Somewhere, something incredible is waiting to be known.—Blaise Pascal (1623-1662)
Why should I? Everyone knows me there
(upon being told by his wife to dress properly when going to the office)
Why should I? No one knows me there
(upon being told to dress properly for his big conference)—Albert Einstein,
Quoted from Ehlers, Liebes Hertz! (1994)Alliance, n. In international politics, the union of two thieves who have their hands so deeply in each other’s pocket that they cannot separately plunder a third.—Ambrose BierceHere richly, with ridiculous display
The politician’s corpse was laid away.
While all of his acquaintances sneered and slanged,
I wept; for I had longed to see him hanged.—Hilaire Belloc
The meek shall inherit the earth, but not the mineral rights.—J. Paul GettyAbove quotes from The Big Curmudgeon, ed. Jon Winokur, Black Dog & Leventhal Publishers, New York, (2007)
Vitamin C Treatment Reduces C-Reactive Protein but Only in Those Who Have High CRP
A new study1 reveals more evidence to support the view that the reason antioxidants have “failed” to show benefits in intervention studies is that (1) target biomarkers (such as levels of oxidative stress) have not been established at baseline, and (2) that antioxidants should not have been expected to improve people with low levels of oxidative stress or other biomarker of disease risk.
In the new study, researchers examined the effects of vitamin E or vitamin C for two months on CRP levels in 396 healthy nonsmokers. The findings showed that, while vitamin E had no effect, vitamin C (1000 mg/day) reduced the median CRP by 25.3% (as compared to placebo) but only in those participants with elevated CRP (≥1.0 mg/L), indicative of elevated cardiovascular disease risk. When all participants were included in the analysis, no vitamin C effect on CRP was seen because the median baseline CRP was only 0.85 mg/L and no significant treatment effect was seen in those with CRP <1.0 mg/L.
Why Earlier Antioxidant Intervention Trials “Failed”
As the authors conclude, “[t]hese results may shed light on the mixed results found in large ‘antioxidant’ trials with clinical endpoints. No such trials have limited participants to persons with elevated CRP or oxidative stress, and most have not characterized participants with respect to those factors.” “If one mechanism of potential antioxidant treatment is reduction in CRP, inclusion of subjects without elevations in CRP would tend to have weakened such studies, particularly those involving healthy volunteers. In the present study of healthy volunteers, 55% of participants had CRP values <1.0 mg/L.”
Vitamin C as Effective as Statins in Reducing CRP
The authors also discuss several trials of statins on CRP levels and the effectiveness of vitamin C in this study as compared to statins. The 0.25 mg/L (16.7%) reduction of CRP by vitamin C found in the participants with baseline levels of CRP >1.0 mg/L “suggests an effect of vitamin C on CRP that is similar to that observed with statins.”
- Block et al. Vitamin C treatment reduces elevated C-reactive protein. Free Radic Biol Med 46:70-7 (2009).
A recent meta-analysis of 12 randomized controlled trials for short term L-arginine oral supplementation reported an improvement in flow-mediated dilation (FMD, the ability of blood vessels to relax in response to appropriate stimuli, such as acetylcholine release), but only when baseline FMD is low.1 Note the similarity to the result with Vitamin C and CRP where CRP was reduced only when baseline CRP was high.* In this instance, L-arginine supplementation significantly increased FMD when the baseline levels were <7%, but had no effect when baseline FMD was >7%.1 In the pooled analysis (in which all subjects were included), L-arginine supplementation significantly increased the FMD by 1.98%, whereas the FMD was increased by 2.56% in those with baseline FMD <7% and was not significantly changed (–0.27%) in those whose baseline FMD was >7%. In order to understand the effects of L-arginine on FMD as reported in these trials, it is necessary to know the baseline levels of FMD and to analyze the effects on the basis of changes in baseline FMD. The trials included in the meta-analysis had 10 to 36 subjects, doses of L-arginine ranged from 3 g to 24 g per day, and subjects included children with chronic kidney failure, patients with heart failure, healthy young men, healthy individuals older than 70 y, clinically asymptomatic elderly subjects, patients with peripheral artery disease, or hypercholesterolemic patients.
*See “Vitamin C Treatment Reduces C-Reactive Protein but Only in Those Who Have High CRP” at:
We both take L-arginine supplements (our InnerPower Plus™ formulation contains 6 grams of L-arginine per serving). Though neither of us has to our knowledge a low baseline FMD, there are many other healthful properties of L-arginine (which we have written about in earlier newsletters), such as its action as a precursor of nitric oxide, important for numerous functions, including proper endothelial vasodilation. As another example, arginine supports healthy kidney function by reducing the age-associated increase in the basement membrane (thickening of which reduces the kidney’s ability to filter blood) as well as being part of the urea cycle that allows the body to excrete ammonia formed during the metabolism of amino acids.
- Bai et al. Increase in fasting vascular endothelial function after short-term oral
L-arginineis effective when baseline flow-mediated dilation is low: a meta-analysis of randomized controlled trials. Am J Clin Nutr 89:77-84 (2009).
Phenolic Compounds Rutin and o-Coumaric Acid
Ameliorate Obesity Induced by High Fat Diet in Rats
One of us (Sandy) has two pet female rats (Queen and Tiny) and, while it is a lot of fun (because they are soooooo appreciative) to feed them a cafeteria style diet (lots of variety), you have to be very careful about limiting fat and calories so they do not become fat. Rats have short lifespans as it is and obesity will just increase the risk that they die young (probably of cancer). We report here a study showing that increasing the dietary content of the phenolic compounds rutin or o-coumaric acid reduced the obesity-inducing effect of a high fat diet in rats. Take note, Queen and Tiny . . . (munching in the background).
A number of studies have reported beneficial effects of phenolic compounds in reducing the risks of high fat diets in humans and other animals, including red wine polyphenols, antioxidants, and gallic acid.1 A recent in vitro study1 showed that the phenolic compounds rutin and o-coumaric acid inhibited intracellular triglyceride and glycerol-3-phosphate dehydrogenase (GPDH) activity the best among the 15 phenolic acids and 6 flavonoids that were tested. “GPDH occupies a central position in the triglyceride synthesis
In their in vivo study,2 the researchers divided Wistar rats into normal and obese groups. The obese rats were then prefed a high fat diet (HFD, 40% beef tallow) for four weeks following which the normal rats received the ND (normal diet) for 8 weeks, while the obese group was fed the HFD plus R (rutin, either low or high dose), or HFD plus oCA (o-coumaric acid, either low or high dose) for 8 weeks. The high doses were 100 mg/kg, while the low doses were 50 mg/kg (roughly equivalent to 1 to 2 grams per day for an adult human).
Body weight was significantly increased (21%) in the HFD group as compared to the ND group, whereas there was a significant decrease in the increased body weight in the HFD plus R and the HFD plus oCA groups, both low and high doses, as compared to the HFD group. The weights of the liver and adipose tissue in the HFD plus R and HFD plus oCA were significantly decreased as compared to those of the HFD group.
Oxidative stress was significantly reduced in the HFD plus R and the HFD plus oCA as compared to the HFD group. For example, by the end of the study, the reduced glutathione level, an important marker of antioxidant capacity, was 3.38 ± 0.20 µmol/mg protein (ND) as compared to 0.77 ± 0.07 µmol/mg protein (HFD), but was 3.03 ± 0.35 (HFD plus R, high dose), 2.6 ± 0.3 (HFD plus oCA, high dose), 1.54 ± 0.15 HFD plus R, low dose), and 1.4 ± 0.1 (HFD plus oCA, low dose). MDA (malondialdehyde, a product of lipid peroxidation) was increased by the HFD (13.1 ± 1.7 nmol/mg protein) as compared to the ND (2.8 ± 0.9 nmol/mg protein), but that increase was attenuated by the addition of rutin or o-coumaric acid to the HFD: HFD plus R (low dose), 12.0 ± 1.2; HFD plus R (high dose), 7.7 ± 0.9; HFD plus oCA (low dose), 11.3 ± 1.1; HFD plus oCA (high dose), 9.7 ± 0.9.
Antioxidant enzymes, such as glutathione peroxidase, glutathione reductase, and glutathione-s-transferase showed 35, 57, and 40% reductions, respectively, in the HFD group as compared to the ND group, whereas those enzymes in the HFD plus R and HFD plus oCA groups were significantly increased as compared to those in the HFD group.
We highly recommend increased dietary intake and supplementation of rutin. While we do not yet offer o-coumaric acid in any of our supplements, the figures in this study suggest an overall better result with the rutin supplement (at least in Wistar rats) and we do get a substantial amount of rutin from our Durk & Sandy’s® AGEless which, in the recommended 6 capsules a day, contains 499.8 mg. of rutin. As we explained in our article on AGEless (see the Feb. 2008 issue of Life Enhancement), rutin has powerful protective effects against the formation of AGEs (advanced glycation endproducts), part of the aging process in which sugars (including glucose, fructose, ribose, and others) in the bloodstream chemically react (glycosylation) with proteins.
- Hsu and Yen. Effect of flavonoids and phenolic acid on the inhibition of adipogenesis in 3T3-L1 adipocytes. J Agric Food Chem 55:8404-10 (2007).
- Hsu, Wu, et al. Phenolic compounds rutin and o-Coumaric acid ameliorate obesity induced by high fat diet in rats. J Agric Food Chem 57:425-31 (2009).
Acute Treatment of Stroke with Red Wine Polyphenols
A very interesting study1 reports highly protective effects of red wine polyphenols in a rat stroke model (90 minutes of middle cerebral artery occlusion). The results suggest that red wine polyphenolic compounds (RWPC) might be an effective therapy if administered during an acute stroke to reduce the resulting damage, though they were administered intravenously rather than orally in the study. Oral dosage would very likely still be protective, but not necessarily to the same extent as IV due to the faster delivery to the damaged tissues with IV.
The authors compared the resulting damage after 90 minutes of middle cerebral artery occlusion between untreated rats and those receiving a bolus of RWPC at the onset of cerebral ischemia. (The dose used in the study was 0.1 mg/kg—corresponding to the RWPC found in 1/20 glass of red wine—which is comparable to the lowest dose of Provinol™ (a branded RWPC product) applied orally (0.2 mg/kg/day) in a previous study that the researchers report had shown a potent protective effect in a model of leg ischemia.
The protective effects they document include decreasing toxicity elicited by the release of excitatory amino acids (glutamate and aspartate), increasing the release of free radical scavengers (uric acid and ascorbic acid), and improving blood flow restoration as well as improving cerebral energy metabolism (by increasing the availability of the energy substrates glucose and lactate).1
An earlier study reported by the same authors2 found that chronic pre-treatment (before middle artery occlusion) with RWPC was able to “increase brain perfusion during occlusion by inducing a remodeling of the cerebral arteries and to completely block the release of EAA [excitatory amino acids].” In contrast, the acute administration of RWPC did not have a significant effect on blood flow during occlusion, but only during reperfusion.
- Ritz et al. Acute treatment with red wine polyphenols protects ischemia-induced excitotoxicity, energy failure and oxidative stress in rats. Brain Res 1239:226-34 (2008).
- Ritz et al. Chronic treatment with red wine polyphenol compounds mediates neuroprotection in a rat model of ischemic cerebral stroke. J Nutr 138:519-25 (2008).
Superoxide Radicals, NO, and cGMP:
Links Between Erectile Dysfunction and Cardiovascular Disease
There is a known correlation between erectile dysfunction and the risk of cardiovascular disease. For example, in a recent paper1 researchers commented “[t]he importance of PDE5 [phosphodiesterase 5, which is inhibited by Viagra, Cialis, and certain other ED therapies] in the aetiology of vasculopathies [endothelial dysfunction] is exemplified by the proven and postulated therapeutic use of sildenafil [Viagra] and other PDE5 inhibitors to treat an array of cardiovascular diseases and syndromes. These include erectile dysfunction, pulmonary hypertension, angina pectoris, myocardial infarction, ischaemia reperfusion injury, vein graft disease and heart failure. In turn, the overproduction of O2- [superoxide radicals] and an upregulation of NOX [NADPH oxidase, which generates superoxide radicals] have been demonstrated in these clinical conditions.”1 See, for example,2 in which daily treatment with sildenafil reversed endothelial dysfunction and oxidative stress in an animal model of insulin resistance.
As discussed in this paper1 and others,3 NADPH oxidase is a major source of superoxide radicals, which interact with nitric oxide (NO), destroying the vasodilative properties of NO and producing a potent oxidant, peroxynitrite. As explained1 by the researchers, “[t]he biological actions of NO are mediated principally by the activation of GC [guanyl cyclase], which generates cGMP [cyclic guanosine monophosphate]. In turn, cGMP activates PKG [protein kinase G] which then phosphorylates other proteins that elicit vasculoprotection. The biological effects of the NO-PKG system are reduced by type 5 PDEs (PDE5) [type 5 phosphodiesterase], which hydrolyze cGMP to inactive GMP. In this way, the type 5 PDEs are able to reduce the ability of NO to dilate blood vessels by inactivating cGMP. The ability of drugs like Viagra to restore erectile function is due to their anti-PDE5 activity. Their cardiovascular protection is, as is clear from the above, also due to inhibition of PDE5, leading to protection of the NO-cGMP system pathway that supports normal endothelial function.
As an examination of the metabolic pathway leading to erectile dysfunction shows, however, there are more ways to improve erections than to directly decrease PDE5 activity. For example, increasing SOD (superoxide dismutase) activity directly decreases the presence of superoxide radicals by scavenging them. This helps prevent the induction of PDE5 by the superoxide radicals, as well as protecting NO from destruction by superoxide radicals. In this particular paper,1 scientists found they could inhibit the upregulation of type 5 phosphodiesterase in human vascular smooth muscle cells with either iloprist, a prostacyclin mimetic drug, or NONOate, an NO donor drug. (These drugs were chosen on the basis that prostacyclin and NO inhibited NOX expression and activity as reported in a separate paper.3)
Some recently identified natural products that also inhibit PDE5 are found in Ginkgo biloba4 and red grapes.5 There are, in addition, many natural products that improve endothelial dysfunction that, theoretically, could be expected at an appropriate dose to improve erectile function. See, for example, papers on L-arginine,6 strawberry extract,7EGCG,8 and cocoa.9,10 The flavonoid quercetin* has been shown to downregulate NADPH oxidase in spontaneously hypertensive rats.11 Quercetin and catechin were shown to synergistically enhance platelet nitric oxide by inhibiting protein kinase C-dependent NADPH oxidase activation in vitro.11b Other papers on NADPH oxidase12,13identify it as having a causative role in erectile dysfunction and as a therapeutic target for cardiovascular diseases, in support of the discussion above.
*Our Personal Radical Shield™ contains, in the recommended 12 capsules per day, 130 mg of quercetin.
1. Muzaffar et al. Superoxide from NADPH oxidase upregulates type 5 phosphodiesterase in human vascular smooth muscle cells: inhibition with iloprost and NONOate,” Br J Pharmacol 155:847-56 (2008).
2. Liming Jin et al. NADPH oxidase activation: a mechanism of hypertension-associated erectile dysfunction. J Sex Med 5:544-551 (2008).
3. Behr-Roussel et al. Daily treatment with Sildenafil reverses endothelial dysfunction and oxidative stress in an animal model of insulin resistance. Eur Urol 53:1272-81 (2008).
4. Dell’Agli et al. Inhibition of cGMP-phosphodiesterase-5 by biflavones of Ginkgo biloba. Planta Med 72:468-70 (2006).
5. Dell’Agli et al. In vitro inhibition of human cGMP-specific phosphodiesterase-5 by polyphenols from red grapes. J Agric Food Chem 53:1960-5 (2005).
6. Bai et al. Increase in fasting vascular endothelial function after short-term oral L-arginine is effective when baseline flow- mediated dilation is low: a meta-analysis of randomized controlled trials. Am J Clin Nutr 89:77-84 (2009).
7. Edirisinghe et al. Strawberry extract caused endothelium-dependent relaxation through the activation of PI3 kinase/Akt,” J Agric Food Chem 56:9383-9390 (2008)
8. Kim et al. Epigallocatechin gallate, a green tea polyphenol, mediates NO-dependent vasodilation using signaling pathways in vascular endothelium requiring reactive oxygen species and Fyn. J Biol Chem 282(18):13736-45 (2007).
9. Schroeter et al. (-)-epicatechin mediates beneficial effects of flavanol-rich cocoa on vascular function in humans. Proc Natl Acad Sci USA 103(4):1024-9 (2006).
10. Steffen et al. (-)-epicatechin elevates nitric oxide in endothelial cells via inhibition of NADPH oxidase. Biochem Biophys Res Commun 359:828-33 (2007).
11. Sanchez et al. Quercetin downregulates NADPH oxidase, increases eNOS activity and prevents endothelial dysfunction in spontaneously hypertensive rats. J Hypertens24:75-84 (2006).
11b. Pignatelli et al. Polyphenols enhance platelet nitric oxide by inhuibiting protein kinase C-dependent NADPH oxidase activation: effect on platelet recruitment. FASEB J 20:1082-9 (2006).
12. Cai et al. The vascular NAD(P)H oxidases as therapeutic targets in cardiovascular diseases. Trends Pharmacol Sci 24(9): 471-8 (2003).
13. Liming Jin, Arthur L. Burnett. NADPH oxidase: recent evidence for its role in erectile dysfunction. Asian J Androl 10:6-13 (2008).
DNA Damage, Repair, and Aging
As reported in a recent issue of Cell,1 scientists have now discovered a new mechanism by which DNA damage accelerates aging. The researchers found that, like Sir2 in yeast, which has been shown to regulate gene activity (whether genes are turned on or off) and to repair DNA breaks in yeast cells, the mammalian version SIRT1 does the same in mouse cells. Moreover, the new study reports, when DNA damage occurs, the Sir complex relocalizes to the site of the damage to promote repair (so as to prevent genomic instability), but in doing so it abandons its prior location, allowing transcriptional changes to take place there that are characteristic of aging. When mice were administered extra copies of the SIRT1 gene, or fed with the sirtuin activator resveratrol, they had more efficient DNA repair without deterioration in the control by SIRT1 of gene expression; moreover, their mean lifespan was extended by 24% to 46%.
- Oberdoerffer et al. SIRT1 redistribution on chromatin promotes genetic stability but alters gene expression during aging. Cell 135:907-918 (2008).
Indole-3-Carbinol, a Potent Tumor Suppressor Example:
Human Breast Cancer
A growing body of evidence documents the potent inhibition of the growth and invasion of a number of human cancers by indole-3-carbinol, a natural constituent of cruciferous vegetables such as broccoli and cauliflower. Its effects have been particularly well studied in its suppression of human breast cancer. For example, one study reported that indole-3-carbinol is a negative regulator of estrogen receptor alpha signaling in human breast cancer cells by inhibiting its binding to its cognate DNA responsive element, thereby suppressing breast cancer cell proliferation.1
In another study,2 researchers report that indole-3-carbinol inhibits cell proliferation and in vitro markers of angiogenesis (expansion of tumor blood supply) in human endothelial cells in the presence of phorbol myristate acetate, a potent angiogenesis stimulator.
A very interesting new study3 further reports that indole-3-carbinol is a natural elastase enzymatic inhibitor that disrupts cyclin E protein processing, causing a G1 cell-cycle arrest (stopping progression of cell division) of human breast cancer cells. As the authors explain, “I3C [indole-3-carbinol] acts as a specific and potent noncompetitive enzymatic inhibitor of human neutrophil elastase activity, which is highly expressed in breast cancer cells and has been shown to be a prognostic marker for reduced survival rates of primary breast cancer patients.”
- Meng et al. Indole-3-carbinol is a negative regulator of estrogen receptor-alpha signaling in human tumor cells. J Nutr 130:2927-31 (2000).
- Hsiao-Ting et al. Inhibition of cell proliferation and in vitro markers of angiogenesis by indole-3-carbinol, a major indole metabolite present in cruciferous vegetables. J Agric Food Chem 53:5164-9 (2005).
Nguyen et al. The dietary phytochemical indole-3-carbinol is a natural elastase enzymatic inhibitor that disrupts cyclin E protein processing. Proc Natl Acad Sci USA 105(50):19750-5 (2008).
Elastase Inhibitors Could Be a Useful Therapy for COPD
COPD (chronic obstructive pulmonary disease) is a common and generally slowly progressive disease that results in chronic morbidity (such as breathlessness during exercise or even, in advanced cases, at rest) and shortened lifespan. It is characterized by excessive elastase activity in the lung (secreted by neutrophils that infiltrate the lungs, generally as part of an immune response to the chronic lung infections that are a problem for COPD patients). Elastase is responsible for the loss of elasticity that occurs in the COPD lung. Unfortunately, current treatments for COPD (such as bronchodilators) treat symptoms but do not prevent the loss of elasticity and other aspects of the progression of the disease.
The report (see last paragraph of article above) that indole-3-carbinol is a natural inhibitor of elastase suggests that it might be useful in the treatment of COPD, possibly slowing the progression of the disease. Since indole-3-carbinol is a safe dietary substance that is readily available as a constituent of cruciferous vegetables or as an indole-3-carbinol supplement, it could be worthwhile trying it if you have COPD. Whether decreased elastase expression in the lungs would allow for improvement (repairing) of elasticity or just slow the progressive loss of elasticity remains to be seen. Beneficial effects would probably not be immediately noticeable but might, in the long run, provide significant protection. Restoration of the natural anti-elastase compound alpha1-antitrypsin in COPD patients does slow progression. Indole-3-carbinol has not been clinically tested for this purpose.
“The concentration of I3C [indole-3-carbinol] required to inhibit elastase activity in vitro has relevance to the dietary intake of the indole. In a typical western diet (European or American) an individual ingests ~10 mg of I3C per day where in a typical Asian diet intake is ~10-fold higher. Clinical studies have established that ingestion of 400 mg of I3C twice daily is the maximum tolerated dose of this indole that alters estrogen metabolism and other cellular pathways. Importantly, under these conditions, the concentration of indole metabolites in the plasma was reduced to 15 ng/ml after 12 h (the lower limit of detection), strongly suggesting that because I3C is rapidly cleared from the system it is likely to be active at target sites at much lower concentrations that are closer to the observed concentrations needed to inhibit elastase activity.”1
“Furthermore, elastase acts on a variety of other intracellular and extracellular substrates, such as elastin [found in, for example, the lungs, arteries, and skin] . . . and the direct I3C inhibition of elastase activity implicates this natural phytochemical as a potential therapeutic not only for certain cancers but also for other physiological disorders associated with alterations in the levels of elastase activity.”1
- Nguyen et al. The dietary phytochemical indole-3-carbinol is a natural elastase enzymatic inhibitor that disrupts cyclin E protein processing. Proc Natl Acad Sci USA 105(50):19750-5 (2008).
Liar, Liar, Pants on Fire or Where Global Warming is Really Coming From
According to a report in Environment & Climate News,1 James Hansen, astronomer and director of NASA’s Goddard Institute for Space Studies (GISS) has been caught doctoring temperature data from California “to make a long-term cooling trend look like a warming trend.” The article explains that the temperature history (as reported by the U.S. Historical Climatology Network (USHCN) for Santa Rosa, California) was examined by California meteorologist Anthony Watts and found to show a long-term decline, especially since the 1930’s. Watts then examined the temperature history for the same town as reported by GISS; the GISS report was completely different, reporting a long-term increase in Santa Rosa temperature. “USHCN reports a decline of nearly one-half degree Celsius during the twentieth century, while GISS reports a temperature increase of one-half a degree.”1
The article goes on to explain that the USHCN measures temperature by “taking daily readings from an immobile temperature station,” while GISS collects the USHCN temperature readings and then subjects them to adjustments (using methods which Hansen will not reveal), allegedly to correct for artificial influences such as land-use changes. The urban heat island effect (as Santa Rosa’s population increased from slightly more than 10,000 in 1905 to about 158,000 today) would have been expected to result in warmer temperatures (unrelated to global influences) and, hence, to adjust for the urban heat island effect would require the long-term temperature record should be adjusted downward, not upward. Yet, GISS is adjusting the raw temperature data upward instead of downward.
In the article on the facing page,2 it described how “[i]n 2007, statistical scientists showed GISS had been artificially inflating U.S. temperatures by 0.15 degrees Celsius since the year 2000.” Furthermore, “[i]n 2008 statistical scientists showed GISS had falsely reported October 2008 was the warmest October on record when, in fact, it was a quite normal temperature month.” NASA later admitted that the “Warmest October” claim had been wrong.3
- Taylor JM. GISS, Hansen Caught Doctoring More Data. Environment & Climate News Feb. 2009
- Taylor JM. GISS, Hansen Frequently Report False Warming. Environment & Climate News Feb. 2009
- ‘Warmest October’ Claim Was Wrong, NASA Admits. Environment & Climate News Jan. 2009