AGINGSCIENCES™ – Anti-Aging Firewalls™

Little Jane asks “Daddy, where will they get the stem cells to make me better?”

Daddy’s answer now “They will get some from your bone marrow.  They stick in a big needle to get it.”

Previous posts on this blog have discussed applications of adult stem cells for tissue regeneration, for example the recent post Simple but powerful non-invasive adult stem cell cures.  Many of these feature the use of autologous mesenchymal stem/progenitor cells extracted from a patient’s own bone marrow.  Mesenchymal stem/progenitor cells are an important category of stem cells which self-renew and are capable of differentiating into bone, adipose and cartilage tissue.  Traditionally these cells have been extracted from bone marrow although they have been known to exist also in placenta, lung and other tissues.  Recently there has been increasing interest in Dental Pulp Stem Cells (DPSCs) for dental tissue restoration, including regeneration of dental pulp and dentine(ref)(ref)(ref)(ref). 

Beyond dental applications, there may be a number of other applications of DPSCs for repair or regeneration of other body tissues.  DPSCs appear to be functionally equivalent to mesenchymal stem/progenitor cells, are relatively easy to collect, have an extensive differentiation capability and offer several other advantages(ref)(ref).  DPSCs “have been demonstrated to answer all of these issues: access to the collection site of these cells is easy and produces very low morbidity; extraction of stem cells from pulp tissue is highly efficiency; they have an extensive differentiation ability; and the demonstrated interactivity with biomaterials makes them ideal for tissue reconstruction. SBP-DPSCs are a multipotent stem cell subpopulation of DPSCs which are able to differentiate into osteoblasts, synthesizing 3D woven bone tissue chips in vitro and that are capable to synergically differentiate into osteoblasts and endotheliocytes. Several studied have been performed on DPSCs and they mainly found that these cells are multipotent stromal cells that can be safety cryopreserved, used with several scaffolds, that can extensively proliferate, have a long lifespan and build in vivo an adult bone with Havers channels and an appropriate vascularization(ref).”  In other words, for many tissue repair and regeneration applications DPSCs might offer a better choice that bone-marrow extracted mesenchymal stem cells.  And there are likely to be a lot of such applications.

In the future little Jane asks: “Daddy, where will they get the stem cells to make me better?”

Daddy’s answer:  “The tooth fairy collects it.  Just leave your baby tooth under your pillow when it comes out.”

Since I started taking 600mg a day of trans-resveratrol, my bowel movements have become reliably punctual, sometimes almost overly so.  If you are taking large doses of resveratrol and are experiencing the same phenomenon or worse, it may be due to emodin, an impurity commonly found in commercial resveratrol supplements.  Among other properties, it is a laxative.  Emodin is a phyto plant substance present in rhubarb and in aloe vera leaves.  Emodin is also present in Japanese knotweed (Polygonum cuspidatum), the source ingredient of most  resveratrol   supplements.  It may not all be removed in refining resveratrol and can represent more than 5% of the content of a marketed resveratrol supplement capsule.  Discussion of this impurity has appeared in an Imminst.org forum on resveratrol. 

The amount of the impurity varies significantly by product.  Most popular suppliers of resveratrol (including Swanson Vitamins, NSI, Country Life, Biotivia, Life Extension InstituteJarrow Formulas, source Naturals and Puritan’s Pride) do not disclose emodin content in their resveratrol labels.  Revgenetics is an exception and has posted laboratory analyses showing emodin is .1% present in their “99% pure” resveratrol product(ref) and .7% present in their “50% pure” product(ref).  Some grape-based resveratrol products avoid emodin completely(ref). 

Despite its laxative effects, emodin may offer health benefits of its own such as a cancer-preventative ones. It may inhibit cell growth and angiogenesis in human colon cancers(ref), It “could be a useful chemotherapeutical agent for treatment of hepatocellular carcinoma(ref),”  “Emodin affects the expression of genes involved in various cellular functions and plays important roles in cell apoptosis, tumor metastasis and chemotherapy-resistance, which suggests emodin might become an effective chemopreventive or chemotherapeutic agent for small cell lung cancer(ref),” “Emodin induces apoptosis in human promyeloleukemic HL-60 cells—(ref).”  Further, Emodin has unique antibacterial properties, “taking into account its unique cytotoxicity profile and mode of action, aloe-emodin might represent a conceptually new lead antitumor drug(ref).” And emodin is “a potential lead compound for further anti-bacterial drug discovery(ref).”  To sum it up, emodin appears to be another phyochemical whose possibly important health properties are just-now being systematically explored.

Personally I am taking a resveratrol supplement with less than 1% emodin content and find the laxative effect definitely present but tolerable.  If you are experiencing diarrhea due to taking a large amounts of resveratrol, you might want to switch to a different brand with lower emodin content. 

The longevity research literature citations that appear in this blog or in my treatise are increasingly likely to refer to recombinant DNA laboratory analysis techniques like Western blot analysis, PCR, Alkaline lysis, Column chromatography, Sanger sequencing, Agarose gel electrophoresis, Radio-immune precipitation, and Sodium dodecyl (lauryl) sulfate-polyacrylamide gel electrophoresis.     If you have not studied these, after repeatedly coming across some of them you may wonder what in the dickens they are, how they work and what they are good for.  I recently came across Mama Ji’s Molecular Kitchen, a web site that explains a number of these basic techniques in simple language.  Further, it tells you in a cookbook fashion how to go about doing each of them if you are so inclined.  The site also covers a few key molecular genetic entities like Plasmids and Restriction Enzymes.

My basic message is that if you are reading a research report of otherwise great interest but come across entities you know nothing about, you don’t necessarily have to let yourself be thrown.  In many cases the concept behind an arcane technical term is quite simple: 

·        Take PCR (polymerase chain reaction) for example.  “Let’s say you have a biological sample with trace amounts of DNA in it. You want to work with the DNA, perhaps characterize it by sequencing, but there isn’t much to work with. This is where PCR comes in. PCR is the amplification of a small amount of DNA into a larger amount. It is quick, easy, and automated. Larger amounts of DNA mean more accurate and reliable results for your later techniques(ref).”  

·        Restriction Enzymes, another example, are used as scissors for cutting DNA.  “Restriction enzymes, also known as restriction endonucleases, are enzymes that cut a DNA molecule at a particular place. They are essential tools for recombinant DNA technology. The enzyme “scans” a DNA molecule, looking for a particular sequence, usually of four to six nucleotides. Once it finds this recognition sequence, it stops and cuts the strands. This is known as enzyme digestion(ref).”

I have created a 39-slide PowerPoint presentation THE SCIENCE OF AGING AND PRACTICAL ANTI-AGING INTERVENTIONS that provides an overview of the content in my treatise ANTI-AGING FIREWALLS – THE SCIENCE AND TECHNOLOGY OF LONGEVITY.  It is a good introduction to my anti-aging work and is recommended for readers who have not read the treatise.  The presentation also contains hyperlinks to multiple topics covered in more detail in the treatise and to selected items in this Blog.

Online PowerPoint presentations use scripts.  Because of this feature in PowerPoint, you may have to give your browser security permission to view the presentation.  If a warning bar comes up on your browser, click on it.  And then you might be asked “Install active X control” and/or “Allow blocked content.”  You will then get another security warning and if you click “Yes” the full presentation will come up.  Unfortunately, you will probably have to do at least one of these steps every time you navigate away to a hyperlink and then return to the PowerPoint presentation. 

Like both the Blog and treatise, this presentation is a work in progress.  As time goes on I expect to update its contents and I may add additional multimedia features. 

While most popular discussion has centered around the disease-curing potential of embryonic stem cells and induced pluripotent stem cells, small-scale experiments are beginning to show the power of very simple, inexpensive and non-invasive techniques using a patient’s own (autologous) adult stem cells to clear up formerly intractable disease conditions.  I briefly describe two examples of preliminary but promising applications here.

The first application involves clearing up Scleradactyly, a condition featuring chronic fingertip or toetip hardening and often accompanied by lack of circulation and painful  infections.  Scleradactyly often occurs in people with auto-immune disorders, particularly with scleroderma, CREST syndrome and mixed connective tissue disease.  Up until now, there has been no known way to clear it up.  Dr. Vincent Falanga, working at the Roger Williams Medical Center, in Providence, Rhode Island has developed a procedure that uses mesenchymal stem cells derived from a patient’s bone marrow. “ The cells are cultured ex vivo and their numbers are expanded greatly. A solution of the stem cells and fibrinogen is placed in 1 chamber of a double-chambered syringe, and the second chamber is filled with a solution of dilute thrombin.  “The 2 solutions combine when ejected from the syringe as a spray over the wound. The mix begins to polymerize, and that “clotting” helps to hold the stem cells in place in the wound. The wound is then covered with 2-layer bioengineered skin, containing a layer of keratinocytes and a layer of fibroblasts(ref).”  Except for extracting the cells from bone marrow, there is nothing invasive about the procedure. “This proof-of-concept procedure has been applied to 9 patients with nonhealing ulcers without scleroderma and to 3 patients with nonhealing wounds related to scleroderma. After 6 months of follow-up, 4 patients healed completely and the others improved significantly, “with improved quality of life.”  Dr. Falanga is following up with additional patients and is refining his technique.

The second application is cure of blindness caused by corneal disease.  (Thanks to blog participant Brian Hart who put me onto this one via a comment.)  The technique again seems ridiculously simple for the amazing results achieved.  Work so far in Australia has been only with three patients that were blind in one eye with one healthy eye.  The process is very simple.  “The researchers extracted stem cells from the working eye, cultured them in contact lenses for 10 days, and gave them to the patients to wear. Within 10 to 14 days of use, the stem cells began recolonizing and repairing the cornea(ref).”  Apparently, the stem cells migrate readily into the cornea.  Of the three patients, two were legally blind but can now read the big letters on an eye chart, while the third, who could previously read the top few rows of the chart, is now able to pass the vision test for a driver’s license.”  The procedure can most likely be used to repair a variety of forms of corneal damage due to infections, burns or chemotherapy.  See this web site for a short video on the process.

I think simple autologous stem cell therapies like these will proliferate and soon become part of mainstream medicine significantly contributing to health.  As to their contribution to extraordinary longevity, however, effectiveness of these techniques depends on a supply in the body of healthy adult stem cells and the techniques will not work without them.  In the recent post More research insight on gray hair and adult stem cell reproduction, for example, I cited research indicating that gray hair was ultimately due to depletion of melanocyte stem-cells(MSCs) which in turn is the result of DNA damage.  Other approaches must be used to assure continuing active supplies of adult stem cells, such as taking antioxidants and telomerase activation.  In the longer run, induced pluripotent stem cells might be induced to differentiate in a controlled manner into specific lines of adult stem cells.  See the discussion for the Decline is Stem Cell Differentiation theory of aging. 

I cite a sampling of research studies here related to the quality of human male semen as a function of age and the possible impacts of dietary supplementation. This topic is increasingly important because of the trend in our society for men to have children at more advanced ages. 

First of all, there is a general inverse relationship between sperm quality and age of the donor.  A study report on The effects of male age on sperm DNA damage in healthy non-smokers looked at 80 non-smoking men (mean age: 46.4 years, range: 22–80 years).  The researchers concluded: “Our findings indicate that (i) older men have increased sperm DNA damage associated with alkali-labile sites or single-strand DNA breaks and (ii) independent of age, men with substantial daily caffeine consumption have increased sperm DNA damage associated with double-strand DNA breaks. DNA damage in sperm can be converted to chromosomal aberrations and gene mutations after fertilization, increasing the risks of developmental defects and genetic diseases among offspring.”  Being both old and a coffee drinker this would be bad news for me if I were planning to have additional children, which I am not.

Another study Advancing age has differential effects on DNA damage, chromatin integrity, gene mutations, and aneuploidies in sperm brings additional bad news for older would-be fathers.  The study looked at 97 nonsmoking men aged 22-80 years.  The researchers report:  “Our findings predict that as healthy males age, they have decreased pregnancy success with trends beginning in their early reproductive years, increased risk for producing offspring with achondroplasia mutations, and risk of fathering offspring with Apert syndrome that may vary across cohorts, but with no increased risk for fathering aneuploid offspring (Down, Klinefelter, Turner, triple X, and XYY syndromes) or triploid embryos.”

Implicated in deterioration of sperm quality are some of the same factors that affect cell health and drive aging, for example Oxidative Damage and Cell DNA Damage.  One study entitled Antioxidant intake is associated with semen quality in healthy men based on study of sperm from 97 men reports: “–, higher antioxidant intake was associated with higher sperm numbers and motility.”  For another study of 114 subjects (60 infertile patients and 54 age-matched healthy workers), the researchers report: “Data from this study thus indicate that oxidative damage to sperm DNA may be important in the etiology of male infertility – .“  This review article looks more broadly at the relationship between oxidative stress and male infertility. “ — oxygen ions, free radicals and peroxides (ROS) — produce infertility by two key mechanisms. First, they damage the sperm membrane, decreasing sperm motility and its ability to fuse with the oocyte. Second, ROS can alter the sperm DNA, resulting in the passage of defective paternal DNA on to the conceptus.” 

A very recent study looked at the Effect of parental age at birth on the accumulation of deficits, frailty and survival in older adults.  “Data was collected on individuals aged 65 (or greater) using a Self-Assessed Risk Factor Questionnaire and screening interview. In this secondary analysis, 5112 participants had complete data for parental age, frailty status and 10-year survival. Parental age was divided into three groups, with cut-offs at 25 and 45 for fathers and at 25 and 40 for mothers. Frailty was defined by an index of deficits.” The researchers reported “There was no effect of maternal or paternal age on survival for either sons or daughters. Similarly, there was no association between parental age and subject frailty in old age.” This time the news was good.  On the other hand since all the participants had already made it to age 65, the study does not tell us what effect parental age at time of birth might have on survival to 65.

The activities of DNA repair mechanisms seem to be age-dependent.  This study Age-Dependent Usage of Double-Strand-Break Repair Pathways looks at DNA repair in the premeiotic germ cells of Drosophila (fruit flies) as a function of age.  The results suggest that at least one repair pathway gets more active with age “We used Rr3, a repair reporter system in Drosophila  to show that the relative usage of DSB repair mechanisms can change substantially as an organism ages. Homologous repair increased linearly in the male germline from 14% in young individuals to more than 60% in old ones, whereas two other pathways showed a corresponding decrease. Furthermore, the proportion of longer conversion tracts (>156 bp) also increased nearly 2-fold as the flies aged.”  I speculate that a similar situation exists for humans: the older the person the more there is a need for repair and actual repair of germline cells.

Among the most widely recognized genetic disorders is Aneuploidy, a condition where there are missing or extra chromosomes.  Testing for aneuploidy may be warranted for men with a serious record of infertility or where his mate has experienced unexplained recurrent pregnancy loss(ref).  

This study investigated the impact of supplementation by zinc, folate, vitamin C, vitamin E and beta-carotene on aneuploidy using sperm samples from 89 healthy, non-smoking men.  Interestingly, the researchers concluded:  “Men with high folate intake had lower overall frequencies of several types of aneuploid sperm.”  There did not seem to be a correlation of use of the other supplements with aneuploidy.

The research literature related to male fertility is extensive and growing and the nine citations above just scratch the surface.  The most obvious implications for normal older men planning to have children are 1.  Protect against oxidative stress and cell DNA damage.  See the protective firewalls for the Oxidative Damage  and the Cell DNA Damage theories of aging, 2.  Include folic acid in your supplement regimen.  And one more point: 3.  Keep yourself young; in today’s world having a father around can be very useful until a child is 30 or older. 

I will return to this topic it at a later time.  It is bound to become more important when extraordinary longevity becomes possible and there will be a demand for interventions that make it possible, safe and easy for a man to father children at the age of 100, 150 or later.

In my earlier post How am I doing I said “All of this is just a start though.  I not only want a full head of hair; I want it to be black instead of gray.”   Also, in an earlier post Why does your hair turn gray? I described research that pointed to hydrogen peroxide as the culprit for hair turning gray.   Newly-reported research looks in a different direction and deeper, shedding light on the underlying cell-level cause of graying hair. Melanocytes are cells that live in hair follicles of mammals that produce melanin, the pigment responsible for hair color. Normally, the melanocytes gives hair its characteristic color of youth – black in my case.  (Melanin is also responsible for skin color and the color in moles, freckles, suntan and the cancer melanoma.  Older people may develop light or dark patches on their skin due to too-little or too-much melanin.)

The new research suggests that gray or white hair is due to age-related depletion of melanocytes which is a direct result of depletion of melanocyte stem-cells(MSCs) which in turn is the result of DNA damage.  It has been known for some time that “ – hair graying is caused by defective self-maintenance of MSCs(ref).”  These stem cells, also living in hair follicles, can normally both reproduce making new stem cells and differentiate into mature color-producing melanocytes. The new research based on experimentation with mice suggests that DNA damage to MSCs causes them to stop reproducing and instead terminally differentiate into melanocytes.  As the melanocytes in hair follicles die off, there are no new melanocytes to replace them because there are no more MSCs to make them.  The result is loss of hair color, in other words, white or gray hair.  The researchers discovered that when mice were exposed to intense radiation, MSCs stopped self-reproducing and terminally differentiated into melanocytes.  Consequently, the fur on the mice turned from brown to gray.  It is thought that the cessation of self-reproduction of genetically damaged MSCs could be an evolutionary protection against cancers.

Wanting black hair, this poses a challenge for me.  If my hair follicle MSCs have died off, how do I get them back?  There seems to be no short term answer though there may be one in the longer term.

It appears that there is a larger issue at stake here when it comes to aging.  The reader may want to review my discussion of the 14^th theory of aging Decline in Adult Stem Cell Differentiation.  What the new research says with respect to that theory is 1. that there is not only the issue of decline in differentiation to be concerned with as part of aging but also an issue of stem cell self-renewal, and 2.  At least some stem cells stop self-renewal in the presence DNA damage.

Both immediate and long-term anti-aging interventions appear to be suggested.  In the immediate outlook the obvious approach is to use antioxidants to minimize DNA damage.  Millions of people are already doing this.  See the Cell DNA Damage theory of aging in my treatise as well as the associated firewall discussion.  For the longer term, it may be possible to induce Pluripotent Stem Cells (iPSC) to selectively differentiate in a controlled manner into adult stem cells, including MSCs.  See the earlier post on this Blog Rebooting cells and longevity.  If a practical iPSC approach could be found to generating MSCs in hair follicles, I might get my black hair back.  Meanwhile I stay tuned for more research in this area.  I do not plan to use shoe polish.

Personally I love spicy foods, and ginger, curcumin and garlic have long been parts of my Anti-Aging Firewalls dietary supplement regimen.  There is an extensive body of literature supporting the health and potential anti-aging effects of spices.  Sage (salvia officinalis), thyme (thymus vulgaris), oregano (oreganol) and rosemary (rosmarinus officinalis) all have antioxidant properties(ref)(ref).  But the basic “reasoning for seasoning” appears to be inhibition of NF-kappaB(ref).  Control of expression of NF-kappaB is of course a major strategy for longevity proposed in the firewall for the Programmed Epigenomic Changes theory of aging.

To start off, hot chili peppers (capsaicin), ginger (gingerol) and turmeric (curcumin) are all inhibitors of NF-kappaB, and thereby regulate COX-2 and inflammation(ref)(ref)(ref)(ref).  The same general statements can be made for black pepper (piperine); it inhibits NF-kappaB expression, is an anti-inflammatory, etc.(ref).  The list goes on to include cloves, anise, cumin, fennel and garlic (ref).  Many of the active ingredients in these spices are also thought to be chemopreventative of cancers(ref) and have numerous other health benefits, curcumin being an example(ref).  “Curcumin, a yellow pigment present in the Indian spice turmeric (associated with curry powder), has been linked with suppression of inflammation; angiogenesis; tumorigenesis; diabetes; diseases of the cardiovascular, pulmonary, and neurological systems, of skin, and of liver; loss of bone and muscle; depression; chronic fatigue; and neuropathic pain(ref).”

So, in general I feel free to spice-up my foods as much as I want.  If you haven’t already read it, see the blog post Red wine, hot peppers and my uncle Gigi.

If you are used to reading research abstracts full of abbreviations for genes and proteins, how about this one?  Do you think you get the general idea?  Or do you just tune out on anything that sounds so technical?  The title actually relates to the Battle of Britain (BOB) during the early 1940s, particularly how US-built fighter planes intercepted German bombers over the English Channel when the Luftwaffe was mounting daily raids on London.  The P38, P39 and P40 were US WWII fighter planes, the BF-110, HE111 and HE177 were German bomber planes and (SC)1000 was a one-ton German Sprengbombe Cylindrich general demolition bomb.  The fact reported had to do with the longevity of hundreds of thousands of people and the fate of Western Civilization was possibly at stake.  The message is that when reading biomolecular-genetic research reports, you can be badly mistaken if you think you get the general idea but don’t really understand what is going on. 

Increasingly, researchers are investigating genetic fixes for otherwise intractable conditions.  For example, see the recent blog entry A genetic fix for obesity?  Now, a possible genetic fix is suggested that addresses HIV, a fix that undoes an ancient mutation present in humans.   

The work is reported in a recent online publication Reawakening Retrocyclins: Ancestral Human Defensins Active Against HIV-1.  The good news is that primates including us humans have a gene that produces retrocyclin, antiviral peptide proteins in the defensin family of proteins.  Retrocyclin is a powerful anti-viral substance and “was recently shown to strongly inhibit HIV entry into human cells by blocking the interaction of viral proteins with their cellular receptors(ref).”  The bad news is that way back in evolutionary history our branch of primates, including gorillas and chimps, experienced a  “nonsense mutation” in the retrocyclin gene, so it fails to do its job and produce the retrocyclin proteins.  A nonsense mutation is one where a codon encoding an amino acid is changed into a premature stop codon.  This means the protein-making machinery in the cell ribosomes reading mRNA instructions as if they were on a tape stops before it should resulting in an incomplete protein being made or no protein at all.  

The research challenge was to see if repairing the 7 million-year old nonsense mutation in the human retrocyclin gene could restore retrocyclin production and block entry of HIV into human cells like it does in Old World monkeys. “To determine whether human cells have retained the capacity to make retrocyclin protein, Venkataraman et al. corrected the premature stop codon mutation in a copy of the human retrocyclin gene. Next, they inserted the corrected gene into human promyelocytic cells, and looked to see if protein was produced from the gene. They found that cells harboring the corrected gene could make a protein similar to the monkey version of retrocyclin. But could human retrocyclin block HIV infection? Indeed, extracts made from cells containing the corrected gene could reduce HIV growth, and so could the retrocyclin protein purified from these extracts. Collectively, these results suggest that human cells have a potentially important—but latent—mechanism to protect against HIV(ref).” 

So, there is a possibility that a genetic fix could be created to mobilize retrocyclin to protect against HIV.  But the researchers identified an alternative and much easier approach, and that is to use aminoglycosides instead to override the erroneous stop signals in human retrocyclin protein production. “An aminoglycoside is a molecule composed of a sugar group and an amino group.  Several aminoglycosides function as antibiotics that are effective against certain types of bacteria. They include amikacin, arbekacin, gentamicin, kanamycin, neomycin, netilmicin, paromomycin, rhodostreptomycin, streptomycin, tobramycin, and apramycin(ref).”  Aminoglycosides “– don’t block protein creation in human cells but cause ribosomes to make occasional errors—like missing stop codons. The authors found that treating human cells with aminoglycosides allowed the cells to make retrocyclin at sufficiently high levels to inhibit infection by HIV(ref).”

Hmm.  What this seems to say is that taking one or several of those antibiotics might help protect against HIV infection.  The research says “– we exploited the ability of aminoglycoside antibiotics to read-through the premature termination codon within retrocyclin transcripts to produce functional peptides that are active against HIV-1(ref).”  One possibility would be to make topical creams containing such antibiotics to inhibit sexual transmission of HIV.  Of course there are likely to be other disease risks associated with ignoring protein-making stop instructions and taking these antibiotics.  In any event, human activation of retrocyclin seems to be a promising avenue of research for prevention of HIV infections and AIDS.  If it can be done safely and economically, tens of millions of lives and billions or trillions of dollars could be saved.