APPETIZERS

In an honest search for knowledge you quite often
have to abide by ignorance for an indefinite period.
— Erwin Schrodinger

Being a scientist requires having faith in uncertainty, finding
pleasure in mystery, and learning to cultivate doubt. There is no surer
way to screw up an experiment than to be certain of its outcome.
— Stuart Firestein, Ignorance: How It Drives Science (Oxford Univ. Press, 2012)

When you’re over the hill, you pick up speed.
— Satchel Paige (D&S: But that’s only true if you’re not too far over the hill.)

Any man who thinks he can be happy and prosperous by letting the
Government take care of him better take a closer look at the American Indian.
— Henry Ford

Governments specialize in creating problems that they then generously solve for the people, creating yet more serious and more systemic problems in the process.
— George Gilder, Winter 1989 Cato Journal

Libenter homines id quod volunt credunt.
(People easily believe that which they want to believe.)
— Julius Caesar

I wondered why the baseball was getting bigger. Then it hit me!
— from the Casey Daily Dispatch (Doug Casey, not Casey at the Bat)

I know a guy who’s addicted to brake fluid. He says he can stop any time.
— from Casey Daily Dispatch

HYDROGEN THERAPY – HYDROGEN PROTECTS AGAINST LIPID
METABOLISM DISORDERS AND ATHEROSCLEROSIS

One of the signaling pathways that causes excessive fatty acid uptake in obesity results from the scavenger receptor CD36. Platelet quantities of CD36 are found at elevated levels in the plasma of hyperlipidemic mice and in the plasma of humans with low levels of HDL. These higher levels of CD36 were found to promote platelet activation and, hence, the risk of blood clot-induced heart attacks.¹ “Thus, interactions of platelet CD36 with specific endogenous oxidized lipids play a crucial role in the well-known clinical associations between dyslipidemia, oxidant stress, and a prothrombotic phenotype.”¹ “CD36 is implicated in a variety of pathological conditions, including atherosclerosis, diabetes and innate immunity.”¹

It was known that oxidized LDL increased platelet activation but the mechanism was not understood. The authors of paper #1 found that oxidized choline glycerophospholipids, generated as a result of LDL oxidation and then bound to the scavenger receptor CD36 in platelets caused the increased platelet activation. Moreover, these oxidized choline glycerophospholipids promote foam cell formation, which are key factors in the development of atherosclerotic plaques.²

A new paper in the developing field of hydrogen therapy³ has reported that molecular hydrogen treatment of HepG2 human liver cells downregulated the protein expression of CD36 and decreased the cellular uptake of fatty acids and decreased lipid accumulation in the cells. “Our results suggest that downregulation of CD36 expression by hydrogen pretreatment may be the primary mechanism against hepatic steatosis [fatty liver] in this in vitro model.”³

As we have written before, certain prebiotics can be converted by hydrogen-producing gut microbiota into gaseous hydrogen, a very selective antioxidant that is mostly active against the highly toxic hydroxyl radical and the oxidant peroxynitrite. Hydrogen then diffuses throughout the body (even into mitochondria) before being exhaled from the lungs or passing out of the body via the skin or as farts. (See “Hydrogen Therapy” in the June 2012 issue of Life Enhancement).

Hydrogen Reported to Downregulate CD36

As this new paper³ shows, hydrogen is also able to downregulate CD36 and this may be a new way to prevent platelet hyperactivity that can lead to blood clot-induced heart attacks and strokes.

References

1. Podrez et al. Platelet CD36 links hyperlipidemia, oxidant stress and a prothrombotic phenotype. Nat Med. 13(9):1086-95 (2007).
2. Jackson & Calkin. The clot thickens—oxidized lipids and thrombosis. Nat Med.13(9):1015-1016 (2007).
3. Iio et al. Molecular hydrogen attenuates fatty acid uptake and lipid accumulation through downregulating CD36 expression in HepG2 cells. Med Gas Res. 3:6 (2013).

Hydrogen, a Selective Antioxidant, May Interfere Less with
Physiological Radical Signaling Than Other Antioxidants Do

One of the things we explained about hydrogen acting as a selective antioxidant is that it is a potent scavenger of the toxic hydroxyl radical, not known to be a radical engaged in physiological signaling, but to be much less of a scavenger of those radicals that ARE known to act in physiological signaling, such as superoxide or nitric oxide. Here is an example recently reported in the journal Free Radical Biology And Medicine¹ that shows how hydrogen, acting as a selective antioxidant, could, hypothetically, not interfere with the physiological signaling of very low dose superoxide radicals, while still scavenging undesired hydroxyl radicals and peroxynitrite, the potent oxidant resulting from the chemical reaction between superoxide radicals and nitric oxide.

The new paper¹ reported that very low doses of superoxide (acting at the picomolar level) can stimulate angiogenesis after the induction of ischemia (reduced blood flow) in a mouse model of peripheral artery disease. Under these conditions, angiogenesis is a beneficial compensatory mechanism for restoring blood flow in the ischemic area. An antioxidant that blocked the signaling of the very low dose superoxide would interfere with the induction of angiogenesis and, hence, be undesirable. As the authors¹ said, the induction of angiogenesis by VEGF (vascular endothelial growth factor) observed in this study, “verif[ied] that slightly elevated levels of ROS [reactive oxygen species] restore blood flow by stimulating endothelial cell proliferation through a VEGF-dependent pathway.” Hydrogen is an antioxidant that would not be expected to interfere with these slightly elevated levels of physiological radical signaling.

Reference

1. Bir et al. Control of angiogenesis dictated by picomolar superoxide levels. Free Rad Biol Med. 63:135-142 (2013)

More Beneficial Effects Reported for Increased Intake of Oleic Acid 

Two new papers report additional beneficial effects of dietary intake of oleic acid (a monounsaturated fat) as a substitute for saturated fat. (Though olive oil is an excellent source of oleic acid, high oleic sunflower oil contains 85% oleic acid, a far higher percentage.) The first paper¹ reports that substituting monounsaturated fats (oleic acid) for saturated fats (palmitic acid) in the diet of 12 human volunteers increased daily physical activity, increased resting energy expenditure, and (very interestingly) was associated with LESS ANGER AND HOSTILITY.

The subjects participated in a cross-over design in which they received for three weeks a high palmitic acid diet similar to a Western diet that contained 40.4% of calories as fat (palmitic acid, 16.0% calories, oleic acid 16.2% of calories), followed (separated by a another week of the control diet) a diet similar to the Mediterranean diet in fat composition (40.1% of calories as fat with 2.4% of calories as palmitic acid and 28.8% of calories as oleic acid). The oils were provided at room temperature as blends that were not used for cooking, but were mixed with food. There were two cohorts of 18 and 14 young adults as subjects.

The second paper² reports that oleic acid, but not other long chain fatty acids, signals to the SIRT1-PGC-1alpha complex to increase rates of fatty acid oxidation, thus improving the mitochondria energy metabolism of skeletal muscle cells. In this way, oleic acid provides protection against inflammation, dyslipidemias, and insulin resistance.²Subjects took a Profile of Mood States questionnaire to assess mood. Ten of 12 men and women exhibited a significantly lower anger-hostility score with the HOA (high oleic acid) than with the HPA (high palmitic acid). The authors note that they could not tell from their data if this result was due to increased oleic acid or decreased palmitic acid. They also found that 27 of 29 subjects in the 2 cohorts increased their physical activity during the low saturated fat (reduced palmitic acid) diet. (Reduced physical activity while consuming more saturated fat could be a result of increased inflammatory activity induced by consuming saturated fats, such as increased secretion of TNF-alpha (tumor necrosis factor alpha) associated with saturated fats as compared to monounsaturated fats.)

References

1. Kien et al. Substituting dietary monounsaturated fat for saturated fat is associated with increased daily physical activity and resting energy expenditure and with changes in mood. Am J Clin Nutr. 97:689-97 (2013).
2. Lim et al. Oleic acid stimulates complete oxidation of fatty acids through Protein Kinase A-dependent activation of SIRT1-PGC1alpha complex. J Biol Chem.288(10):7117-26 (2013).

PEPPERMINT OIL CAN IMPROVE EXERCISE PERFORMANCE IN HUMAN ATHLETES

A couple of issues ago in this newsletter we reported on a study of menthol¹(administered by subcutaneous injection in young mice). Treatment with menthol for 10 successive days improved spatial memory (as reflected in the animals’ performance in the dreaded Morris water maze. In addition, pre-treatment with menthol in both young and old mice reversed the amnesia induced in the animals by treatment with beta-amyloid on the tenth day. In other studies cited in the paper, menthol exhibited acetylcholinesterase inhibitory activity, thus increasing cholinergic nervous system activity, which may explain at least in part the improvement in memory in the Morris water maze and in response to beta-amyloid-induced amnesia.

Here, we report on another new study,² showing that functional parameters of exercise significantly improved in 12 healthy male students after receiving peppermint oil treatment (one bottle of 500 ml. of mineral water per day containing 0.05 ml peppermint oil for ten days). Tests were administered after treatment for ten days and compared to the initial test results before receiving peppermint oil. The results strongly supported the effectiveness of peppermint oil on exercise performance, respiratory function variables, systolic blood pressure, heart rate, and respiratory gas exchange parameters.² There were significant differences exhibiting improved performance between pre- and post-test results for resting heart rate, maximum heart rate, systolic blood pressure, diastolic blood pressure, chest circumference at maximum inhale, and chest circumference at maximum exhale. There have been inconsistencies in the results shown by peppermint oil in other studies, which the authors attribute at least in part to lack of good-quality evidence.

References

1. Bhadania et al. Protective effect of menthol on beta-amyloid peptide induced cognitive deficits in mice. Eur J Pharmacol. 681:50-4 (2012).
2. Meamarbashi and Rajabi. The effects of peppermint on exercise performance. J Int Soc Sports Nutr. 10:15 (2013) Open Access.

EAT LESS BY PAYING ATTENTION TO YOUR FOOD
YUP, IT’S OK TO PLAY WITH YOUR FOOD

A new paper¹ suggests that paying more attention to your food as you eat it will increase the neurological activity that signals satiety and awareness of food intake, thus having a beneficial effect on limiting how much you eat. The only will power required here is to pay attention to your food as you eat. You don’t try to force yourself to limit your intake, but to increase the focus your brain has on what you’ve eaten and, with that information, your brain will regulate your intake without any effort on your part.

The authors reviewed 24 papers of “neurologically intact” adults 18 years of age or older that took part in studies of the effect of distractions, awareness, or attention to food intake and food memory. (Food memory is important because it is generally recognized that in assessing food intake from memory, it is common to underestimate how much was eaten.)

Interestingly, the scientists found evidence that distraction produced a larger increase on later food intake than it did on immediate food intake. The six studies examining distractions while eating and later food intake were all performed by the same group.

The overall findings suggested four principles for reducing food intake:

1. Eat while avoiding distracting stimuli, such as watching television.
2. Prepare for eating by remembering similar meals before beginning the meal. Imagine reruns of similar meals.
3. Pay attention to the food you’re eating.
4. Remember the parts of the meal as you eat.

The authors suggest, for example, that food wrappers and other cues of eating (such as empty plates) not be removed from the eating area until the meal is completed, so as to constantly remind you of what and how much you’ve eaten. Forgetting all the food you’ve eaten impairs the brain’s ability to maintain a running total of all that you’ve consumed and hence to generate the proper regulatory signals.

5. D&S: We would also add to these other suggestions that playing with your food, as in “sculpting” carrots into shapes with your teeth and tongue as you eat the carrots, may be a good way to interact with the food, increasing the attention you pay to the sensory stimuli as you eat. Remember when your mother admonished you not to play with your food? Maybe, in that case, her advice was not so good. (It probably had more to do with your mother’s concern that you would dawdle and end up wasting food. Way back then, there were actually a lot of children starving in China.)

References

1. Robinson et al. Eating attentively: a systematic review and meta-analysis of the effect of food intake memory and awareness on eating. Am J Clin Nutr. 97:728-42 (2013).

BUT WHEN YOU’RE NOT EATING—OR ABOUT TO EAT,
IT MAY BE BEST NOT TO PAY ATTENTION TO FOOD

To get the most out of food and possibly eat less as a result, you should try to increase the signals that the food induces in your nutrient-sensing systems by paying close attention to your food when eating and avoiding distractions. However, as reported in an article in the December 28, 2007 Cell, Drosophila melanogaster (fruit flies) that are on a regime of dietary restriction but exposed to food-derived odorants don’t get the full benefit from the life extending effects of the diet restriction (DR). The food odorants alone decrease the expected lifespan increase, attenuating the longer lifespan the flies would otherwise derive from DR. This effect is not observed in fully fed flies.

In the Cell article,¹ flies with severe olfactory defects are reported to live significantly longer than wild-type flies. They are of normal size and have a normal metabolic rate but increased triglyceride storage and are resistant to both starvation and hyperoxia.²

What the fly results might mean for humans is that constant exposure to the sights and smells of food could induce the signals of nutrient-sensing systems that interfere with the “stop eating” metabolic pathways that would otherwise be active. Or, as hinted at in the article,¹ reducing sensory responses to food when one is not eating might be a way to mimic DR. The commentary authors point out that there is a “‘cephalic phase response’ which is an assemblage of physiological changes elicited by the sight, smell and taste of food, a sort of small scale “preview” of the full eating program. It involves preabsorptive release enzymes and hormones including pancreatic polypeptides, insulin, and glucagon and is generally viewed as a mechanism to optimize nutrient absorption and metabolism. A study of how this preview program is initiated and how it is terminated in the absence of actual eating might reveal some clues on how the nutrient-sensing pathways regulate the drive to eat.

References

1. Libert and Pletcher. Modulation of longevity by environmental sensing. Cell.131(7):1231-4 (2007).
2. Libert et al. Regulation of Drosophila life span by olfaction and food-derived odors. Science. 315:1133-37 (2007).

LIFE EXTENDING NATURAL PRODUCTS CAN GIVE DIFFERENT RESULTS WHEN FED OVER AN ANIMAL’S ENTIRE LIFETIME AS COMPARED TO FED AT JUST SPECIFIC PARTS OF THE LIFESPAN

The researchers report that there is significant change (~23%) in genome wide transcript profiles with age in Drosophila, suggesting that gene-induced changes by treatments with putative life extending materials may not have the same effects at all life stages.¹ They found that curcumin had beneficial effects in the normal lived Ra strain of Drosophila in that it increased longevity when administered in the developmental or health span stages, which (they suggest) could be due to systemic effects on the TOR complex, where reduced expression has been associated with life extending effects in studies with model organisms, but that curcumin had a negative effect on lifespan when administered over the entire adult lifespan or over the senescent stage only. They propose that this could be due to “an apparent curcumin-dependent accelerated neuromuscular degeneration observed in the legs of mid- and late-life treated animals.”A new study¹ reports that curcumin acts as a dietary restriction mimetic by increasing lifespan when administered during specific periods of the lifespan of Drosophila but find that whole life intervention is not always the most effective intervention. Basically, they report here1 that 4-phenyl butyrate (4PB), a life extending natural product, administered during the first 12 days of the adult lifespan was not as effective as feeding during the time period from 12 days to the end of life. The researchers identified the first 12 days as including a portion of the adult healthspan, while the period of 12 days to the end of life included the transition and senescent periods. The results of this and other studies looking at treatment with life extending materials at only specific periods of the lifespan are pointing to additional complications in interpreting how increases in lifespan are produced.

The authors conclude that life-time feeding may be responsible for the failure of curcumin to significantly extend the healthspan and delay the onset of senescence by studies in other labs.

In a different paper,² researchers found that increased oxidative resistance induced in human MEF cells and in Drosophila by exposure to acute, chronic, and repeated oxidative stress had different effects on lifespan. “Regrettably, regimens of adaptation to both repeated and chronic stress that were successful for short-term survival in Drosophila nevertheless also caused significant reduction in life span for the flies.”²

The authors sum up their findings: “repeated MEF cell oxidant exposures seem to prevent adaptive stress responses if the interval between exposures is too short and/or if each exposure is at too high an oxidant level. In contrast, repeated cell exposures at 12-h intervals, especially at lower oxidant concentrations, seem to potentiate positive stress adaptations and extend the period of protection. Chronic oxidant exposure at low oxidant levels can actually potentiate and extend adaptive responses, but chronic exposure to higher oxidant levels prevents adaptation.” For the Drosophila melanogaster, repeated oxidative stress adaptation at intervals of 1 or 3 days is toxic but intervals of 7 days are tolerated in terms of [improved] short-term survival. Life span however, seems to be negatively influenced by all repeated stress adaptation regimens. Short-term survival is also negatively influenced by chronic oxidative stress adaptation at all levels.”² “All repeated treatments appeared to have a negative long-term effect, because flies that were repeatedly exposed to H2O2 [hydrogen peroxide] had a shorter life spans than untreated flies. We also observed a progressive decline in Drosophila life span with frequency of H2O2 treatment.”²

These results indicate that there is a great deal yet to learn about the dose and timing of hormesis that results not only in short term resistance to stresses such as oxidative stress but increased lifespan, which would appear to require different processes that work over a long period of time.

Nevertheless, the results of these studies must be reconciled with others that found mild stresses to have anti-aging hormetic effects and appeared to show an effective approach to modulate aging.³ For example, in paper #3. researchers reported that a single heat-shock early in life extended longevity by 20% or more in C. elegans by decreasing initial mortality only, with the rate of increase of subsequent mortality (the Gompertz component) being unchanged. Worms were also exposed to multiple heat shocks³ in the study with each heat shock having a hormetic effect on lifespan but with the magnitude of each additional single heat-shock decreasing as the age of the animal increased.

References

1. Soh et al. Curcumin is an early-acting stage-specific inducer of extended functional longevity in Drosophila. Exp Gerontol. 48:229-39 (2013).
2. Pickering et al. Oxidative stress adaptation with acute, chronic, and repeated stress. Free Radic Biol Med. 55:109-18 (2013).
3. See, for example, Wu et al. Multiple mild heat-shocks decrease the Gompertz component of mortality in Caenorhabditis elegans. Exp Gerontol. 44:607-12 (2009).

DETERMINING WHETHER CLIMATE CHANGE IS CAUSED BY HUMANS, THE EASY WAY

A study reported in the 1 Feb Science reveals that, when asked whether humans are causing climate change, Demo­crats were likely to say yes, Republicans likely to say no, but Independents were likely to say yes on unseasonably warm days but to say no on unseasonably cold days. No surprise but it just goes to show that very short term events rather than consideration of the totality of the evidence can be a deciding factor in how people perceive the reality of human-induced global warming (or probably anything else, for that matter). This is an excellent way for those looking for the “right” response to a survey to plan when to ask people if they think humans are causing climate change to get the answers they want.

EPIGENETIC CHANGES IN EXPRESSION
OF GENES VIA DNA METHYLATION PART II

In the May 2013 issue of this newsletter, we published an article on “Methylation, a major mechanism regulating gene expression, changes with age, revealing dynamic landscape.” We continue that here with new data. In our May article, we discussed how methyl donors (such as s-adenosylmethione (SAMe), folic acid, choline, and methionine) are part of a system that, under the regulation of DNA methyltransferases either adds a methyl group to or removes a methyl group from a specific site on DNA histones which controls whether a gene can be transcribed (turned on) or silenced (turned off). With the discovery of this process, much exciting new research has already been and continues to be published.

Changes in Age-dependent Epigenetic Parameters by Exercise in Rat Hippocampus

New research on the effect of exercise on DNA methylation are being published. One paper¹ reports on the effects in male Wistar rats at ages 3 and 20 months old of engaging in two exercise protocols on a treadmill, one for a single session of 20 minutes of exercise and the other chronic treadmill exercise (20 minutes running each day for 2 weeks). The results showed that the 20 month old (and 20 months qualifies as aged if you’re a rat) had lower histone H3-K9 methylation levels (about 50%) as compared to the 3 month old rats. Also, the levels of DNMT1 (DNA methylatransferase 1) were significantly diminished in the aged animals by about 25%. The level of DNMT3b (DNA methyltransferase 3b) was not modified by age.

These results are consistent with the data from other studies as noted by the authors.¹They explain that there is a “genome-wide tendency to DNA hypomethylation in multiple vertebrate organs during aging process. In addition, the age-related global hypomethylation is related to DNMT1 deficits in senescent human fibroblasts. However, studies reporting DNMT content in the brain during aging process are lacking.”

Further results¹ showed that “[w]hen measured 1 h after [single] session ended, young adult exercised rats exhibited lower levels of DNMT3b (about 30%) when compared to its sedentary group [young adult rats not exercised] (p=0.042); while no delayed (18 h) effects of exercise were observed.” This change was not observed in the aged rats. There were no changes in DNMT3b as a result of chronic exercise in all groups. Likewise for DNMT1, the acute (single session) of exercise acutely diminished this DNA methyltransferase by about 45% in the hippocampi of 3 month old rats (p<0.001) without any changes 18 hours after exercise. Again, this change was not observed in the aged rats. Meanwhile, there was no significant effect of the chronic exercise regimen on DNMT1 in either young adult or aged rats.

The transient effect of decreased DNMT3b after acute exercise implies that, as suggested by the authors of a different (earlier) study,² DNA hypomethylation is an early event in contraction-induced gene activation. The second study² also found that the promoter of genes with exercise-induced increased expression (PGC-1alpha, PDK4, PPAR-delta) were hypomethylated (had decreased methylation after as compared to before exercise) in human healthy sedentary subjects following acute exercise. Hypo­methylation supports the notion that there was greater expression of those genes. Note that PGC-1alpha is a “master” gene that controls mitochondrial biogenesis.

The results¹ also support the idea that there is a decrease in histone methylation in many genes with an increase in others occurring as part of the aging process.

DNA Methylation Deficits in Memory with Age Can Be Reversed by Increasing a DNA Methyltransferase

A recent paper³ reports that aged mice have decreased expression of a DNA methyltransferase (DNMT3a2) that results in memory deficits and that restoration of the expression of this enzyme can reverse the memory deficits.

One of the tests used by the researchers to detect the memory defect was by exposing the mice to a tone followed by a foot shock. When exposed to the tone in a different context 24 hours later, the old mice froze with fear much less often than the young mice, indicating that the memory of the old mice functioned less well (e.g., they were less likely to remember the warning tone). Using a recombinant adeno-associated virus to deliver the gene for DNMT3a2, the scientists were able to restore the memory of the old mice.

The authors³ hypothesized that the loss of the DNA methyltransferase resulted in decreased methylation of and less expression of the enzyme’s target genes. (In this case, the DNA methylation acted as a transcriptional activator, increasing gene expression, although DNA methyl­ation can in other circumstances act as a transcriptional suppressor, decreasing gene expression—yup, it’s complicated.) See also reference #4, which is the commentary on paper #3.

Methylation Changes Found in Night Shiftworkers

Finally, we have another new paper⁵ that reports on a study of DNA methylation changes in 10 randomly selected women from an initial group of 17 with long histories of night shiftwork that were compared to 10 female dayworkers. The researchers found widespread methylation changes at imprinted genes in the night shiftworkers. (Imprinted genes are the alleles of genes, which are expressed depending upon the parent of origin. “Loss of monoallelic expression at imprinted genes, known as loss of imprinting (LOI), has been associated with various cancer types and may play a role as an early driver in tumor development.”⁵) Among the genes with methylation changes, the paper reported, were many with roles in circadian clock pathways that have been linked to cancer risk.

The authors propose that, although it has not been proven, long-term shiftwork may induce changes in methylation by perturbing the normal circadian exposure to light. “Interestingly, the magnitudes of methylation changes observed are comparable to methylation changes which have been attributed to occupational exposures to low-dose benzene and polycyclic aromatic hydrocarbons.” Such exposures can be carcinogenic at high enough doses.

Hypomethylation to Resensitize Diffuse Large B-Cell Lymphoma to Chemo­therapy; Restores Tumor Suppressor Gene Activity

We have written about the exciting new finding that DNA hypomethylation has been shown in some cancers to restore the activity of tumor suppressor genes that have been silenced by hypermethylation.⁶ Hypomethylating drugs such as azacytidine are being used in early trials to restore the sensitivity of cancers that have become chemotherapy resistant. In a new paper,⁶ researchers used diffuse large B-cell lymphoma cell lines to study the use of the hypomethylation drug azacytidine to reactivate the SMAD1 gene that resensitized the cells to standard chemotherapeutic treatment after they had become resistant.

In another very recent paper,⁷ the authors review the new work on DNA hypermethylation that silences tumor suppressor genes and the discovery of agents that can reactivate these silenced genes, such as drugs (for example, azacytidine) or natural compounds acting via different mechanisms to inhibit DNA methyltransferases, thus decreasing methylation, that includes curcumin, EGCG, resveratrol, parthenolide, and others).

References

1. Elsner et al. Exercise induces age-dependent changes on epigenetic parameters in rat hippocampus: A preliminary study. Exp Gerontol. 48:136-9 (2013).
2. Barres et al. Acute exercise remodels promoter methylation in human skeletal muscle. Cell Metab. 15:405-41 (2012).
3. Oliveira et al. Rescue of aging-associated decline in Dnmt3a2 expression restores cognitive abilities. Nat Neurosci. 15(8):1111-3 (2012).
4. Su & Tsai. DNA methylation in cognition comes of age. Nat Neurosci. 15(8):1061-2 (2012).
5. Jacobs et al. Jacobs et al. Methylation alterations at imprinted genes detected among long-term shiftworkers. Environ Mol Mutagen. 54:141-6 (2013).
6. Clozel et al. Mechanism-based epigenetic chemosensitization therapy of diffuse large B-cell lymphoma. Cancer Discov. 3(9):1-18 (2013).
7. Singh et al. DNA methyltransferase inhibitors as epigenetic therapy for cancer. Curr Cancer Drug Targets. 2013 Mar. 18 [Epub ahead of print].

YOU’RE INVITED TO A PARTY! NOT JUST ANY PARTY—
HOW THE TASTE OF FOOD IS AFFECTED BY THE WEIGHT, SIZE, SHAPE,
AND COLOR OF THE CUTLERY USED TO EAT IT AND THE PLATES IT IS SERVED ON.

Taking a break from the usual health oriented contents of this newsletter, we found a new open access journal called Flavour at @FlavourJournal which contains a wide variety of papers on the scientific basis of flavor with ideas tested on improving the taste and flavor or limiting the amounts of food eaten by changing the shape and weight of cutlery and the color of plates food is served on. Sounds like a great source of information for the small restaurant trying to get a one-up on the competition without spending much money or for people who like to throw parties and want to have attendees really remember your party.

How Mere Cutlery—the Weight, Size, Shape, and Color of It—Can Change How You Perceive Your Food

This new paper¹ tested variations on the cutlery people were given to eat food on the ratings people gave on various flavor components of that food. For example, yogurt was perceived as denser and more expensive when tasted from a light colored plastic spoon as compared to a spoon made to appear heavier (with artificial weights). Food was rated as tasting saltiest when eaten from a knife as compared to a spoon, fork, or a toothpick. In their description of other studies, the authors¹ mentioned that drinking from a “cold” colored glass (a dark color as compared to a yellow, orange, or red color) may result in the perception that the drink is more thirst-quenching. In another study they described, researchers found that food eaten from bowls artificially weighted to make them appear heavy resulted in food perceived as thicker and denser (e.g., heavier), which people preferred to the same food eaten from a light bowl. (Do you suppose this could improve the eating quality of stone soup?) In yet another study (the results haven’t been published), cheddar cheese was reported to taste “sharper” when sampled after viewing pointy figures as compared to that tasted after viewing round images. (We aren’t making this up, folks.)

The researchers brought up the issue of the expectations of food consumers and suggested that food would taste better when cutlery met tasters’ expectations based on the sort of cutlery that tasters had previously used in eating better quality food.

Other nifty little tidbits reported in this paper1 include: a blue colored bowl resulted in unsalted popcorn that tasted salty. (The researchers¹ mentioned that in the UK blue packaging is often associated with salty snack products.) White yogurt tasted more expensive when tasted from a white spoon as compared to a black spoon. Strawberry mousse was perceived as sweeter when sampled from a white plate as compared to a black one. In a separate study, researchers found that people would eat less when a snack was served on a red plate or a drink that has a red label.

Merely Changing the Color of a Plate Can Give People a New Taste Perception

Another paper² reported on the effects of the color of a plate on how people perceive the taste of a dessert.

You are at a disadvantage here because, although pictures of these lovely desserts are provided in the paper, obviously you cannot see them (unless you download the free paper). They were listed as dessert A (fraisier), dessert B (fraicheur of raspberry and vanilla) and dessert C (vacherin glace with vanilla, raspberry, and basil), presented on white or black plates.

A few of the results: participants in the low-contrast condition (white pasta sauce on a white plate or red pasta sauce on a red plate) served themselves significantly more pasta than those in the high contrast situation (white pasta sauce on a red plate or red pasta sauce on a white plate). Diners rated the lunch desserts uniformly more appetizing when served on a white plate while the dinner desserts were found to be similarly appetizing no matter what the color of the plate. As to overall liking of the desserts (rated after desserts had been tasted), desserts A and C resulted in similar scores no matter what the color of the plate while the pattern for dessert B was said to be the “reverse. though the meaning of “reverse” here is unclear. No hypothesis is offered for why these differences between the desserts were seen, but there is certainly something there that requires explanation since it was a consistent effect.

So the next time you go to a party, remember that your taste might be affected by a number of different tricks for serving food without actually changing the food. If you think that your latest party had some of the best food you ever had at a party, don’t be TOO impressed, it may be somewhat illusory (though still fun) and, once you innocently walked in the door, you didn’t stand a chance!

Or, the next time you give a party, consider the use of some special effects to bring out a larger WOW effect in your party crowd when they eat your food.

WE ATE AND WE ATE
WHILE WE TASTED THE PLATE
AND THE NEXT COURSE THEY BROUGHT
THE SPOONS WERE PART OF THE PLOT.
YOU WEREN’T SUPPOSED TO GUESS
WHY THE FOOD WAS A SUCCESS
AND THE PARTY SURE TO IMPRESS.
THE SCIENCE WAS THERE
BUT YOU DIDN’T KNOW WHERE
AND THE NEXT STUDY WE DO
WILL BE INSIDE OF YOU:
CAN YOUR GUT
TASTE THE DIFFERENCE, TOO.^3,4

Oh, and one other thing, if you serve alcoholic drinks at your party, be sure to also serve Party Pill.™ We never go to a party without it!

References

1. Harrar and Spence. The taste of cutlery: how the taste of food is affected by the weight, size, shape, and colour of the cutlery used to eat it. Flavour. 2:21 (2013). http://www.flavourjournal.com/content/pdf/2044-7248-2-21.pdf
2. Piqueras-Fiszman, Giboreau, and Spence. Assessing the influence of the color of the plate on the perception of a complex food in a restaurant setting. Flavour.2:24 (2013). http://www.flavourjournal.com/content/pdf/2044-7248-2-24.pdf
3. Surprisingly, taste buds that can literally taste sweet and bitter via the same receptors as the taste buds on your tongue have been found in gastrointestinal cells (and also elsewhere, such as in the lungs). See, for example, Mayer, Gut feelings: the emerging biology of gut-brain communication. Nature Reviews Neuroscience. 12:453-466 (2011).
4. Untitled poem by Sandy Shaw.

WHY ARE THE NUMBERS OF INDIVIDUALS RENOUNCING
THEIR AMERICAN CITIZENSHIP INCREASING SO FAST?

We don’t think there is any great mystery about why people might be interested in renouncing their American citizenship when the costs imposed upon citizens (taxes and regulations) are increasing so rapidly. Like everything else, citizenship provides benefits and imposes costs, and there is a limit to what people are willing to accept as a reasonable “deal.” The numbers are not yet large, but the rate of increase is pointing in a steeply upward direction. These numbers are for the last quarter, data that will be shortly officially released but has been obtained and publicized by a reliable media figure. Last quarter, 1,131 individuals renounced their U.S. citizenship as compared to the same quarter last year in which 188 people renounced their U.S. citizenship.

Incredibly, the government is now charging an exit tax to prevent some high net worth individuals from taking their wealth with them. This applies to anyone whose average U.S. tax liability over the last five years was about $150,000 (roughly $500,000 in taxable income in 2012 dollars) and/or has a net worth of at least $2 million on the date of their expatriation. The net worth figure is NOT adjusted with inflation. Compare this exit tax outrage with the “Act of July 27, 1868” in which the U.S. Congress declared that “the right of expatriation is a natural and inherent right of all people, indispensable to the enjoyment of the rights of life, liberty, and the pursuit of happiness.”

We are fed up with the incredible GREED of the government. An average taxable income of $150,000 over the last five years is hardly enough to consider someone rich. In fact, this is well under the $250,000 that Obama promised was an income level that would not, under his Administration, be subject to higher taxes. Just another LIE in a growing output of government communication that is almost all LIES. Don’t get used to it, folks, then they’ve really gotcha.

References

http://www.sovereignman.com/expat

https://s3.amazonaws.com/public-inspection.federalregister.gov/

See also:

http://www.sovereignman.com/expat

APPETIZERS

The following short excerpt is from Freehold by Michael Z. Williamson (Baen, 2004):

Background: UN vs. FREEHOLD, a former Earth colony, now an independent planet being invaded by the UN, which expects the yokels to roll over and play dead. But, unfortunately for the UN, FREEHOLD isn’t Earth.

The convoy was four vehicles with UN markings. One was a Mk 17 Infantry Light Armored Wheeled Assault Vehicle, the others simply multipurpose vehicles with heavy weapons mounted. They stopped in front of the farm and several people dismounted. They approached the door and met Dak at the steps as he came out.

“Yes?” he asked, bluntly and without any friendship.

The one in civilian clothes spoke, “I am Lynet Krishnamurti with the United Nations Readjustment Task Force. I am here to give you an informational package on the recent improvements we are implementing.”

“Thanks, but we don’t need any improvements. I have the latest gear I can afford.” Dak replied. He wanted them to leave quickly.

“Well, that’s the point,” Krishnamurti said, “One of the benefits the UN offers is investment capital to buy better equipment. We also guarantee reparations not covered by insurance, accident insurance … many benefits. This package is on hard copy and on datachip, compatible with most systems.”

“And what does this cost me?” Dak asked, trying to sound like a suspicious bumpkin.

“It’s free. The UN provides it as a service to all agrobusiness operations.”

“Well, if I need it, I’ll call you. Thanks. Is that it?”

“I’m also here to assess your hectareage,” Krishnamurti admitted.

“Not sure. Probably six or seven thousand.” He knew to the millimeter what he planted, but he wasn’t about to admit it. “Why?”

“We need an accurate measure to assess commercial property taxes. The package also contains information on tha—.”

“Property tax?” Dak acted confused. “It’s my property and a gift from the Lord. Why should I pay tax on it?” Krishnamurti looked exasperated. Was every one of these peasants utterly ignorant of basic principles? They wrangled for long segs, while the troops looked amused. They’d seen it all before.

“So let me get this straight,” Dak was trying desperately but successfully to avoid hysterical laughter at his guest’s discomfort. “In exchange for taxing the property the Lord gave me to clear and use and taking a whopping chunk of my income from said property, and dictating what I grow, how I grow it, what equipment to use, and how to wipe my nose most likely, you’ll grant me a ‘free’ loan at interest to buy the equipment I wouldn’t need without your regulations? And I’ll have to spend an extra four days a week, unpaid, doing bookkeeping to prove it to you?”

“Un, put that way it sounds stupid,” she said.

To find out how this all ends up, you’ll need to read the book. We both read it and highly recommend it for those who see where the American political class is heading, don’t like it, and would enjoy reading fiction where people like us and (hopefully) you fight back and WIN!

Politics is the art of looking for trouble, finding it whether it exists or not, diagnosing it incorrectly, and applying the wrong remedy. .
— Ernest Benn

TThen you add two forkfuls of cooking oil.
— Julia Child

He who joyfully marches in rank and file has already earned my contempt. He has been given a large brain by mistake, since for him the spinal cord would suffice.
— Albert Einstein

… the human gut microbiome contributes 36% of the small molecules that are found in human blood …
— Leroy Hood, cofounder and president of the Institute for Systems Biology (Science 8 June 2012)

The National Park Service, administered by the U.S. Department of the Interior, asks us, “Please Do Not Feed the Animals.” Their stated reason for the policy is: “The animals will grow dependent on handouts and will not learn to take care of themselves.”
— Attributed to The National Park Service as reported in Casey’s Daily Dispatch 6/28/13

CARDIOVASCULAR PROTECTION

Lipid Accumulation in Vascular Endothelial Cells Induced by Palmitate, a Saturated Fatty Acid, Markedly Reduced by Green Tea Polyphenol EGCG

A new paper¹ reports another potentially important beneficial effect of the major polyphenol in green tea, epigallocatechin gallate (EGCG). EGCG is shown to stimulate autophagy (a form of self-disassembly or “self-eating” that is required to maintain homeostasis) that degrades lipid droplets induced by palmitate in primary bovine aortic endothelial cells (BAEC). The accumulation of lipids in vascular endothelial cells is a major part of the process promoting atherosclerosis.

A large epidemiological study in Japan² with an 11-year followup reported in a 2006 paper that green tea consumption was associated with reduced mortality due to cardiovascular disease but not with mortality due to cancer.

In the new study, the authors investigated whether EGCG could normalize the palmitate impairment of autophagy flux that prevents lipid accumulation in vascular endothelial cells by degrading lipid droplets and, if so, to identify the mechanism for this action. As the authors describe their findings:

“Excess intake of lipid causes obesity and ectopic lipid accumulation, which is implicated as one of the causes for cardiometabolic syndrome. Fatty acid overload increases intracellular lipid droplets, and the presence of lipid droplets in non-adipose tissue [such as muscle] plays a role in various pathophysiologies.” Their experimental results suggest, the authors say, that “the effect of EGCG in reduction of lipid droplets is mainly dependent on degradation but not inhibition of formation. These results suggest that EGCG decreases accumulation of lipid droplets through facilitation of lysosomal degradation, which may contribute to prevention of lipotoxicity [toxicity to cells by fatty acid overload] in vascular endothelial cells.”

Intensive investigation of possible mechanisms led the authors to conclude that EGCG induces autophagy through a CA2+/CaMKKbeta/AMPK mediated mechanism, which reduced palmitate-induced accumulation of lipid droplets in endothelial cells. Autophagy is a major mechanism by which old organelles (such as mitochondria) and damaged molecular debris are degraded and the basic components made available for reuse. It is also a way to regulate the turnover of supplies of resources for more efficient use. In the case of lipids, autophagy may have “important implications for human diseases with lipid over-accumulation such as those that comprise the metabolic syndrome.”⁴“Decreased autophagy in the liver with ageing may contribute to hepatic [liver] lipid accumulation that occurs along with an increased incidence of the metabolic syndrome in aged humans.”⁴

Earlier papers that led up to this new paper1 include references #2 and #3 listed below, where two scientific groups reported an inhibitory effect of EGCG on lipid accumulation in adipocytes (fat cells). The new findings explain how autophagy plays a key role in EGCG inhibition of lipid accumulation in vascular cells which links the EGCG protection to a reduced risk of cardiovascular disease, whereas the two earlier papers reported that EGCG decreased lipid accumulation in adipocytes (fat cells), which is particularly associated with reduction of the risk of metabolic syndrome that includes obesity and insulin resistance.

Whichever way you look at it, EGCG has become something of a phenomenon if you want to maintain a healthier life (healthspan) for longer. It has earned a place as a basic component for your cabinet of medical supplies. Don’t leave home without it!

References

1. Kim et al. Epigallocatechin gallate stimulates autophagy in vascular endothelial cells: a potential role for reducing lipid accumulation. J Biol Chem. 288:22693-705 (2013).
2. Moon et al. Inhibitory effect of (-)-epigallocatechin-3-gallate on lipid accumulation of 3T3-L1 cells. Obesity. 15(11):2571-82 (2007).
3. Lee et al. Inhibitory effects of green tea catechin on the lipid accumulation in 3T3-L1 adipocytes. Phytother Res. 23:1088-91 (2009).
4. Singh et al. Autophagy regulates lipid metabolism. Nature. 458:1131-5 (2009).

MECHANISM OF SYSTEMIC AGING
CONTROLLED BY THE HYPOTHALAMUS

​A new paper¹ reports the important discovery that activation of innate immunity in the hypothalamus plays a key role in aging. The researchers found that “through activating or inhibiting immune pathway IKK-beta and NF-kappaB in the hypothalamus of mice, we were able to accelerate or decelerate the ageing process, leading to shortened or increased lifespan.”

The researchers found that there was a direct link between IKK-beta and NF-kappaB activation and gonadotropin-releasing hormone (GnRH) decline. GnRH regulates the release of sex hormones and, hence, controls reproduction. The researchers found that GnRH release decreased in GT1-7 cells, a cell line of GnRH neurons, after IKK-beta and NFkappaB activation, but increased after IKK-beta and NFkappaB inhibition. Focusing on the effect of GnRH on aging, the scientists found that delivering the hormone into the third ventricle of the hypothalamus of old mice caused increased adult neurogenesis, whereas neurogenesis normally declines with age. In fact, administration of GnRH via peripheral injections (that is, outside of the brain) ameliorated age-related cognitive decline. (The authors note that certain areas of the brain that are sensitive to GnRH are actually located outside of the blood-brain barrier, thus permitting GnRH to reach the brain from the periphery. The mechanism for the peripheral anti-aging effects of GnRH requires additional study.)

The scientists found, further, that reduced hypothalamic release of GnRH also contributed to other aspects of aging, including declining muscle strength, skin atrophy, bone loss, memory impairment, in addition to (as noted above) adult neurogenesis.

The researchers discovered that while NFkappaB signaling increased in many regions of the brain in aging mice, this increase was greatest in the hypothalamus. Part of their earlier work that led up to this latest discovery revealed that “infection-unrelated inflammatory changes in the mediobasal hypothalamus contribute to the development of various metabolic syndrome components, and the molecular basis is mediated crucially by NF-kappaB and its upstream IKK-beta.”¹

Earlier work has also reported NFkappaB involvement in aging processes. For example, another recent paper² found that the mechanism by which DNA damage drives aging is due in part to NFkappaB activation. A recent review³ reported that transcriptional activation of NFkappaB is increased in a variety of tissues with aging and associated with age-related degenerative diseases such as Alzheimer’s disease, diabetes, and osteoporosis. The review also reports that inhibition of NFkappaB in mouse models has been shown to delay onset of age-related symptoms and pathologies.

Moreover, NFkappaB has been implicated in aging in much earlier work. One paper from 1997⁴ reported that “NFkappaB … exists in a constitutively activated state in cells obtained from the major lymphoid organs of aged animals.” One earlier paper⁵ found that overnutrition activated hypothalamic IKKbeta/NF-kappaB at least in part by elevated endoplasmic reticulum stress, a mechanism that has also been identified in diabetes. Yet another paper⁶ reports that depressive-like behaviors in mice subjected to chronic unpredictable stress are mediated by NFkappaB signaling in the adult hippocampus.

Hydrogen Modulation of NF-KappaB Reported In Three Papers

Research on the mechanisms of hydrogen therapy is in early stages, but three papers have reported it to modulate NF-kappaB activation. In one paper,⁷ hydrogen-rich saline was tested as a treatment in a rat model of amyloid-beta-induced Alzheimer’s disease. Amyloid beta has been found to induce neuronal cell death via ROS (reactive oxygen species) mediated by NF-kappaB activation. In this study,⁷ NF-kappaB activation in the hippocampus was inhibited by the hydrogen-rich saline. In another paper,⁸ hydrogen-rich saline prevented neointima (atherosclerotic plaque) formation that followed carotid balloon injury by suppressing ROS and the TNF-alpha/NF-kappaB pathway. In another paper,⁹ NF-kappaB activation increased early and transiently in response to ventilator-induced lung injury in mice, providing protection against apoptosis and inflammation. This is an example of how timing can be a key factor in determining the effect of a powerful molecule such as NF-kappaB. As the authors comment, “[t]he function of NF-kappaB activation during VILI [ventilator-induced lung injury] has not been fully elucidated and conflicting roles for NF-kappaB, protective and injurious, have been proposed.”⁹ On the one hand, NF-kappaB triggers upregulation of genes involved in inflammation, infection, and stress responses and, on the other hand, it mediates a cellular survival mechanism against apoptotic cell death. Hence, an early transient increased activation of NF-kappaB might provide a protective effect against the early stages of induced cell death and, if not prolonged, not increase inflammatory and stress responses. Further research is required to unravel the complex timing process.

Finally, a new paper¹⁰ reported that treatment of TNF-alpha-induced cell injury in neonatal rat osteoblasts (bone forming cells) with hydrogen dissolved in vehicle inhibited the TNF-alpha-induced activation of the NF-kappaB pathway. This resulted in reduced oxidative stress, preservation of mitochondrial function, suppression of inflammation, and enhancement of nitric oxide availability. The authors note that circulating NO level is reduced and correlated with osteoporosis in aged rats and ovariectomized rats, suggesting (they propose) that hydrogen might be a useful way to treat osteoporosis.

Natural Products That Inhibit IkappaB Kinase (IKKbeta)

Other natural products have been reported to inhibit IKKbeta and/or NFkappaB. For example, the anti-inflammatory parthenolide, a major component of the medicinal herb Feverfew has been reported to directly bind to and inhibit IKKbeta¹¹ and that this results in the inactivation of NFkappaB.

As NFkappaB is involved in many functions, including importantly beneficial activities (such as cellular pro-survival pathways), the key to the use of NFkappaB suppression for optimal anti-aging is to target it to reduce its activity in areas such as the hypothalamus, where overactivity links it to aging-induced changes. Hydrogen easily passes the blood-brain barrier to enter the hypothalamus.

Toll-Like Receptors Play a Crucial Role in the Signaling Pathways Which Lead to Nf-Kappab Activation

Toll-like receptors are molecules that, as part of the innate immune system, recognize structures of microbes (such as LPS, lipopolysaccharide, a component of Gram-negative bacterial cell walls, viral or bacterial nucleic acids, and proteins unique to microbes), thereby activating immune system response. Importantly, “Toll-like receptors (TLRs) play a crucial role in the signaling pathways which lead to NFkappaB activation. TLR4 is considered the lipopolysaccharide (LPS) receptor.”¹² A new paper¹² ­reports that molecules that activate the mu opioid receptor (such as morphine) decreases TLR4 in mouse macrophages, thus acting as a powerful immunesuppressive agent. In fact, the paper¹² reports, “[a] series of studies have demonstrated that MOR [mu opioid receptor] activation is responsible for most of the immunosuppressive effects of opioids.”

We report this interesting and important link between toll-like receptors, opioids, and NF-kappaB, but do not propose that mu opioid agonists (such as morphine) be used to reduce the activation of TLRs for the purpose of decreasing NF-kappaB signaling and reducing immune system inflammation. The risk there is, aside from opioid addiction, that too much suppression of TLRs could increase the risk of infection. As the authors¹²note, excessive immunosuppression is a risk when using opioids such as morphine for pain control. We think that the use of hydrogen therapy or other natural products that modulate the effects of NF-kappaB, such as parthenolide (from the herb feverfew),¹³offer a safer approach to decreasing excessive activation of NF-kappaB.

References

1. Zhang et al. Hypothalamic programming of systemic ageing involving IKK-beta, NF-kappaB and GnRH. Nature. 497:211-6 (2013).
2. Tilstra et al. NFkappaB inhibition delays DNA damage-induced senescence and aging in mice. J Clin Invest. 122(7):2601-12 (2012).
3. Tilstra et al. NF-kappaB in aging and disease. Aging Dis. 2(6):449-65 (2011).
4. Spencer et al. Constitutive activation of NF-kappaB in an animal model of aging. Int Immunol. 9(10):1581-8 (1997).
5. Koo et al. Nuclear factor-kappaB is a critical mediator of stress-impaired neurogenesis and depressive behavior. Proc Natl Acad Sci USA. 107(6):2669-74 (2010).
6. Zhang et al. Hypothalamic IKKbeta/NF-kappaB and ER stress link overnutrition to energy imbalance and obesity. Cell. 135:61-73 (2008).
7. Wang et al. Hydrogen-rich saline reduces oxidative stress and inflammation by inhibit of JNK and NF-kappaB activation in a rat model of amyloid-beta-induced Alzheimer’s disease. Neurosci Lett. 491:127-132 (2011).
8. Qin et al. Hydrogen-rich saline prevents neointima formation after carotid balloon injury by suppressing ROS and the TNF-alpha/NF-kappaB pathway. Atherosclerosis. 220:343-50 (2012).
9. Huang et al. Hydrogen inhalation reduced epithelial apoptosis in ventilator-induced lung injury via a mechanism involving nuclear factor-kappa B activation. Biochem Biophys Res Commun. 408:253-8 (2011).
10. Cai et al. Treatment with hydrogen molecule alleviates TNF-alpha-induced cell injury in osteoblast. Mol Cell Biochem. 373:1-9 (2013).
11. Kwok et al. The anti-inflammatory natural product parthenolide from the medicinal herb Feverfew directly binds to and inhibits IkappaB kinase. Chem Biol. 8:759-66 (2001).
12. Franchi et al. Mu opioid receptor activation modulates Toll like receptor 4 in murine macrophages. Brain Behav Immun. 26:480-8 (2012).
13. Dai et al. The NF-kappaB inhibitor parthenolide interacts with histone deacetylase to induce MKK7/JNK1-dependent apoptosis in human acute myeloid leukaemia cells. Br J Haematol. 151(1):70-83 (2010).

CHRONIC RED WINE CONSUMPTION BY MIDDLE AGED MEN REDUCES PLASMA LPS, AN IMPORTANT SOURCE OF INFLAMMATION

The researchers were investigating the effect on LPS concentrations and on the gut microbiota of chronic red wine consumption and acute red wine intake in middle-aged men on a diet high in fat. It has been shown that even a single high fat meal can result in a significant increase in absorption of LPS from the gastrointestinal tract in a small trial of 12 healthy men.^2B Studies in animal models of high fat diet-induced obesity have reported increased plasma LPS concentration as an early event in the development of diabetes.³We have written before on how LPS, lipopolysaccharide (a component of bacterial cell wall), is a major source of inflammatory stimulation of the immune system and probably a key element in the aging process.¹ A new paper² reports that chronic red wine consumption by middle aged men increases certain gut microbes and that this increase was correlated negatively with LPS concentrations. The resident gut microbiota appears to play an important role in supplying LPS to plasma in metabolic endotoxemia2 and could, therefore, be a major supplier of inflammatory immune system stimuli.

While the investigators didn’t find a significant difference between LPS or LPS-binding protein (LBP) concentrations between subjects who were taking chronic red wine, dealcoholized red wine, or gin for 20 days,* they did observe significant increases in the amounts of Bifidobacterium and Prevotella gut microbes in subjects consuming chronic red wine, where these increases were correlated negatively with LPS concentration. An earlier paper⁴ had also reported that increases in bifidobacteria in mice fed a high fat diet supplemented with a prebiotic, oligofructose, resulted in reduced inflammation and decreased endotoxemia (LPS).

---

* The subjects drank 272 ml/day of red wine (250 ml is about a cup), dealcoholized red wine (272 ml/day), or gin (100 ml/day) during the 20 day study.

Interestingly, the researchers² found that there was an enhanced postprandial lipid response to a meal induced by red wine but, despite that, a moderate dose of red wine prevented the increase in NF-kappaB activation ordinarily induced by a high fat meal. As suggested by the paper reported above, aging is associated with increased expression of NF-kappaB and decreased expression of this molecule (particularly in the hypothalamus) may be an effective way to slow aging.

References

1. Finch and Crimmins. Inflammatory exposure and historical changes in human life-spans. Science. 305:1736-9 (2004).
2. Clemente-Postigo et al. Effect of acute and chronic red wine consumption on lipopolysaccharide concentrations. Am J Clin Nutr. 97:1053-61 (2013).
2B. Erridge et al. A high-fat meal induces low-grade endotoxemia: evidence of a novel mechanism of postprandial inflammation. Am J Clin Nutr. 86:1286-92 (2007).
3. Cani et al. Metabolic endotoxemia initiates obesity and insulin resistance. Diabetes. 56:1761-72 (2007).

EXTRACELLULAR SUPEROXIDE DISMUTASE (EC-SOD) PROTECTS AGAINST ATHEROSCLEROSIS AND SUPPORTS HIPPOCAMPAL NEUROGENESIS

EC-SOD May Be Induced by Natural Hypo­methylation Agents Such as Curcumin and EGCG

Extracellular superoxide dismutase (EC-SOD) is part of the superoxide defense system present in the vascular wall that plays an important role in normal redox homeostasis by regulating superoxide activity there, for example, by reducing atherogenic superoxide oxidation of low density lipoprotein (LDL). A new paper¹ now reports that EC-SOD can be induced by 5-azacytidine, a DNA methyltransferase inhibitor commonly used in experimental research. Curcumin (along with other curcuminoids, major active components of turmeric root) and EGCG have also been identified as DNA methyltransferase inhibitors (hypomethylation agents)^1A,1B and, hence, might also be able to induce EC-SOD activity.

EC-SOD has become the subject of increasing numbers of research publications as a result of its important anti-inflammatory action against superoxide in the vascular system. In fact, EC-SOD acts as an important anti-inflammatory and anti-oxidative stress agent in monocytes and macrophages that are observed to accumulate in atherosclerotic plaques and to act as promoters of atherosclerosis.¹

A large literature is evolving to explain the regulation of genetic expression via epigenetics, a modification of histone proteins associated with DNA that alters the histones without changing the chemical structure of the DNA itself. The researchers¹here observed changes in DNA methylation in the promoter and coding regions of EC-SOD that “suggest the strong possibility that DNA methylation within promoter and coding regions might regulate cell-specific expression of EC-SOD in human monocytic cells.”¹

Another epigenetic modification, histone acetylation, is also involved in gene regulation. Here the researchers studied the effects of histone acetylation/deacetylation, mediated by histone acetyltransferase (HAT) and/or histone deacetylase (HDAC) on EC-SOD expression. They found that TPA (12-0-tetra-decanoylphorbol-13-acetate) treatment significantly decreased the expression of EC-SOD in U937 cells but increased it in THP-1 cells, determining the effect of histone modification, especially H3 and H4 acetylation, in TPA-inducible EC-SOD expression in THP-1 cells.

The most interesting findings here were the ones related to increased EC-SOD expression by decreasing methylation of its promoter and coding regions because of the possibility of using easily available and safe natural products curcumin (found in turmeric root with related curcuminoids) and EGCG as hypomethylating agents. As the authors^1® explain, “the presence of a high level of EC-SOD throughout the vessel walls can contribute to the regulation of nitric oxide-derived vasodilation by preventing the formation of peroxynitrite.” Moreover, they say, “it has been shown that EC-SOD protects arteries from ischemia-induced cytotoxicity and reduces infarct sizes.” Note that hydrogen selectively destroys peroxynitrite.

EC-SOD Promotes Long-Term Survival of Newborn Neurons

A paper from last year² reported that EC-SOD was important in a mouse model of neurogenesis by comparing mice in which EC-SOD was either knocked out, was normal in wild type (WT) mice, or was overexpressed in other mice. EC-SOD was important in progenitor cell proliferation, dendritic development, and long-term survival of newborn neurons. The study also “suggested that maintenance of the dendritic system following cranial irradiation was important for preservation of neurocognitive functions.”²

In a 2013 review paper,³ researchers noted that “… increasing SOD3 [EC-SOD] levels in various experimental disease models, e.g., chemically induced diabetes, hypertension, and inflammatory arthritis, reduced oxidative stress and improved disease state, thereby placing SOD3 as a central therapeutic target.”

References
1. Kamiya et al. Epigenetic regulation of extracellular-superoxide dismutase in human monocytes. Free Radic Biol Med. 61:197-205 (2013).
1A. Liu et al. Curcumin is a potent DNA hypomethylation agent. Bioorg Med Chem Lett. 19:706-9 (2009).
1B. Fang et al. Tea polyphenol (-)-epigallocatechin-3-gallate inhibits DNA methyltransferase and reactivates methylation-silenced genes in cancer cell lines. Cancer Res. 63:7563-70 (2003).
2. Zou et al. Extracellular superoxide dismutase is important for hippocampal neurogenesis and preservation of cognitive functions after irradiation. Proc Natl Acad Sci USA. 109(52):21522-7 (2012).
3. Frank et al. Targeting the redox balance in inflammatory skin conditions. Int J Molec Sci. 14:9126-67 (2013).

GENE EXPRESSION PROFILING REVEALS SOME DIFFERENCES BETWEEN SODIUM SELENITE, SELENOMETHIONINE AND YEAST-DERIVED SELENIUM FOLLOWING INGESTION IN THE MOUSE

Gene expression profiling has become a valuable tool in uncovering mechanisms of action of genes and nutrients that affect their expression. In this new study,¹ scientists examined the effects of three forms of selenium used to supplement a base diet at 1 mg/kg diet of added selenium as compared to the base diet deficient in selenium (<0.01 mg/kg diet). The subjects were weanling mice. They received either the base diet without added selenium or the selenium supplemented base diet for 100 days.

The three forms of selenium were reported to be equally effective in activating standard measures of selenium, such as expression of genes encoding selenoproteins. The gene expression profile revealed that sodium selenite and yeast-derived selenium had similar profiles and different from that of selenomethionine. The most interesting/revealing result of the study was that sodium selenite changed the expression of far more genes than the other two forms in the cerebral cortex and gastrocnemius (muscle) and slightly more genes in the intestine than the other two forms.

In the liver, the yeast-derived selenium changed the expression of more genes than the other two forms. For example, in the gastrocnemius, sodium selenite changed the expression of 1082 genes while yeast-derived selenium changed the expression of 637 genes and selenomethionine changed the expression of 148 genes. In the cerebral cortex, the genes changed were 1166, 832, and 421 for selenite, yeast-derived selenium, and selenomethionine. We were quite surprised, wondering why there would be such a large difference in the number of expressed genes (particularly in muscle). Moreover, the DNA damage response gene Gadd45b had a lowered expression in response to yeast-derived selenium in all tissues, while sodium selenite reduced its expression in cortex and gastrocnemius, and selenomethionine reduced its expression only in cortex. The authors also point out that the overlap of changes in gene expression that were changed in the same direction was greater for sodium selenite and yeast-derived selenium than any other pair of treatments. There is a complete list of the individual genes changed in expression by more than one diet for all four tissues in Online Resources 1–4, which we have not reported here.

The authors point out that the results of this study clearly show that research based upon different forms of selenium cannot be considered equivalent and that “the published data regarding the effect of selenium should be re-evaluated with respect to the source of selenium that was administered.”¹

Note that the most common form of natural dietary selenium is sodium selenite. Importantly, the prostate cancer killing effect of selenium was strongest for sodium selenite as reported in a recent paper.²

References

1. Barger et al. Gene expression profiling reveals differential effects of sodium selenite, selenomethionine, and yeast-derived selenium in the mouse. Genes Nutr. 7:155-165 (2012).
2. Olm et al. Extracellular thiol-assisted selenium uptake dependent on the xc- cystine transporter explains the cancer-specific cytotoxicity of selenite. Proc Natl Acad Sci USA. 106(27):11400-5 (2009).

PROTECTIVE EFFECTS OF ASTAXANTHIN ON CAPILLARY BLOOD FLOW IN MUSCLES OF MOUSE MODEL OF COUCH POTATOES

Despite the proven benefits of exercise, there are still an awful lot of people who don’t exercise, including those who don’t think they have the time or others that simply don’t enjoy exercising. For those people (and we include ourselves among those who would rather spend our time reading, writing, or thinking than doing boring exercises), protecting our muscles from atrophy due to being sedentary is an important priority in maintaining good health. A new paper¹ reports protective effects against decreased capillary blood flow in the muscles of rats as a result of disuse by the carotenoid astaxanthin.

The researchers explain that capillary regression in skeletal muscles is associated with decreased muscular activity as a result of increased oxidative stress and this results in reduced blood flow in the affected muscles. The researchers hypothesized that astaxanthin, a powerful antioxidant that is stronger than vitamin E under a number of stress conditions, might reduce this oxidative stress and ameliorate the capillary regression associated with muscle disuse.

They used a common experimental model of muscular disuse where the hindlimbs of the rats were suspended by the tail so that there was no weight bearing on the hindlimbs and, hence, those muscles were not subject to contractions that activate pathways induced by exercise. The effects are very similar to extended bedrest. The researchers explain that these “unloaded” hindlimbs have been shown to have lower levels of VEGF (vascular endothelial growth factor) that supports the blood vessels that, in active muscles, maintain blood flow.

The results showed that the VEGF protein level was lower in the hindlimb suspended group than in both control groups (in one of the control groups, rats were not hindlimb suspended and in the other control group, rats were not hindlimb suspended but did receive astaxanthin supplementation), but that the VEGF protein level in the hindlimb suspended rats supplemented with astaxanthin, the VEGF level was not different from the two control groups. In fact, the authors found that all angiogenic (blood vessel growth promoting) factors, except ANG-2, were higher in the the hindlimb suspended + astaxanthin group than the hindlimb suspended (no astaxanthin) group. The decreased angiogenic factors in the hindlimb suspended group that did not receive astaxanthin resulted in regression of capillaries.

In this study, astaxanthin treatment prevented the decrease in capillary volume and shift towards smaller diameter capillaries that occurred in the disused hindlimb muscles but did not prevent the decrease in muscle mass that resulted from the decreased hindlimb loading. Therefore, the mechanisms responsible for the decreased capillary capacity and the decreased muscle mass must be different.

Though the researchers did not examine the effects of astaxanthin on blood vessel capacity in the heart in this experiment, it would be of considerable interest to see whether astaxanthin would help maintain capillary volume in the heart of sedentary animals or people. Exercise is known to provide beneficial results to heart capillary blood flow (collateral blood flow) and this may very well be due to similar angiogenic factors as astaxanthin maintained in the hindlimb suspension model.

Reference

1. Kanazashi et al. Protective effect of astaxanthin on capillary regression in atrophied soleus muscle of rats. Acta Physiol (Oxf). 207:405-15 (2013).

ASTAXANTHIN IMPROVES SWIMMING ENDURANCE IN MICE AND DECREASES FAT ACCUMULATION, POSSIBLY BY INCREASING UTILIZATION OF FAT AS AN ENERGY SOURCE

The carotenoid astaxanthin has been reported to increase endurance capacity in male mice exercised to exhaustion.¹ The mice were supplemented with astaxanthin by stomach intubation in doses of 1.2, 6, or 30 mg/kg body weight 5 days a week for 5 weeks or they received vehicle (olive oil). In another experiment, the mice received the same amount of astaxanthin or vehicle by the same route and had a weight attached to their tails, swimming to exhaustion. In experiment 3, the mice had to swim for a predetermined amount of time and were assessed for blood concentrations of exercise-induced metabolites.

In the control group, plasma glucose was decreased by 15 minutes of swimming exercise, but in the astaxanthin groups (receiving 6 mg/kg or 30 mg/kg) the plasma glucose was significantly higher than in the control group. This may suggest that the animals receiving astaxanthin used more fat as an energy fuel, thus sparing glycogen. The researchers in fact observed significantly higher levels of liver and muscle glycogen in the astaxanthin groups than in the control group after swimming for 15 minutes. That, and the lower levels of blood lactate in the astaxanthin groups also suggested a decrease in glucose utilization.

Reference

1. Ikeuchi et al. Effect of astaxanthin supplementation on exercise-induced fatigue in mice. Biol Pharm Bull. 29(10):2106-10 (2006).

COENZYME Q10 MAY BE AN EFFECTIVE
TREATMENT FOR OSTEOPOROSIS

A new paper¹ reports findings for the first time on the beneficial effects of CoQ10 on bone, in which it enhanced bone-forming osteoblast differentiation while inhibiting osteoclast (bone resorbing cells) differentiation. These findings, while discovered in in vitro cell culture experiments, could be of particular value to postmenopausal osteoporosis that develops as a result of estrogen deficiency if the effects prove to work similarly in vivo.

At a dose of 100 μM, CoQ10 decreased the number of osteoclasts in cell culture to approximately 87.3% of controls (didn’t receive CoQ10).

The CoQ10 was found to work by its effects on specific bone signaling pathways. For example, RANKL (receptor activator of nuclear factor kappa B ligand) is released by preosteoblasts, which activates its receptor RANK that is expressed on osteoclasts with the monocyte/macrophage colony-stimulating factor (M-CSF) for osteoclastogenesis. Osteoclasts are bone cells that demineralize bone, hence decrease bone mineral density. The inhibition of the RANKL-induced osteoclast differentiation is one of the mechanisms for CoQ10 increase of bone formation. Another natural product that is reported to suppress RANKL-induced osteoclast differentiation is alpha lipoic acid.¹

CoQ10 increased osteoblast differentiation by enhancing alkaline phosphatase (ALP) activity. “CoQ10 enhanced not only early osteoblastic biomarkers like ALP and Col1, but also late osteoblastic biomarkers such as BSP and matrix minralization through transcription factors Runx2 and OSX. In this way, CoQ10 acts as an enhancer for all stages of osteoblast differentiation.”¹

The researchers suggest that further study be done to determine what the optimal dose of CoQ10 for bone formation should be. In the meantime, CoQ10 is quite safe. Sandy, for example, takes 100 mg. CoQ10 a day.

The authors sum up the results of their study: “… CoQ10 may have great therapeutic implications in treating osteoporosis and other bone diseases.”

Reference

1. Moon et al. Coenzyme Q10 regulates osteoclast and osteoblast differentiation. J Food Sci. 78(5):H785-891 (2013).

SURPRISE! KAPPA OPIOID RECEPTORS MEDIATE THE
ANTIDEPRESSANT-LIKE ACTIVITY OF HESPERIDIN IN MICE

Who would have thought it? That hesperidin could exhibit anti-depressant effects in mice subjected to the forced swim test and do so by interacting with the kappa opioid receptor. But that’s one thing about biomedical science—there are always lots of surprises.

The kappa opioid receptor is one of the three currently known opioid receptors, that also include mu and delta opioid receptors. Hesperidin, a naturally occurring flavonone glycoside, is found in rich quantities in citrus fruits and has been reported to have antioxidant, neuroprotective, and anticancer effects. An earlier paper² reported that in traditional Mexican medicine, infusion of flowers of the Citrus genera are used as a sedative to treat insomnia. This paper² explored the sedative effects of Citrus; the authors reported that, in an earlier study, hesperidin in the methanol extract of Citrus sinensis was identified as the sedative active principle of the plant. Others had described the involvement of opioid receptors to the behavioral and antinociceptive (anti-pain) effects of hesperidin.

The immobility period of the mice subjected to the forced swim test (freezing as a result of fear) was significantly reduced by treatment with hesperidin at doses of 0.1, 0.3, and 1 mg/kg.¹ Pretreatment with naloxone (a nonselective opioid receptor antagonist) reversed the antidepressant-like effect of hesperidin in that study.

References

1. Filho et al. Kappa-opioid receptors mediate the antidepressant-like activity of hesperidin in the mouse forced swimming test. Eur J Pharmacol. 698:286-91 (2013).
2. Guzman-Gutierrez and Navarrete. Pharmacological exploration of the sedative mechanism of hesperidin identified as the active principle of Citrus sinensisflowers. Planta Med. 75:295-301 (2009).

NEGATIVE EFFECT OF MARKETS ON MORAL VALUES

Of Mice And Men And Markets

A new paper studied the effect of markets on moral values by setting up a moneymaking experiment in trade whereby it would cost people money to avoid harms to third parties. The researchers found that the more the number of people involved in a market trade, the less money people would forego to avoid harms to third parties. They concluded, therefore, that markets had a negative effect on moral principles.

The experiment went like this: a buyer and seller, each in possession of a mouse, could divide a sum of money and keep the proceeds. (The mouse was left over from an experiment and, in the usual course of events, would be euthanized at that point, but for the sake of this experiment would be allowed to live if the owner of the mouse were willing to forego a certain amount of a sum of money (the price for allowing the mouse to live).) The idea was to see what effect being in a moneymaking trade arrangement, a market, would have on the price people were willing to pay to allow the mouse to live.

The experimenters obviously considered it moral to be willing to forego some money to keep the mouse alive, but immoral to simply keep the money and let the mouse die. (As part of the experiment, the participants watched a short movie of how a mouse would die, so nobody had any illusions. For the mouse, the outcome was a matter of life or death.) We would have preferred it if the researchers had called the decision to keep a mouse alive a personal value rather than a matter of morality, as we do not agree that allowing a mouse to die is immoral. (Such values are intensely subjective. In the Jaine moral system, for example, it would be considered immoral to kill even an ant.)

In the individual decision option, Option A, a mouse would be allowed to live but the participant would receive no money. Option B meant that the mouse would die but the participant would receive 10 euros. There were 124 participants in the individual decision option, of which 45.9% were willing to allow the mouse to die in order to get 10 euros. In the bilateral market (2 participants traded for prices), 72.2% of sellers were willing to let the mouse die for prices below or equal to 10 euros. This difference was statistically significant from the individual decision option. In the multilateral market (where there were 7 bidders), 75.9% of sellers were willing to kill a mouse for less than or equal to 10 euros. This, too, was statistically significantly different from the individual decision option.

The more bidders, the lower the average price offered in exchange for allowing the mouse to live. (In a separate experiment, people could either keep and spend a coupon or take a certain amount of money. The effects of numbers of bidders in this condition revealed no significant price trend.) The authors conclude, therefore, that the market had the effect (and the larger the market, the greater the effect) of reducing the price paid for a moral outcome (allowing the mouse to live).

The authors sum it up: “We therefore agree with the statement quoted at the beginning that we as a society have to think about where markets are appropriate—and where they are not.”

We offer the following comments:

Power Law

The decreasing price paid as the number of bidders increases looks a lot like a power law, which is a characteristic of self-organizing systems as described in the book Ubiquity by Mark Buchanan (Three Rivers Press, 2000). As explained in Ubiquity, for example, if you look at a sandpile on which grains of sand are being dropped one by one, eventually at some point you will have an avalanche. If you double the amount of energy released by the avalanche, the avalanche becomes four times less likely. You see the same pattern with earthquakes: the larger the earthquake, the less likely that size event will occur, consistent with the mathematical power law you see with avalanches.

Thus, we propose that the higher the number of bidders in the market, the lower the price paid to avoid harms to third parties, is a power law, something that is mathematically built into markets as self-organizing systems. It is not that people become less moral as the number of bidders increases; it is that the likelihood of the decision of each individual determining the average price becomes less and less as the number of bidders increases.

Political Decision Making vs. Markets

The researchers wonder, as indicated in their summation, whether under some circumstances decisions should be made by a different mechanism than markets. Whether a political process would result in a more “moral” outcome than a market is we believe questionable. The more voters, the less likely that your decision will affect the outcome of an election and, hence, the less likely that voters will concern themselves with harms to third parties. Moreover, the less likely that your decision will decide the outcome of an election, the less likely you will have an incentive to be well informed before you vote, the “rational ignorance” problem. The authors note that, “it is a pervasive feature of market interaction to impose costs on uninvolved third parties.” We agree. Yet, there can be no doubt that the production of externalities (costs to uninvolved third parties) is a pervasive feature of political processes.

Moreover, there may be costs in the use of politics as a way of making decisions by large numbers of people that go beyond that of markets. Even as the authors themselves noted, not all the participants chose to take part in the bidding of their experimental market.* In politics, you can refuse to vote but the outcome of the election (the decisions of other people) will be imposed upon you anyway, whereas refusing to enter the bidding in a market transaction allows you to opt out, at least in part, of the costs of whatever other people decide. In the usual trade, opting out means you can keep your money and use it for something else.

 

---

* “22.2% of all traders in the bilateral market never traded, while in the multilateral market, 24.1% never traded. … 27.1% of subjects [in the individual decision option] were unwilling to kill their mouse even for the maximum offered monetary amount of 50 euros.”

 

Reference

1. Falk and Szech. Morals and Markets. Science. 340:707-11 (2013)

APPETIZERS

The problem with doing nothing is not knowing when you’re finished.
— Benjamin Franklin

Time is what keeps everything from happening at once.
— Ray Cummings, The Time Professor (1921)

If any student comes to me and says he wants to be useful to mankind and go into research to alleviate human suffering, I advise him to go into charity instead. Research wants real egotists who seek their own pleasure and satisfaction, but find it in solving the puzzles of nature.”
— Albert Szent-Gyorgyi

And they began to deal with one another as foreigners.
— Montesquieu, The Greatness of the Romans and Their Decline

I’m not here as a serf or a vassal. I’m not begging my lords for mercy. I’m a born-free American woman, wife, mother, and citizen. And I’m telling my government that you’ve forgotten your place.
— Becky Gerritson, founder and president of the Wetumpka Tea Party in testimony before the House Ways and Means Committee 6/4/2013 on IRS abuses against her group

The important thing in science is not so much to obtain new facts as to discover new ways of thinking about them.
— Sir William Lawrence Bragg

FOR THOSE USING METFORMIN, GOOD NEWS

BETTER RESULTS WHEN METFORMIN IS COMBINED WITH THE PREBIOTIC OLIGOFRUCTOSE THAN WHEN EITHER IS USED ALONE IN THE TREATMENT OF OBESITY—A RAT STUDY

A defect in glucagon-like peptide 1 (GLP-1) secretion, a hormone that increases insulin secretion and induces satiety, is believed to be one of the causes of obesity. Overeating is known to blunt the GLP-1 response to food ingestion, whereas prebiotics can improve the response to the hormone. Both oligofructose (a form of the prebiotic inulin) and metformin are known to improve GLP-1 sensitivity. Researchers decided to test whether a combination of these two treatments would result in a greater improvement in obese rats on a high fat high sucrose diet as compared to the result of treatment by metformin or oligofructose alone. They assessed the results by measuring peripheral glucose clearance through the phosphorylation (activation) of AMPK, decreasing gluconeogenesis (glucose release by the liver, which is supposed to be inhibited by insulin but in diabetics is not effectively suppressed) and increasing peripheral glucose uptake in liver and skeletal tissue.

The researchers observed a decrease in energy intake, percent body fat, and blood glucose with both oligofructose and metformin. The combination resulted in improvement as compared to the individual treatments.

“The interaction between OFS [oligofructose] and MET [metformin] affected fat mass, hepatic TG [liver triglycerides], secretion of glucose-dependent insulinotropic polypeptide (GIP) [a satiation hormone] and leptin, and AMPKalpha2mRNA and phosphorylated acetyl CoA carboxylase (pACC*) levels (p<0.05). “The rats in the OFS group had lower fat mass than the control rats, and rats in the MET group and the OFS+MET group had lower fat mass than those in the control and OFS groups. Thus, MET or the combination of OFS + MET resulted in a superior outcome by reducing fat mass as compared to that of OFS alone, while OFS alone reduced fat mass as compared to controls.

Thus, the authors conclude, metformin (already the most utilized treatment for type 2 diabetes) when combined with oligofructose has “the potential to improve metabolic outcomes associated with obesity.”

This is very good news for those taking metformin for diabetes, obesity, or obesity-related metabolic defects (such as insulin resistance), assuming that the metformin-oligofructose combination works similarly in people as it did in the obese rats. (Current data imply that it does.) Simply take supplemental oligofructose (we use long chain inulin) 2 or 3 times a day along with your usual dose of metformin.

* “ACC, the downstream target of AMPK … once phosphorylated, increases oxidation and suppresses fatty acid synthesis … The hepatic [liver] pACC [phosphorylated ACC] level was higher in rats in the OFS + MET group compared to all other groups.”

Reference

1. Pyra et al. Prebiotic fiber increases hepatic acetyl CoA carboxylase phosphorylation and suppresses glucose-dependent insulinotropic polypeptide secretion more effectively when used with metformin in obese rats.J Nutr.142:213-220 (2012).

METFORMIN INCREASES LIFESPAN OF C. ELEGANS VIA ITS EFFECTS ON THE C. ELEGANSRESIDENT BACTERIUM

The implications of this finding (assuming that it is verified) could be enormous when considering the human-microbiota system as a whole. Germ-free rodents weigh less than rodents with a normal gut microbiota, which is believed to be due to decreased extraction of nutrients from food, a form of dietary restriction. Also, rodents with a normal microbiota given a microbiota transplant from fat rodents themselves become obese on a high fat diet. The implication is that, just as the C. elegans gets less nutrients from its E. coli microbiome as a result of metformin’s antibiotic effect on the E. coli, metformin might (by a similar antibiotic effect) reduce the nutrients derived from food (and thus made available to us) by our normal gut microbiota. Indeed, metformin induces weight reduction, among other things, in diabetics that receive it as a treatment.We may be on the verge of a new expanded analysis of life extension that includes the effects of the microbiota living within us starting with a new paper¹ in Cell. There, the researchers found that metformin, said to be the most commonly used treatment for diabetes worldwide, extended lifespan in the nematode Caenorhabditis elegans by acting as an antibiotic that altered the metabolism of the E. coli that are the worm’s resident bacterium. The E. coli, the primary food source of the worm, had a decreased production of nutrients such as folate and methionine as a result of the antibiotic effect of metformin, resulting in nutritional restriction in the worm and THAT was what increased the lifespan of the worm. If this analysis is correct, then the lifespan effects of metformin on C. elegans have to be considered from the point of view of both C. elegans and its microbiome.

Interestingly. “[i]t was recently discovered that C. elegans live longer on an E. colimutant with reduced folate levels (aroD). Moreover, metformin can decrease folate levels in patients.”¹ The authors draw no conclusions from these curious facts, which may or may not be relevant to the effect of metformin in humans. We know of no studies in humans that have provided evidence for a life extending effect of metformin. Humans have such a long lifespan that such studies would be impractical. However, studies of the gene modulating effects of metformin in humans as compared to gene microarrays showing changes of gene expression with age that occur in the absence of metformin supplementation can be used to infer anti-aging gene expression changes with metformin.

It was noted by the authors of the C. elegans-metformin paper in Cell¹ that, while metformin activates AMPK (one of its antidiabetic effects), it does not increase lifespan in Drosophila and suggest that this might reflect the presence of metformin-resistant microbiota in Drosophila. This prediction shouldn’t be difficult to test.

A recent paper² reported that, using a sophisticated analysis of 184,094 sequences of the microbial rRNA genes from 9 individuals, 3 in each of the categories of normal weight, morbidly obese, and post-gastric-bypass surgery, they “detected significantly higher numbers of H2 [hydrogen]-utilizing methanogenic Archaea in obese individuals than in normal weight or post-gastric-bypass individuals.” These researchers therefore propose that “interspecies H2 transfer between bacterial and archaeal species is an important mechanism for increasing energy uptake by the human large intestine in obese persons.”² One implication of this paper’s results would be that reducing the number of methanogens in the gut would be a way to decrease energy uptake for the purpose of reducing weight.

In another paper,³ researchers tested the hypothesis that modification of the gut microbiota with two antibiotics could cause antidiabetic effects, e.g., improvement in whole body glucose tolerance. They thought that this might be the case because of prior work that showed that (for example) an infusion of a low dose of lipopolysaccharide, a component of gram-negative cell wall, leads to excessive weight gain and insulin resistance in mice concommitant with the production of low-grade inflammation. In their study,³ they found that treatment of the ob/ob diabetic mouse and diet-induced obese mice with a combination of two antibiotics, norfloxacin and ampicillin, improved whole body glucose tolerance and reduced liver accumulation of fat.

References

1. Cabreiro et al. Metformin retards aging in C. elegans by altering micorbial folate and methionine metabolism. 153:228-39 (2013).
2. Zhang et al. Human gut microbiota in obesity and after gastric bypass. Proc Natl Acad Sci USA.106(7):2365-70 (2009).
3. Membrez et al. Gut microbiota modulation with norfloxacin and ampicillin enhances glucose tolerance in mice. FASEB J.22:2416-2426 (2008).

 

GLYCINE PROTECTS BRAIN TISSUES UNDERHYPOXIC (OXYGEN DEFICIENT) CONDITIONS

Glycine is being used as a therapy for ischemic stroke² and has been reported to increase the lifespan of cortical neurons under hypoxic conditions.³ A new paper¹ now explains a mechanism underlying this protective effect that has advanced glycine to patient treatment in the clinic. Hypoxia in brain tissues was induced in rats by ligation of the common carotid artery. The resulting hypoxia reduced respiratory control in brain cortex mitochondria from 7.7 ±0.5 to 4.5 ±0.3, indicating impaired respiration. Preliminary oral administration of glycine at 40 mg/kg 4 times with 1 hour intervals almost completely prevented this decrease.The amino acid glycine, which among other things, is part of the regulatory pathways for sleep and has been shown to improve sleep quality, has been reported in a recent paper¹ to protect the brain against energetic disturbances arising under hypoxic (oxygen deficient) conditions. We think that glycine may be a useful supplement for neurological protection in those with sleep apnea.

The authors¹ report that published data “suggest that hypoxia is associated with activation of peroxidation processes in the brain tissue.” The authors, therefore, measured hydrogen peroxide accumulation in mitochondria after short-term exposure to hypoxia and found that the oxidation of succinate was accompanied by a rapid generation of hydrogen peroxide. Hydrogen peroxide formation was significantly lower after incubation of brain cortex slices with glycine. Oxidative phosphorylation (energy generation in mitochondria) was preserved in mitochondria in the presence of 5 mM glycine in the incubation medium. The researchers suggest, therefore, that mitochondria are the target of glycine in the brain cortex under hypoxic conditions. This is particularly interesting because of the fact that substances, such as many antioxidants, that are protective under the oxidative conditions induced by hypoxia frequently are unable to enter mitochondria and are, therefore, unable to provide much, if any, protection there. (Hydrogen is one antioxidant that does reach mitochondria.)

References

1. Selin et al. Mechanism underlying the protective effect of glycine in energetic disturbances in brain tissues under hypoxic conditions. Bull Exp Biol Med. 153(1):44-7 (2012).
2. Gusev et al. Neuroprotective effects of glycine for therapy of acute ischaemic stroke. Cerebrovas Dis.10(1):49-60 (2000).
3. Zhao et al. GABA and glycine are protective to mature but toxic to immature rat cortical neurons under hypoxia. Eur J Neurosci.22(2):289-300 (2005).

CHRONIC INFLAMMATION IMPAIRS ADULT HIPPOCAMPAL NEUROGENESIS PROTECTION AGAINST RADIATION DAMAGE BY HYDROGEN

The researchers also studied the effect of LPS (a bacterial cell wall component that induces a strong immune response) injected into adult female rats to produce an inflammatory cascade. They measured a 240% increase in the density of activated microglia in the dentate gyrus that was accompanied by a 35% decrease in hippocampal neurogenesis. This effect was completely blocked by systemic treatment with indomethacin. (Indomethacin under normal conditions had no effect on neurogenesis.)Using a cranial radiation injury model in adult rats, researchers found that using a dose of 10 Gray cranial irradiation with x-rays, the radiation spared roughly 30% of the neuron progenitor cells proliferative capacity but completely eliminated the production of neurons (neurogenesis).¹ As the scientists reported, “irradiation caused a striking inflammatory response characterized by the persistence of activated microglia relative to the minimal levels in normal control animals.” “If inflammation were the primary cause of the lack of neurogenic signaling within the dentate subgranule zone (SGZ), then inflammatory blockade would be expected to restore neurogenesis. Treating the irradiated animals with indomethacin (a non-steroidal antiinflammatory drug) during and after radiation exposure “partially restored the relative proportion of proliferative cells adopting a neuronal fate relative to untreated, irradiated animals (37% versus 15%, respectively).”¹

The scientists found that activated microglia produced potent inflammatory cytokines including IL-1beta, TNF-alpha (tumor necrosis factor alpha), interferon gamma, and IL-6.¹ Progenitor cells were allowed to differentiate in the presence of each of these cytokines with the result that IL-6 or TNF-alpha decreased in vitro neurogenesis by about 50%, whereas under these test conditions IL-1beta and interferon gamma had no significant effect.

This may be a mechanism by which NSAIDs such as indomethacin protect against Alzheimer’s disease, which does involve inflammation. It has been known for many years that there is a negative correlation between the use of NSAIDs and the incidence of Alzheimer’s.

Another paper² also reports increased inflammation and impaired neurogenesis as a result of exposure to radiation.

Radiation Protection By Hydrogen

Hydrogen is an effective radioprotective antioxidant,^2A as it potently scavenges hydroxyl radicals, which are the source of most tissue damage caused by radiation. Of particular importance, too, is the fact that hydrogen reaches mitochondria, thus providing protection against hydroxyl radicals there as well as other areas of irradiated tissue. As reported in a paper on targeted delivery of radioprotective agents to mitochondria,³mitochondria are the principle sources of the reactive oxygen and nitrogen species, especially hydroxyl radicals and peroxynitrite, that are importantly involved in irradiation-induced apoptosis (programmed cell death); hydrogen is a potent scavenger of both hydroxyl radicals and peroxy­nitrite.

Another natural product providing protection against the oxidative stress resulting from radiation is epicatechin⁴ (found in cocoa, grapes, tea, and apples).

References

1. Monje et al. Inflammatory blockade restores adult hippocampal neurogenesis. Science.302:1760-5 (2003).
   2. Rola et al. High-LET radiation induces inflammation and persistent changes in markers of hippocampal neurogenesis. Radiat Res.164:556-560 (2005).
   2A. Qian et al. Radioprotective effects of hydrogen in cultured cells and mice. Free Rad Res. 44(3):275-82 (2010).
   3. Zabbarova and Kanai. Targeted delivery of radioproective agents to mitochondria. Mol Interv. 8(6):294-302 (2008).
   4. Sinha et al. Epicatechin ameliorates ionising radiation-induced oxidative stress in mouse liver. Free Rad Res. 46(7):842-9 (2012).

ALPHA LIPOIC ACID PROTECTS AGAINST GASTRIC MUCOSA
DAMAGE CAUSED BY NON-STEROIDAL-ANTIINFLAMMATORY
DRUGS: INDOMETHACIN

As we have written before, the amino acid L-arginine taken at the same time as a non-steroidal antiinflammatory drug (NSAID) helps prevent the damage to the gastric mucosa that these drugs cause. A recent paper¹ now reports that alpha lipoic acid (ALA), an important natural antioxidant, is also protective against gastric mucosa damage by interfering with the prooxidant effects of NSAIDs, that includes lipid peroxidation and interference with the antioxidant systems of the mucosa. ALA is known to be an effective antioxidant because it can bind free radicals directly or indirectly and enhances the effectiveness of other antioxidants.¹

In the study, scientists treated rats with the NSAID indomethacin (IND) five minutes after receiving one of seven doses of ALA (50, 75, 100, 150, 200, and 300 mg/kg). Controls got water to a similar amount as that received by the experimental animals. Another group received IND without ALA. “All doses of ALA showed gastro­protective effect against IND-induced gastric damage in rats.”¹ In fact, the 50, 75, 100, 150, 200, and 300 mg/kg doses of ALA reduced the ulcer areas at rates of 88.2%, 89.9%, 91.4%, 93.5%, 95.1%, and 96.1%, respectively compared to the IND only group.

The researchers found that ALA increased the levels of glutathione, glutathione S-transferase, and superoxide dismutase, while decreasing lipid peroxidation, myeloperoxidase activity, and catalase in gastric tissues.¹

Incidentally, if you have ever wondered why protective substances such as ALA and L-arginine are not included in tablets or capsules of NSAIDS, it is all due to the regulations of the FDA. You’re not allowed to do it and you would also not be able to inform product users of the beneficial effects (reduced damaging effects to the stomach lining) even if you could add these protective nutrients. Just another example of how the FDA literally kills people—in fact, at least 16,500 Americans die every year of gastric ulcers brought on by the use of NSAIDS. Does this make the FDA an agency committing crimes against humanity? You bet. The FDA is far more dangerous to Americans than Al Qaeda.

Reference

1. Kaplan et al. Alpha-lipoic acid protects against indomethacin-induced gastric oxidative toxicity by modulating antioxidant system.J Food Sci. 77(11):H224-30 (2012).

THE ROLE OF COMT IN IMPATIENCE:

CHOOSING BETWEEN A SMALL PAYOFF RIGHT AWAY OR
WAITING FOR A LARGER PAYOFF LATER

A molecular genetics study in 2007² identified a particular polymorphism (Val158Met) of the COMT gene as being associated with a steeper DD, that is, linked to a stronger preference for immediate as compared to later rewards. “The Val allele (compared to the Met allele) is associated with higher enzymatic activity and thus lower prefrontal cortex dopamine levels.”¹ The heritability of DD has been reported in a recent twin study to explain up to 50% of the individual differences in DD.¹A new paper¹ reports a reason why people differ in how much they prefer an immediate small payoff rather than waiting for a larger payoff later. This is called the discount rate and, as the researchers note,¹ has a large effect on whether one leads a healthy and successful life. They identify disorders such as substance abuse, pathological gambling, overeating, relationship infidelity, and Attention Deficit Disorder as examples of how impulsively choosing short-term rewards can often lead to costly consequences. Their new research1 focused on the effect of COMT (catechol-O-methyltransferase, a gene regulating the enzyme that degrades dopamine, thus controlling the level of dopamine especially in the prefrontal cortex) on the individual’s DD (delay discounting, the tendency to prefer immediate over future rewards

These researchers used an EEG technique to determine neural baseline (what the authors called a “neural fingerprint”) to establish a stable resting EEG, which they could use for observing COMT influences on the neural baseline activation. The seventy-three healthy Swiss men (mean age = 25.7 years, SD = 5.0 years) had their EEGs recorded in a protocol of 20 seconds eyes open followed by 40 seconds eyes closed, repeated five times. Only data from the eyes closed condition were analyzed. Then they had buccal epithelial cells collected for DNA.

A greater number of Val alleles was associated with a lower level of baseline activation in the left dorsal prefrontal cortex (DPFC), which in turn leads to a steeper DD (greater preference for immediate as compared to delayed rewards), which the authors called “impatient choice.”

An increase in EGCG/green/white tea consumption by a population might well have very substantial long-term effects on the general level of prosperity by increasing savings and investment versus immediate consumption gratification. The populations of both China and Japan save and invest a much higher percentage of their incomes than Americans; perhaps their higher levels of tea drinking has something to do with this.As the paper described in the article before this one explains, EGCG has been found to be a natural inhibitor of COMT, decreasing the degradation of L-Dopa, which increases the availability of L-Dopa to enter the brain to be converted to dopamine. This is consistent with the results of other studies that have reported a reduced risk of Parkinson’s disease in those who drink tea regularly. (Green and white teas contain much higher levels of catechins, including EGCG, as compared to black tea.) It is therefore conceivable that tea drinking may affect delay discounting (DD) of individuals by modulating the activity of COMT to mimic the effects of having a COMT polymorphism that fosters a less impulsive DD.

References

1. Gianotti et al. Why some people discount more than others: baseline activation in the dorsal PFC mediates the link between COMT genotype and impatient choice. Front Neurosci.6:54. doi: 10.3389/fnins.2012.00054. (May 2012).
2. Boettiger et al. Immediate reward bias in humans: frontoparietal networks and a role for the catechol-O-methyltransferase 158(Val/Val) genotype. J Neurosci.27:14383-91 (2007).

BENJAMIN FRANKLIN ON POVERTY

Our new Constitution is now established, and has an appearance that promises permanency, but in this world nothing can be said to be certain except death and taxes …

I am for doing good to the poor, but I differ in opinion of the means. I think the best way of doing good to the poor is not making them easy in poverty, but leading or driving them out of it. In my youth I traveled much, and I observed in different countries, that the more public provisions were made for the poor, the less they provided for themselves, and of course became poorer. And, on the contrary, the less was done for them, the more they did for themselves, and became richer.
— Benjamin Franklin

OUT OF THE GOODNESS OF THEIR HEART

“In July 2003, the FDA began allowing qualified health claims for food in an effort to make it easier for food manufacturers to make health benefit claims for their products. Even if the scientific evidence in support of the claims are conclusive, the FDA has been helping the consumers to obtain accurate, up-to-date science based information about the health effect of these products.”

(Sorry, folks. This quote taken from Functional Foods, Nutraceuticals, and Degenerative Disease Prevention, edited by Paliyath, Bakovic, and Shetty, published by Wiley-Blackwell (2011) is not true. The reason the FDA began allowing qualified health claims is that they lost a Court decision (Pearson v. Shalala) that prohibited the agency from censoring truthful, non-misleading information on labels and in adverts for dietary supplements and foods. If they hadn’t lost this landmark decision, the FDA would still be censoring all truthful health claims in violation of the First Amendment’s guarantee of free speech.)

U.S. SUPREME COURT DECISION IN U.S. V. JONES: SEARCH
WARRANT REQUIRED FOR ATTACHING GPS DEVICE TO
CAR, DECIDED JAN. 23, 2012

In a decision limiting the government’s surveillance powers, a recent U.S. Supreme Court decision (majority opinion written by Scalia) determined that, on the basis of trespass as well as the expectation of privacy, attaching a GPS device to a vehicle was a search within the meaning of the Fourth Amendment at the time it was adopted and, hence, a warrant was required.

The Court agreed 9–0 that this was a search requiring a warrant. Perhaps most interestingly, however, there was a division on the Court as to the reason that this came under the Fourth Amendment’s prohibition on unreasonable searches and seizures. There was the majority opinion in which Scalia was joined by Roberts, Kennedy, and Sotomayer. Sotomayer also filed a separate concurring opinion, while Alito (joined by Ginsburg, Breyer, and Kagan) filed an opinion concurring in the judgment. The Alito concurrence did not agree on a trespass basis for the Fourth Amendment violation, but relied on the “expectation of privacy” argument. The big surprise, however, was the concurring opinion by Sotomayer, which agreed with the majority opinion on trespass and “expectation of privacy” arguments for a Fourth Amendment requirement for a warrant. Moreover, Sotomayer said, “I would also consider the appropriateness of entrusting to the Executive, in the absence of any oversight from a coordinate branch, a tool so amenable to misuse, especially in light of the Fourth Amendment’s goal to curb arbitrary exercises of police power to and prevent ‘a too permeating police surveillance.’”

Sounds as though Sotomayer has (maybe) declared her independence from Obama, at least with respect to the Fourth Amendment.

Though this SCOTUS decision favored Fourth Amendment protections against unreasonable searches and seizures, a more recent 5–4 decision did the opposite by allowing police to take DNA samples without probable cause or a warrant from people who have been arrested but not convicted of anything. Scalia was incensed by the decision and insisted on reading his dissent word for word before the entire court. BRAVO!

Coercively taking a DNA sample from a mere suspect is NOT like taking a photograph or fingerprints! It is forcibly taking a piece of your body containing your internal blueprint, which is a search or seizure going much farther than a mere warrantless search/seizure of your home. Moreover, it is far easier to salt a crime scene with an innocent person’s DNA than with fake fingerprints. Want to imprison someone you hate? Swab their driver’s side car door handle, smear it on a piece of child porn, and send it to the FBI …

Who Needs Data? Just Stick Your Finger in The Air

A study reported in the 1 Feb Science reveals that, when asked whether humans are causing climate change, Democrats were likely to say yes, Republicans likely to say no, but Independents were likely to say yes on unseasonably warm days but to say no on unseasonably cold days. No surprise but it just goes to show that very short term events rather than consideration of the totality of the evidence can be a deciding factor in how people perceive the possibility of human-induced global warming. This is an excellent way for those looking for the “right” response to a survey to plan when to ask people if they think humans are causing climate change.

HAPPINESS RESEARCH

THE EFFECT OF PERIPHERAL CIRCULATING PRO-INFLAMMATORY CYTOKINES AND PERCEIVED HAPPINESS IN MIDDLE-AGED ADULTS IN JAPAN

A recent study¹ reports on a possible connection between the level of circulating proinflammatory cytokines and perceived happiness, where higher amounts of one of these cytokines, interferon gamma, was correlated with lower levels of happiness.

Experiment 1 involved 160 healthy volunteers (77 males and 83 females, ranging in age from 19–40 years). The researchers screened the subjects for their perceived level of happiness by taking the Japanese version of the subjective happiness scale (JSHS), using the results to divide the group into high happiness respondents and low happiness respondents. Statistical analysis of the two groups found that the high happiness group had significantly higher scores on the Japanese translated version of the Short-Form 36 Health Survey in general health, vitality, emotional role, and mental health as compared to the low happiness group. “Interestingly, serum concentrations of IFNgamma [interferon gamma] were significantly lower in the high happiness group … than in the low happiness group.”

Experiment 2 involved 7 romantic couples; 7 males and 7 females aged 21–38 years. They were requested to answer a number of questions concerning their feelings of romantic love for their partners using the Passionate Love Scale (PLS), which contained questions such as “Sometimes I can’t control my thoughts; they are obsessively focused on __________.” “I would rather be with __________ than anyone else.” The participants were considered to have relatively passionate love relationships. The couples then spent 1 hour in a room with closed doors in which they could kiss and hug but not have intercourse. (This study gets more interesting the longer it goes on.) This was called a warm contact session, after which the couples were asked questions such as “Did you kiss and hug your partner very much?” and “Did you sense your partner’s love?” The results indicated that there was a lot of warm contact in the warm contact session.

Before the warm contact session, however, participants were requested to evaluate their present happiness and blood samples were drawn. Then, with 1 partner remaining in the closed room, the other partner was moved to a second room where he or she read a book (not a book about romance) for 1 hour with closed doors. After that, a second blood sample was drawn. There was at least a 2 week interval between the two conditions and the order of the two conditions was counterbalanced across couples. Finally (yes, there is a finally) the participants were asked to evaluate their feelings of happiness on a scale of 1 (not at all) to 7 (Yes, extremely): Do you feel happy at present?

The IFNgamma (interferon gamma) concentration decreased significantly after the warm partner contact but did not change in the control condition (reading a book). Moreover, the IFNgamma concentration was lower and health related QOL was higher in individuals with self-perceived high happiness than in those with self-perceived low happiness.

How Interferon Gamma Relates to Happiness

Interferon gamma is a pro-inflammatory cytokine that has been shown to reduce circulating serotonin levels by increasing the activity of indoleamine 2,3-dioxygenase, an enzyme that degrades tryptophan, hence reducing tryptophan available for the brain to convert to serotonin. Reduced brain serotonin levels can result in depression. “… previous studies indicated that circulating pro-inflammatory cytokine levels in individuals with depressive symptoms were higher than those in individuals with no depressive symptoms.”¹

The researchers also report that their previous study2 found that “the short form of the serotonin transporter gene-linked polymorphic region, which increases serotonin secretion from presynaptic neurons through reduced serotonin reuptake, also enhances amygdala response to desired persons. When IFNgamma levels decrease, the reactivity to positive stimuli increases, and consequently, the evocation of happiness is enhanced.”

The only question remaining, then, is what would have happened to the happiness score and the IFNgamma levels if the couples had been allowed to have intercourse behind those closed doors. We assume that followup experiments will tie up that loose end. In the meantime, we note that since the couples were left in privacy behind closed doors, there is no way to actually know whether they obeyed the instruction to not have intercourse. We leave it to the reader to consider the implications.

References

1. Matsunaga et al. Association between perceived happiness levels and peripheral circulating pro-inflammatory cytokine levels in middle-aged adults in Japan. Neuro Endocrinol Lett. 32(4):458-63 (2011).
2. Matsunaga et al. Genetic variations in the serotonin transporter gene-linked polymorphic region influence attraction for a favorite person and the associated interactions between the central nervous and immune systems. Neurosci Lett. 468:211-5 (2010).

GETTING L-DOPA INTO THE BRAIN

EGCG: SIGNIFICANT ADVANCE IN TREATMENT OF PARKINSON’S DISEASE

The most common treatment for Parkinson’s disease (PD) is a combination of levodopa (L-Dopa) and carbidopa. The L-Dopa serves as the precursor for production of dopamine in the brain where its deficiency (in the striatum) can result in severe movement disorders typical of PD. The carbidopa is a dopa decarboxylase inhibitor and is combined with the L-Dopa because it restricts the peripheral conversion of L-Dopa to dopamine, allowing the L-Dopa to enter the brain and be converted there where it is needed, while reducing the peripheral effects of increased dopamine, such as lowered blood pressure.

A new paper¹ now reports that taking EGCG along with L-Dopa and carbidopa results in a significantly increased beneficial effect of therapy by modestly inhibiting the methylation of L-Dopa by the enzyme catecholamine-O-methyltransferase (COMT), thus preventing the rapid conversion of L-Dopa by the liver to inactive 3-O-methyldopa. “Studies have shown that the use of a COMT inhibitor is particularly helpful in controlling the wearing-off phenomenon in PD [Parkinson’s disease] patients by prolonging the half-life of L-Dopa and improving its brain entry.”¹

The researchers got the idea for testing the effectiveness of EGCG as a COMT inhibitor as part of Parkinson’s disease therapy from the fact that catechol-containing bioflavonoids and tea catechins have been identified as good substrates for COMT. In addition, these compounds have also been found to be strong inhibitors of liver COMT-mediated O-methylation of endogenous catechol estrogens. “Among these dietary compounds, EGCG was found to be the most potent inhibitor, with an IC50 [concentration required to inhibit the enzyme by 50%] of approximately 0.1 μM when 2-hydroxyestradiol was used as substrate.” The authors of the paper¹ also note that “[t]he findings of this study may also shed a mechanistic light on the recent epidemiological observation suggesting that regular tea drinking is associated with a reduced risk of PD.”

Male Sprague-Dawley rats were the experimental animal in the study. The researchers note that an earlier study reported that EGCG given at a daily oral dose of 500 mg/kg for 13 weeks was not toxic in rats. The doses used on the rats in this study¹ were 100 and 400 mg/kg, given two hours before L-Dopa/carbidopa. Results showed that the rats treated with 400 mg/kg had levels of 3-OMD in circulation and in the striatum that were reduced by approximately 30%, revealing the decrease in L-Dopa methylation.

The authors report that earlier human studies showed that “circulating concentrations after oral administration of 800 mg EGCG are higher than its effective IC50 concentrations required for inhibiting L-Dopa methylation in vitro. Therefore, it is possible that oral administration of EGCG may readily reach therapeutically-effective concentrations needed for inhibiting L-Dopa methylation in PD patients.”¹ Moreover, the authors suggest, although daily intake of EGCG through tea drinking may not yield the therapeutically effective concentration, there are other polyphenolic components contained in tea and coffee that also contain the same catecholic structures as does L-Dopa, thereby making them good substrates of and thus inhibitors of COMT.

Interestingly, the authors¹ suggest that the reduction in striatal 3-OMD level may aid in reducing side effects associated with L-Dopa/carbidopa therapy in PD. One study cited by the authors found that the plasma levels of 3-OMD in patients with dyskinesia were significantly higher than those in patients without dyskinesia. Also, they note that it has been suggested that 3-OMD accumulation following long-term L-Dopa treatment might contribute to progression of the neurodegeneration in PD patients.

Reference

1. Ki Sung Kang et al. Dual beneficial effects of (-)-epigallocatechin-3-gallate on levodopa methylation and hippocampal neurodegeneration: in vitro and in vivo studies. PLoS One.5(8):e11951 (Aug. 2010).

PIMA INDIANS: FROM A HEALTHY DIET TO A DIABETES-PROMOTING ONE

When the white men arrived in North America, the Pima Indians were eating a considerable amount of purple corn that they grew themselves. Purple corn is, of course, chock-full of anthocyanins. After the white men put the Indians on reservations and provided “free” food (only free if you disregard the value of the Indian lands seized in the process) that didn’t include purple corn, the Pima Indians became a population plagued by diabetes and hyperobesity. They have been frequently studied as populations with a genetic predisposition to diabetes, but their main problem may be a switch from an evolved diet that inhibited the development of diabetes (with purple corn-derived anthocyanins likely to be a major factor in that protection^1–4) to one (given to them for “free”) that did the opposite.

The moral of this sad story could be to beware of governments bearing free gifts after they have stolen much of your wealth. At a different level, the moral is to beware of making dramatic changes in your diet from the one your forebearers had, over thousands of years, evolved to consume and thrived on.

References

1. Li et al. Purple corn anthocyanins dampened high-glucose-induced mesangial fibrosis and inflammation: possible renoprotective role in diabetic nephropathy. J Nutr Biochem.23(4):320-31 (2011 May 2).
2. Tsuda et al. Dietary cyanidin 3-O-beta-D-glucoside-rich purple corn color prevents obesity and ameliorates hyperglycemia in mice. J Nutr. 133:2125-30 (2003).
3. Regulation of adipocyte function by anthocyanins: possibility of preventing the metabolic syndrome. J Agric Food Chem.56:642-6 (2008).
4. Tsuda et al. Microarray profiling of gene expression in human adipocytes in response to anthocyanin. Biochem Pharmacol.71:1184-97 (2006).

NEUROGENESIS

LITHIUM RESTORES NEUROGENESIS IN MOUSE MODEL
OF DOWN’S SYNDROME, CORRECTING COGNITIVE DEFECTS

A remarkable feat of pharmacological treatment to induce adult neurogenesis in a mouse model of Down’s syndrome¹ suggests the possible correction of cognitive impairments in humans with the disorder.

Cognitive defects are the most prominent features of human Down’s syndrome, resulting from three copies of Chromosome 21 rather than the normal two copies. One would think that to correct the features of the disease it would be necessary to administer genetic therapy. However, researchers publishing a new paper¹ found that treatment with the mood stabilizer lithium markedly increased adult neurogenesis in the subventricular zone (SVZ) of the brain as well as restoring brain volume in several areas where it is smaller than in normal mice.

In individuals with Down’s syndrome, brain hypotrophy is particularly evident in the cerebral hemispheres, frontal lobe, temporal cortex, hippocampus, and cerebellum and shows up early in development.¹

The Down’s syndrome mouse is produced by creating three copies of a distal region of the mouse chromosome 16—“a region that shows perfectly conserved linkage with human chromosome 21.”¹ Treatment with lithium resulted in an 85% increase in the pool of proliferating cells in the Down’s syndrome mice, while the pool was increased in the normal mice by 54%. “In the [Down’s syndrome mouse] the number of Brdu+ [label indicating proliferation] cells became similar to that of the untreated euploid [normal] mice.” DOSAGE: The DS female mice were, starting from the age of 12 months, treated with lithium contained in their food pellets at 2.4 g of Li2CO3/kg of food for one month. This dose scales to approximately that used to treat mania and bipolar disorder in humans. Since renal excretion varies, doses would need to be individualized based on readily available blood tests.

The authors conclude in a carefully worded summation: “If such a treatment will be proven to be effective [in humans], this might open the way to possible therapeutic intervention practicable by human subjects, aimed at correcting defects in brain development in DS individuals.”

LITHIUM AS AN ESSENTIAL NUTRIENT

A 2012 paper² provided detailed information on the presence of lithium in drinking water and vegetation and on its essentiality in a large number of animal and human studies.

The review reported that there is a wide variation in the amount of lithium found in drinking water, spring water, spa water and bottled water. One study of 132 brands of bottled water from 28 countries found lithium concentrations ranging over 5 orders of magnitude (from 0.057 to 5,460 μg/l. “Li concentrations of 8.7 mg/l and of >9 mg/l have been reported for the Friedrich-Quelle, a famous spring in Baden-Baden/Germany, and for the spa water in Pompeya/Argentina, respectively.”²

“The long-term consumption of low-Li drinking water is regarded as a risk factor for higher incidences of suicides, homicides, and crimes as shown in several studies. For example, in 24 counties in Texas/USA with a total population of 6 million (>60% of the state population), during a 2-year study (1967-1969), a significantly inverse relationship between 1) Li concentration of local drinking water, 2) Li concentrations of the urine among the respective residents, and 3) mean annual rainfall amounts in the respective county sites and 1) state mental hospital admission rates, 2) admitting rates for four major mental disorders (psychosis, neurosis, schizophrenia, personality problems), 3) homicide rates, and 4) road distance from resident county to nearest state mental hospital was found (p ≤ 0.05 to ≤ 0.001). The Li levels in drinking water were classified into 4 groups: <11.0; 11.0–29.9, 30.0–69.9; >70.0 μg/l.”²

A second study reported in the review² included 27 counties and lasted for 10 years (1978–1987) and found that that the incidence rates of suicide, homicide, and rape were significantly higher in counties whose drinking water contained low (0–12 μg/l) and medium (13–60 μg/l) lithium concentrations than in counties whose drinking water had high Li concentrations (79–160 μg/l)(p<0.01). They also found associations with the rates of robbery, burglary, and theft significant at p<0.05. These are really remarkable differences that suggest improved mental function with low dose lithium.

On the basis of these and many other studies, the basic requirement of lithium in humans has been assessed at 1 ug/kg. body weight/d in humans derived from intake data in Germany, thus, the lithium requirement for an adult of 70 kg body weight would be 70 μg/d. In another assessment study, Schrauzer suggested a provisional recommended dietary allowance (RDA) of 1 mg lithium/day for an adult of 70 kg body weight.³

References

1. Bianchi et al. Lithium restores neurogenesis in the subventricular zone of the Ts65Dn mouse, a model for Down syndrome. Brain Pathol. 20:106-18 (2010).
   2. Schafer. Evaluation of beneficial and adverse effects on plants and animals following lithium deficiency and supplementation, and on humans following lithium treatment of mood disorders. Trace Elem Electrolytes. 29(2):91-112 (2012).
   3. Schrauzer. Lithium: occurrence, dietary intakes, nutritional essentiality.J Am Coll Nutr. 21:14-21 (2002).

GENOME-WIDE METHYLATION PROFILES OF 656 INDIVIDUALS
REVEALS EPIGENETIC CHANGES IN AGING

CHANGES IN METHYLATION PREDICT AGE
AND ARE ASSOCIATED WITH COMPLEX DISEASES

At a completely different level than the human genome, the human methylome represents a map of the methylation of DNA, which determines how genes are expressed. Consequently, it helps reveal how alterations in the regulation of DNA transcription (whether a gene is turned on or off) takes place throughout life. For example, a new paper¹ discusses the fact that “epigenetic drift” takes place in identical twins, where the methylation marks increasingly differ with age, causing them to “drift” from their starting point with identical DNA at birth. The DNA remains the same (except for mutations) but is expressed differently as a result of methylation-induced changes.

As the authors¹ put it (and we couldn’t have put it better): “the idea of the epigenome as a fixed imprint is giving way to the model of the epigenome as a dynamic landscape that reflects a variety of chronological changes.”

In this fascinating paper, the researchers have determined DNA methylation at more than 450,000 CpG markers (where the methylation takes place) from the whole blood of 656 human individuals, aged 19 to 101. Their model, based upon the data they got from these measurements, can be used to determine the rate at which an individual’s methylome ages.¹ They found an optimal model consisting of a selected set of 71 methylation markers; this model was highly predictive of age, with a correlation between age and predicted age of 96% with an error of 3.9 years.¹ “Nearly all markers in the model lay within or near genes with known functions in aging-related conditions, including Alzheimer’s disease, cancer, tissue degradation, DNA damage, and oxidative stress.”

Another finding was that the methylome of men appeared to age approximately 4% faster than that of women. The scientists also compared the methylome derived from whole blood to that of breast, kidney, lung, and skin samples and found strong predictive power for chronological age for these tissues as well, although noting some linear offset from the expected age prediction. Another finding was that “tumors appear to have aged 40% more than matched normal tissue from the same individual. Accelerated tumor aging was apparent regardless of the primary tissue type.” Moreover, the researchers compared the methylation changes of tumors to normal tissue, examining all 70,387 age-associated markers, where 44% tend to increase and 56% tend to decrease with age. The tumor markers coincided with older values by 74% of the markers regardless of the trending direction (e.g., whether they increased or decreased).

The scientists even determined (using a mathematical model developed by Shannon and Weaver²), the increase in entropy (loss of information content) in the methylome over time. As the researchers explain it, “[a]n increase in entropy of a CpG marker means that its methylation state becomes less predictable across the population of cells (i.e. its methylation fraction tends toward 50%).” The researchers found a highly significant increase in methylome entropy over the sample cohort. “Furthermore, extreme methylome entropy for an individual was highly correlated with accelerated aging rate …”¹

The scientists suggest that this model could be used to assess the aging rate of an individual and to determine whether diet or environmental factors can accelerate or or retard the aging process and diseases of aging.

This is a remarkable new development that is just now beginning to be used in aging research. Better yet would be if such measurements were to become inexpensive enough for individuals such as ourselves to monitor our own aging.

Reference

1. Hannum et al. Genome-wide methylation profiles reveal quantitative views of human aging rates. Mol Cell.49:359-67 (2013).

THE NEW ECONOMICS OF ORPHAN DISEASES

Another article¹ on hot research areas in the pharmaceutical industry notes that the search for orphan drugs has attracted a lot of interest because of smaller clinical trial size, shorter trial time, and commercial benefits such as fast track FDA approval, tax credits, and fee waivers. Not mentioned in the article, though, is what may prove to be the most important incentive to develop orphan drugs: after getting approval for an orphan drug, off-label uses may be the entryway to much larger markets involving common diseases. Importantly, recent court decisions have provided much support of the First Amendment rights of companies to provide truthful, non-misleading scientific information on off-label uses of drugs approved for other purposes. This is a very effective way to increase the availability of therapies for both rare and common diseases and to improve physician (and public) awareness of these therapies.

“Orphan status is granted to drugs for which the costs of developing and marketing a therapeutic are unlikely to be recovered, and in the U.S., to diseases that affect fewer than 200,000 people …” (The reason why the costs would be unlikely to be recovered is, of course, the huge developmental costs imposed by FDA’s rules and regulations for getting approval.) The article further notes that since the Orphan Drug Act of 1983 was passed, the FDA has approved 350 drugs and biologics for approximately 200 orphan diseases. These treatments are said to account for about 22% of current drug sales, with a total global value of about $50 billion, according to the article.¹

We believe that these beneficial court decisions resulted from applying the First Amendment arguments that won our landmark decision in Pearson v. Shalala (U.S. Court of Appeals for the District of Columbia, 1999) (that protects truthful non-misleading speech on the health effects of dietary supplements and foods) to prescription drug marketing.

Reference

1. The new economics of orphan diseases. Genetic Engineering & Biotechnology Newspp. 12-13 (Jan. 1, 2013).

LATEST BLOCKBUSTER TREATMENT FOR LIFE THREATENING BACTERIAL INFECTION: FECES TRANSPLANTATION!

Recurrent Clostridium difficile infection is reportedly difficult to treat. The responsible bug, an enterocolitis bacteria, recurs after antibiotic treatment in about 25% of patients.¹Those who suffer a recurrence are more likely to have a second recurrence after treatment, with the subsequent recurrence rates surpassing 50%.¹

Amazingly, about 50 years ago, doctors tried an infusion of donor feces (fecal microbiota transplantation) for these these life-threatening resistant C. difficile infections and found an “immediate and dramatic” response. Yet it appears the treatment has not won favor with medical practitioners despite what the New England Journal of Medicinecalls an “alarming increase in the incidence and severity of this disorder.”¹ According to the Journal, only the most desperate patients are likely to want feces transplantation. The reasons for the failure of this treatment to catch on are given in the article as: “it is aesthetically unappealing, it is logistically challenging (in terms of harvesting and processing suitable donor material), and there is a lack of efficacy data from randomized, controlled trials.” The problem of lack of data from randomized, controlled trials has been solved by the publication² of a clinical trial of the treatment.

The results of the trial² showed that fecal microbiota transplantation (FMT) was effective in 81% of patients. As the authors of the paper declared, “[t]he results of this study represent a clear precedent in which planned therapeutic manipulation of the human intestinal microbiota can lead to demonstrable, clinically important benefits, thereby bringing FMT to the mainstream of modern, evidence-based medical practice.” The author of the commentary on the clinical trial study¹ suggests similar trials of this sort of therapy for other indications, such as inflammatory bowel disease, irritable bowel syndrome, prevention of colorectal carcinoma, and metabolic disorders, as examples. “As such, it heralds the delayed adolescence of a broad and exciting new branch of human therapeutics.”¹

The authors of the trial² noted that patients receiving the feces transplantation had increased fecal bacterial diversity, similar to that in healthy donors, with an increased quantity of Bacteroidetes species and clostridium clusters IV and XIVa and a decrease in Proteobacteria species.

There is something very amusing about this, as one visualizes the source of the therapeutic medicine. And especially nice to see a disorder that was killing people reined in, even by the likes of resident bacteria in shit. Now we wonder how the FDA will generate rules and regulations to make sure the shit is under their control and, hence, very expensive. To start with, note that the shit is regulated by them as a drug (biological).

SHIT WARS

THE FDA AGAINST MEDICINAL POOP

Now, however, in a more recent report on feces transplantation published this year in Nature,³ we learn that the FDA has declared feces transplantation to be under its authority and has begun issuing rules that will slow down this development and even risk the lives of critically ill people who will be made to wait for the FDA to decide whether to grant their doctor an Investigative New Drug Application before the doctor can proceed with the treatment (the FDA has 30 days to stop an experiment). In typical FDA fashion, the agency simply decreed its regulatory authority over feces transplantation without waiting for Congress to grant them the authority via statute; hence, the FDA has no legitimate authority over medicinal shit.

The June 20, 2013 GenomeWeb Daily News reports that the FDA’s attempt to take over regulation of feces transplants has run into an outcry by doctors pointing out that seriously ill patients don’t have time to wait for the FDA to grant an Investigative New Drug Application. In response, the FDA is reported to be exercising enforcement discretion (e.g., they are still claiming to be in CONTROL of feces transplantation but will (maybe) not prevent (on a case by case basis) a doctor from performing the procedure, depending on whether the agency bureaucrats feel magnanimous that day or not. The rule of law means nothing when all decisions are made at the discretion of bureaucrats.

There is some good news, however. The article¹ points out that, interestingly, “[r]esourceful individuals can [ ] get in on the act at home, by following step-by-step enema instructions from online videos.” The FDA is likely to find it a great deal more difficult to stop the production of how-to-do-it videos, as the First Amendment has the authority (last we heard) over free speech. It might be safer (as well as a lot less messy), however, if people could get the procedure done by a qualified doctor rather than doing it themselves.

One concern is that the fecal material being transplanted is not “standardized” and that there may be considerable variation in their contents. We think that attempting to standardize feces transplants at this point is a mistake because of the limited amount of information on what are the important components (including the probiotics) that produce positive results. More experimentation with different varieties of feces makes sense to us than deciding by decree, in the absence of a lot more experimental data, what would be appropriate in a “standardized” version of feces. One researcher, infection-disease specialist Trevor Van Schooneveld of the University of Nebraska Medical Center in Omaha was reported in the article as having performed about 20 feces transplants since 2011, but in the past few weeks he has had to turn down patients while he submits his Investigative New Drug application. “Van Schooneveld questions whether the agency should preside over an organic, personal substance, rather than a drug. ‘How the FDA plans to regulate human feces is a mystery to me,’ he says.”

We couldn’t agree with Dr. Van Schooneveld more. But the most serious problem of FDA meddling is what happens to people facing a potentially fatal infection and having to wait 30 days while the FDA decides whether it is going to allow their doctors to proceed with a feces transplantation. Is the FDA going to pay for the damages resulting from the delay, including the possible death of the patient? Of course not. Too bad if you die while they are playing games with your brother’s, sister’s, mother’s, or father’s shit. We truly can’t understand how a large majority of the public actually believes that the FDA is protecting the public health!

References

1. Fecal microbiota transplantation – an old therapy comes of age. N Engl J Med. 368(5):474-5 (Jan. 31, 2013).
2. van Nood et al. Duodenal infusion of donor feces for recurrent Clostridium difficile. N Engl J Med.368:407-15 (2013).
3. FDA gets to grips with faeces. Nature.498:147 (13 June 2013).

MINOR SCIENTIFIC MYSTERY SOLVED

IF YOUR UNDERARMS DO NOT SMELL BAD WHERE DO YOU SUPPOSE THE BACTERIA RESPONSIBLE FOR UNDERARM MALODOR HAVE GONE?

A recent paper¹ reports that there is a recessive gene that makes the armpits of a lucky 2% of the European population and most people of Asian descent free of underarm malodor. Those lucky people have two copies of the recessive gene for a transporter that fails to transport the necessary excreta to reach the bacteria living under your armpits that usually feed on this stuff, making a stink while they’re at it. (It is not clear from the article² describing the paper where the excreta actually ends up. Could the underarm microbiota be altered as a result of the rare transport glitch?) As the authors of the article put it people who don’t have smelly armpits may have a malfunctioning transporter gene.

The thing that is weird about all this is that of the people who probably don’t need to use underarm deodorants because of their screwed up stink transporter, 78% still do according to the article. In an attempt to explain the unexplainable, the article authors offer the following: England is still dominated by people with smelly armpits, a constant reminder to people to put on something to suppress what they may not have. In a very informative statistical analysis, the authors point out that since being stink-free is so rare in the U.K., both parents of an “odorless” child are probably heterozygous, e.g., they carry a dominant allele for the stink and a recessive allele for no stink. They themselves do stink. Only ¼ of these parents’ kids will be stink-free. This may explain why people take no chances and put on underarm smell protection. Moreover, they suggest that ubiquitous advertising has overwhelmed people’s thinking abilities to where they naturally assume they must be in need of protection against the socially dangerous stench. (Sandy occasionally uses rubbing alcohol, which kills any bugs lurking around waiting for dinner to chemically convert into a stench. Works like a charm. Durk takes a lot of showers.)

References

1. Rodriguez et al. Dependence of Deodorant Usage on ABCC11 Genotype: Scope for Personalized Genetics in Personal Hygiene. J Invest Dermatol.133(7):1760-7 (2013 Jul).
2. Fear of stink, driven by pheromones. Chemical & Engineering News.Volume 91, Issue 8, p. 48 (Feb. 25, 2013).

KEEP A SUPPLY ON HAND FOR …

When You Need It But Can’t Get It

When we appeared at the recent Life Enhancement Symposium in Las Vegas, one of the things we discussed was how to plan ahead for upcoming shortages due to rationing under government health care programs. In this short supplement to our regular monthly newsletter, we report what might be lifesaving information on a readily available, safe, and inexpensive over the counter cough medicine that has been shown to be effective in rats and mice for the treatment of sepsis (systemic infections with a high mortality rate). With some anti­biotics already becoming difficult for doctors and hospitals to get for patients, this is potentially important information, assuming that it works similarly in humans. We wanted to let you know.

Dextromethorphan, an Effective Treatment of Endotoxic Shock in Mice, Rats

Endotoxic shock (or sepsis) is a severe systemic infection with a high mortality rate. Often occurring in hospitals where large areas of the body can be exposed to bacterial invasion (as in surgical procedures and large area burns) or as a result of catheters or other devices that enter the body through the skin, allowing bacteria to enter.

It has been reported¹ that the readily available and non-prescription cough medicine dextromethorphan provided significant, very impressive protection against endotoxin shock in mice. Endotoxin shock was induced in the animals by administering a single intraperitoneal dose of LPS/GalN (lipopolysaccharide and galactosamine). Control mice received injections of saline. Pretreatment with dextromethorphan (30 minutes prior to LPS) at 25 and 12.5 mg/kg, subcutaneously, significantly increased the survival rate up to 90%, while even at the lower dose, survivorship was increased to 67%.

The researchers found that TNF-alpha (tumor necrosis factor-alpha), a powerful proinflammatory cytokine associated with sepsis, was significantly decreased by dextromethorphan in both liver and serum. Interestingly, TNF-alpha is increased in people with rheumatoid arthritis, suggesting that dextromethorphan might be a useful treatment for that disease as well. And unlike the TNF-alpha monoclonal antibodies currently in use in the treatment of rheumatoid arthritis, dextromethorphan would not have the severe off-target effects (particularly the risk of infection and leukemia) associated with those treatments.

The authors¹ note in their abstract that “DM [dextro­methorphan] may be a novel compound for the therapeutic intervention for sepsis.” Our comment: Don’t wait for FDA approval. Dextromethorphan is already an FDA approved over-the-counter (no prescription required) inexpensive cough medicine.

Another paper² reported that pretreatment with dextromethorphan (1, 5, and 10 mg/kg i.v.) attenuated the deleterious effects (e.g., hypotension and tachycardia) in rats treated with LPS (lipopolysaccharide, a component of bacterial cell walls). The researchers suggested that, “DM can possibly be used as a prophylactic agent for sepsis in the future.”

The recommended human dose for cough control is 60 mg dextromethorphan every 12 hours, which is likely to be effective for sepsis, too, based on the mouse and rat experiments (scaling the mouse dose to human dose on the basis of food consumption—similar to scaling via body surface area—the approximate human dose would be 37.5 mg to 75 mg).

Then, in 2011, another paper on the antimicrobial and antisepsis effects of dextromethorphan was published.³ In this study of gram-positive Group A streptococcus (GAS) infections, dextromethorphan was found in mice to increase survival, enhance bacterial clearance, and reduce systemic inflammatory response and organ injury. The animals were inoculated in the air pouch with a lethal dose of GAS NZ131 microbes with or without dextromethorphan (DM) treatment (12.5 mg/kg) 30 minutes before and 1, 12, and 24 hours after bacterial inoculation.

After 14 days, the survival rate of GAS-infected mice without DM treatment was only 10%, while the DM treated GAS infected mice showed approximately 72% survival.³Liver damage (as shown by fatty degeneration of liver cells, in which the cytoplasm was filled with foamy vacuoles of fat) was inhibited by DM, which was also reflected by lower serum levels of AST (a liver enzyme increased under conditions of liver injury) in the GAS infected, DM-treated mice as compared to the GAS infected (but no DM) mice. The researchers explain: “Although infecting bacteria can be taken care of by antibiotics, the bacterial components released from dead bacteria, such as peptidoglycans, lipoproteins, lipoteichoic acids, and LPS, may be responsible for systemic inflammation and cause organ failure and sepsis.”³ By contrast, DM reduces the inflammatory response. With respect to dosage, the authors say, “[c]ombined with antimicrobial agents, the dosage of DM may be reduced comparably to the recommended doses as a cough suppressant, but this issue needs to be tested.”

The bacteria used in this experiment are one of the infamous “flesh eating bacteria” that rapidly devour human flesh and often require limb amputation.

Interestingly, one of the papers³ reports that DM has been shown in previous studies to decrease ROS at least in part through the inhibition of NADPH oxidase, a major source of oxidative stress.

This is a medicine that could be stockpiled for use when you or a loved one are threatened by sepsis and either the infection is antibiotic resistant (a rapidly growing problem due to out-of-control drug approval costs) or supplies of antibiotics are limited due to health care system rationing or other problems. Dextromethorphan is stable and can be stored for at least a few years.

Just as we were wrapping up this article, we found an earlier paper4 that adds to the remarkable anti-inflammatory effects of dextromethorphan. In this earlier study, scientists found that the death of dopaminergic neurons in neuron-glia cell culture by inflammation induced by activated microglia (as occurs in Parkinson’s disease) could be decreased by dextromethorphan at the unbelievably low levels of micromolar and even femtomolar concentrations. This is far lower than the dose used in the control of cough. The authors of this paper suggest that their findings offer a “novel therapeutic concept of using ‘ultra-low’ drug concentrations for the intervention of inflammation-related neurodegenerative diseases.”⁴ (Note: ultra-low dose treatment cannot be assumed to work for any antiinflammatory substance; dextromethorphan in this study has been tested for and found effective at that low dose in cell culture.)

References

1. Li et al. Protective effect of dextromethorphan against endotoxic shock in mice. Biochem Pharmacol. 69:233-40 (2005).
2. Wang et al, “Dextromethorphan prevents circulatory failure in rats with endotoxemia. J Biomed Sci. 11:739-47 (2004).
3. Li et al. Dextromethorphan efficiently increases bactericidal activity, attenuates inflammatory responses, and prevents Group A streptococcal sepsis. Antimicrob Agents Chemother. 55(3):967-73 (2011).
4. Li et al. Femtomolar concentrations of dextromethorphan protect mesencephalic dopaminergic neurons from inflammatory damage. FASEB J. 19:489-96 (2005).

APPETIZERS

There is a certain relief in change, even though it be from bad to worse; as I have found in traveling in a stage-coach, that it is often a comfort to shift one’s position and be bruised in a new place.
— Washington Irving

When you came to examine the American Constitution, you found that it was not really a constitution, but a Charter of Anarchism. It was not an instrument of government: it was a guarantee to the whole American nation that it never should be governed at all. And that is exactly what the Americans wanted.
— George Bernard Shaw, address in New York, 11 Apr. 1933

I will be brief. But not nearly as brief as Salvador Dali, who gave the world’s shortest speech. He said, “I will be so brief I am already finished,” and he sat down.
— E. O. Wilson

John Major relates a serious conversation with Boris Yeltsin on Russia’s economy: “Tell me, Boris, in one word, how is Russia doing? “Good.” “In two words, Boris, how is Russia doing?” “Not good.”
— from What Is Your One Sentence?
by Mimi Goss, a marketing book

The girl said she recognized me from the vegetarian club, but I’d never met herbivore.
— from the 3/29/13,
Casey Daily Dispatch

A fanatic is one who can’t change his mind and won’t change the subject.
— Winston Churchill

 

ARGININE UPDATE ARGININE POTENTLY PREVENTS FORMATION OF AGES PROTECTS AGAINST ENDOTHELIAL DYSFUNCTION, OXIDATIVE STRESS, AND METABOLIC SYNDROME

A new paper¹ reports on highly protective effects of L-arginine (and also D-arginine) against endothelial dysfunction, increased AGE (advanced glycation endproduct) formation, and other features characteristic of type 2 diabetes by scavenging of methylglyoxal, a reactive dicarbonyl molecule produced during glucose, fatty acid, and amino acid metabolism that is a major precursor in the formation of AGEs.

The authors note that there is currently a lack of specific scavengers for methylglyoxal (MG), but that MG has been reported to have a high affinity for arginine. Hence, they tested the protective effects of arginine on the vascular smooth muscle cells and isolated aortic rings derived from rats and then incubated in high glucose (25mM) or methylglyoxal (MG) 100 mM) and tested for relaxation in response to acetylcholine.

Results showed that D-arginine and L-arginine prevented high glucose-induced elevation of MG levels in vascular smooth muscle cells and rat isolated aorta. MG and high glucose increased the expression and activity of the enzyme arginase (an enzyme in the urea cycle that catalyzes the formation of urea and ornithine) which competes with NOS (nitric oxide synthase) for L-arginine. Hence, increase in arginase can reduce the availability of arginine for conversion to nitric oxide by NOS, thus inducing endothelial dysfunction.

Another result of the study was the finding that D-arginine, L-arginine, and N-acetylcysteine prevented MG and high glucose-induced formation of the methylglyoxal-specific advanced glycation endproduct (AGE) N-carboxyethyl lysine (CEL). Coincubation of vascular smooth muscle cells with D-Arg (300 μM), L-Arg (300 μM) or NAC (N-acetylcysteine) (600 μM) attenuated the formation of CEL.

The authors note that, because L-Arg, but not D-Arg, is a precursor for NOS (nitric oxide synthase) and because D-Arg as well as L-Arg prevented MG- and high glucose-induced reduced relaxation, they conclude that “it strongly suggests that arginine, especially D-Arg, prevents MG- and high glucose-induced reduced relaxation by an eNOS-independent mechanism.” “The therapeutic potential of arginine against MG and high-glucose-induced pathology merits further investigation.”¹

Reference

1. Dhar et al. Arginine attenuates methylglyoxal- and high glucose-induced endothelial dysfunction and oxidative stress by an endothelial nitric-oxide synthase-independent mechanism. J Pharmacol Exp Ther. 342(1): 196-204 (2012).

THE METHYLOME DNA METHYLATION CHANGES WITH AGE REVEALS DYNAMIC LANDSCAPE

Changes in DNA methylation (the methylome) with time are being studied for insights into aging and age-related diseases. A recent study reported some initial findings of a quantitative model of aging using measurements at more than 450,000 CpG markers (hot spots where DNA methylation takes place) from the whole blood of 656 people.¹These subjects were aged 19 to 101 years.

The model was designed to “measure [ ] the rate at which an individual’s methylome ages.” The authors report that differences in aging rates were affected by gender and genetic variants and that, very interestingly, tumors reveal an accelerated aging rate.^®

Changes in methylation are one reason that identical twins, which start out with the same DNA blueprint, become different with age (for example, becoming divergent in their susceptibility to various diseases), a phenomenon called “epigenetic drift.” Environmental factors, such as the dietary supply of methyl donating nutrients (e.g., choline and methionine) can also modulate gene expression via changes in DNA methylation.

The authors found that their model was able to predict the age of most individuals with high accuracy, with a correlation between age and predicted age of 96% and an error of 3.9 years. “The methylome of men appeared to age approximately 4% faster than that of women, even though the overall distributions of age were not significantly different between the men and women in the cohort (p > 0.05, KS test).”¹ On the other hand, BMI did not contribute significantly to aging rate.

The authors report that “[n]early all markers in the model lay within or near genes with known functions in aging-related conditions …” However, “none of the genetic variants were significant predictors of age itself,” indicating that there was a pattern of changes in methylation that predicted age, but that methylation changes in a single gene did not do so. Examining all 70,387 markers of global methylation levels showed that 44% tend to increase and 56% tend to decrease with age. “[T]umors coincide with older values for 74% of the markers regardless of the trending direction.”¹

“Interestingly, use of our aging model indicated that tumors appear to have aged 40% more than matched normal tissue from the same individual (Wilcox test, p<10^-41).”¹

One other interesting finding¹ was that there was a loss of information (increase in entropy) in the methylome over time. “An increase in entropy of a CpG marker means that its methylation state becomes less predictable across the population of cells, i.e., its methylation fraction tends toward 50%. Indeed, over all markers associated with a change in methylation fraction in the sample cohort, 70% tended toward a methylation fraction of 50%.” “Furthermore, extreme methylome entropy for an individual was highly correlated with accelerated aging rate …”¹

Another paper on DNA methylation changes with age reported on changes in development and aging of the human prefrontal cortex.² “The human prefrontal cortex (PFC) plays a critical role in complex cognitive behaviors, personality, decision making, and orchestration of thoughts and actions and thus has been referred to as the CEO of the brain.”²

In this study, the researchers “investigated the genome-wide temporal dynamics of DNA methylation in a large cohort of well-characterized human PFC specimens from the second trimester of gestation until old age …” “DNA methylation at CpG dinucleotides has long been considered a key mechanism of transcriptional regulation and a critical factor in embryonic development and in cancer.”²

The researchers divided the PFC specimens by age into the fetal period, the childhood period, and those over ten years of age. The overall methylation changes were much greater during the fetal period, but involved fewer loci than in other life stages. During childhood and in later life, methylation changes occurred at a much slower rate (2–3 orders of magnitude slower).

One interesting finding was that many of the cancer-related genes began showing methylation changes during childhood and continuing into old age. Important tumor suppressor genes were found to have increased methylation levels during aging of adults. Hypermethylation silences the tumor suppressor genes, thereby increasing the risk of cancer.

We are beginning to see a deluge of papers on methylation changes with age. For example, another paper3 reported that age-dependent decreases in DNA methyltransferase and low transmethylation micronutrient levels (methyl donors) synergize to promote increased expression of genes implicated in autoimmunity and acute coronary syndromes. Generally, hypomethylation increases gene expression while hypermethylation silences genes by decreasing expression. Here, the researchers found that the age-associated decrease in DNA methyltransferase was synergistic with low folate, low methionine, or high homocysteine levels that demethylate and activate methylation-sensitive genes. Hypomethylation has also been shown to reactivate hypermethylated (silenced) tumor suppressor genes and that curcumin and EGCG are two natural materials that act as hypomethylating agents.^5A,5B An additional paper⁴reported that axonal regeneration in the repair of the adult central nervous system in rodents was mediated at least in part through DNA methylation and that folic acid, as a methyl donor, promotes methylation.

References

1. Hannum et al. Genome-wide methylation profiles reveal quantitative views of human aging rates.Mol Cell.49:359-67 (2013).
   2. Numata et al. DNA methylation signatures in development and aging of the human prefrontal cortex. Am J Hum Genet. 90:260-72 (2012).
   3. Li et al. Age-dependent decreases in DNA methyltransferase levels and low transmethylation micronutrient levels synergize to promote overexpression of genes implicated in autoimmunity and acute coronary syndromes. Exp Gerontol. 45:312-22 (2010).
   4. Iskandar et al. Folate regulation of axonal regeneration in the rodent central nervous system through DNA methylation. J Clin Invest. 120(5):1603-16 (2010).
   5A. Liu et al. Curcumin is a potent DNA hypomethylation agent. Bioorg Med Chem Lett. 19(3):706-9 (2009).
   5B. Fang et al. Tea polyphenol (-)-epigallocatechin-3-gallate inhibits DNA methyltransferase and reactivates methylation-silenced genes in cancer cell lines. Cancer Res. 63(22):7563-70 (2003).

HYDROGEN THERAPY STROKE

Here we continue our series on the use of hydrogen gas in the prevention and treatment of medical conditions, largely having to do with disorders induced by increased oxidative stress. As you may recall when we began this series of articles nearly a year ago (see “Hydrogen Therapy” in the June 2012 Life Enhancement magazine), we explained the special effectiveness of hydrogen as a selective antioxidant that scavenges the highly toxic hydroxyl radical, while having much less effect on radicals that play a role as signaling agents in normal physiology (such as superoxide and nitric oxide), and also scavenges the powerful oxidant peroxynitrite formed by the chemical reaction between superoxide and nitric oxide. We also pointed out that, unlike many other antioxidants, hydrogen is able to enter the mitochondria and scavenge free radicals there where most of the ROS (reactive oxygen species) are produced. Next, we explained the remarkable fact that hydrogen is produced in your lower intestine by gut bacteria, especially when they get larger quantities of fermentable dietary fiber called long chain fructooligosaccharides (LCFOS). From there, the hydrogen diffuses throughout your body, providing protection against hydroxyl radicals and peroxynitrite and in other ways, until leaving your body by exhalation from the lungs, diffusion from the skin, and from farts.

We have been following the development of this new field of medicine, hydrogen therapy, and writing up some of the studies in our newsletters. The one that follows shows how hydrogen was able to provide protection in Wistar rats against TIAs (transient ischemic events), where a section of the brain is briefly deprived of oxygen but not long enough for a complete stroke to take place.

In this study,¹ researchers used a common model of these transient reductions of blood flow to the brain by occluding two blood vessels that enter the brain for ten minutes. The experimental animals received hydrogen gas for 3 hours (at a concentration of 2%) inhalation immediately after the operation that was done to occlude the vessels. As a result of this treatment, many neurons die in the targeted CA1 region of the brain. The cause is not entirely understood but oxidative stress is believed to be the main cause. Increases in oxidative stress create molecules that can be measured to follow this damage.

In this study, the transient ischemia in the rats that did not receive hydrogen resulted in increased lipid peroxidation (as indicated by increased levels of malondialdehyde) and the product of oxidation 8-iso-PGF2alpha, whereas the amounts of these ROS (reactive oxygen species) were significantly reduced by hydrogen in the rats treated with hydrogen. Moreover, the hydrogen significantly reduced neuronal death in the CA1 brain region. In a test of cognitive function, hydrogen-treated rats performed better than the rats subjected to ischemia but not receiving hydrogen in the Morris water maze, where animals have to find a submerged platform in a container of water in order to climb onto the platform to discontinue treading water. It took the hydrogen-treated rats a shorter length of time to find the hidden platform.

As the researchers of paper #1 report, there have been earlier studies finding that inhalation of hydrogen gas could decrease the infarct size (the volume of dead cells) in animal models of stroke, limit the size of heart infarction in animal models of heart attack, and protect against generalized inflammation and improve survival in animal models of poly-microbial sepsis.

While the animals in this study received hydrogen gas by inhalation, we have written earlier in this series (see, especially, our introductory article in the June 2012 issue of Life Enhancement magazine) on how hydrogen gas can be obtained conveniently (a gift of certain gut bacteria) in appropriate quantities and periods of time by consuming the LCFOS (long chain fructooligosaccharides) type of dietary fiber.

Reference

1. Ge et al. Inhalation of hydrogen gas attenuates cognitive impairment in transient cerebral ischemia via inhibition of oxidative stress. Neurol Res. 34(2):187-194 (2012).

A KEY ENZYME IN YOUR CHOICE BETWEEN A REWARD NOW OR A BIGGER REWARD LATER IS PART OF THE DOPAMINE REWARD SEEKING SYSTEM

COMT—catechol-O-methyltransferase—if you haven’t heard of it before, then here is a bit of an introduction to an important part of your dopamine reward seeking system that is appearing in a considerable amount of recent research. It is important because it is the enzyme that degrades dopamine, thus having a major impact on the dopamine level in the brain, especially in the prefrontal cortex, a major site of higher cognitive processes.¹ You make a lot of decisions in the prefrontal cortex, including assessing the tradeoffs between getting a reward now or waiting a while and possibly getting a larger reward later. Some researchers call COMT a factor in impatient choice (impulsivity).¹

A specific variant (allele) of the COMT gene has been identified as associated with having a steep delay discounting, “the tendency to strongly discount future rewards as a function of their delay and thus choose tempting SSs [sooner-smaller rewards] over even substantially larger LLs [later-larger rewards].” Researchers in one paper¹cite papers that show suboptimal life outcomes in financial, academic, and health domain and in different psychiatric or behavioral disorders such as substance abuse, overeating, relationship infidelity, and pathological gambling in people who have the steep delay discounting. The Val allele (compared to the Met Allele) of COMT is associated with a higher level of enzymatic activity and, as it degrades dopamine, lowers prefrontal cortex dopamine levels.¹ COMT has a big effect on your life.

Using a sophisticated noninvasive EEG measurement system, the researchers exposed participants to a choice paradigm between SSs (sooner-smaller rewards) and LLs (later-larger rewards) and were able to calculate the intracortical electrical sources that generated the scalp-recorded activity of each of seven frequency bands. They were able, for example, to identify brain regions whose baseline activation correlates with DD (delay discounting), separately for each EEG frequency band.

The researchers concluded that: “Our research suggests that the Val allele predisposes individuals to a low level of baseline activation in the left DPFC [left dorsal prefrontal cortex] which then biases them toward impatient choice.”

There was another study, however, that found the Met allele to be associated with significantly steeper discounting rates, i.e., more impulsive choices, as compared with the Val allele (reported in #4). It was suggested that the discrepancy may be due to the fact that this study¹ had adults as participants whereas the other study’s participants were adolescents. “Numerous aspects of PFC [prefrontal cortex] function, including the expression and activity of COMT, change over the course of adolescence.”⁴

But that isn’t the end of the story because there are natural products, such as EGCG, that can inhibit COMT, thus decreasing the degradation of dopamine, leading to higher levels of dopamine and, hence, potentially avoiding the disadvantage of impatient choice.

Inhibiting COMT To Decrease Dopamine Degradation in the Brain

EGCG ((-)-epigallocatechin-3-O-gallate, the major catechin found in green (or white) tea), has been recently identified as a high-potency inhibitor of the ubiquitous human enzyme catechol-O-methyltransferase (COMT).² As of 2010 when another paper was published on EGCG’s effects on COMT,³ there were two drugs approved as COMT inhibitors: tolcapone and entacapone. “However, the use of tolcapone is only limited to fluctuating [Parkinson’s disease] patients who are refractory to other therapies, and requires heightened monitoring for the occurrence of hepatotoxicity [liver damage]. Although entacapone is relatively safer, it appears less efficacious than tolcapone.”³

The detailed molecular analysis of the mechanism by which EGCG inhibits COMT shows it to be a potent non-competitive inhibitor that, while able to interfere with the molecular mechanism by which COMT binds to dopamine, renders EGCG itself a poor substrate for COMT methylation.

In one of the papers,³ researchers studied the effect of EGCG both in vitro (human liver samples) and in vivo (modulation of L-Dopa methylation in rats). “When normal rats (treated with L-DOPA + carbidopa) were given an oral administration of EGCG (at 400 mg/kg), their 3-OMD levels [L-Dopa product after interaction with COMT] in circulation and striatum were reduced by approximately 30%, clearly reflecting an in vivo inhibition of L-DOPA methylation.” The breakdown of L-Dopa was reduced, but L-Dopa plasma concentration was increased only slightly (not statistically significant). This was similar to the results of treatment with L-Dopa + carbidopa plus tolcapone or entacapone in rats or humans.³

The authors³ concluded that: “The significant reduction of 3-OMD by EGCG may increase L-DOPA bioavailability in the central nervous system and particularly, reduce potential cytotoxicity associated with elevated levels of 3-OMD.”

Dopamine and Reward Processing

Another recent paper⁴ provided an overview of dopamine and reward processing that included COMT and a lot more, noting (for example) that reward processing takes place in multiple brain regions, including the VStr, midbrain, orbitofrontal cortex, anterior cingulate cortex, prefrontal cortex, ventral pallidum, and the medial dorsal nucleus of the thalamus. No wonder, then, that understanding the entire dopamine-reward system is far from complete and is very complex. One oddity, for example, is that “systemic administration of dopamine antagonists selectively reduces the motivation of animals to pursue rewards, without affecting their preferences for such rewards when they can be obtained without effort.”⁴ (Makes those who prefer getting rewards via the government’s freebies—rewards without effort—sound like they’re on dopamine antagonists.)

EGCG Increases Neurogenesis in the Sub-granular Zone of the Dentate Gyrus in Adult Mice

EGCG does a great deal more than just inhibit COMT, of course. In the area of cognition alone, EGCG also promotes neurogenesis, thus increasing the availability of new youthful adult-born neurons in those areas of the brain where neurogenesis takes place throughout life. One of those areas is the subgranular zone of the dentate gyrus in the hippocampus.

In one study of EGCG and neurogenesis, mice were divided into two groups of 7 mice each, one group of which received vehicle (controls) and one of which was treated with 25 mg/kg of EGCG for four weeks.⁵ Animals were treated with BrdU (5-bromo-2’-deoxyuridine) by injection and the neurons marked by Ki67 and DCX determined (these are markers of neuronal proliferation). “In the EGCG-treated groups, the number of Ki67+-positive cells were increased by 221.3% (p=0.01) compared to that in the vehicle-treated group (11.7 ± 0.17/section).”⁵

Hence, the authors conclude, EGCG enhances the survival of immature neuroblasts in the subgranular zone of the dentate gyrus in adult mice.

EGCG Increases the Number of Neural Stem Cells Around a Damaged Area After Rat Traumatic Brain Injury

EGCG is also potently neuroprotective in the injured brain. In one study,⁶ for example, male Wistar rats were subjected to acute brain trauma while under anesthesia after receiving either water or water with EGCG added to it at 0.1% w/v from 6 to 10 weeks of age. Excitotoxicity elicited under ischemia, such as that occurring during acute brain trauma, is known to cause excessive glutamate release from the injured neurons, with the subsequent influx of increased Ca2+ via glutamate receptors. The increased Ca2+ leads to a substantial increase in free radical generation, with cell degeneration and death following.

The researchers report that, following the initial mechanical insult, damage resulted from blood-brain barrier disruption, excitotoxic damage and free radical production. The scientists then looked for nestin (a marker of neural stem cells) in the cells around the damaged area. They found that the number of nestin-positive cells in the EGCG treatment group showed a significant increase when compared with the number in the water group. There was also a significant increase in the nestin-positive cells at days 3 and 7 following the brain trauma in the EGCG treatment group as compared with the number in the water group (73.0 ±25.8 and 12.4 ±3.3, respectively).⁶

“Therefore, we speculate that EGCG crosses the BBB [blood-brain barrier] and inhibits neuronal and NSC [neural stem cells] death by free radicals in the damaged area following TBI [traumatic brain injury].”⁶

References

1. Gianotti et al. Why some people discount more than others; baseline activation in the dorsal PFC mediates the link between COMT genotype and impatient choice. Front Neurosci. 6:54 (May 2012).
2. Zhu et al. Molecular modelling study of the mechanism of high-potency inhibition of human catechol-O-methyltransferase by (-)-epigallocatechin-3-O-gallate. Xenobiotica. 38(2):130-46 (2008).
3. Kang et al. Dual beneficial effects of (-)-epigallocatechin-3-gallate on levodopa methylation and hippocampal neurodegeneration: in vitro and in vivo studies. PLoS One. 5(8):e11951 (Aug. 2010).
4. Tunbridge et al. The role of catechol-O-methyltransferase in reward processing and addiction. CNS Neurol Disord Drug Targets. 11:306-23 (2012).
5. Yoo et al. (-)-epigallocatechin-3-gallate increases cell proliferation and neuroblasts in the subgranular zone of the dentate gyrus in adult mice. Phytother Res. 24:1065-70 (2010).
6. Itoh et al. (-)-epigallocatechin-3-gallate increases the number of neural stem cells around the damaged area after rat traumatic brain injury. J Neural Transm. 119:877-90 (2012).

QUICK NUTRITIOUS LUNCH: THE REUBEN HALFWICH

Finding you have less time for cooking? Next time you have only a few minutes available and want to eat something tastier than scrambled eggs or (gasp) a frozen meal, try this:

The halfwich is basically a toasted piece of bread topped by meat, cheese, mustard, pickles, sauerkraut, a slice of avocado, and a slice or two of onion, then heated in a microwave until the cheese melts. The whole thing takes just a few minutes. The “secret” ingredient is: CORNED BEEF.

You can get lean corned beef in a convenient refrigerated serving size for 2 or 3 at most supermarkets.

You’ll need: 1 piece of bread, preferably high fiber, toasted corned beef, prepared in microwave, two slices of Swiss cheese, about ½ cup of sauerkraut, your favorite mustard, pickles, sliced so the pieces lie flat on the bread, a slice of avocado, and a slice or two of onion.

Cook corned beef as instructed (4–6 minutes in a microwave). Layer on a microwave-safe ceramic plate: toasted bread, slice of Swiss cheese topped by mustard, sauerkraut, sliced pickles, corned beef cut into thin slices, and slice or two of onion. Top the whole thing with another slice of Swiss cheese, then microwave for a minute or so until the cheese is melted.

That’s all there is to it for a great hot lunch!

Optional: Sprinkle ground caraway seed on sauerkraut.

ATHEROSCLEROSIS PROTECTION AGAINST VASCULAR AGING IN NOX-2-KNOCKOUT MICE

At the start of a new paper,¹ the authors note that patients with advanced atherosclerosis have severe deficiencies in the ability to generate new blood vessels to adapt to inadequate blood flow. Moreover, advanced age is a major risk factor for coronary and peripheral arterial diseases and is associated with impaired ability to generate new blood vessel formation to increase blood flow around areas of arterial occlusion. In addition, “the number and/or the functional activities of EPCs [endothelial progenitor cells] have been shown to be impaired by aging in both animals and in humans.”¹

The researchers, searching for a mechanism to explain the defective neovascularization in the context of aging, have discovered that mice lacking NOX-2 (a subunit of the enzyme NADPH oxidase) were protected against age-associated impairment in reparative neovascularization following ischemia, as well as protecting EPCs against functional impairments associated with aging.

NADPH oxidase is a major source of ROS (reactive oxygen species) critical for neutrophil antimicrobial function.²

As part of the experimental protocol,¹ the scientists studied the effect of aging on the expression of NADPH oxidase subunit NOX-2 in mouse ischemic hindlimb muscles. They found NOX-2 expression to be significantly increased in old as compared to young wild type animals. (NOX-2 knockout animals, of course, do not express the NOX-2 subunit in hindlimb muscles.) Hindlimb ischemia (impaired blood flow) was established in experimental animals with and without NOX-2 by surgery. In wild type mice, aging was associated with a significant impairment of blood flow recuperation [restoration by new blood vessel growth] at day 7, whereas, by contrast, NOX-2-/- [knockout animals] were protected against age-dependent impairment of blood flow recuperation. Likewise, wild type aging animals exhibited a significant increase in oxidative stress in ischemic muscles while NOX-2-deficient mice were protected against the age-associated increase in oxidative stress.

The scientists¹ report that in previous studies (as cited), NOX-2-deficient mice were protected against the effects of ischemic strokes, some forms of hypertension, and aortic atherosclerosis induced by high blood cholesterol. Moreover, reduced neovascularization is associated with increased oxidative stress and some antioxidants (including red wine) have been reported to restore at least in part the ability to regenerate new blood vessels.

In one study,³ apocynin, thought to be an indirect NADPH oxidase inhibitor, increased collateral growth capacity, whether administered prior to, or 7 days following, arterial ligation. We were particularly interested, however, in the reported efficacy of annatto extract and beta-carotene in inhibiting NADPH oxidase and increasing expression/activity of antioxidant enzymes.⁴ Both annatto and beta carotene were able to increase the mRNA levels of SOD (superoxide dismutase) and CAT (catalase) in neutrophils from diabetic rats. As NOX-2 has been reported to be required for the microbiocidal function of neutrophils, it is possible that the annatto/beta-carotene treatments inhibited NOX-2.² Previous papers published by the same group that published this paper³ were reported to show a reduction in ROS (reactive oxygen species) production in neutrophils of diabetic rats treated for 30 days with beta carotene and annatto extract. The researchers note that this study³ is the first (to their knowledge) to report an increased expression of CAT (catalase, an important antioxidant enzyme that protects against hydrogen peroxide) in neutrophils from diabetic rats treated with beta carotene and annatto extract.

The control group and diabetic group of rats³ received a modified standard diet, whereas experimental groups received the diet + annatto (control-annatto), and diet + annatto (diabetic-annatto) or diet + beta carotene (control-beta) and diet + beta carotene (diabetic-beta). The concentration of bixin (a potent carotenoid antioxidant) in the annatto extract was 2.17 g/100 g. As noted by the authors of the annatto/beta carotene study, “bixin, which is present in the extract of annatto, is one of the most effective biological singlet molecular-oxygen quenchers.” Moreover, they note that annatto, which is a natural yellow-red color, is widely used as a food colorant and has a relatively low cost of production and low toxicity, making it a very attractive pigment for the food industry.

The Terpenoid Bixin May Activate PPARgamma, Enhance Insulin Sensitivity and Increase Adiponectin Secretion from Adipocytes

A 2010 paper⁵ evaluated the effects of various terpenoids derived from herbal and dietary plants on the activity of PPARgamma, an important regulator of insulin sensitivity. (The synthetic glitazones that activate PPARgamma are drugs used in the treatment of diabetes). In a graph showing the effects of 24 hours of incubation of a number of terpenoids at 50 or 100 μM as compared to a vehicle control set at 100%, bixin showed the greatest activation of PPARgamma ligand at five-fold induction (compared to the vehicle control).

Another paper⁶ reported that, in mice, bixin also activated PPARalpha, a transcription factor that regulates the expression of genes involved in fatty acid oxidation. Activation of fatty acid oxidation in the liver by PPARalpha improved carbohydrate and lipid metabolism in the subject obese mice. Bixin treatment also improved obesity-induced dysfunctions including hyperglycemia, hyperinsulinemia, and adiponectin deficiency.

One of the effects of PPARgamma is to induce differentiation of preadipocytes to adipocytes. The new adipocytes, being small, have various properties that differ from mature adipocytes, such as being more insulin sensitive and secreting more adiponectin, an insulin-sensitizing adipokine.⁷

References

1. Turgeon et al. Protection against vascular aging in Nox2-deficient mice: impact on endothelial progenitor cells and reparative neovascularization. Atherosclerosis. 223:122-9 (2012).
2. Lamb et al. Endotoxin priming of neutrophils requires endocytosis and NADPH oxidase-dependent endosomal reactive oxygen species. J Biol Chem. 287(15):12395-404 (2012).
3. Miller et al. Antioxidants reverse age-related collateral growth impairment. J Vasc Res. 47:108-14 (2010).
4. Rossoni, Jr. et al. Annatto extract and beta carotene enhances antioxidant status and regulate gene expression in neutrophils of diabetic rats. Free Radic Res.46(3):329-338 (2012).
5. Goto et al. Various terpenoids derived from herbal and dietary plants function as PPAR modulators and regulate carbohydrate and lipid metabolism. PPAR Res. Volume 2010, Article ID 483958, 9 pp. (this is an open access article).
6. Goto et al. Bixin activates PPARalpha and improves obesity-induced abnormalities of carbohydrate and lipid metabolism in mice. J Agric Food Chem. 60:11952-8 (2012).
7. Takahashi et al. Bixin regulates mRNA expression involved in adipogenesis and enhances insulin sensitivity in 3T3-L1 adipocytes through PPARgamma activation. Biochem Biophys Res Commun. 390:1372-6 (2009).

NITRATED FATTY ACIDS NITRATED FATTY ACIDS ARE NATURAL METABOLITES
THAT MEDIATE ANTI-INFLAMMATORY SIGNALS

For some time, there have been papers published on the anti-inflammatory and vasorelaxation activity of nitrated fatty acids.^1–4 Now, a new paper⁵ reports that “conjugated linoleic acid (CLA) is the preferential unsaturated fatty acid substrate for nitration reactions during oxidative conditions and digestion.” It is reported that “multiple enzymatic and cellular mechanisms account for CLA nitration, including reactions catalyzed by mitochondria, activated macrophages, and gastric acidification.”⁵ It is reported here that CLA yields up to 10⁵ greater extent of nitration products as compared to it’s natural competitor bis-allylic linoleic acid.

The nitrated fatty acids act as a circulating reservoir of nitrite for local conversion to nitric oxide. Hence, it is a way to increase nitric oxide availability throughout the body.

“Dietary CLA and nitrite supplementation [which can be derived from nitrate-rich foods such as spinach] in rodents elevates NO2–CLA levels in plasma, urine, and tissues, which in turn induces heme oxygenase-1 (HO-1) [an important antiinflammatory enzyme] expression in the colonic epithelium.” Incredibly, in human coronary artery endothelium, the paper notes, fatty acid nitroalkenes significantly influence the expression of ~400 metabolic and anti-inflammatory-related genes, including important inflammation regulators such as PPARgamma, Keap1/Nrf2, heat shock factor-1, and NF-kappaB. According to the authors, NO (nitric oxide) doesn’t directly nitrate protein or lipids, but must be oxidized to the proximal nitrating species NO2. “Importantly, NO2–[nitrite]-supplemented diets are associated with a variety of beneficial anti-inflammatory and metabolic actions, including the regulation of mitochondrial function, adipogenesis, oxygen delivery to tissues, and blood pressure. Although these events can in part be attributed to the generation of NO [nitric oxide], salutary responses to NO2–-derived oxides of nitrogen may also be transduced by the concomitant generation of electrophilic nitro-fatty acids (NO2–FA).”⁵

The Bottom Line

In this new paper⁵ the researchers show that CLA is the preferential endogenous fatty acid substrate for fatty acid nitration by NO and NO2– and provides tissue-protective and anti-inflammatory actions. Beneficial effects reported in the paper for nitrated fatty acids included: in mouse models of metabolic and inflammatory injury, fatty acid nitroalkene administration at nanomolar concentrations prevents restenosis after blood vessel injury, in mouse models of metabolic syndrome, nitrated fatty acids limit weight gain and loss of insulin sensitivity, nitrated fatty acids protect against ischemia-reperfusion injury, reduces plaque formation in a rodent model of atherosclerosis, and inhibits the onset of chemically induced inflammatory bowel disease. So, although the two of us do not use high-dose CLA for purported weight control, we do consider it’s use at lower, more physiological doses for the formation of nitrated-CLA as reported here⁵ to be potentially important.

References

1. Baker et al. Fatty acid transduction of nitric oxide signaling. J Biol Chem. 280(51):42464-75 (2005).
2. Lima et al. Nitrated lipids decompose to nitric oxide and lipid radicals and cause vasodilation. Free Radic Biol Med. 39:532-9 (2005).
3. Trostchansky and Rubbo. Nitrated fatty acids: mechanisms of formation, chemical characterization, and biological properties. Free Radic Biol Med. 44:1887-96 (2008).
4. Tsikas et al. Nitro-fatty acids occur in human plasma in the picomolar range: a targeted nitro-lipidomics GC-MS/MS study. Lipids. 44:855-65 (2009).
5. Bonacci et al. Conjugated linoleic acid is a preferential substrate for fatty acid nitration. J Biol Chem. 287(53):44071-82 (2012).

BLOOD GROUPS AND THE RISK OF CORONARY HEART DISEASE

A 2012 paper¹ reported on the association between blood groups and the risk of coronary heart disease in two large cohorts: the Nurses’ Health Study (NHS, including 62,073 women) and the Health Professionals Follow-Up Study (HPFS, including 27,428 men).

The frequency for the blood types O, A, B, and AB was 42.9%, 36.0%, 13.3%, and 7.8% in women and 43.0%, 37.2%, 12.3%, and 7.5% in men. These frequencies were similar in both the NHS and HPFS across the four ABO blood groups. The authors determined that during up to 26 years of follow-up, there were 2055 confirmed coronary heart disease cases (1666 nonfatal MI and 389 fatal CHD) in the NHS, while during 20 years of follow-up, there were 2015 CHD cases (with 1420 nonfatal MI and 595 fatal CHD). The incidence of CHD among the blood groups per 100,000 person-years were 125, 128, 142, and 161 for those with blood type O, A, B, and AB in women and 373, 382, 387, and 524 for those with blood types O, A, B, and AB in men. The cumulative incidence of CHD was statistically significantly different among the 4 ABO blood groups in both cohorts (P=0.0048 in NGS and 0.0002 in HPFS, respectively.

“Compared with participants reporting blood group O, those with non-O blood type [A, B, and AB] had an age-adjusted hazard ratio of 1.09 (95% CI, 1.03–1.17).” “Compared with the O blood group, the non-O blood type (A, B, and AB) had a stronger relationship with CHD risk in overweight and obese women than those with BMI<25 kg/m squared. However, this interaction was not confirmed in men …”

The authors suggest that recent studies lend support to the relation between ABO blood type and cardiovascular risk. They had recently discovered that the ABO gene is located on chromosome 9q34 in association with the plasma soluble E-selectin levels in the NHS, as was found in another genome-wide association study. In addition, they say, the ABO locus was related to tumor necrosis factor alpha, a powerful proinflammatory cytokine that has been associated with increased CHD risk.¹ Since most of the participants were white, however, the researchers suggest that the results may not necessarily apply to other races.

Reference

1. He et al. ABO blood group and risk of coronary heart disease in two pro­spective cohort studies. Arterioscler Thromb Vasc Biol. 32:2314-20 (2012).

FENNEL – CHEWING FENNEL SEEDS AFTER A MEAL AS A MOUTH
FRESHENER OFFERS SURPRISING HEALTH BENEFITS

A new paper¹ reports that in the Indian sub-continent and around the world, chewing fennel seeds (Foneiculum vulgare) is a common way to freshen one’s mouth after eating a meal. But surprisingly, there appears to be a lot more to it than that. In the new paper, the researchers explain that they found that fennel seeds contain significantly higher amounts of nitrites and nitrates when compared to other commonly chewed post-meal seeds. Nitrites and nitrates are “known to play crucial roles in maintaining vascular and digestive function”¹ by being chemically reduced in the body to nitric oxide, an important vasoactive compound. In this new study, the researchers show that nitrites derived from fennel seeds can modulate vascular functions. Quite a nice benefit for a little bit of post-prandial chewing. (And, incidentally, if you haven’t tried them, they are tasty, with a sort of licorice flavor.)

The authors¹ cite recent animal studies exploring different potential applications of what they call “nitrite therapy” that included pulmonary hypertension, acute tissue ischemic injuries, cerebral vasospasm, myocardial infarction, and stroke. “Studies have demonstrated that a high intake of nitrite containing fruits and vegetables in one’s diet has protective effects on the cardiovascular system.”¹ The authors even suggest that chewing fennel seeds could be a useful way to maintain nitric oxide levels at high altitudes to help prevent the onset of acute hypoxia.² Sounds like a good method for mountain climbers to use to avoid altitude sickness.

Cautionary note on increasing dietary intake of nitrates: A recent paper³ reports on the risk of esophageal and gastric cancer subtypes in relation to dietary N-nitroso compounds in 120,852 men and women aged 55-69 when they were recruited in the Netherlands Cohort Study in 1986. N-nitroso compounds are naturally occurring compounds containing a nitroso group attached to a nitrogen atom that can be converted in animals to carcinogenic forms in the acidic environment of the stomach or in the lower gastrointestinal tract by contact with heme-iron derived from meat. The Netherlands Cohort Study³ found that high dietary intakes of N-nitroso compounds (NOC) increased the risk of esophageal squamous cell carcinoma in men, though there was no increased risk in women. Importantly, adequate amounts of vitamin C, an inhibitor of endogenous nitrosation, can prevent this increased risk.³

References

1. Swaminathan et al. Nitrites derived from Foneiculum vulgare (Fennel) seeds promotes vascular functions. J Food Sci. 77(12):H273-9 (2012).
2. Erzurum et al. Higher blood flow and circulating NO products offset high-altitude hypoxia among Tibetans. Proc Natl Acad Sci USA. 104(45):17593-8 (2007).
3. Keszei et al. Dietary N-nitroso compounds, endogenous nitrosation, and the risk of esophageal and gastric cancer subtypes in the Netherlands Cohort Study. Am J Clin Nutr. 97:135-46 (2013).

IS EVERYTHING WE EAT ASSOCIATED WITH CANCER?

A paper with this actual title was published very recently.1 Noting that large and increasing numbers of different foods and food constituents are being investigated for positive or negative associations with cancer, scientists investigated the appearance of 50 common ingredients from random recipes in a cookbook, evaluating the conclusions, statistical significance, and reproducibility of published associations with cancer appearing in papers in PubMed.

The authors found that forty ingredients (80% of those that they examined in the literature) had articles published on cancer risk. They found that “[a]ssociations with cancer risk or benefits have been claimed for most food ingredients. Many single studies highlight implausibly large effects, even though evidence is weak. Effect sizes shrink in meta-analyses.”

The authors also state that “[s]tudies may spuriously highlight results that barely achieve statistical significance or report effect estimates that either are overblown or cannot be replicated in other studies.” Overall, they found that 39% of studies concluded that the examined ingredient conferred an increased risk of malignancy, 33% concluded that the ingredient reduced the risk of malignancy, 5% concluded that there was a borderline statistically significant effect, while 23% could not discern a clearly increased or decreased risk. Studied ingredients included veal, salt, pepper spice, egg, bread, pork, butter, tomato, lemon, duck, onion, celery, carrot, parsley, mace, olive, mushroom, tripe, milk, cheese, coffee, bacon, sugar, lobster, potato, beef, lamb, mustard, nuts, wine, peas, corn, cayenne, orange, tea, and rum. You can imagine some of the difficulties in, say, examining the effects of lamb on the risk of cancer. How do you hold everything constant (including subject genetic diversity) while varying only the intake of lamb?

Some of the problems that likely contributed to the inconsistency included variation in definitions of “serving size,” “often,” “never” as well as there being no standardized, consistent selection of exposure contrasts for the reported risks. The authors claim that the vast majority of these claims (increased or decreased malignancy) were based on weak statistical evidence. We would also like to point out that the variation in human responses to the analyzed ingredients imposed a very difficult hurdle to the isolation of effects that can be generalized to large human populations. That is why we like to see considerable research done on mechanistic (cell free or cell culture) studies and animal studies considered together with human studies. The analysis of studies of a particular ingredient’s effects on cancer risk, considered as a whole, still is as much of an art as a science, depending upon the ability to see connections between, say, chemical structure and function of an ingredient. You don’t get definitive answers. You get leads.

See quotes on the next page, on the dangers of scientific bias, which appeared in the editorial commentary on this paper, published in the same issue of The American Journal of Clinical Nutrition.

Reference

Schoenfeld and Ioannidis. Is everything we eat associated with cancer? A systematic cookbook review. Am J Clin Nutr. 97:127-34 (2013).

SCIENTIFIC BIAS

Imagination, on the contrary, which is ever wandering beyond the bounds of truth, joined to self-love and that self-confidence we are so apt to indulge, prompt us to draw conclusions which are not immediately derived from facts; so that we become in some measure interested in deceiving ourselves.

— Antoine-Laurent Lavoisier¹

White hat bias, defined by Cope and Allison as “bias leading to distortion of research-based information in the service of what may perceived as ‘righteous ends’” may be a factor in the overstatement of research find- ings. In addition, overstatement of results can be influenced by confirmation bias, by which the overstated results match preconceived views and hypotheses, leading to acceptance of the results even if the results are weak or nonsignificant.

— Bohan Brown, Brown, and Allison¹

1. Brown et al. Nutritional epidemiology in practice: learning from data or promulgating beliefs? Am J Clin Nutr. 97:5-6 (2013).

 

RECENT COURT DECISIONS RULE COMMERCIAL SPEECH
ON OFF-LABEL USE OF DRUGS IS PROTECTED BY THE FIRST AMENDMENT

The Jan. 10, 2013 issue of The New England Journal of Medicine contained an article reviewing four recent Court decisions supporting the rights (under the First Amendment) of pharmaceutical companies to promote off-label uses of drugs approved by the FDA for a different purpose. Recognizing that the FDA’s authority does not extend to the practice of medicine, the article notes that the agency cannot prohibit physicians from prescribing approved drugs for non-approved uses.

It is widely recognized, though this was not discussed in the article, that a large percentage of drugs prescribed for the treatment of cancer (as an example) are for off-label uses (not the uses the FDA approved them for) and that this constitutes an important part of modern medicine, extending or saving large numbers of lives.

One of the main arguments made against the promotion of off-label uses of drugs approved by the FDA for a different purpose is that without being able to silence pharmaceutical companies from promoting off-label uses, manufacturers can seek approval only for certain limited uses of drugs, then promote that same drug for off-label uses, effectively circumventing FDA’s new drug requirements. In fact, the trial court in Caronia (a case discussed in the article) upheld the constitutionality of the FDA’s regulations prohibiting off-label uses of FDA approved drugs because it (the court) could not identify a less restrictive manner in which to prohibit pharmaceutical companies from circumventing the FDA approval process.

We note that just because the court could not think of any less restrictive manner (than an outright ban on First Amendment protected speech) to prevent the circumvention of the FDA’s approval process is hardly a legitimate basis for throwing out the First Amendment. (It certainly suggests that something is wrong, e.g., unconstitutional, with the FDA’s approval process if it cannot accommodate the First Amendment’s protections of speech and press.) The Constitution does have a procedure for amendment and the proper way to eliminate the First Amendment’s protection of some area of speech would be to pass an amendment to do that. Otherwise, it is just a court deciding on its own that it didn’t approve of the consequences of exercising one’s First Amendment’s rights and declaring the First Amendment doesn’t protect that speech.

The article noted that “[i]n overturning Caronia’s conviction, the three judge panel of the Second Circuit agreed that the FDA regulations were overly broad, specifically noting that nothing Caronia did constituted conspiracy to put a false or misleading or deficient label on a drug product. The court appeared particularly persuaded by the argument that the FDA regulations allow unfettered prescribing of approved drugs for off-label uses but then, through the off-label restrictions, refuse to allow the free flow of information that would result in a full vetting of the uses, limitations, and side effects of the drug. The Second Circuit held that such restrictions violate the principles of the First Amendment.”

The article concludes: “The question now is whether a host of other state and federal regulations can withstand such First Amendment scrutiny.” Thank goodness the First Amendment protection of speech and press still appears to have some teeth!

This case is a logical extension of our First Amendment arguments in Pearson v. Shalala.

Reference

• Marcia M. Boumil, J.D., LL.M. Off-label marketing and the First Amendment.N Engl J Med. 368(2):103-5 (2013).

LINEAR THRESHOLD DOSE RESPONSE MODEL OF
RADIATION DAMAGE MAY BE ABOUT TO EXIT STAGE LEFT

The model that has long been used to predict tissue damage from low dose radiation is based upon extrapolation from high dose radiation exposure. As has been noted in recent scientific studies,^1–5 however, this model poorly predicts the effects in the low dose radiation range, whereas the hormetic model (a biphasic response) provides excellent predictions.

According to Edward J. Calabrese,¹ the biphasic dose response model was originally proposed by Schulz in the 1880s based on his extensive testing of the effects of numerous disinfectants on the metabolism of yeast. Unfortunately, however, Schulz went on to claim that his dose-response findings provided the explanatory principle of homeopathy, leading to marginalization of both Schulz and his theory (due to the conflict between orthodox medicine and homeopathic practitioners). It would be another 60 years before the biphasic response (hormetic) explanation for low dose effects of radiation was to become a respected notion. However, government agencies that regulate toxic substances (such as the EPA) still use the linear threshold dose response model. By so doing, these agencies are costing huge sums of money using data that do not provide accurate estimates of toxicity at low doses, hence, not delivering greater safety in exchange for these immense costs.

We are beginning to see more scientific papers published in support of the hormesis explanation of why extrapolation of high dose radiation damage to tissues is not a proper way to estimate the damage, if any, to tissues derived from low dose radiation.

Dr. Calabrese¹ reports his research and that of his colleagues during the past approximately ten years which, using three data sets, has found that “the threshold and the linear models poorly predicted the effects in the low dose zone whereas the hormetic model made uniformly accurate predictions.” The three data sets included1 publications in the peer-reviewed pharmacological and toxicological literature on a broad range of biological models, endpoints and chemicals, (2) an NCI series of 57,000 dose response involving over 2200 anti-tumor agents that had been tested on 13 strains of years considered models of human tumor cells, and (3) about 2100 agents in E. coli.

Dr. Calabrese notes that the threshold model was never validated and that during this long period during which a poor model was being used in toxicology and government agency regulations, it failed multiple, large objective validation tests. As Calabrese notes, getting the “dose response wrong was not simply an historical curiosity involving a conflict between homeopathy and traditional medicine, but one that has shaped the risk assessment process, profoundly affecting clinical medicine, public health, risk communication messages, personal health choices, the proper allocation of vast public/private resources as well as governmental legislation/regulatory programs.”

A second paper² published in the latest FASEB J. examined the effects of low dose radiation exposure in a mouse model using mice that are very sensitive to low dose radiation, changing coat color in response to it. As the authors note in their introduction, “the human health risks of LDIR [low dose ionizing radiation] are still estimated by extrapolation from the biological effects observed at high doses, according to the linear no theshold (LNT) risk assessment model.” Pointing out that high doses of ionizing radiation can result in epigenetic modifications to DNA in adult mice, they wanted to study whether epigenetic alterations also occur in vivo in response to low dose ionizing radiation.

They used a type of mouse (the A^vy mouse) that, in response to early developmental exposure to methyl donors (genistein, bisphenol A, ethanol) and in vitro culturing develop phenotype changes by altering the epigenome. In this study, the researchers show that low dose ionizing radiation causes both dose- and sex-dependent epigenetically induced adaptive changes at the A^vy locus that in part depend on a cellular oxidative stress response.2

As the authors explain, this mouse is very sensitive to environmental effects on the fetal epigenome. Hypomethylation causes inappropriate Agouti gene expression in all tissues, leading to a yellow coat color but also other effects such as developing obesity, diabetes and cancer. When the promoter of this gene is hypermethylated, however, the animals have a much lower incidence of disease as well as having a pseudoagouti (brown) color. The experimental results showed that low dose ionizing radiation altered DNA methylation such that the animals ended up with a brown color and the reduced levels of disease as an adaptive response. This, the authors say, was due to exposure during pregnancy to doses of radiation equivalent to those received for a chest or head CT scan. “Hypomethylation of DNA at high doses of radiation and hypermethylation at low doses are indicative of a hormetic biphasic radiation dose response effect.”² (The authors note that hypermethyl­ation was beneficial in this particular disorder but is not always beneficial. For example, tumor suppressor genes can become hypermethylated and, as a result, silenced, so that cancer risk is increased.)

The study² “brings into question the assumption that every dose of radiation is harmful.”

References

1. Calabrese. Hormesis: Toxicological foundations and its role in aging research. Exper Gerontol. 48:99-102 (2013).
2. Bernal et al. Adaptive radiation-induced epigenetic alterations mitigated by antioxidants. FASEB J. 27:665-71 (2013).
3. Son et al. Hormetic dietary phytochemicals. Neuromolecular Med. 10(4):236-46 (2008).
4. Hayes. Nutritional hormesis and aging. Dose Response. 8:10-5 (2010).
5. Wu et al. Multiple mild heat-shocks decrease the Gompertz component of mortality in Caenorhabditis elegans. Exper Gerontol. 44:607-12 (2009).

PREDATOR VS. PREY

by Sandy Shaw & Durk Pearson
(originally appeared in Liberty)

A recent paper in Science¹ on what caused the extinction of large North American canids (wolf-like carnivores) is startlingly parallel to what is happening to the government of the United States and its constituent states and leads us to a hypothesis concerning the death of democracies.

The paper reports that during the past 50 million years, successive branches of large carnivorous mammals have diversified and then gone into extinction. The authors¹argue that “energetic constraints and pervasive selection for larger size (Cope’s rule) in carnivores lead to dietary specialization (hypercarnivory) and increased vulnerability to extinction.” They explain that Cope’s rule—the evolutionary trend toward larger size—is common in mammals because larger size makes it easier to evade predators and to capture prey. Moreover, larger size improves thermal efficiency, thus increasing the potential range of habitats into colder areas. As the size of carnivores increases beyond about 45 pounds, the amount of nutrition obtained from small prey becomes inadequate to cover the energy used in capturing them. Thus, the larger carnivores became what the authors call “hypercarnivores,” which hunt only large prey (as large as or larger than themselves). A plot of the index of hypercarnivory (PCI score) against estimated species duration shows that none of the hypercarnivorous species persisted for more than 6 million years, as compared to other, more omnivorous species that lasted as long as 11 million years. Hence, the authors propose, the hypercarnivores are more vulnerable to extinction. The researchers also note that hypercarnivores reverting to a more generalized diet and morphology was rare.

Reliance upon a smaller number of large prey increases the statistical variation in the nutritional intake. Moreover, the larger the carnivore, the lower their population density. Both of these are factors that increase the risk of extinction. It is interesting that the government of the United States and of its constituent states are moving rapidly in the direction of targeting large prey, with an increasing statistical variance in the yearly revenue from these relatively small number of prey. The federal government relies heavily on a steeply progressive income tax (with the well known result that the bottom 50% of income earners pay only 4% of income taxes). Relying upon the fat targets at the highest level of earnings has resulted in greater statistical variation in revenues, as well as increasing demand for government services from those paying little or nothing for them. Worse yet, today’s government is already preying upon the fat targets of the future by rapidly increasing government debt, something canids never had the option to do.

We suggest, therefore, that hypercarnivory may be one reason that democracies don’t last much longer than about 200 years. If we start counting from the passage of the 16th Amendment on Feb. 3, 1913 (which provided the means to target most of the large prey via unlimited progressive income taxation) rather than from 1787, the United States could theoretically last another century or so, though for a number of reasons (the immensely rapid expansion of government debt, for instance) we think it unlikely.

APPETIZERS

Can’t act, slightly bald, also dances.
— RKO Pictures screen test, comments about Fred Astaire

I suppose a knighthood is out of the question now?
— Spike Milligan, in a fax to Prince Charles after he called him a “little groveling bastard.”

A thing moderately good is not so good as it ought to be. Moderation in temper is always a virtue; but moderation in principle is always a vice.
— Thomas Paine, Common Sense

It is hard to imagine a more stupid or more dangerous way of making decisions than by putting those decisions in the hands of people who pay no price for being wrong.
— Thomas Sowell

You can’t possibly hear the last movement of Beethoven’s Seventh and go slow.
— Oscar Levant, explaining his way out of a speeding ticket

THE SLEEPING BRAIN DECIDES WHAT TO REMEMBER AND WHAT TO FORGET

A new paper¹ describes the sleep-dependent memory-processing factory that decides what to do with all that information you encountered during the day. As the paper’s authors point out, only some of that information is consolidated so as to become a part of a long-term knowledge base. The paper sifts through evidence from studies of naps, sleep deprivation, correlations of sleep stages and memory processing, sleep physiology, regional brain activity measured during and after sleep with PET and fMRI studies, cellular firing patterns, and synaptic and intracellular measures of plasticity to conclude that there is convincing evidence of sleep processing of memory with improvement of the overall knowledge base.

The authors discuss the new understanding that not all information is uniformly preserved, but that there is an exquisite selection process of memories underway during sleep. For example, they report that emotional memories can be selectively consolidated, especially during rapid eye-movement (REM) sleep. It has also been found that memories can be selectively maintained when they contain information on potential monetary rewards. Interestingly, when subjects of sleep memory studies have been told that they would be tested on certain areas of information and not on others provided before sleep, they were found after sleep to have retained more of the information they were told they would be tested on. Hence, the brain “knew” what to do to recall the relevant information.

Moreover, the authors explain, it is possible for the brain to generate new information during the processing of the memory-derived information. “Whether consolidation necessarily precedes these integrative processes (serial processing) is not yet known, but no clear cases of integration without consolidation have been observed. We use the term ‘memory evolution’ to reflect both the qualitative changes that can occur during such integrative processing and the extended time course over which they occur.”¹ In gist extraction, the authors refer to the identification of commonalities between items in a collection of memories even when individual item memories are forgotten. Some studies have been found to show that during sleep subjects can extract overarching rules that govern recently studied sets of information, such that understanding of relationships between the sets is improved following sleep.

One study reported by the authors dealt with subjects taught a rote method for solving a class of mathematical problems for which there was a shortcut solution (about which subjects were not told). After a night of post-training sleep, however, subjects were found to be 2.6 times more likely to discover this shortcut than after an equal period of time awake (59.1 versus 22.7% of subjects).¹ But, interestingly, even those who did not discover the shortcut benefitted from sleep by improving the speed with which they were able to perform the rote method of solving the problems. Those who became faster without discovering the shortcut improved their speed (using the rote method) three times more than either those who discovered the shortcut or those who remained awake.

This study examines important sleep processes at a systems level rather than at a neurotransmitter level. Understanding sleep involves comprehending its mechanisms from the micro-level details (neuro­transmitters and synapses) to the overarching system architecture. For more about the neurotransmitter level, see our article describing how our SleepScape formula works here: (www.life-enhancement.com/magazine/ article/2807-introducing-sleepscape) and the first of its ingredients (glycine) here:(www.life-enhancement.com/ magazine/article/2813-ingredients-in-sleepscape) , the second of its ingredients (GABA) here: (www.life-enhancement.com/magazine/article/2836-ingredients-in- sleepscape)

Life Enhancement magazine and Durk Pearson & Sandy Shaw’s Life Extension Newsletter therein have been providing details on SleepScape’s ingredients, continuing this month on page 17 with choline (a precursor to the neurotransmitter acetylcholine).

Reference

1. Stickgold and Walker. Sleep-dependent memory triage: evolving generalization through selective processing. Nat Neurosci. 16(2):139-45 (2013).

HYDROGEN GAS PROTECTS BEAGLE DOG HEARTS FROM
ISCHEMIA-REPERFUSION INJURY-INDUCED INFARCTION

Researchers investigated a possible mechanism to explain how the inhalation of hydrogen gas following ischemia-reperfusion injury in rat hearts limits the infarct size.¹Noting that the effect of hydrogen on such heart injury in large animals has not been studied, they focused their study on beagle dogs that had the left anterior descending coronary artery occluded for 90 minutes, followed by reperfusion for 6 hours. The animals were treated with either 1.3% hydrogen or control gas (70% nitrogen and 30% oxygen) ten minutes prior to reperfusion until 1 hour of reperfusion had taken place.

The area of killed heart cells (infarct) was reduced by hydrogen gas as compared to control gas (and normalized by risk area) 20.6 ± 2.8% vs. 44.0 ± 2%; p<0.001. (In other words, the area at risk due to the occlusion following by reperfusion was reduced by 20.6 ± 2.8% by the hydrogen gas treatment as compared to 44.0 ± 2% by the control gas treatment.) They also identified the opening of mitochondrial K-ATP channels followed by inhibition of the mitochondrial permeability transition pores as accompanying the protective effects of limiting ischemia-reperfusion-induced infarct. The authors note that “recent accumulated evidence regarding cardioprotection afforded by ischemic pre- or post-conditioning has culminated in the idea that opening of mitochondrial ATP-sensitive K+ channels followed by inhibition of mitochondrial permeability transition pores plays a central role in limiting infarct size.”¹

The researchers used drugs that prevent the opening of the mitochondrial K-ATP channel as a way to test whether the protective effects of hydrogen gas was mediated by the opening of these channels. The administration of either of two such drugs abolished the infarct size-limiting effect of hydrogen gas.

The researchers also observed that inhalation of hydrogen gas tended to reduce the incidence of lethal ventricular arrhythmia, although they did not investigate the mechanisms responsible for that.

Reference

1. Yoshida et al. H2 mediates cardioprotection via involvements of K-ATP channels and permeability transition pores of mitochondria in dogs. Cardiovasc Drugs Ther. 26:217-226 (2012).

IMPORTANT ADVANCE IN CANCER TREATMENT:
RESTORING ACTIVITY OF TUMOR SUPPRESSOR GENES
NO FDA APPROVAL REQUIRED!

A very recent paper¹ reports the exciting news that curcumin, a yellow pigment derived from the curry spice turmeric, has been shown to reactivate several hyper­methyl­ated (turned off) tumor suppressor genes in lung, colon, prostate, and breast cancer cells. One of the most common causes of cancer is loss of one or more key tumor suppressor genes, which can occur as a result of mutation, but also by another process, DNA hyper­methyl­ation.

Too little DNA methylation (hypomethylation) can result in the activation of oncogenes that promote cancer, whereas excessive DNA methylation (hypermethylation) can silence tumor suppressor genes that would otherwise inhibit the development of cancer. When tumor suppressor genes are hypermethylated, they are prevented from being expressed. Unlike mutations, however, DNA hypermethyl­ation can be reversed by dietary methylation inhibitors for chemoprevention and also chemotherapy.¹

A new paper¹ now reveals in vitro and in vivo evidence in mice that curcumin can reactivate the tumor suppressor gene RASSF1A in breast cancer cells, resulting in significant activity against tumor growth. Another paper² reports that “RASSF1A methylation has the potential to be an ideal cancer biomarker as it occurs at moderate to high frequency in a very wide range of tumour types, yet is comparatively rarely found in normal tissues.” The same paper² also reports that “RASSF1A promoter methyl­ation has been demonstrated in epithelial hyperplasia and intraductal papillomas of the breast, as well as cancerous epithelium, suggesting that RASSF1A methylation is an early event in breast tumourigenesis.” RASSF1A methyl­ation has also been suggested to be an early event in thyroid tumouri­genesis, childhood neoplasia, and endometrial carcino­genesis.² In lung cancer, RASSF1A methylation as a diagnostic marker has been investigated and found to occur in ~34% of NSCLC (non-small cell lung cancer) tumors, accompanied by methylation in the corresponding serum.²

Another paper³ describes curcumin as a “potent DNA hypomethylation agent” and explains how the authors understand curcumin to restore the activity of deactivated tumor suppressor genes. The authors of this paper³ note that two nucleoside analogs (azanucleosides) with hypomethylating activity (decitabine or 5-azacytidine) have been approved by the FDA for the treatment of myelodysplastic syndrome, but that, although the results are encouraging, these drugs can be very toxic (by suppressing bone marrow). Curcumin, on the other hand, has been shown to be well tolerated in three different phase 1 clinical trials of up to 12 grams a day.^3B The authors³ found that curcumin and one of its major metabolites, tetrahydrocurcumin, are effective in inhibiting hypermethylation induced by DNMT1 (a DNA methyltransferase). Using a leukemia cell line, the global DNA methylation of MV4-11 cells remained unchanged at 1.0 μM curcumin, but decreased about 15–20% at 3.0 and 30.0 μM curcumin compared to the untreated basal methylation level of the cell line.

A paper⁴ with the same lead author as paper #3 who is also the last author of paper #1 reports that the sesquiterpene lactone parthenolide, the principal bioactive sesquiterpene lactone of the herb feverfew, reactivated the tumor suppressor gene HIN-1 by inhibiting DNA methyltransferase 1 (DNMT1) and thus is an agent of DNA hypomethylation.

Finally, an earlier paper⁵ reported that EGCG, a major polyphenol in green tea, was effective in inhibiting DNA methyltransferase and reactivating methylation-silenced tumor suppressor genes in cancer cell lines.

What these results suggest is that cancer prevention could be enhanced, perhaps very significantly, by taking hypomethylation agents such as these in safe doses and that their use (possibly together with standard chemotherapy, which is often effective in cancer patients that do not have silenced tumor suppressor genes) could markedly improve chances of survival for these patients who, in the absence of reactivating their silenced tumor suppressor genes may have a dismal prognosis.

Another advantage of using safe natural products for the treatment of cancer is that, provided no claims are made for their efficacy in the treatment of cancer, they do not have to receive FDA approval for sale in the marketplace and, hence, do not require a doctor’s prescription and are generally relatively inexpensive. However, that makes you responsible for your own health in a really major way, so be sure to work with a knowledgeable physician if you choose to use these substances to treat your cancer, as there may be very little in the way of clinical data on their use (correct dosages are usually difficult to predict from animal studies). There is nothing that can replace a physician with a lot of experience, who follows the natural products scientific literature, and who has good diagnostic skills. Also avoid, as much as possible, doctors who are tied up in knots by bureaucratic political rules and regulations (e.g., Medicare, Medicaid, Obama­care) that may (though they may not want to admit it even to themselves) make it impossible for them to use their best judgment in treating their patients.

HYPERMETHYLATION ISN’T JUST A MECHANISM AFFECTING
THE REGULATION OF TUMOR SUPPRESSOR GENES: AGE-
RELATED NUCLEAR CATARACTS

A new paper⁶ reports that the gene (CRYAA) critical for the maintenance of eye lens transparency can be repressed by hypermethylation in age-related cataract, the leading cause of blindness among older adults. The paper mentions DNA methylation inhibitor drugs, zebularine and 5-Aza-C as possible treatments, but refers to complications related to their toxicity, though mentioning that zebularine has much lower toxicity than 5-Aza-C. However, we refer to the above description of nutritional hypomethylating agents that are almost certainly to be far lower in toxicity. Apparently, these natural substances have not yet been tried as treatments for age-related cataract or we would have expected to see this mentioned in the paper. We hope to see such research soon, though we note that these natural substances cannot be patented and, hence, are unlikely to receive research funding by pharmaceutical companies. In fact, Big Pharma has heretofore been likely to use its huge political lobbying machine to prevent competition by natural products, in part by engaging the FDA on their behalf.

If dietary supplement companies could make truthful claims concerning their effects on cataracts (let alone in the treatment of cancer), we would see substantial money invested by dietary supplement companies to establish data to support truthful health claims for diseases. If you can’t tell people about the results of your research, what’s the point of doing it? The FDA’s censorship of truthful information results in the suffering and, in the case of cancer, premature deaths of large numbers of Americans. Incredibly, most Americans believe the Big Lie that the FDA protects people from the lies of dietary supplement companies who will supposedly say anything to sell you their pills when, as we see it, the FDA’s suppression of truthful health claims are causing far more harm. No system can entirely eliminate false claims, but the current one is throwing out most truthful claims (and freedom of speech along with it) in order to eliminate false claims. Unfortunately, most of what is published in major media as “news” on health claims largely parrot back what is contained in FDA press releases.

References

1. Du et al. Reactivation of RASSF1A in breast cancer cells by curcumin. Nutr Cancer.64(8):1228-35 (2012).
2. van der Weyden and Adams. The Ras-association domain family (RASSF) members and their role in human tumourigenesis. Biochim Biophys Acta. 1776(1):58-85 (2007).
3. Liu et al. Curcumin is a potent DNA hypomethylation agent. Bioorg Med Chem Lett.19:706-9 (2009).
3B. Anand et al. Bioavailability of curcumin: problems and promises. Mol Pharm.4(6):807-18 (2007).
4. Liu et al. Modulation of DNA methylation by a sesquiterpene lactone parthenolide. J Pharmacol Exp Ther. 329(2):505-14 (2009).
5. Fang et al. Tea polyphenol (-)-epigallocatechin-3-gallate inhibits DNA methyltransferase and reactivates methylation-silenced genes in cancer cell lines. Cancer Res. 63:7563-70 (2003).

STUCK IN AN APATHETIC RUT? CHOLINESTERASE INHIBITION
AMELIORATES APATHY AND LOSS OF MOTIVATIONAL DRIVE IN MICE

Scientists studying neural mechanisms for apathetic behavior report1 that a cholinesterase inhibitor (phenserine was used in this study) reversed many of the signs of apathy in male C57BL/6 mice) subject to chronic restraint stress. As the authors explain, apathy shares some overlapping features with depression, but is not the same. Apathy “can be distinguished [from depression] by lack of dysphoric symptoms including sadness, hopelessness and guilt.”¹ Instead, apathetic animals exhibit extreme withdrawal of involvement and a paucity of emotion, a state characterized by a lack of motivation in relation to goal-oriented and self-initiated activity.

The authors wanted to test whether facilitating the cholinergic system could ameliorate the apathetic state induced by chronic restraint stress. It is not clear why they used the particular cholinesterase inhibitor (phenserine) that they did, but it is interesting to note that the final author of the paper (who is usually the scientist who runs the lab where the work is done) is listed as a paid employee of Johnson & Johnson Pharmaceutical Research under “Competing Interests” at the end of the paper. This MAY (just a speculation on our part) indicate that the cholinesterase inhibitor is a product of Johnson & Johnson Pharmaceutical Research.

Several features of the apathetic behavior exhibited by the control animals were ameliorated in the experimental animals treated with the cholinesterase inhibitor. “Overall, CRS [chronic restraint stress]-exposed mice displayed significant decreases in exploratory behavior such as rear-up and sniff, as well as in locomotor behaviors including walk slowly, remain low, and hang cuddled for several hours directly prior to the onset of the dark cycle and before the onset of the light cycle, suggesting a decrease in initiation of non-essential activities. However, directly prior to dark cycle onset, inactive behaviors such as twitch and rest increased in the CRS mice.”¹ The researchers observed accumulation of deltaFosB in the MS/vDB, said to be one of the major basal forebrain cholinergic nuclei, suggesting that this area had undergone significant changes in gene expression as a result of CRS.

The authors note that two other papers reported that the cholinesterase inhibitors galantamine and donepezil lead to increased dopamine release in the NAcc (nucleus accumbens), an area associated with motivation and reward and speculate that the behavioral effects of phenserine in this paper may have resulted from activating cholinergic interneurons in brain areas associated with motivation and reward.

Reference

1. Martinowich et al. Acetylcholinesterase inhibition ameliorates deficits in motivational drive. Behav Brain Funct. 8:15 (2012)

POWERFUL PUNCH OF EVIDENCE-SUPPORTED
AROMATHERAPY: MENTHOL PROTECTS AGAINST BETA
AMYLOID PEPTIDE INDUCED COGNITIVE DEFICITS IN MICE

A surprising new paper¹ reports that menthol, a constituent of essential oils (e.g., peppermint and other mint oils) that have been used since the time of ancient Egypt and are now considered part of the aromatherapy wing of alternative medicine, provided significant protection in aged and beta-amyloid protein-treated mice against cognitive decline. Sniff, baby, sniff!

One particularly nice thing about menthol is that it is readily available, safe and inexpensive, and easy to use. In fact, menthol (frequently in combination with eucalyptus, camphor, and thymol) is a commonly used and effective bronchodilator that, unlike bronchodilator drugs such as beta-adrenergic agonists, does not cause cardiac arrhythmias or other adverse reactions.

The researchers reported that menthol administered successively for 10 days significantly increased the amount of time spent in the target quadrant of the dreaded Morris water maze in young mice, thus showing improved spatial learning and memory. Moreover, “pre-treatment with menthol reversed the amnesia induced in animals upon treatment with beta-amyloid on 10th day.” Menthol improved working memory in both aged and young mice, with the potentiation of working memory more profound with 100 mg/kg s.c. menthol. Other studies were cited in which menthol exhibited anticholinesterase inhibitory activity and increased intracellular ATP in nerve cells (which the researchers proposed may have an inhibitory effect on ATP gated potassium channels).

Beta-amyloid peptide treatment administered to young mice on day 7 reduced glutathione content (increased oxidative stress).

Pretreatment with menthol showed a significant increase in glutathione content, reversing the increased oxidative stress seen in the animals not pretreated with menthol.

“Higher dose of menthol also improves the brain function through decrease in malondialdehyde level [lipid peroxidation product] in aged and beta-amyloid treated mice. Lower dose of menthol (100 mg/kg s.c.) showed improvement in spatial working memory as compared to higher dose.”¹

One easy way to use menthol is to buy it as a spray, spray a couple of spritzes into a Kleenex^® tissue, then hold over your nose and breathe deeply a few times.

Whether enough menthol can be obtained by inhalation of its vapor was not determined by this experiment since it was administered by subcutaneous injection. Never­theless, peppermint oil can certainly be taken orally, though we recommend diluting it with a vegetable cooking oil to prevent irritation. Note that menthol delivered by either peppermint hard candy or smoking (though we do not recommend smoking to obtain menthol) is directly absorbed by the oral and nasal mucosa and efficiently transported to the brain.

Reference

1. Bhadania et al. Protective effect of menthol on beta-amyloid peptide induced cognitive deficits in mice. Eur J Pharmacol. 681:50-4 (2012).

WAKE UP AND FEEL YOUR JUICES FLOW WITH DURK & SANDY’S MORNING SMOOTHIE

A delicious, nutritious fun smoothie—you will need:

• 8–10 ounces milk
• two or three handfuls of fresh or frozen blueberries (frozen are great because they help chill your smoothie); fresh or frozen strawberries are great, too
• 40–60 grams (4–6 tablespoons) of whey protein
• two to three tablespoons of natural cocoa (NOT dutched—dutching destroys much of the healthful polyphenols)
• two tablespoons of erythritol
• 1⁄2 tsp. vanilla

Plus (as desired):

• 1 tablespoon Hydrogen Power™ (prebiotic for hydrogen producing gut microbes)
• 1⁄2 tsp. taurine (tasteless)
• 1⁄2 tsp. trehalose (adds no taste)
• 1⁄2 tsp. glutamine (adds no taste)
• 1 tsp. Memory Upgrade III™ (contains 1 gram choline)
• 1 tbsp. InnerPower Plus™ (contains 6 g. arginine)

Also, if desired, add 1⁄4 cup ice cream. You can add more, but remember there is a lot of fat and calories in the ice cream.

Process for about 25 seconds until blended. Enjoy!

We drink one of these smoothies nearly every morning and sometimes have one again in the afternoon. Durk also adds a single serving of the French Roast Bold coffee by Starbucks to his (only in the morning).