro- and anti-inflammatory properties. A serious problem associated with glycosylation is the production of what are called “ADVANCED GLYCATION END PRODUCTS” (AGES). This occurs when the sugars bind to proteins causing a host of problems as can be seen in the following diagram. Advanced glycation end products (AGEs) are proteins or lipids that become glycated as a result of exposure to sugars. They are a bio-marker implicated in aging and the development, or worsening, of many degenerative diseases, such as diabetes, atherosclerosis, chronic kidney disease, and Alzheimer's disease. Animal-derived foods that are high in fat and protein are generally AGE-rich and prone to new AGE formation during cooking. In particular, grilling, broiling, roasting, searing, and frying propagate and accelerate new AGE formation. The following diagram shows the variety of the clinical manifestations of glycosylation. Getting back to the GlycanAge test, only a few drops of blood are needed. Since environmental factors heavily influence glycosylation and since glycan patterns can change relatively quickly (compared to DNA methylation), this might be an even better test for assessing whether your new workout regimen or favorite supplements are working their best for you. The information on the difference between a person's biological and chronological age, enriched with other data (clinical parameters, dietary info, exercise regime etc.) and targeted interpretation can serve a wide spectrum of applications. For instance, one application is “Lifestyle Assessment – a way to healthy ageing. After having received the information on the general health status by GlycanAge® test, the subject has an opportunity to make necessary changes in their lifestyle (regarding exercise, diet, sleep, tobacco exposure, stress etc.) with the aim of improving their general health status. Another important application is “Sports Diagnostics” - GlycanAge offers the users (amateur and professional athletes, sports coaches, athletic trainers etc.) an insight into the health aspect of training (microtraumas, overtraining etc.) thereby enabling the creation of a more appropriate personalized training plan. When all is said and done, the best method to assess one’s biological age is to use a combination of both the TruAge test for checking on DNA methylation and the GlycanAge test to check glycosylation levels. The more information we have the more potential we have to improve your health. We are very excited about these new tools. Thanks, Dr. P

As a physician and scientist, I always like to put politics aside. I must give accolades to Operation Warp Speed. It is difficult for people outside the biomedical research circuit to grasp the magnitude of creating a working vaccine for any disease, within a year. To put that in perspective, the fastest vaccine development cycle on record was four years, for mumps. To have a vaccine developed, found to be relatively safe, and available for wide distribution in such a short period of time is unheard of. This dedication and mind set reminds me of back in the 1960s when we were involved in the space race for the moon. It took us 10 years to go from dream to reality. For the vaccine race it took us 10 months to go from dream to reality. These two achievements are not that dissimilar. The space race gave our world many different gifts of technology ranging from different types of plastics to computer technology. Our society would be much different without the “technological gifts” from the space race. I suspect we are going to see similar “gifts” from the vaccine race that will forever change our society. First let’s talk about the implications for the here and now about the vaccine. One big question everyone has if it will still be necessary to wear a mask after receiving the Covid-19 vaccine? The answer for this is yes. For a couple reasons, masks and social distancing will still be recommended for some time after people are vaccinated. First off remember that the protection the vaccine affords will take a few weeks to develop. We will need to get two shots and effective immunity may take a few weeks after the second injection. The still unknown out there is if the vaccine protects us from the infection or does it just prevent symptoms? Potentially, vaccinated people might still be able to get infected and pass the virus on, although it would likely be at a much lower rate. Lastly, the sheer numbers of people that will require vaccination will take several months to complete. So, for now I do not see masks going away until sometime late in the second half of 2021. WHAT ABOUT THE VACCINE ITSELF? Any vaccine has controversy to it, yet by and large, they have changed the course of history in a positive way. Just read about the struggles with the Polio virus until an effective vaccine was produced. The Polio Virus was a scourge that would cause paralysis in many otherwise healthy people. Also, unlike the Covid-19, the Polio Virus had a predilection for younger patients actually it was also known as “infantile paralysis” WHAT IS THE VACCINE MADE OF? THE ANSWER IS MESSENGER RNA (mRNA) Every strand of mRNA is made up of four molecular building blocks called nucleosides. But in its altered, synthetic form, one of those building blocks, like a misaligned wheel on a car, can throw everything off by signaling the immune system. Thus, scientists simply subbed it out for a slightly tweaked version, creating a hybrid mRNA that could sneak its way into cells without alerting the body’s defenses, a giant step. That was an aha moment. It changed the course of history. Looking at things further, the above diagram is an excellent representation on how the Covid-19 vaccine works by using mRNA also known as messenger RNA. In molecular biology, messenger RNA is a single-stranded molecule of RNA that corresponds to the genetic sequence of a gene. One can consider mRNA as a blueprint. The mRNA is read by a ribosome which subsequently synthesizing a protein in the cell. In step 1 of the above diagram we see the protein spike on the virus outer shell. What the scientists do is make a mRNA sequence that codes (blueprints) for the virus spike protein. Once the spike is produced it is mixed with a lipid coating and is prepared for injection into the patient. Once inside the patient the mRNA prompts the cells to produce millions of spike proteins. The spike proteins than stimulate the immune system to produce antibodies to protect the cell when a virus attacks the cells. Although this is a simplistic view it is gives a good idea on how the vaccine works. What I like about the next diagram is that it shows more specifics about the immune system. We see that the immune system is making use of what are called T-helper cells. The T-helper cells recognize the virus and activate either cytotoxic T cells which kill virus infected cells or B cells which launch antibodies which will block the virus from infecting healthy cells. Although the studies don’t clarify whether people who have had a clear a SARS-CoV-2 infection or receive the vaccine can ward off the virus in the future, both identified strong T cell responses to it. In another words this bodes well for the development of long-term protective immunity. The Antibodies that are produced may not last that long but the T cells should give us long lasting immunity. This may explain why a large portion of the population may be able to deal with the Covid 19 virus due to previous exposure to common cold viruses which indeed have some cross over to Covid-19. I feel comfortable that the immunity we receive from the vaccine will be long lasting. I feel the same about the immunity received from an infection. SOME COMMON QUESTIONS ABOUT THE VACCINE One important question people are asking is how the new vaccine is different from traditional vaccines. The above diagram shows the major differences on how the vaccines are produced. Traditional vaccines use bits of injected proteins to train the immune system to take down future viruses displaying those same proteins. Manufacturing them takes months, a timescale too slow to combat emerging epidemics. mRNA vaccines, on the other hand, simply encode these protein fragments in a single mRNA strand. As mRNA companies optimize and scale up their enzymatic production of mRNA, scientists anticipate these vaccines could be made in a matter of weeks. A number of questions arise concerning the vaccines. One question is if the mRNA will incorporate into our DNA. The simple answer to this is NO. mRNA is designed to transcribe proteins etc. Its job is not to act as an enzyme and affect the structure of the DNA. mRNA is not able to alter or modify a person’s genetic makeup (DNA). The mRNA from a COVID-19 vaccine NEVER ENTERS THE NUCLEUS OF THE CELL, WHICH IS WHERE OUR DNA IS KEPT. This means the mRNA does not affect or interact with our DNA in any way. The vaccine will not cause you to test positive on the viral tests however if you are tested for antibodies that test may be positive. Another question is if a person has had the virus should they get the vaccine? There is no simple answer to this question since we really do not know. Some studies show that natural immunity may not last long while others say it conveys the best immunity of all. I suspect that if one was infected with the Covid- 19 virus they would have long lasting immunity. However, we will need to view the landscape in a few months to see where we are on this question. It may very well turn out that both natural immunity and vaccine induced immunity may provide the best protection of all. When will there probably not be a need for the vaccine? The goal of the vaccination program is to achieve heard immunity. What percentage of the population that needs to receive the vaccine or have had a previous infection is still not known to achieve herd immunity. MANY PEOPLE ASK ME IF I WILL TAKE THE VACCINE. THE ANSWER TO THIS QUESTION IS YES. I suspect that a vaccination will be required to travel to many different countries. Also, airlines such as Qantas will probably start requiring passengers to be vaccinated to travel on their planes. As a matter of fact, I reserved the domain name Immunityvisas.com for this very reason. I would suspect that the cruise industry may also require a vaccination to take a cruise. The bottom line is for the world to return to some semblance of normal we will have to achieve herd immunity either by immunization or actually having the disease or both. ANYTHING ELSE WE WILL BENEFIT FROM WITH THE COVID-19 VACCINE BESIDES IMMUNITY? THE ANSWER IS mRNA THEURAPEUTICS The two preceding illustrations give the essence of mRNA therapy. This is a therapy that already has and will continue to disrupt medicine. Here is a quote that I read in an article that was written in early 2019 before we realized the pandemic was here: “When the first mRNA therapeutic crosses the finish line, we’ll all be winners, suggests CureVac, a biopharma that anticipates an mRNA stampede”. Like the space race of the 1960s, there will be much in the way of spinoffs from the Covid-19 vaccine. The first vaccines against Covid-19 aren’t just a landmark in the fight against the pandemic. They’re also the stepping stone for an unconventional technology that could one day defeat other ailments that have eluded doctors, from cancer to heart disease. What I am talking about here is the new field of mRNA therapeutics. This is possibly at the very heart of Stem Cell and Regenerative Medicine therapies. This is one of the basic mechanisms that can allow stem cell to help regenerate tissue. We can see how the Big Pharma and the Biotech industry has embraced this new technology. Perhaps they can come up with a vaccine for many types of cancer. There are already important achievements in the field of immunology and cancer with some promising new therapies. Remember that the Covid-19 virus was not the first kid on the block. Every year influenza kills at least 650,000 people. Perhaps with mRNA technology we can make one vaccine to essentially eliminate influenzas. This is not as far-fetched as it sounds. There is also a possibility that mRNA technology may be used for reprograming stem and other cells in the future. There are still problems that we will encounter with mRNA therapy but these will be overcome. I think the appropriate way of ending this piece on the vaccine and mRNA therapy is to take some poetic license on the quote from Neil Armstrong as he took his first step on the surface of the moon: That’s one small step contributed by many people, but one giant leap for mankind. Thanks, Dr. P

The above diagram shows the difference between normal bone and osteoporotic bone. We can see that the osteoporotic bone has much bigger holes and in general is much weaker bone. When we talk about Osteoporosis I can wear both of my hats. The hat of an orthopedic surgeon and that of a Regenerative Medicine stem cell doctor. For 35 years I have seen first hand the ravages of osteoporosis. I have operated on thousands of fractures that were caused by osteoporosis. Many of these fractures were quite challenging. The fixation of the fractures required different surgical solutions including plates, rods, and screws and different combinations. These fractures were the precursor of many other serious problems such as blood clots, strokes, pneumonia and other serious medical problems. The next slide shows the incidence of osteoporosis in the USA. There is not any widely accepted treatment for osteoporosis. In the above diagram we can see some of the methods that are used to treat osteoporosis. Unfortunately, these treatments are at best band aid approaches. Last year a meta-analysis of no fewer than 33 studies was published in the prestigious BMJ (the former British Medical Journal). This study confirms what had been suspected. Namely, that bisphosphonates are totally ineffective at preventing fractures. Bisphosphonates are a class of osteoporosis drugs such as Boniva (ibandronate), Fosamax (alendronate), Actonel (risedronate) and Reclast (zoledronic acid). Their mode of action is to disrupt normal bone remodeling, which is ultimately detrimental to bone integrity and fracture resistance. They do not increase bone density but slow down bone loss. Teriparatide also called Forteo is the only osteoporosis medicine which has been shown to have the potential to rebuild bone. It has to be given as a daily injection and one of its risks is that it can cause a bone cancer called osteosarcoma. As can be seen none of these medicines are a bargain. Either they do not work or have a significant risk. The interesting new aspect concerning Osteoporosis is the use of stem cells to treat osteoporosis. The following slides give a good idea of what the stem cells are doing. From this slide we get a better idea of the concept of how stem cells can treat osteoporosis. The cell we are talking about is not any stem cells but what we call an Adult Pluripotent Embryonic-Like Stem Cell. Also called a VSEL stem cell. These cells are revolutionary. We are fortunate in that we are one of the few facilities that are utilizing these cells. I describe these cells as an emergency stem cell supply. They are found in everyone but typically they are not activated. One of the common threads on the studies with these cells is that they seem to be activated by cold temperatures. The propriety information here includes the methods to dramatically increase the number of cells released, how to activate them and finally how to keep them going. We are fortunate to be able to utilize this technology. The technique is relatively non-invasive. It requires an injection or two of some propriety compounds and then a harvesting of some blood. We have utilized these cells for a few years. They work exceptionally well with auto-immune diseases (a type of condition where the body can attack itself) such as Rheumatoid Arthritis and Ankylosing Spondylitis. We also use them for anti-aging as well as for musculoskeletal conditions with good success. When we talk about osteoporosis we think osteoporosis is somewhat like an autoimmune disease. We have seen some of our preliminary results with using these cells for osteoporosis. The results are astounding. The slide below shows one of the patients we have treated. We have had similar results in other patients we have treated for osteoporosis. As can be seen this patient has received two injections of the pluripotent cells. They were spaced about one year apart. Our recommendations are to perform two of these treatments a year, spaced approximately 6 months apart. We are also placing the patients on some propriety growth factors and supplement mixes. To make things somewhat easier I have included the patient's most recent bone density report below. I think that this is easier to follow. This report shows the patient's bone density INCREASED 14% IN LESS THAN 6 MONTHS!! This increase is exceptional. There is currently no treatment for osteoporosis that I am aware of that would even approach this amount of improvement. Unlike many things in regenerative medicine we see very specific improvement in results in a test (bone density) that is completely objective. The implications of this treatment are profound. If we are able to turn the tide of osteoporosis it will save our health care system billions of dollars. The amount of money spent on a fractured hip is astounding. More importantly this treatment can dramatically improve the quality of life of the patient. Fractures take a toll on the patient and their families. The problem with osteoporosis is that it is a silent problem. Most of the time you have osteoporosis or its less severe counterpart which is called osteopenia there are initially no symptoms. The symptoms occur when fractures start happening. Other manifestations include some wedging of the vertebral bodies (back bones) which can cause spinal deformities. As far as other manifestations of the VSELs we have seen significant improvements in the treatment of auto-immune diseases. We have had a number of patients go off their medications for these diseases. Some of the side effects for these medications include developing a lymphoma. There also seems to be some manifestations of some anti-aging properties but I will leave this for another blog. We are very high on these cells. I will be attending and lecturing at a meeting in Kuala Lumpur on Dec. 17 and 18. This meeting will present some of the newest information on VSELs by those experts in the world involved with these cells. Some of the topics will include infertility, treatment of various inflammatory diseases including COPD, addiction problems. We are quite excited about these cells and have a published paper coming out about these cells. We will now be offering an osteoporosis program that will include two of these VSEL treatments a year. Incorporated into these treatments we will utilize a proprietary blend of cytokine growth factors and a supplement program. Both the growth factors and the supplements will also target osteoporosis. Also we will pay for one bone density so that we can track our results. So the warning should go out to Osteoporosis and Osteopenia. We have targeted them in our gun sites and we are going to assassinate them. More to come Thanks Dr. P