CLnA, the "Omega-3 Variety" of CLA from Pomegranate & Co, Has Potent Anti-Obesity Effects and the Potential to Become More Than Just Another Anti-Diabetes Drug.

Image 1: Pomegranate - I loved to eat them even before I realized that their seeds are the #1 dietary source (83%) of punic acid.

While more and more people are beginning to grasp the notion that with (naturally occurring) fats - as with everything else in life - there is no simple "good" and "bad", no clearcut "black" and "white" and no definite "beneficial" and "detrimental". The number of different fatty acids and their respective effects on the human metabolism is so vast that it is pretty hard to keep track of all those varieties of saturated and unsaturated carboxylic acids. I would thusly not be surprised if you simply assumed that the "n" in the headline of this blogpost was a type that had slipped in because poor Dr.Andro is chronically stressed from Christmas shopping... well, while the latter is actually correct, the former is not: CLnA is actually the omega-3 variety of the famous conjugated linoic acid (CLA), which in and out of itself is not a single but a group of 28 different trans- and cis-isomers that occur in our diet mainly in the shape of high and full-fat meat and dairy products.

CLnA - Conjugated Linolenic Acid is not a typo ;-)

Within the last couple of years even the medical establishment has come to realize that the chronic omega-6 (n6: linolic acid) overload in our diet is killing us. The "heart-healthy" PUFAs have now become the more and less heart-healthy PUFAs with the totally healthy *rofl* omega-3s and the not just as healthy omega-6s - both, of course, still totally "essential" and WAY better than saturated fats,... (attention: the afore statements are full or irony! Saturated fats are of course NOT the bad guys. Sorry, David if that lead to confusion)... but I am getting derailed, here. So let's get to the point. What every reasonable person appears to agree on, these days, is that we have to lower the ratio of n6:n3 fatty acids in our diets. Now, I am asking you: Has it ever occured to you that CLA essentially is an omega-6 fatty acid? I mean its conjugated linoleic acid - "linoleic" as in omega 6 = linoleic acid! Probably not, right? The reason for that is yet (hopefully ;-) not that you are dump, but simply that the existence of an omega-3 "variety of CLA", namely conjugated linolenic acid, or short, CLnA, is something about which you will only hear, when you read blogs (such as the SuppVersity ;-), which do not stick to copying, pasting and commenting the stuff the authors have read on one of the major news-portals.

Table 1: CLnA isomer content in natural sources (data adapted from Hennesey. 2011)

From a molecular perspective,  CLnA isomers combine the conjugated double bond system of the classic conjugated linolic acid, you know, with the octadecatrienoic fatty acid (C18:3) structure of omega-3s, i.e. linolenic acid. Interestingly, this make-up confers these fatty acids with a high bio-active potential. Now, while this may sound like one of the frankenfood test-tube results of the gene-technology laboratories of Monsanto, we know at least 10 CLnA isomers which occur naturally in foodstuff or as byproduct of fermention processes (cf. table 1).

Adiposity, hyperlipidemia, cancer - CLnAs could help with all!

Image 2: Even if CLnAs would just prevent obesity, this illustration I borrowed from multiplemyelomalifeexpectancy.tk, shows that not being / getting obese alone would prevent a plethora of related maladies. Such as kidney failure, arthritis, gallbladder disease, infertility, asthma, fatty liver disease, sleep apnoea, depression, heart disease, hyperlipidemia, diabetes,... basically every major ailment the increasingly obese convenience society of the Western hemisphere is suffering from.

Due to their anti-adipogenic (meaning preventing the accumulation of body fat) effects CLnA fatty acids have been investigated as potential candidates for the treatment of the obesity epidemic for quite some time, now (Hennesey. 2011). In a 2002 article that was published in the Journal of Applied Biochemistry and Biotechnology, Nishimura et al. report that CLnA isomers exert apoptotic effects on mouse preadipocyte 3T3-L1 cell - or, in plain English, incubation with CLnA did not only hinder the "pubertal" fat cells from becoming mature adipocytes, it actually killed them. In vivo studies with rodents, such as Arao et al. (2004), where the administration of a diet that was enriched with 1% pomegrenate seed oil lead to a 27% reductin in omental white adipose tissue, were able to confirm the "rodent-real world signficance" of these test-tube results.

Other studies showed a normalization of hyperlipidemia in rodent models of the metabolic syndrome and a hand full of studies have explored the usage of CLnA isomers as cytotoxins in the treatment of cancer. In their concise review of the literature, Hennesey, et al. thusly rightly conclude that with their "potent inflammatory and immune modulating properties", their ability to "reduce the risk of obesity, improve cardiovascular health, and mediate strong anti-carcinogenic activity", the use of CLnA isomers or dietary enrichments could offer treatment strategies for pathologies, which "represent some of the greatest mortality risks to humans in the Western world and have been inextricably linked with diet" (Hennesey. 2011).

Adding diabetes to the list of potential targets for CLnA

For today, we are however going to focus on the most recent result from the research front: The effects of CLnAs on diabetes, or, to be precise, the increases in blood glucose, and decreases in anti-oxidant capacity that go hand in hand with the latter. In a recently published study (Saha. 2011), Siddhartha S. Saha and Mahua Ghosh from the Department of Chemical Technology at the University College of Science and Technology of the University of Calcutta (I don't have to tell you that this is in India, do I?) injected male albino lab rats with 60mg/kg streptozotocin (STZ) - this is a common and well-established method to induce a metabolic state that serves as a model of type II diabetes - and fed them diets that contained either no, or 0.5% of the total fat in the form of alpha-eleostearic acid (from bitter gourd, cf. table 1) or punic acid (which was in this case taken from snake gourd oil, but could as well have been extracted from the eponymous pomegrenate, cf. table 1).

Figure 1: Relative blood glucose levels vs. non-STZ injected control in streptozotocin injected rats over the course of the dietary intervention (data calculated based on Saha. 2011)

As you can see in figure 1, this 100% natural "food additive" had a more than pronounced effect on the +300% (vs. non STZ-injected control) elevated blood glucose levels of the "type-2 diabetic" rodents.

Figure 2: Relative level of lipid peroxidation (left) and total antioxidant capacity (right) levels vs. non-STZ injected control in streptozotocin injected rats after the 28-day dietary intervention (data calculated based on Saha. 2011)

And while the glucose levels were still 150% above those of the healthy control levels, the streptozotocin-induced lipid peroxidation in plasma, pancreas and erythrocytes of the lab animals was ameliorated by the snake gourd oil treatment (remember that is the stuff from pomegranate) and even reversed by the bitter gourd diet. Judged by the standardized FRAP assay, the "diabetic animals" that were fed a diet that contained 0.1% alpha-eleostearic acid (of the total diet, which had 20% fat) even exhibited a 10% greater total antioxidant capacity than the totally healthy control!

Figure 3: Relative expression of inflammatory cytokines, TNF-alpha and interleukin 6 in plasma capacity (right) levels vs. non-STZ injected control in streptozotocin injected rats after the 28-day dietary intervention (data calculated based on Saha. 2011)

Snake gourd oil, on the other hand, exhibited more profound effects on the elevated TNF-alpha, interleukin-6 and NF-kappaB levels of the STZ-treated rodents (cf. figure 2) and thus, at least this is my humble opinion, render punic acid the overall more promising agent with respect to the treatment of all sorts of inflammatory (or related diseases). After all, disturbances in the regulation of the nuclear factor kappa-light-chain-enhancer of activated B cells  (NF-kappaB) and the downstream over-expression of TNF-alpha and IL-6 are hallmark features of allmost all the aforementioned ailments of the increasingly obese western convenience society. This is also why I am quite certain that we are going to hear much more about the CLnAs in the month to come... and I guess, I don't have to tell you that right here, at the SuppVersity, is where you will read about respective studies first!

Some Things Fishy: Oxidized Fish Oil Totally Benign!?Plus: The Inflammatory Side of EPA and Peroxide & Alkenal Levels in Commercial Fish and Vegetable Oils.

Image 1: Surströmming, a Swedish delicates is essentially rancid fish and it stinks exactly like that. Now, the results of a recent study show that the rancidity does probably not compromise the health benefits of the fish... so if you like it, go for it!

You know that whenever something is so (over-)hyped like fish oil or vitamin D that rings an alarm with me and when I hear "experts" on popular podcast say things along the lines of "as long as you take your fish oil that can compensate for a whacky diet", this is totally burning me up. Yes, there is conclusive evidence that for someone who has damaged his/her body by years and years of omega-6 over-consumption the inclusion of even "high" dose (I consider 5-6g high!) fish oil supplements can make sense, but NO, it will neither allow you to keep eating the same crap that has brought you to where you are at now, nor (and I think this is even more important for most of the SuppVersity readers) is there conclusive evidence that a healthy, active and lean human being is not way better off by limiting his total PUFA intake instead of popping grams of highly oxidizable n-3 fatty acids from fish oil caps.

Highly oxidizable? Yes! Dangerous? Surprisingly not!

A pros pos "highly oxidizable", the argument that polyunsaturated fatty acids (PUFAs) are readily oxidized not only in your body, but even at the shelves of your nutrition store, is one of the few possible caveats of fish oils supplementation even fish oil enthusiasts will acknowledge. After all previous animal studies have shown that diets rich (5%) in rancid (=oxidized) fish oils lead to increases in thiobarbituric acid-reactive substances (TBARS) levels and elevate liver specific transaminases, as well as the alkaline phosphatase (ALP) levels in the plasma of rats (detrimental effects which can by the way be ameliorated by taurine supplementation, cf. Hwang. 2000). The results of a recent study by Inger Ottestad and colleagues from Norway may thusly surprise the "pro-fish oil"-faction about as much as they surprised me (Ottestad. 2011): The ingestion of 8g of oxidized (peroxide value: 18mEq/kg; ansidine value: 9) fish oil (1.6g EPA+DHA) did not have any unfavorable short term-effects in previously healthy individuals.

Figure 1: Serum (left, 8-iso on secondary axes was measured in urine) and erythrocyte (right, GPx on secondary axes) markers of oxidative stress in 68 healthy subjects who were randomly assigned to ingest 8g of "fresh" fish oil, oxidized fish oil or high oleic-acid sunflower oil per day for before (pre) and after (post) the 7 week intervention (data adapted from Ottestad. 2011)

If you take a closer look at the measured levels of serum (4-hydroxy-2-hexenal: 4-HHE, 4-hydroxy-2-nonenal: 4-HNE, alpha-tocopherol,  high-sensitive C-reactive protein: hsCRP and 8-iso-PFG2a, the latter in urine) as well as erythrocyte (total GSH, 4-hydroxy-2-nonena: GR, CAT and glutathione peroxidase: GPx) markers of oxidation before and after the 7-week intervention (cf. figure 1), it is quite obvious that there were no statistically significant oxidation-related changes in the concentrations of the measured markers of oxidative stress, of which the scientists state that they are the current, yet debatable, "gold standard" for in vivo studies.

Figure 2: Changes in n-3 and n-6 levels and the n-6/n-3 ratio (small graph) in the course of the study period (data calculated based on Ottestad. 2011)

It is thus not really surprising that both fish oil groups experienced virtually identical (and highly favorable) -50% reductions in the ratio of omega-6 (n-6) to omega-3 (n-3) fatty acids. Moreover, ...

[a]fter 3 and 7 weeks of intervention, the plasma level of EPA, docosapentaenoic acid and DHA were significantly increased in both fish oil groups compared to the HOSO group, but no significant difference in EPA, doc-osapentaenoic acid and DHA between the FO and oxFO groups was observed.

The scientists are thusly right to conclude that their results do not support the often-heard hypothesis that higher intakes n-3 long-chain fatty acids could increase in vivo lipid peroxidation and more importantly, that ...

[...] the content of hydroperoxides in fish oil supplements, even with a PV that exceeds the European Pharmacopeia for marine n-3 oils, does not apparently influence the plasma level of n-3 FA.

With regards to the obvious differences to previous animal studies, the scientists state that secondary oxidation of hydroperoxides, which are then absorbed in the intestine has until now been observed in animal and cell studies. In view of the relative short duration of the study and the reliance on healthy subjects, it is also questionable whether identical results would have been achieved, when sick patients (the usual customer group at least for the pharma-grade n-3 supplements) had been treated with the same product for years.

Oxidized fats in fish oil and beyond

It is also worth mentioning that Ottestad et al. are not sure, whether their "aritifically oxidized" fish oil (oxidation was achieved by sparkling pure oxygen through the oil for 20 min twice a day for 21 d) was an appropriate model for commercially available (oxidized) fish oils. After all, there could be major differences in the composition of the oxidation products, when the oils go rancid over months or get damaged by heat etc. While I obviously cannot answer this question without setting up my own lab, I can however tell you that another recent study by Halvorsen et al. who examined the peroxide and alkenal (one of the major products of secondary oxidation) content of fish and vegetable oils, found average peroxide levels in 33 commercially available fish oil products (mean PV: 3.61mEq/kg) that were ~500% below the ones of the oxidized fish oil (18mEq/kg) in the Ottestad study.

Figure 3: Mean peroxide and alkenal values of 33 commercially available fish and 35 vegetable oils (Halvorsen. 2011).

In this regards, fresh vegetable oils, obviously are way in front, as the data from a study by Bente Lise Halvorsen and Rune Blomhoff clearly shows, that they have lower peroxide and much lower alkenal levels than fish oils (cf. figure 3). Interestingly, vegetable oils are also less prone to being oxidized during storage, something Halvorsen and Blomhoff conclude based on the absence of the "negative correlation (r=−0.557, p<0.001) [...] between the number of days until expiry and the PV [peroxide value]" they observed in the marine omega-3 oils.

Figure 4: Peroxide (PV in mEq/kg) and alkenal (in nM/ml) levels in fresh vegetable oils and after being heated for 25 minutes at 225°C in an oven (data adapted from Halvorsen. 2011); solid red line - maximal peroxide value for olive oils, dotted red line - maximal peroxide values for fish oils as suggested by Turner et al. (Turner. 2006)

Contrary to fish oils, which are usually taken "fresh" and in a capped form, the main fate of vegetable is however to be (ab-)used as cooking / frying oils. During the heating process, the amount of secondary lipid oxidation products, the alkenals, doubles or quadruples depending on the type of oil (cf. figure 4). In that, it may at first seem counterintuitive that, when the scientists heated the samples for 25 minutes at 225°C in an oven, the amount of primary oxidation products was slightly reduced in most, but not all (e.g. soy bean oil) of the 11 vegetable oils. If you do yet take into consideration that the latter are the "raw material" for the secondary oxidation products, it becomes quite clear that this is not a desirable process ;-)

Image 3: Extra virgin olive oils (EVOOs) have generally higher peroxide values than the cheap refined stuff, and yet, EVOOs and not refined oils have been shown to exhibit numerous health benefits.

Putting peroxide values (PV) into perspective: All potential health hazards aside, it may be interesting to know that the general "rule of thumb" says that a fat is rancid when the PV is about 10 meq/kg (the fish oil in the study with PV=18 was thusly "rancid"). A fresh and refined product on the other hand should have PV below 1 meq/kg (Gunstone. 1996). That being said, it may surprise you that for high quality extra virgin olive oils, the PV limit is 20meq/kg, while for "regular" olive oil it is only 10 meq/kg. If you know look at studies related to the health benefits of refined vs. extra virgin olive oil, you will have to admit that - quite obviously - fish oil apparently is not the only oil, where increased peroxide levels do not negate the beneficial health effects of the oil.

So, if pure vegetable oils are generally "fresher" than fish oils does that mean that as long as you do not heat them, they are the better choice? No, they are not! I mean, look at the research that is out there... the abundance of n-6 fatty acids in the "healthy" vegetable oils that are getting pimped especially by the US government, is at the heart of an epidemic of which the authorities still claim that it was caused by high cholesterol levels. Instead banning all saturated fats from YourPlate (which should never look like the governments MyPlate ;-), you should rather incorporate more coconut oil and saturated fats from butter, beef etc. into your diet. Select (vegetable) oils that are relatively high in mono-unsaturated fatty acids, like extra virgin olive oil (don't care about its high peroxide value, cf. red box above) and try to reduce the amount of n-6 fats you ingest - you will get more than enough even from grass-fed meats, olive and other oils and any processed foods that may still be part of your diet, anyway.

Image 4: Not all Omega-3 are created equal. We know for some time that DHA (not EPA) is what your brain needs and a recent study from Norway suggest that eicosapentaenoic acid (EPA) is actually pro- not anti-inflammatory at a cellular level. It may yet well be that this in turn triggers a beneficial hormetic response which would support my "fish oil = exercise in a pill hypothesis"

Although this is not directly related to the topic of oxidation I still want to add that another study appears to confirms my long-cherished skepticism towards EPA (most fish oils have a 2:1 EPA to DHA ratio), which, as a recent study from Norwegian scientists shows (Myhrstad. 2011), is not really "beneficial super-antioxidant" people are led to believe. In their trial the scientists fed 14 healthy female volunteers test meals. The cakes the participants ate were enriched with either flaxseed, cod liver or coconut oil and the intention of the study was to elucidate differential effects of meal fatty acid composition on inflammatory markers. Not to my, but probably to the scientists surprise the "evil" saturated fat from the coconut oil turned out to be similarly benign as the flaxseed cake. Only the EPA-laden cod liver oil cake produced a statistically significant increase in IL-8 mRNA levels 6h post ingestion. Similarly, incubation of peripheral blood mononuclear cells with EPA, yet not ALA lead to >3x increase in IL-8 and >2x increases in IL-6 mRNA expression.

While I am not quite sure what to make of these observations, these results stand in line with previous studies reporting differential effects of EPA vs. DHA rich fish-oils, where across the board, the DHA appeared to be the major driving force of the beneficial health effects people hope to be getting from their fish oil caps (e.g. brain health, Engström. 2009).

Fish oil caps can be a good addition to this regimen specifically for those who are just about to start out on a low omega-6 diet to offset the skewed n-6 to n-3 ratio (something that takes its time). They are yet by no means obligatory for someone who eats fish on a regular basis and invests the extra bucks into grass-fed beef and eggs from pastured chicken. If you thusly satisfy your (anyway low) dietary DHA requirements... and most importantly, taking fish oil will not compensate for eating shitloads of processed foods and lack of exercise, even if the aforementioned pro-inflammatory effects of EPA support my previously uttered hypothesis that fish oil has some resemblance to "exercise in a pill".

Artichoke Leaves Diabetes No Chance: Thistle Qualifies as Anti-Oxidant, Carb Blocker, Digestive Help & Diabesity Drug

Boiled Artichokes slow down the absorption of glucose and minimize the insulin + glucose surges (Nomikos. 2007)

This article has made quite a remarkable transformation. It started as a Facebook Short News Item, was upgraded to a mini-article for the SuppVersity Short News and, when I realized that those green thistles have surprisingly diverse beneficial effects on human health, eventually became the regular SuppVersity Article you are about to read, right now.

Artichoke leaves the future of diabesity treatment!?

I have to admit, I am not exactly and artichole expert, but I was still surprised that the paper Joanna Magielse and her colleagues from the University of Antwerp are about to publish in the scientific journal Food & Function claims to be the first to investigate the beneficial health effects of artichoke leave extracts on diabetes-induced oxidation in vivo.

I mean, we all have seen the broad range of artichoke supplements on the shelves; and my brief review of the literature yielded studies from the mid 1920s discussing how beneficial artichoke can be for patients with type II diabetes (Root. 1925).

When I kept digging through the currently available literature, though, I had to realize that the vast majority of the in-vivo studies dealt with the anti-cholesterol (Wider. 2013) and digestive benefits (Marakis. 2002) of artichokes and their leaf extracts (ALE). The proven anti-oxidant effects of its major polyphenolic constituents (Zapolska-Downar. 2002;  Betancor-Fernandez. 2003; Jimenez-Escrig. 2003; Menghini. 2010), on the other hand, haven't been tested in either animals or humans with type II diabetes. The rodent-experiment the Belgian researchers describe in the paper at hand is thus probably in fact the first to investigate the anti-oxidative effects ALE in an in-vivo scenario.

More about chlorogenic acid.

What are the active ingredients in artichoke leaves? Mono- and dicaffeoylquinic acids were the major polyphenolic constituents: The total caffeoylquinic acid (CQA) content was 1.5%, with chlorogenic acid (CGA) being most abundant (0.30%), next to cynarin (0.12%) and several isomerization products of CGA (the sum of which was 0.75%), including neo- and crypto-CGA and dicaffeoylquinic acids. Luteolin-7-O-glucoside was the major flavonoid present.

Over the course of 3 weeks the rodents who had been injected with streptozotocin (STZ) to induce a diabetic phenotype that is commonly and successfully used as a model for type II diabetes in man consumed either

• Why 0.2g/kg BW? This is the rodent equivalent of a well-tolerated amount of 2g/day that has been used for treatment of hypercholesterolemia and digestive complaints (Englisch. 2000; Holtmann. 2003; Barnes. 2007).
  0.2g/kg BW of artichoke extract (CYN1),
• 1g/kg BW of artichoke extract (CYN2),
• 50mg/kg BW alpha-tocopherol-acetate (VIT E),

or no supplement at all. To make sure to have an appropriate reference, a fifth non-streptozotocin injected group served as a healthy control. In contrast to their their com-rats (sorry, I could not resist) who had blood glucose levels of 250mg/kg, the control rats started into the 3-week treatment phase with normal (=healthy) blood glucose levels and inflammation.

Learn About Artichoke Alternatives for Glucose Management:

Lifestyle Changes

ALA, GABA, Taurine & Co.

Berberine, Banaba & Co.

Cinnamon, Curcumin & Co.

Lemon, Starch, Coffee & Co.

Chlorogenic acid, fucoxanthin & Co.

To determine the effects the provision of artichoke leaves would have on the baseline inflammation of the diabetic rats, the researchers measured the plasma malondialdehyde (MDA) and urinary 8-hydroxydeoxyguanosine (8-OHdG). The status of plasma coenzyme Q9(CoQ9, necessary for the biosynthesis of coq10 increases indicate lower coQ10 levels) and erythrocyte reduced glutathione (GSH), on the other hand, were used as indicators of the state of the endogenous antioxidative defense system.

Figure 1: MDA, 8-OHdG, CoQ9 and GSH levels expressed as rel. difference to control (Magielse. 2013)

With the information from the previous paragraph, even a rocket scientist without a clue of biology will recognize that the buffered MDA,8-OHdG and coQ10 and GSH levels (the further away from the x-axis the more "disturbed" they are compared to the CON group), indicate that the artichoke leave extract did what the were not 100% sure it would to: It survived the passage through the gastrointestinal tract and wasn't extensively metabolized by colonic microflora or liver phase II enzymes, as it is the case for other potent in-vitro antioxidants like reseveratrol.

What's the mechanism and what do we already know about artichoke?

With respect to the underlying mechanism that could explain the improvements the researchers observed in the study at hand, Magielse et al. point out that...

"...it should be emphasized that not only direct antioxidant actions, but also indirect mechanisms affecting gene expression of inflammatory pathways and modulating antioxidant enzyme synthesis have been reported for polyphenols and may contribute to the reduction of oxidative stress." (Magielse. 2013)

In that, the scientists are referring to the previously established beneficial effects on lipid metabolism and glucose absorption other researchers have observed in previous studies:

• Boiled wild artichoke reduces postprandial glycemic and insulinemic responses in normal subjects by inhibiting the glucose absorption (100g of boiled plant on 50g of glucose in complete meal). Interestingly, this trick doesn't work in patients with type II diabetes (Nomikos. 2007)
• Significant increase in bile secretion and thus improved fat digestion.  1.92g of artichoke extract lead to increase of up to ~140% in a randomised placebo-controlled double-blind cross-over study published in Phytomedicine (Kirchhoff. 1994). 
• Beneficial improvements in endothelial function (-21% VCAM-1; - 17% ICAM-1 brachial +37% FMV) in hyperlipidemic patients with only 20ml of "self-made"* artichoke juice per day (*juiced by the researchers; Lupattelli. 2004).
• Provides "food" for probiotic Lactobacillus paracasei (LMGP22043) that have beneficial effects on faecal bacteria and biochemical parameters in human subjects (Valerio. 2011).
• Has UV protective effects, when applied to the hair (Fernandez. 2012) - not that important for your overall  health, but still quite telling, right?)
• 2x 200mg of ALE per day decrease total cholesterol and LDL and increase HDL in subjects with existing hypercholesterolaemia (Rondanelli. 2013)

I know that you'd probably love another 200 references to studies that support the beneficial health effects of artichoke and artichoke extracts, but you know what? I think that's enough for a preliminary conclusion.

Diabetes or Not-Diabetes - That's not just about the foods you eat, but also about what your body does with them: "Dietary Fructose vs. Endogenous Fructose Production: Is The Aldose Reductase Mediated Production of Fructose to Blame for Diabesity & NAFLD? Could Amla Help? " | more

Bottom line: With the existing evidence for its acute anti-hyperglycemic effects, its beneficial effects on lipid digestion and metabolism and reliable evidence from a 2005 study by Wittemer that the antioxidant caffeoylquinic acids and flavonoids are orally bioavailable in humans (Wittemer. 2005), it appears as if artichokes would make a a highly underrated, yet profoundly beneficial addition to everyone's diet.

With commercially prepared extracts, on the other hand, you do not just run the risk of buying not / incorrrectly standardized products without active ingredients, but could also be missing out on the actue anti-hypoglycemic effects, which are probably mediated by the fructan-content of the whole "fruit"; not the caffeoylquinic acids and flavonoids you can buy in capsule-, pill- or tablet-form (cf. Rumessen. 1990; the dosage of fructans that did the trick here was 20g!)

Reference:

• Barnes, J., Anderson, L.-A., Phillipson, J.-D. (Eds.), Herbal Medicines, Pharmaceutical Press, London 2007. 
• Betancor-Fernandez, A., P´ erez-G´ alvez, A., Sies, H., Stahl, W., Screening pharmaceutical preparations containing extracts of turmeric rhizome, artichoke leaf, devil’s claw root and gar lic or salmon oil for antioxidant capacity. J. Pharm. Pharma col. 2003,55, 981–986. 
• Englisch, W., Beckers, C., Unkauf, M., Ruepp, M. et al., Effi cacy of artichoke dry extract in patients with hyperlipopro teinemia. Arzneimittel-Forschung. 2000,50, E260–E265. 
• Fernandez, E., Martínez-Teipel, B., Armengol, R., Barba, C., & Coderch, L. Efficacy of antioxidants in human hair. Journal of Photochemistry and Photobiology B: Biology.  2012
• Holtmann, G., Adam, B., Haag, S., Collet, W. et al., Efficacy of artichoke leaf extract in the treatment of patients with functional dyspepsia: a six-week placebo-controlled, double blind, multicentre trial.Aliment. Pharmacol. Ther. 2003,18, 1099–1105.
• Jimenez-Escrig, A., Dragsted, L.-O., Daneshvar, B., Pulido, R. et al., In vitro antioxidant activities of edible artichoke (Cynara scolymusL.) and effect on biomarkers of antioxi dants in rats.J. Agric. Food Chem. 2003,51, 5540–5545.
• Kirchhoff, R., Beckers, C. H., Kirchhoff, G. M., Trinczek-Gärtner, H., Petrowicz, O., & Reimann, H. J. Increase in choleresis by means of artichoke extract. Phytomedicine. 1994, 1(2), 107-115.
• Lupattelli, G., Marchesi, S., Lombardini, R., Roscini, A. R., Trinca, F., Gemelli, F., ... & Mannarino, E. Artichoke juice improves endothelial function in hyperlipemia. Life sciences. 2004, 76(7), 775-782.
• Marakis, G., Walker, A. F., Middleton, R. W., Booth, J. C. L., Wright, J., & Pike, D. J. (2002). Artichoke leaf extract reduces mild dyspepsia in an open study. Phytomedicine, 9(8), 694-699.
• Menghini, L., Genovese, S., Epifano, F., Tirillini, B. et al., Antiproliferative, protective and antioxidant effects of arti choke, dandelion, turmeric and rosemary extracts and their formulation.Int. J. Immunopathol. Pharmacol. 2010,23, 601– 610. 
• Nomikos, T., Detopoulou, P., Fragopoulou, E., Pliakis, E., & Antonopoulou, S.. Boiled wild artichoke reduces postprandial glycemic and insulinemic responses in normal subjects but has no effect on metabolic syndrome patients. Nutrition Research. 2007, 27(12), 741-749.
• Root, H. F., & Baker, M. L. Inulin and artichokes in the treatment of diabetes. Archives of Internal Medicine. 1925, 36(1), 126.
• Rondanelli, M., Giacosa, A., Opizzi, A., Faliva, M. A., Sala, P., Perna, S., ... & Bombardelli, E. (2013). Beneficial effects of artichoke leaf extract supplementation on increasing HDL-cholesterol in subjects with primary mild hypercholesterolaemia: a double-blind, randomized, placebo-controlled trial. International Journal of Food Sciences and Nutrition, 64(1), 7-15.
• Rumessen, J. J., Bodé, S., Hamberg, O., & Gudmand-Høyer, E. Fructans of Jerusalem artichokes: intestinal transport, absorption, fermentation, and influence on blood glucose, insulin, and C-peptide responses in healthy subjects. The American journal of clinical nutrition. 1990, 52(4), 675-681.
• Valerio, F., et al. Role of the probiotic strain Lactobacillus paracasei LMGP22043 carried by artichokes in influencing faecal bacteria and biochemical parameters in human subjects. Journal of applied microbiology 2011, 111(1): 155-164.
• Wider, B., Pittler, M. H., Thompson-Coon, J., & Ernst, E. (2013). Artichoke leaf extract for treating hypercholesterolaemia. status and date: New search for studies and content updated (no change to conclusions), published in, (3).
• Wittemer, S. M., Ploch, M., Windeck, T., Müller, S. C., Drewelow, B., Derendorf, H., & Veit, M. Bioavailability and pharmacokinetics of caffeoylquinic acids and flavonoids after oral administration of Artichoke leaf extracts in humans. Phytomedicine. 2005, 12(1), 28-38.
• Zapolska-Downar, D., Zapolski-Downar, A., Naruszewicz, M., Siennicka, A. et al., Protective properties of artichoke (Cynara scolymus) against oxidative stress induced in cultured endothelial cells and monocytes .Life Sci.2002,71, 2897–2908. 

The Pistachio Manifesto: Antioxidant, Metal Chelator, DNA Protector, Anti-Cancer Agent, Bug Killer (incl. H. Pylori & Herpes Simplex) & More. Have You Been Missing Out?

This is not exactly what I was talking about, when I said "going nuts", but in this case it would actually qualify as "going pistachios" ;-)

Walnuts, almonds and Brazil nuts, these are the stars among the hard-shelled fruits people tend to go nuts about (all puns intended ;-). Pistachios, on the other hand, get very little love. I have in fact written about their surprisingly low effective energy content and their highly bioavailable phenolic content before (learn more), but what the myriad of phenols in these small nutritional powerhouses the ancient Egyptians used as incense, preservative and breath sweetener, while their Iranian neighbors in the North East already knew about their beneficial effects on digestive, hepatic and kidney health (Avicenna. 2008) can do for our health has not been covered here at the SuppVersity.

So what is it pistachios can do for you?

With their traditional use as a remedy for digestive, liver and kidney issues, you already have an idea where this could be heading. The shelled fruits of which we know that they have been part of the human diet for at least 9,000 years and that's cultivated in the Middle East, United States and Mediterranean can however do more for you:

• 

  Histidine happens to be an excellent chelator and potential weight loss adjuvant, as well (learn more)

  Potent antioxidants: Different parts and constituents from P. lentiscus  have been shown in vitro radical scavenging properties They protect your LDL molecules from being oxidized and will thus have direct beneficial effects on your atherosclerosis and overall heart disease risk (Holvoet. 2004).
• Metal chelators: A 2011 study by Orhan et al reports that pistachios (P. terebinthus fruits) are potent metal chelators (on par with EDTA) and have an impressive radical scavenging activity. Interestingly engough, the Antioxidant activity of the fruits actually seems to increase, when they are roasted (Orhan. 2012).
• DNA protection: More or less a downstream effect from the high content of antioxidant compounds, specifically gallic acid, digallic acid and 1,2,3,4,6-pentagalloylglucose, and polyphenols have direct protective effects on cellular DNA and pro-carcinogenic mutations.
• Antimicrobial activity (incl. anti H. Pylori): Certainly among the most interesting effects are the anti H.Pylori effects of α- pinene, a compound from the essential oils in pastachios (speficifally P.atlantica var. kurdica). Other ingredients, like verbenone, rterpineol, and linalool showed high antibacterial activity against Escherichia coli, Staphylococcus aureus and Bacillus subtilis (Koutsoudak. 2005).
  
  

  

  You can battle H. pylori with probiotics, as well. If you want to learn more about this, I'd suggest you go back a couple of months and read the full story in the SuppVersity Short News from October 2012 (go for it)

  Sakami et al. report similar beneficial effects against pathologic bacteria (Porphyromonas gingivalis and Prevotella melaninogenica) antiplauqe activity on teeth by inhibiting bacterial growth in saliva (Sakagami. 2009). Özçelik et al. add noticeable anti-viral effects to the list of anti-microbial activities of pistachio species (Özçelik. 2005). The viruses tested in the study were Herpes simplex (DNA) and Parainfluenza viruses (RNA). The effects were significant for both Kernel and seed extracts.
• Anti-inflammatory activity: Extract of the resin of P. lentiscus var. Chia and its isolated phytosterol tirucallol exert direct anti-inflammatory effects on human aortic endothelial cells and inhibit the activity of adhesion molecules that express the inflammatory cytokine TNF-α (Tzakou. 2007). Tzakou et al. ascribe the effects to phytosterol that goes by the name if tirucallol - never heard of it? Me neither, but who knows on which supplement label you may find it in the future ;-)
• Digestive health: I already mentioned this in the introduction. One of the most important traditional uses of gums from Pistacio species is the management of gastrointestinal disorders; and that has been confirmed by several studies (Rahimi. 2009 & 2010; Farzaei. 2013). Resin of P. lentiscus has been shown to significantly reduced the intensity of gastric mucosal damage induced by pyloric ligation, aspirin, phenylbutazone, reserpine and restraint with cold stress  via its  antisecretory and cytoprotective activities (Al-Said. 1986)
  

  

  Figure 1: Improvements of acid regurgitation and heartburn in forty eight patients fulfilling Rome II criteria for functional dyspepsia were randomly assigned to receive either Pistachio var chia mastic gum 350 mg three times daily or placebo after three weeks (Dabos. 2009)

  A double- blind  placebo controlled trial, P. lentiscus gum lead to significant improvement of the symptoms of patients with functional dyspepsia  (Dabos. 2009). Extracts have been successfully tested in experimental models of acute colitis and IBS (Rahimi. 2013), and there is supporting evidence for beneficial effects of P. lentiscus var. chia resin in patients with established mild to moderate active crohn’s disease (CD) after 4 weeks of supplementation (Kaliora. 2007a,b).

• 

  Suggested read: Supplements to Preserve and Restore Insulin Resistance (read more)

  Antidiabetic activity: This is not about the nuts, but about a leaf extract, which has demonstrated significant acute postprandial  antihyperglycemic activity comparable to metformin and glipizide in starch-fed rats, in which it also lead to significant overall improvements in glucose tolerance (Kasabri. 2011). Unfortunately, a study by Kasibri et al. does not support these results - at least not for normoglycemic and streptozocin-induced hyperglycemic rats (Kasibri. 2004) on a regular diet.
  
  This would suggest that there is a direct correlation with carbohydrate intake, and voilá a 2007 human study confirmed that the gum, not the leaf extract can effectively, lower blood serum glucose levels in men... however, there is another "on the other hand attached here": this did not work for the female study participants (Triantafyllou. 2007).

• 

  The Protective Hull of These 61 Super Fruits Can Ward Off Cancer (more)

  Anti-cancer effects: In a relatively recent review of the literature, Giaginis & Theocharis call pistachio mastic gum a "conglomeration of effective anticancer drugs" (Giaginis. 2011) - probably not without reason, after all they cite a plethora of scientific data from peer-reviewed studies to support the anticancer activities of mastic gum and its major constituents and highlighting the various molecular mechanisms through which the triterpenoids work their anti-cancer magic.
  
  Rezaei et al., for example were able to show that the fruit extract of P. atlantica sub. kurdica exerts direct inhibitory effects on human colon carcinoma cells that was comparable to the drug Doxorubicin (Rezaei. 2012). Another example? Well what about the anti-breast cancer, anti liver-, anti cervical and anti skin-cancer effects of oleoresin (Almehdar. 2012)
• Hypolipidemic effects (=lowering high blood lipids): Extracts from P. vera  fruits have shown beneficial effects on HDL and LDL level in rabbit model of atherosclerosis, they exert positive effects on the lipid levels of patients with moderate hypercholesterolemia (Edwards. 1999). And have several animal studies to support their anti-artherogenic effects (e.g. Bakirel. 2003) 

Impressed? Well I'd hope so, after all this turned out to be more work than I initially thought, when I started to "just write a brief overview of the health effects of pistachios" ;-)

---

Brazil nuts & selenium: "How much is too much?" A weighty question I addressed in a July 2013 SuppVersity article: "Brazil Nuts & Selenium: Are You Nuts If You Have More Than One Per Day?" (more)

Bottom line: Ok, I have to admit there is no scientific evidence that all the good things mentioned above are going to happen to you, if you have a handful of pistchios every other day, but you know what? It may still be a piece in the puzzle people often refer to as "healthy diet" - so if you are into nuts and can "afford" their relatively high energy content, go for it! After all, the above are only the "confirmed" effects.

Note: Actually everybody can "afford" eating nuts, it's more a question of being able to stop before the whole 500g family pack is annihilated... and trust me, if you love nuts, this can happen pretty fast, even if you have to shell them as it is the case with pistachios.  

Last but not least, in traditional Iranian medicine, certain ingredients in pistachios are known to exhibit various additional pharmacological activities incuding diuretic, lithontripic, anti-tussive, anti-rheumatic, anti-asthmatic, anti-hypertensive, and aphrodisiac effects of which Bozorgi et al. point out that the "are not [yet] supported by any current scientific documents and so, they could be considered  for investigating by researchers." (Bozorgi. 2013)

References:

• Almehdar H, Abdallah HM, Osman AM, Abdel-Sattar EA. In vitro cytotoxic screening of selected Saudi medicinal plants. J Nat Med. 2012 Apr;66(2):406-12.
• Al-Said MS, Ageel AM, Parmar NS, Tariq M. Evaluation of mastic, a crude drug obtained from Pistacia lentiscus for gastric and duodenal anti-ulcer activity. J Ethnopharmacol. 1986 Mar;15(3):271-8.
• Avicenna.The canon. Translated by: A . Shrafkandi. Soroush Press, Tehran. 2008.
• Bakirel T. The Investigation of the Effects of Pistacia terebinthus L. Upon Experimentally
  Induced Hypercholesterolemia and Atherosclerosis in Rabbits. Turk. J. Vet. Anim. Sci. 2003; 27: 1283- 1292.
• Bozorgi M, et al. Five Pistacia  species (P. vera, P. atlantica, P. terebinthus, P. khinjuk and P. Lentiscus): A review of their traditional uses, phytoche mistry and pharmacology. The Scientific World Journal. 2013.
• Dabos KJ, Sfika E, Vlatta LJ, Frantzi D, Amygdalos GI, Giannikopoulos G. Is Chios mastic gum effective in the treatment of functional dyspepsia? A prospective randomised double-blind placebo controlled trial. J Ethnopharmacol. 2010 Feb 3;127(2):205-9.
• Edwards K, Kwaw I, Matud J, Kurtz I. Effect of pistachio nuts on serum lipid levels in patients with moderate hypercholesterolemia. J Am Coll Nutr. 1999 Jun;18(3):229-32.
• Farzaei R et al. An evidence-based review on medicinal plants used for the treatment of peptic ulcer in traditional Iranian medicine, Int J Pharmacol. 2013 [ahead of print]
• Giaginis C, Theocharis S. Current evidence on the anticancer potential of Chios mastic gum. Nutr Cancer. 2011 Nov;63(8):1174-84.
• Holvoet P. Oxidized LDL and coronary heart disease. Acta Cardiol. 2004 Oct;59(5):479-84. Review.
• Orhan IE et al. Neuroprotective potential of some terebinth coffee brands and the unprocessed fruits of Pistacia terebinthus L. and their fatty and essential oil analyses. Food Chemistry. 15 February 2012; 130(4):882–888.
• Özçelik B, Aslan M, Orhan I, Karaoglu T. Antibacterial, antifungal, and antiviral activities of the lipophylic extracts of Pistacia vera. Microbiol Res. 2005;160(2):159-64. 
• Kaliora AC, Stathopoulou MG, Triantafillidis JK, Dedoussis GV, Andrikopoulos NK. Chios mastic treatment of patients with active Crohn's disease. World J Gastroenterol. 2007a Feb 7;13(5):748-53.
• Kaliora AC, Stathopoulou MG, Triantafillidis JK, Dedoussis GV, Andrikopoulos NK. Alterations in the function of circulating mononuclear cells derived from patients with Crohn's disease treated with mastic. World J Gastroenterol. 2007b Dec 7;13(45):6031-6.
• Kasabri V, Afifi FU, Hamdan I. In vitro and in vivo acute antihyperglycemic effects of five selected indigenous plants from Jordan used in traditional medicine. J Ethnopharmacol. 2011 Jan 27;133(2):888-96.
• Koutsoudaki C, Krsek M, Rodger A. Chemical composition and antibacterial activity of the essential oil and the gum of Pistacia lentiscus Var. chia. J Agric Food Chem. 2005 Oct 5;53(20):7681-5.
• Rahimi R, Mozaffari S, Abdollahi M. On the use of herbal medicines in management of inflammatory bowel diseases: a systematic review of animal and human studies. Dig Dis Sci. 2009 Mar;54(3):471-80.
• Rahimi R, Shams-Ardekani MR, Abdollahi M. A review of the efficacy of traditional Iranian medicine for inflammatory bowel disease. World J Gastroenterol. 2010 Sep 28;16(36):4504-14. Review. 
• Rahimi R, Baghaei A, Baeeri M, Amin G, Shams-Ardekani MR, Khanavi M, Abdollahi M. Promising effect of Magliasa, a traditional Iranian formula, on experimental colitis on the basis of biochemical and cellular findings. World J Gastroenterol. 2013 Mar 28;19(12):1901-11. 
• Rezaei PF, Fouladdel S, Hassani S, Yousefbeyk F, Ghaffari SM, Amin G, Azizi E. Induction of apoptosis and cell cycle arrest by pericarp polyphenol-rich extract of Baneh in human colon carcinoma HT29 cells. Food Chem Toxicol. 2012 Mar;50(3-4):1054-9.
• Sakagami H, Kishino K, Kobayashi M, Hashimoto K, Iida S, Shimetani A, Nakamura Y, Takahashi K, Ikarashi T, Fukamachi H, Satoh K, Nakashima H, Shimizu T, Takeda K, Watanabe S, Nakamura W. Selective antibacterial and apoptosis-modulating activities of mastic. In Vivo. 2009 Mar-Apr;23(2):215-23. 
• Triantafyllou A, Chaviaras N, Sergentanis TN, Protopapa E, Tsaknis J. Chios mastic gum modulates serum biochemical parameters in a human population. J Ethnopharmacol. 2007 Apr 20;111(1):43-9.
• Tzakou, O., Bazos, I. and Yannitsaros, A. (2007), Volatile metabolites of Pistacia atlantica Desf. from Greece. Flavour Fragr. J., 22: 358–362.

Green Tea Dose-Dependently Sheds 7%-20% Body Weight in Mouse Model, But Higher Doses Turn Out to Be Pro- not Anti-Inflammatory In Recent Tufts Study

Image 1: A cup of freshly brewed
green tea (photo by Wikimol)

Green tea is healthy, it gets you going, will make you lose fat and calm down inflammation... wait! Is it possible that all that was just another marketing scam? Scientists from the Tufts University in Boston (Pae. 2011) have now found that at least in case of the capped green tea extracts, health (over-)conscious GNC-consumers are popping like candy, these days, the latter, i.e. the anti-inflammatory effect is absent.

And, it even gets worse, higher doses of a standardized EGCG extract (TEAVIGO; 95% EGCG content) even increased inflammation (measured via proinflammatory cytokines in the blood of the animals) in a group of 6-9 months old pathogen-free male mice who had been fed a diet enriched with 1% of the extract for 6 weeks:

Contrary to the assumption that EGCG would reduce inflammatory response, mice fed 0.15% and 0.3% EGCG diet exhibited no change while those fed 1% EGCG diet produced more proinflammatory cytokines tumor necrosis factor-α, interleukin (IL)-6, and IL-1β and lipid inflammatory mediator prostaglandin E2 in their splenocytes and macrophages (MΦ) and less IL-4 in splenocytes.

In that, it is quite interesting that the cytotoxicity of the green tea extract didn't hinder it from unfolding its unquestionably more marketable potential to reduce body weight.

Figure 1: Body weight of ECGC treated in the course of the 6 week treatment period; expressed relative to the weight of the unsupplemented control group (data calculated based on Pae. 2011)

As the data in figure 1 indicates, all three levels of EGCG enrichment produced statistically significant and surprisingly instantaneous reductions in body weight. The mean weight difference to the control group were -7%, -12%, -20% for the 0.15%, 0.3% and 1% EGCG groups, respectively. Based on previous research on the weight loss effects of EGCG extracts, Pea et al. hypothesize that the underlying mechanisms "involve decreased energy/lipid absorption and lipogenesis",  "increased fat oxidation" and improved glucose tolerance. Since the authors of respective studies were interested mainly in the weight loss effect, they failed to measure inflammatory status, as well, which is why Peat et al. are unable to tell, whether these beneficial metabolic effects are associated or even stem from EGCG's effect on inflammation status.

To evaluate the potentially detrimental effects of the wide-spread use of commercially available green tea extracts, the authors conduct the following dose-equivalent calculation:

Based on the average mice's consumption of 3 g/days, diets containing the medium dose (0.3%), or 3 g EGCG /kg diet, provided mice with a daily intake of 9 mg EGCG, or 300 mg/(kg BW/d) for a mouse of 30 g (average BW in the current study). When this dose is converted from mice consuming 12 kJ/day to humans consuming 2000 kJ/day by using isocaloric calculation, it is roughly equivalent to 22 mg/(kg BW/day) in humans, or 1540 mg EGCG per day consumed by a 70-kg person. Likewise, the low dose (0.15%) and high dose (1%) of EGCG for mice in this study are equivalent to human daily consumption of 770 and 5040 mg, respectively.

In view of the fact, a cup of tea contains only 150−180 mg EGCG, it is unlikely that habitual green tea (~3-5 cup = 450-900mg EGCG) consumption will increase daily intake to the "critical" level of  commercially available EGCG - it could, on the other hand, suffice to induce beneficial effects on body weight, glucose management and fatty acid metabolism.

Did you know that in many studies concentrations of 50µmol/L, i.e. roughly 230,000mg of epigallocatechin-3-gallate per liter, were used to elicit the often cited health benefits of the green tea phytochemical? In animal models, however, even a dose of 2,000mg/kg body weight incresed plasma concentrations to levels slightly below 1/5th (9µmol/L) of what has been studied in the petri dish.

In case of the consumption of commercially available EGCG caps, which contain anywhere from 150-900 mg EGCG/tablet (depending on capsule size and purity of the extract), the more-is-more mentality of our society along with the desire to lose weight as quickly as possible could however lead to epigallocatechin-3-gallate intakes at levels equivalent to those that were achieved in the study and would thus leave consumers slim, but heavily inflamed. So, if you want to avoid that, you better stick to the suggested serving size (in case of the NOW Foods EGCG product, for example, this practice would deliver ~600mg in three divided doses per 24h), instead of going through your cost-effective 250g bottle of bulk 90% EGCG extract within a single month :-)

Mitochondrial Proanthocyanidin-Tuning: Fruit & Seed Extracts from Grapes Cheer Up Tired Mitochondria.

Image 1: Grapes - mitochondrial super food.

While I am still not sold on the idea that the unfortunate and unhealthy fate of being a fat slob is in your genes, it is a commonly accepted scientific fact that lifestyle factors (and thus epigenetics) contribute to obesity related mitochondrial dysfunctions. Defective cellular powerplants, on the other hand, make it increasingly difficult to lose weight and may be one of the major causes of the weight(re-)gain the majority of dieters experience after ardeous weeks of (un-)successful calories-in-vs-calories-out dieting.

The results of two very studies published in the latest issue Journal of the American Chemical Society (Kang. 2011) and the Journal of Food and Chemical Toxicology (Arola. 2011) indicate that both the proanthocyanidins from grapeseeds, as well as the mixture of bioactive substances from whole red grapes may ameliorate the impairment in oxidative capacity and decrease the oxidative stress due to the increased superoxide dismutase (SOD) production in defective mitochondria. Thusly "tuned" mitochondria could not only use energy more effectively, the experimentally established negative correlation of metabolic waste products and insulin sensitivity (more "waster" = more SOD = less insulin sensitivity) would suggests that the restoration and optimization of mitochondrial function turn could trigger a whole cascade of beneficial health effects which improve metabolic health and facilitate weight loss.

Luis Arola and his colleagues from the Departament de Bioquímica i Biotecnologia at the Universitat Rovira i Virgili in Tarragona, Spain, investigated the effects supplementation of Zucker obese rats (commonly used model for type II diabetes and the metabolic syndrome) with 35mg/kg grape seed extract [GSPE; human equivalent: 5.7mg/kg; 454mg for an 80kg human] for 6 weeks had on the skeletal muscle energy metabolism of the animals.

Note: If you intend to buy a similar extract you want to know the exact proanthocyanidin composition. For the extract in the study the ratio of mono to polymers was as follows: monomers (21.3%), dimers (17.4%), trimers (16.3%), tetramers (13.3%) and oligomers (5-1391 units) (31.7%).

In that, it is interesting to note, that the scientists based their hypothesis that GSPE would exert beneficial effects on the metabolically deranged rodents, on previous observations in healthy male Wistar rats, where the proanthocyanidin content of GSPE increased the activity of key enzymes of energy metabolism in muscle and brown adipose tissue five hours after oral administration. Contrary to what has been observed in those healthy animals, however,

chronic GSPE administration decreased citrate synthase activity [which had been increased by GSPE administration in healthy rats], the amount  of oxidative phosphorylation complexes I and II, and Nrf1 gene expression, without any effects on the mitochondrial
oxidative capacity.

The scientists argue that this decrease of citrate synthase may be related to "a lower mitochondrial density in the [gastromenicus] muscle" of the GSPE treated rats; at first sight, a highly undesirable effect, which - and this was confirmed by gene essays - was yet not mediated by apoptosis, i.e. cell-death, due to a pro-apoptotic activity of GSPE - such stress-related phenomena have been observed with high doses of resveratrol (Pan. 2008) or the green tea polyphenol EGCG (Weinreb. 2003; Quanungo. 2005) in cancer cells. Though, whatever the reasons for the reduced critrate synthase, the initially step of the energy production in the Krebs cycle, may be, the

high-resolution respirometry results, using a combination of carnitine and palmitoyl-CoA as substrates, revealed that proanthocyanidins caused higher state 2 respiration levels [which in turn suggests] an improvement in fatty acid transport into the mitochondria and/or an increased capacity for fatty acid oxidation by beta-oxidation.

Coupled with the observed increase in pyruvate influx into the crebs cycle and the anti-inflammatory effect derived from the 163% increase in the cyclooxygenase (COX) to citrate synthase activity (cf. figure 1) this effect may have (over-)accomodate the potential reduction in the number of mitochondria; despite a small decline in triglycerides, it could yet not completely inhibit the slight, yet statistical significant weight gain (7.4%) in the GSPE group, which in itself was the consequence of a 15.1% increase in food intake.

Figure 1: Citrate synthase (CS) and COX/citrate synthase ratio (primary axis), ATPase/CS ratio (secondary axis) in Zucker obese rats after 8 weeks of GSPE supplementation at 36mg/kg (data adapted from Arola. 2011)

In view of the beneficial effects of exercise training on mitochodrial biogenensis (e.g. Balakrishnan. 2010), it goes without saying that athletes and dieters, alike, could benefit from the addition of GSPE to a reasonably planned weight training regime. While the GSPE will ramp up mitochondrial efficiency (as indicated by the increase in ATPase/CS increase, cf. fig. 1), the resistance training will add new mitochondria to the trained muscle - or as Carl Lenore would have it: You build metabolic currency, while practicing currency revaluation at the same time; better than any building loan contract, isn't it ;-)

Figure 2: Effect of 8 weeks on control, high fat and high fat RISC (resveratrol, genistein and carnitine) supplemented diets on fat pad weight of mice (data adapted from Kang. 2011)

If that ain't not enough and you got further funds to spend on supplements, you may literally take the RISC and add some isoflavones, and l-carnitine to the mix. And although I am convinced that the resulting mixture was quite aptly (yet certainly unconsciously) labeled RISC (R is from resveratrol from red grape extract, IS is from isoflavone and C is from L-carnitine), due to its soy isoflavone content, Jong Song Kang (Kang. 2011) and his/her (?) co-workers were able to show that 400 mg/kg or 800 mg/kg of the mixture of resveratrol, genistein and l-carnitine fed to mice at a ratio of 4:1:2.5...

substantially inhibited high-fat diet (HFD)-induced increase in body weight in a dose-dependent manner in C57BL/6 mice; [decreased] the amount of subcutaneous and mesenteric fat [...] significantly [and] significantly increased the plasma level of high density
lipoprotein cholesterol without affecting the level of low density lipoprotein
cholesterol and triglycerides.

The 'RISCy' treatment also reduced lipid accumulation in the livers of the high-fat-fed mice and returned the elevated plasma levels of glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase into the normal range.

Did you know? Grapes are not the only food rich in proanthocyanidins, pistacios are also a dietary source of ligomer/polymer proanthocyanidin fractions and have shown to exert anti-inflammatory effects on macrophages (immune cells) only, very recently (Gentil. 2011). The proanthocyanidin content of wine and sparkling wine, on the other hand, depends on its origin and, as a study by Jardao et al. on the polyphenol and proanthocyanidin content of Portugues wines suggests, vineyard (Jardao. 2010). While the highest concentrations were in the range of 55-130mg/L the majority of samples taken from wines from Portuguese sparkling wines from Bairrada contained no proanthocyanidin, at all.

In view of the results from the RISC trial, it could be speculated that l-carnitine (write-up for Amino Acids for Super Human Series coming soon!) was the rate limiting factor in the Arola study and additional supplementation of the "fat-carrying amino acid" at what would correspond to 133mg/kg (low dose RISC) or 266mg/kg(high dose RISC) for the mice, 66mg/kg or 133mg/kg for rats and 11mg/kg or 22mg/kg for humans (i.e. 870mg or 1649mg of l-carnitine for an 80kg human being / take in at least 3 doses divided across the day) would eventually turn the 5.7mg/kg of grape seed extract into a true "fat burner"... and if this does not work, I'd suggest the addition of resveratrol (dose in the RISC trial would correspond to 17mg/kg /low dose or 34mg/kg high dose in human beings) and carnitine. In view of the highly devious effects of genistein on both the male, as well as female endocrine system I would pass on the soy-isoflavone if your hormonal health is dear to your heart - the slight decreases in fat accumulation (cf. fig.2) certainly ain't worth it. Ah,... and I do not have to remind you of the importance of complementary restiance training, do I?