Maintain & Increase Your Insulin Sensitivity - Wrap-Up: Overview + 3 Stacks. Plus: AMPK & PPAR-γ Revisited

It's not about exercise or nutrition, it's about both of them. And as long as it is not about the former it should not be about supplements either.

It's Sunday and finally time for the wrap up of the "Maintain & Increase Your Insulin Sensitivity" Series (read all previous installments). It took me quite some time to get through all the compounds on my and your list and the outcome is far from being a "complete" list of all the agents that can have beneficial effects on your insulin sensitivity. Still, if there is at least one agent, one fact or one take home message for each of the 6 previous articles of which you would say: "Ok, I guess that was worth wasting some of my valuable time on", I would be happy.

For myself it was by the way yet another occasion to learn more  about many of the agents, I reviewed in this series.

And if you listened to the SuppVersity Science Round-Up on Thursday, you will probably remember that I said that I was at least a cautiously excited about the potential of berberine... but I am wasting your time here. Let's start with the wrap up.

Lifestyle changes are more powerful than supplements: So don't forget that it's imperative to realize the lifestyle changes described in the first installment of this series. Otherwise the best you can hope to achieve with supplements (and drugs) is to slow the progression from insulin resistance to full-blown diabetes.

Before we take a look at the three stacks I have compiled, I do just briefly want to get you up to speed as far as the major underlying mechanisms of the supplements are concerned:

• AMPK: Some people in the medical establishment think of it as the good counter-part to mTOR. The fairy that will put an end to the pro-carcinogenic reign of the evil mTOR-witch, so to say. Others, who are mostly part of the "muscle head" community, think of AMPK as their fiercest enemy and mTOR as their brother in crime... It goes without saying that neither of these views is accurate. AMPK and mTOR are not even necessarily antagonistic - at least not for people like you - people who work out regularly. The mTOR expression after a workout does after all occur in the presence of increased an expression of AMPK. What else could you possibly ask for, if you want to "recomp" (i.e. lose fat and gain muscle to improve your physical appearance)?
  
  

  

  Few people know that there are two different iso-forms of AMPK. The alpha-2 isoform is the one that's expressed during a workout and it works hand in hand with mTOR and not against it (learn more)

  What may at first look like a paradox is actually easily explained if we don't look at the characteristic downstream effects of mTOR and AMPK, but rather at the circumstances in which they are activated. Outside of the previously mentioned exercise context those are in fact antagonist. With mTOR being triggered by the abundance of nutrients - specifically protein, and even more specifically leucine - you would not expect to see increases of AMPK at the same time. The latter is after all, expressed, when a cell senses a lack of nutrients in form of an increase in ADP (~used ATP) and a decrease in ATP levels. The reaction, i.e. an increased expression of AMPK will then have downstream effects on the uptake of glucose and the oxidation of fatty acids, both of which contribute to a restoration of normal ATP levels in the cell.
  
  Both AMPK and mTOR act highly localized. The exercise induced glucose uptake is thus muscle-specific - that should be obvious, since exercise will raise the ADP levels only in the muscle. Supplemental agents that mimic this effect, on the other hand, act systemically. Agents like alpha lipoic acid will therefore increase glucose uptake in both muscle and adipose tissue (Moini. 2002). After a workout and at other time points, where the glucose uptake is already high and, more importantly, muscle specific, it is thus not necessarily the best idea to try to "escalate" the effects by using a class of supplements that is often mislabeled as "insulin mimetics". 
• A note on the PPAR-effects of CLA, fish oil, TTA & co: I guess as a well-read SuppVersity reader you will know that CLA is a supplemental non-starter. In human studies the outstanding results from rodent trials have never been successfully replicated. This could be - at least in parts - a result of the dosage and the ratio of t10c12 CLA to it's c9t11 counterpart, which has the exact opposite effects on the PPAR-gamma receptor (Toomey. 2005).
  

  

  Some of you will probably also remember my articles about the pertinent effects of TTA and fish oil in the. Of those only the latter has a significant PPAR-gamma activity. TTA, on the other hand is predominantly a PPAR-delta and -alpha agonist and thus more a "true fat burner" than a general "fat handler", which is probably the best way to describe fish oil (learn more).
  PPAR-gamma: If you listened to the Science Round-Up last Thursday you will have heard me say that blocking PPAR-gamma will inhibit the storage of energy in the adipose tissue. That's true, but by no means as beneficial as you may be thinking. It is, for example, pretty likely that the CLA induced PPAR-gamma blockade is also responsible for the increased propensity to develop non-alcoholic fatty liver disease in CLA fed rodents.
  
  The results Fedor et al. present in a 2012 paper, show that these effects can be ameliorated if the PPAR-gamma suppressor (i.e. trans-10, cis-12-conjugated linoleic acid) is combined with a supplement that exerts the opposite effects in the liver - a supplement you all know pretty well: DHA, as in "fish oil". It is, just like many of the "older" diabetes drugs, an (allegedly less) potent PPAR-G agonist (=activator; cf. Neschen. 2006).
  
  Whether the blockade of PPAR-gamma is a good or a bad thing does thus obviously depend on the scenario we are talking about. For the lean individual, who is working out regularly and wants to defend his leanness in the absence of (un-)wanted eating orgies and "mass building regimen" it is probably a good thing. He or she will not have an energetic surplus that could end up clogging up the liver. And while the same goes for the average obese individual who has finally gotten his act together, PPAR-gamma inhibitors would seem clearly counter-indicated for sedentary individuals on the high calorie, high sugar, high fat diet so many people consider "normal", these days.
  
  As paradox as it may seem, the anti-diabetic effects of PPAR-gamma activating thiazolidinediones (TZDs) which are still used to "treat" (I should rather write "manage") diabetes mellitus and other diseases that feature insulin resistance will thus come at the expense of increased body fat storage. The latter can be pretty pronounced,as the data from pre-diabetic individuals Bray et al. published only recently goes to show you (figure 1).
  
  Despite their anti-inflammatory effects and their (limited) use in highly inflamed type II diabetics you will therefore not find any of the herbal PPAR-gamma agonists like pomegranate, pumpkin, mellisa officinalis, morus alba, artemisia capillaris, bitter melon, guggul, banaba or mulberry (cf. Huang. 2009) in the stacks below. And that despite the fact that they have anti-inflammatory activities.

Ok, I guess you are either fed up of theoretical details by now, or were so before and just scrolled down to the "stacks" in the first place.

There is a reason there is no "athlete's stack" here: Since I know you will be asking, I thought I will say it right away. The reason that there is no "athlete's stack" in the list is that athletes are either "normal-weight insulin resistant" or they are insulin sensitive and don't need any supplements on top of the lifestyle changes from episode I. You got your workouts, folks: There is no better muscle specific insulin sensitizer out there! If you are looking for agents to stay lean try berberine, it's anti-ppar-gamma effects may help and if you want more, stack it with taurine, of which you should by now know that its benefits go well beyond "glucose sensitizing" / You don't? Well, then take a look at the previous articles about taurine.

I guess I will not make you wait any longer, then... I have come up with three scenarios, the "insulin resistant obese / overweight individual", "the normal-weight insulin resistant individual" and the "cheater" (explanations follow below the stacks):

• 

  Table 1: List of the most important supplements discussed in the series; more details on each of them, as well as on those I did not include in this overview can be found in the individual installments.

  The insulin resistant obese / overweight individual will have to target weight and fat loss and increases in insulin resistance; a stack that could facilitate all three would contain.
• Metformin - 3x 300-500mg (not lipoic acid, this is why)
• Berberine - 3x 200-400mg
• Fucoxanthin - 3x 5mg
• Taurine - 3x 2-3g
• Chromium - 1x 200mcg (*)
• The normal-weight insulin resistant individual will have to take care of inflammation, (usually) a beginning fatty liver and not taking the next step to the obese diabetic.
• Berberine - 3x 200-400mg 
• Chlorogenic acid - 3x 200-300mg
• Taurine - 3x 2-3g
• Milk thistle - 3x 200-400mg
• The cheater can be either of the former or a healthy perfectly insulin sensitive individual who wants to reduce the sudden rise in blood glucose after a meal.
• Cinnamon - 1-6g (Ceylon cinnamon)
• Vinegar - 2x tablespoons
• Green tea - 1-2 cups

The insulin resistant obese / overweight individual needs relieve most urgently, therefore he will also trial fucoxanthin, which has some impressive, but not exactly reliable weight loss data. He will either have a script for metformin or will use alpha lipoic acid (ALA) as a substitute and he will make sure that he gets adequate chromium by taking 200mcg of chromium picoliante or niacin-bound chromium per day *if this is not already in a multi he or she is taking.

The normal-weight insulin resistant individual has slightly different needs than his overweight comrade. He or she is almost certainly suffering from chronic inflammation and beginning or existing NAFLD (the obese will have that, as well, but for him it's only part of the problem). With it's effects on both AMPK and PPAR-gamma berberine will make sure that the body fat levels remain low. Just like taurine (read more), chlorogenic acid (Panchal. 2012) and milk thistle (read more), it will also help "revive" the liver and sooth the inflammation by promoting the bodies own antioxidant defense system.

The cheater, on the other hand, could be everyone who wants to undo (or fore-do, if you will) a high GI carb meal. The ingredients of this stack will ameliorate the blood sugar response and could thus potentially reduce any damage you could do to your pancreas... but let's be honest. With the occasional cheat you are not going to do any damage and if cheating becomes common practice you violate the "lifestyle-changes first!" principle and won't get away healthily no matter how much supplements you take.

Browse previous articles:

Lifestyle Changes

ALA, GABA, Taurine & Co.

Berberine, Banaba & Co.

Cinnamon, Curcumin & Co.

Lemon, Starch, Coffee & Co.

Chlorogenic acid, fucoxanthin & Co.

References:

• Fedor DM, Adkins Y, Mackey BE, Kelley DS. Docosahexaenoic acid prevents trans-10, cis-12-conjugated linoleic acid-induced nonalcoholic fatty liver disease in mice by altering expression of hepatic genes regulating fatty acid synthesis and oxidation. Metab Syndr Relat Disord. 2012 Jun;10(3):175-80.
• Huang TH, Teoh AW, Lin BL, Lin DS, Roufogalis B. The role of herbal PPAR modulators in the treatment of cardiometabolic syndrome. Pharmacol Res. 2009 Sep;60(3):195-206.
• Moini H, Tirosh O, Park YC, Cho KJ, Packer L. R-alpha-lipoic acid action on cell redox status, the insulin receptor, and glucose uptake in 3T3-L1 adipocytes. Arch Biochem Biophys. 2002 Jan 15;397(2):384-91.
• Neschen S, Morino K, Rossbacher JC, Pongratz RL, Cline GW, Sono S, Gillum M, Shulman GI. Fish oil regulates adiponectin secretion by a peroxisome proliferator-activated receptor-gamma-dependent mechanism in mice. Diabetes. 2006 Apr;55(4):924-8. 
• Panchal SK, Poudyal H, Waanders J, Brown L. Coffee extract attenuates changes in cardiovascular and hepatic structure and function without decreasing obesity in high-carbohydrate, high-fat diet-fed male rats. J Nutr. 2012 Apr;142(4):690-7.
• Toomey S, Harhen B, Roche HM, Fitzgerald D, Belton O. Profound resolution of early atherosclerosis with conjugated linoleic acid. Atherosclerosis. 2006 Jul;187(1):40-9. Epub 2005 Sep 22.

The Quest For the Best N6:N3 Ratio. Swine Study Suggests: 5:1 is Healthy, 1:1 Will Also Cut Fat and Build Muscle

 Yes, this study is about omega-3s, but it is not about the beneficial effects of fish oil.

Ok, I see you are totally excited, so I am not going to beat around the bush for long: According to a soon-to-be-published study from the Hunan Provincial Engineering Research Center of Healthy Livestock, Institute of Subtropical Agriculture at the Chinese Academy of Sciences  a 5:1 ratio of n-6:n3 (as in omega-6 to omega-3) is good enough to keep pigs healthy.

Kicking out another couple of grams of omega-6 fatty acids, on the other hand, had pretty intriguing effects on the body composition of the ninety-six male cross-bred (Large White£
Landrace) pigs who happened to weigh about as much as an average human being (another advangate, of swine - HED calculations are not necessary; learn more).

Did you know that pigs are opportunistic omnivores just like us and provide a better model of human metabolism than our little, furry remote cousins with the big round eyes and the long tails who are populating the laboratory cages of scientists all around the world (cf. Miller. 1987)?

Lineseed or soybean - that makes all the difference

I hope you did not already start popping fish oil, while you are reading this. After all, in this case the glorified residual waste from the fishery industry did not contribute to either the health or weight loss benefits Duan et al. observed in their pigs who were fed one of the four isoenergetic diets with n6:n-3 PUFA ratios of 1:1, 2·5:1, 5:1 and 10:1 for 2 months.

Table 1: Ingredients, nutrient and fatty acids composition of the diets the pigs received; † vitamin premix (Duan. 2013)

As the ingredient profiles of the different diets (table 1) tell you the major source of omega-3 fatty acids was lineseed oil. The latter is basically devoid of long-chain omega-3 fatty acids (EPA & DHA). Instead, it contains the short-chain variety aka alpha-linolenic acid that is often laughed at in the health and fitness community as being the useless precursor to the powerful "fish oils" EPA and DHA (note: the level of DHA was identical in all diets!).

So, no fish oil, just ALA

Although the allegedly more potent LC-PUFAs were missing, the changes in body composition and the overall improvement (=reduction) of the activity of the potentially pro-carcinogenic PI3K-alpha gene and the fat storage genes FATP-1 and PPAR-gamma (learn more about PPAR-gamma) were still impressive.

If you take a closer look at the data in figure 1, though, you will realize that the effects on body composition require a reduction to 2.5:1, better 1:1 to become significant.

Figure 1: Feed conversion rate, muscle mass, adipose tissue mass, lean to fat mass ratio expressed relative to the levels of the pigs in the 5:1 n6:n3 group after 2 months on the different diets (Duan. 2013)

At the same time there is a clear relationship between increased adiposity and the amount of soybean oil in the pig diets. Thank god that the USDA in their incredible wisdom lists it among the top dietary sources for fat *sarcasm*.

Ah, I'd better not get political here, but let me point out one thing: In view of the currently available scientific evidence it borders physical injury resulting from negligence that the "guidelines" do not put an emphasis on the reduction of the the crazily high n-6:n-3 ratio of the Standard American Diet (16-17:1).

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Unlike this pig study, a previous rodent study suggests you should pick EPA over DHA over ALA for weight loss purposes (learn more)

Bottom line: This is one of the most convincing well-controlled animal studies we have that would support that the purportedly "paleolesque" 1:1 ratio of n-6:n-3 fatty acids entails highly significant health benefits,  even if those omega-3 fatty acids don't come from fish, but from lineseed oil.

One thing we must not forget, though, is the fact that the beneficial changes in health and body composition were brought about by the concomittant reduction in omega-6 and increase in omega-3 fatty acids. Simply drinking a bottle of lineseed or cod liver oil everyday could thus do more harm than good, because with ~3g of oil per 100g the diet was also low in total fat and almost devoid of saturated fats and whether the same results would occur in a high fat scenario is beyond what the study at hand can tell us.

References:

• Duan Y, Li F, Li L, Fan J, Sun X, Yin Y. n-6:n-3 PUFA ratio is involved in regulating lipid metabolism and inflammation in pigs. Br J Nutr. 2013 Aug 15:1-7. [Epub ahead of print]
• Miller ER, Ullrey DE. The pig as a model for human nutrition. Annu Rev Nutr. 1987;7:361-82. Review.

Amorfrutins: Plant-Derived Selective PPAR-Modulator Outperforms Regular Diabetes Drug and Exhibit Significant Weight Loss, Insulin and Leptin Sensitizing Effects

Image 1: Amorpha fruticosa (photo R. Ott) is a deciduous shrub growing to 4.5 m, the fruits of which contain about 500mg of amorfrutins per 1kg raw material.

One major argument I have been bringing forward against the use of diabetes "medications" for quite some time, now, is that most of them are not "treatment" strategies, in the sense that they help people lose body fat to naturally restore insulin sensitivity and get off their drugs, but rather the opposite. Drugs like rosiglitazone, for example, allow for the further expansion of the adipose tissue and reduce blood glucose levels by storing the excess glucose in those new or expanding fat cells. A recent paper by Weidner et al. does now suggest that there may be a natural alternative (Weidner. 2012), which - despite acting on the same PPAR pathways lacks the fattening effects of synthetic PPAR-gamma ligands (molecules in drugs that interact with the peroxisome pro-liferator-activated receptor gamma) - lacks those highly undesirable fattening effects of thiazolidinediones.

Glycyrrhiza foetida & Amorpha fruticosa amorfrutins - the future of blood sugar management?

With the so-called "amorfrutins" from the edible parts of the two legumes Glycyrrhiza foetida (roots) and Amorpha fruticosa (fruits), the former being related to the "licorice plant" Glycyrrhiza glaba and the latter a brush that is native to the east of the USA, a group of scientists from Germany and the UK have thus identified yet another potent plant-component that outperforms its synthetic competition pretty easily.

Figure 1: Body weight (in g; left) and phosphorylated / non-phosphorylated PPAR-gamma (data from densitometric analyisis) of diet-induced obese mice on high fat diet (HFD) + placebo (vehicle), HFD + rosiglitazone or HFD + amorfrutin 1 (data adapted from Weidner. 2012)

Administered at a dosage of 100mg/kg per day (human equivalent 8mg/kg), the amorfrutins, which have a 2x higher binding affinity for the PPARγ receptor (236 to 354nM) than the aforementioned diabetes drug rosiglitazone (aka Avandia, one of the commonly prescribed thiazolidinediones), had a much more favorable effect on the phosphorylated to unphosporylated PPAR-gamma ratio and lead to statistically highly significant reductions in body weight over the 23-day supplementation period (cf. figure 1).

A note on the significance of PPAR-gamma phospohorylation: The phosphorylation of the peroxisome proliferator-receptor gamma is associated with a profound dysregulation of a large number of genes whose expression is altered in obesity. Its prevention is thus currently regarded as one of the most promising treatment strategies for insulin resistance; one that comes without the negative side-effect of increases in body weight for which all the other thiazolidinediones are notorious. It is therefore not surprising that the blockade of the phosphorylation of PPAR-gamma by amorfrutin 1 (the one denotes the first of the 4 amorfrutins the biological activity of which was investigated in the study) was "significantly correlated with improved insulin sensitivity" in the study at hand (Weidner. 2012).

Other than in most synthetic selective PPAR-modulators, such as bezafibrate, for example, the weight loss was also not mediated by a reduction in food intake, but, as Weidner et al. speculate, a direct results of an increase in energy expenditure - a hypothesis that would certainly be supported by the slight, yet likewise statistically significant increase in thyroid hormone concentration (T4) in the amorfrutin group:

Because the complex effects of PPARγagonism on various endocrine systems and downstreamphysiological changes (e.g., change in thermogenesis, fatty acid oxidation, or activity) are not fully understood, it is difficult to probe all potential mechanisms by which the amorfrutins may affect weight regulation. For example, recent studies suggest that complex interaction of brain PPARγ-signaling with peripheral organs may contribute to the physio-logical regulation of energy balance (30, 31). Presumably, the amorfrutins as partial agonists may act on neuronal PPARγby an-tagonising diet-derived endogenous agonists such as fatty acids, thereby leading to relative weight loss.

Against the background that we are apparently dealing with the PPAR equivalent to SERMs (selective estrogen receptor modulators such as clomid or tamoxifen) and SARMs (selective androgen receptor modulators), it appears prudent to mention that Weidner et al. did not find any cross-activities with other receptors, such as the estrogen receptors alpha and beta, the liver x receptor alpha, the constitutive androstane receptor, and the pregnane receptor. Cross-reactions like these are quite common with other xenobiotics (exogenous substances with biological effects that are produced by other organisms) and can lead to unexpected and mostly undesirable side effects (e.g. anti-androgen activity).

Leptin resistant? No problem for amorfrutins!

Contrary to rosgliatazone, the amorfrutins work their antidiabetic magic even in the presence of full-blown leptin resistance. While the former does not just fail to reduce, but actually promotes weight gain in leptin receptor-deficient db/db mice, treatment with amorfrutin 1...

[...] had no significant effects on mouse body weigh [... but] reduced plasma insulin concentrations more strongly than rosiglitazone (36% vs 19% decrease after 24 d) . Amorfrutin 1 treatment also decreased plasma concentrations of glucose, triglycerides, and free fatty acids. Possibly as a result of enhanced insulin sensitivity, amorfrutin 1 also appeared to prevent deterioration of pancreatic function in insulin-resistant mice, as pancreatic insulin levels improved compared to nontreated control mice.

In genetically non-disadvantaged, normal diet-induced obese mice (DIO) and thus purportedly in most obese humans, treatment with amorfrutin 1 lead to identical reductions in the areas under the glucose and insulin curve in an intraperitoneal insulin sensitivity test (IPIST; is similar to an oral test, but the injection into the intraperitoneal cave ensures that 100% of the glucose actually hits the blood stream) and reduced the basal leptin levels to the same extend as rosiglitazone did (cf. figure 2).

Figure 2: Areas under the glucose and insulin curve in an intraperitoneal insulin sensitivity test (left), basal leptin levels and photographs of the livers of the diet-induced obese mice at the end of the 24-day study period (data and images adapted from Weidner. 2012)

Despite almost identical i improvements in insulin and leptin sensitivity,u>only the amorfrutins, not the thiazolidinedione, rosiglitazone, were able to reduce the diet-induced triglyceride accumulation in in the liver of the treated animals (cf. figure 2, upper right corner) and could thusly help prevent, maybe even revert non-alcoholic fatty liver disease; an effect, by the way, that may be ascribed to

• an increase of the PPAR-alpha dependent purported "anti liver-fat" co-factor Tbl1, and
• reduced inflammation in both the liver, as well as the white visceral adipose tissue of the rodents.

In the end, it does thusly come down to the usual suspects, inflammation + insulin resistance and while the amorfrutins from the roots of a certain variety of licorice (Glycyrrhiza foetida) and the fruits of an American shrub (Amorpha fruticosa) share their ability to reduce the former and increase the latter, they don't to it at the expense of further increases in body and organ fat and could therefore help to actually resolve - not just manage and perpetuate - the current diabesity epidemic.

"When are we going to see those amor-thingies on the market?"

Image 2: Amorfrutilean could be a weight loss adjuvant that works.

Yet while they could, just as their inferior, since fattening synthetic counterparts, rosiglitazone (Avandia), pioglitazone (Actos) & co be combined with metformin (which works via a totally different mechanism), it appears questionable that the next generation of Avandamedm which combines metformin and rosiglitazone in one pill, is going to have amorfrutins in it. Not because we still need human trials (which is obviously the case), but rather due to the fact that no pharma company will be willing to pay those trials, when the outcome, a "drug" based on a naturally occurring substance that is easily extracted from a common plant, would not be patentable and may seriously compromise the sales of their current "antidiabetic" (actually I should write "pro-obesity") drugs.

If independent future human trials were yet able to confirm the previously discussed results, I am still pretty confident that we are soon going to see the supplement industry jumping aboard. With Amorfrutiburn, Amorfrutilean or Amorfrutibol being the most likely candidates for the BB.COM topselling "fat burners", weight loss adjuvants or "nutrient partitioners", respectively. And you know what, combined with a couple of lifestyle changes, this stuff could actually work - at least  for the chubbier one's among the soon-to-be physical culturist. Whether leaner folks or even bodybuilders will benefit to the same extend does yet remain to be seen.

Vitamin D3 a "Fat Synthesizer"!? Rodent Study Shows +33% Increased Fat Deposition in Vitamin D3 Supplemented Mice.

Illustration 1: Experts will recognize from looking at these Oil Red O-stained longissimus dorsi slices of mice on a normal and a vitamin D3 supplemented diet that supplemental (! not vitamin D from the sun !) "vitamin D3 can be used a s a fat synthesizer and meat tenderizer in meat-producing animals". (img in illustraton from Choi. 2011)

I have been railing against the current vitamin D hype for months now. In that, I have at no point in time implied that "backfilling" depleted vitamin D levels via supplementation could not be beneficial (or at least not harmful), nor have I at any time excluded that vitamin D3 supplementation (even if you are in the "normal" range) could have its merit (cf. vitamin D3 + HMB). What I have done though, was to point at the lack of controlled studies that would support any of the benefits supplemental vitamin D3 is currently hailed for all over the Internet. This amazes me, because the very same "gurus" who are all over the vitamin D bandwagon have lately (just like me) discarded the data from the Iowa Women's Health Study as "unrealiable" and "non-significant" epidemiological bullshit (which is exactly, what I think, as well). When it comes to vitamin D, however, they throw all their concerns on the validity of epidemiological data over board and worship their vitamin D3 pills like a golden calf.

But let's get to the facts, before I get tarred and feathered, again... In the latest issue of the Journal of the Science of Food and Agriculture Hyuck, Choi and Kyuho Myung published a paper that investigated the use of vitamin D3 supplements to fatten animals (Choi. 2011). Now, you may think "How stupid is that, everyone knows that vitamin D will make you lean out!", but as I've pointed out several times within the last weeks, high vitamin D levels may correlate with a lean body composition; however, studies that would show that supplementation of the latter would induce respective changes in body composition in the absence of prior deficiency (and we are talking about the standard reference range with a lower limit of 10ng/mL, here) simply do not exist... but I am digressing again.

Figure 1: Composition of the diet (large figure) and respective vitamin D3 content (small figure) of the diets of the control and the supplement group in the study.

As you can see in figure 1 both groups (2x N=10) of 6 weeks-old male C57BL/6 mice were fed identical chows (AIN93G; cf. figure 1, large), varying only in their vitamin D3 content (1IU in the control group, 10IU in the supplemented group). In human terms this would be like switching from your common western low vitamin D diet with roughly 800IU to taking a 8.000 IU supplement, each day - something I suppose many of you may have done lately!?

Figure 2: Body fat (in g; large figure) and respective serum 1α,25-(OH)2 -vitamin D3 levels (in µg/mL; small figure) after 3 weeks on control or vitamin D3 supplemented diet (data calculated based on Choi. 2011)

As figure 2 shows, this 10-fold increase in dietary vitamin D would only be advisable if you were a "sumo mouse" who has to make weight for the next competition. A plus of +33% in total, +29% in unaesthetic subcutaneous and +25% in unhealthy visceral fat (all statistically significant with p<0.022, p<0.032 and p<0.043) is not what you would expect of the "greatest vitamin of all time" - would you? And while the vitamin D3 mice also gained some more body weight, those changes were statistically non-significant, so that - as the scientists state - vitamin D3 turned out to be an ideal "fat synthesizer and meat tenderizer".

Figure 3: Cytokines, UCP-2 and PPAR-gamma expression in mice after 3 weeks on control or vitamin D3 supplemented diet (based on Choi. 2011)

In that, vitamin D3 works it "fat synthesizing" magic by increasing the inflammatory cytokines TNF-alpha and IL-6 and decreasing the muscle anabolic (Busquets. 2005) and fat catabolic cytokine IL-15 (Carbo. 2001; Alvarez. 2002), as well as the uncoupling protein UCP-2 while ramping up fat storage via increase PPAR-gamma expression (cf. figure 3).

Now obviously, this is just another rodent study and we cannot say how and if the results will translate to humans, but it is a controlled study and it investigates the effects of supplemental vitamin D3 which is something you cannot say of the "scientific backbone" of the current vitamin D3 craze... and now tar and feather me like a child who has just been bereaved of his favorite toy, if you will ;-)

Ephedra is Back! 'Mahabala', Featuring PEA, Ephedrine, Choline, Betaine & More, Is Nature's Hypolipidemic, Anti-Diabetic and Cardioprotective Fat Burning Stack

Image 1: Sida rhomboidea
leafs contain ephedrine and
other fat loss related alkaloids
(Tan Hoard Exports)

If you are a supplement producer, I suppose you will soon drop me an email to get the phone number of Ranjitsinh V. Devkar from the Division of Phytothrapeutics and Metabolic Endocrinology at the Department of Zoology of the M. S. University of Baroda in Gujarat, India. And, I must admit, if I had not always believed that "Nature knows best!", I would probably have been similarly surprised as some of you will have been, when they read the title of this blogpost - the 'ingredient profile' of Sida rhomboidea (also Sida rhombifolia), or  "Mahabala", a weed that is found in marshy places all across India and that has been used in Ayurvedic medicine for centuries to treat fever, heart disease, ever, heart diseases, burning sensations, urinary disorders, piles and all kinds of inflammation, looks like it had been printed on a non-FDA-approved (and thus potentially effective ;-) fat burner.

Figure 1: Alkaloid content of Sida rhomboidea extract (data adapted from Prakash. 1981)

If you have been following the exponential growth of the supplement market and the allegedly creative ideas the supp-designers had as far as fat burners were/are concerned, you will notice that, vasicinol and vasicinone aside, all the alkaloids, as well as choline and betaine Prakash et al. found in an extract from 5kg of Sida rhomboidea sound vaguely familiar.

Figure 2: Chemical structure
of the alkaloid vasicinone
(extracted from seeds of
Peganum Nigellastrum
by Zhang. 2009)

Vasicinol, vasicinone? What is that? Despite the fact that there is not much reliable data on the pharmacology of vasicinol and vasicinone, the two less well-known alkaloids in Sida rhomboidea, the available scientific data and information on their traditional use in Ayurveda suggests that these compounds, which are also present in other Ayurevedic herbs, exhibit hyopglycemic, as well as weak anti-acetylcholinesterase (Zhang. 2009) and bronchodilatory (Amin. 1959) effects. All three of these, which entail lower blood sugar levels, increased levels of acetylcholine and beta-adrenergic activity (which is the most likely explanation for bronchodilatory effect of the alkaloids), could synergistically help facilitate weightloss.

In view of this potent 'ingredient profile', it is no wonder that the addition of 1% of this natural fat loss wonder to the hypercaloric high fat diet (+58% more energy than low fat chow) of male C57BL/6J mice (6–8 weeks of age) staved off >25% of the 20-week weight gain (cf. figure 3) the unsupplemented HFD group experienced with respect to a low fat fed control group in the most recent of a whole series of rodent studies, Devkar et al. have conducted over the course the last years (Thounaojam. 2011).

Figure 3: Weight [in g] of mice during 20 weeks on control, high fat (HFD) and high fat diet with 1% Sida rhomboeidea extract (data adapted from Thounaojam. 2011)

From a health perspective, it may yet be even more important that the Mahabala supplement also prevented the "HFD induced increment in [...] plasma lipids and leptin, visceral adiposity and adipocyte hypertrophy" the scientists had observed in the unsupplemented group. The authors attribute these effects to the down-regulation of PPARγ2 and leptin gene expression that went hand in hand with an attenuation of food intake in the C57BL/6J mice.

Image 2: Mice on low fat (A), high fat (B)
and high fat diet + 1% Sida rhomboeidea
extract after 20 weeks (Thounaojam. 2011)

The decline in food intake, the scientists had already observed in previous studies , makes it quite difficult to give a definite number on the absolute amount of Sida rhomboeidea extract the mice consumed on a daily basis. It ranges from 25mg at the beginning of the study to ~13mg at the end and would translate into a human equivalent dose of 2mg/kg and 1mg/kg of Sida rhomboeidea extract per day. If the simple mathematical calculation that is solely based on the ratio of body surface to weight would suffice to reliably translate data from a rodent model to humans, an 80kg human being would thus have to consume somewhere between 80mg and 160mg of Mahabala per day to see similar effects.

Now, you may argue that, after all, this "wonder extract" turns out to be just another appetite suppressant. Yet, while Mahabala did affect the appetite of the laboratory animals, its profound in-vitro effects on pre-adipocytes differentiation and leptin release, as well as the previously mentioned changes in PPARγ2 and leptin gene expression, the scientists observed in the rodent model, suggest that the primary mechanisms for Mahabala's preventive effects against weight and fat gain (-56% less abdominal fat, -46% less epidididymal fat) is unrelated to the reduction in food intake.

Figure 4: Effect of Sida rhomboeidea extract (SR) and Rosiglitazone (ROS) on insulin tolerance in C57BL/6J mice after 16 weeks on low fat (LFD) or high fat diet (HFD) containing 1% (SR1) or 3% (SR2) SR or  0.05% ROS (data adated from Thounaojam. 2010)

Add to that its profound (as potent as the anti-diabetes drug Rosiglitazone) effects on insulin sensitivity (cf. figure 4) the researchers had observed in a previous study (Thounaojam. 2010) and remind yourself of the fact that not ephedrine, but its foolish and unnecessary abuse / overconsumption were responsible for the unfortunate deaths of a handful of people, the majority of whom had preexisting health problems (yes, being severely overweight is a health problem), and you will probably agree with Ranjitsinh V. Devkar and his colleagues who conclude that their findings "validate the potential application of SRLE as a therapeutic agent against obesity". And, if you asked me, I bet that it won't take long until you see the first Mahabala supplements hit the highly competitive fat loss market - despite the fact that an ephedrine-containing Ayurvedic herb probably ain't FDA compliant ;-)

Xylitol, a Low-Calorie Sweetener With Unknown Fat-Burning Side-Effects!? Replacing Cornstarch With the Five-Carbon Sugar, Xylitol, Ramps Up Fat Burning Enzymes in Rodent Model of Diet Induced Obesity.

Image 1: Xylitol christals under the micro-
scope (photo taken by Anders
Østergaard Madsen, Denmark 2001)

From a health perspective, the five-carbon sugar alcohol Xylitol has hitherto principally been known for its beneficial effects on dental health. With a caloric value of 3kcal/g the naturally occurring ingredient of a wide variety of plants, citrus fruits (plums, strawberries, raspberries) and vegetables (cauliflower) does have less calories than sugar, nevertheless, according to the fundamentally flawed, but still widely held paradigm that "a calorie is a calorie, no matter where it comes from", the idea of replacing sugar with xylitol to reduce the overall caloric load would make little sense even on a diet that was relatively high in carbohydrates. So, if it were not for the highly marketable statement "sugar free" on chewing gums and candies, xylitol would probably not even have made it to the consumer market. A cursory glance at the abstract of a study (Ama. 2011) that is about to be published in the July issue of the Journal of Clinical Biochemestry and Nutrition (Vol. 49, No. 1) does yet suggest that these research results could renew the interest in a compound many of you may have considered to be nothing but another marketing scam from the (diet-)food industry - and, before this new data came out, you were 100% spot-on with your assessment ;o)

In the course of an 8-week feeding period, Kikoko Amu and his collegues from the University of Tokoshima Graduate School, the University of Shizuoka and the Food and Science Institute in Kanagawa, Japan, fed a group of 18 male Sprague-Dawley rats (initial body weight 290-310g) a high fat diet containing 312.3 g/kg cornstarch (control group), of which, in the experimental groups, X1 and X2, 16% (X1) or 29% (X2) were replaced with 1.0g/100kcal (X1) and 2.0g/100kcal (X2) of xylitol for group X1 and group X2, respectively. At the end of this period the scientists found that

long-term intake of xylitol supressed the accumulation of visceral fat [cf. figure 1, below] and the increase in plasma insulin and lipids concentrations in rats fed a high-fat diet.

Interestingly, at least part of the effects were mediated by xylitol-stimulated "expression of fatty acid oxidation genes in the liver, and lipid degradation and adiponectin genes in the adipose tissue." Furthermore, Amo et al.

found for the first time that xylitol ingestion lowered postprandial hyperglycemia [if administered at] a none-effective dose in causing diarrhea, and within the limits of orally administered physiological amounts (1-4g/kg body weight daily).

The human equivalent dose for this "non-effective dose in causing diarrhea" (nice, how these Japanese try to paraphrase that the dose they used did not cause "the runs", isn't it?) is 0.16-0.65g/kg per day, which would amount to somewhere between 13g and 52g of xylitol for a man who weighs 80kg. A pretty large dose considering the fact that some low-carbers out survive on hardly more than 40g of carbs a day.

Figure 1: Weight [in g/kg body weight] of retroperitoneal, epididymal and mesenteric part of visceral fat in high-fat (HFD) and high-fat xylitol-substituted (X1, X2) fed rats after 8 weeks of treatment (data adapted from Ama. 2011)

Despite being on a high-fat diet (I hope the readers of this blog have already gotten the message that low carb wont work in the absence of fat) its highly questionable, especially for "low-carbers", whether and to which extent healthy human beings would benefit from xylitol substitution. After all, the "high-fat" chow the rats were fat contained a whopping amount of 362.3g carbohydrates from corn and only 200g of fat from lard (75%) and soybean oil (25%). In view of the dietary 'quality' (I hardly dare to use this word in the context of what those poor critters were fed) of the grub the rats were given, it should not come as a surprise that despite having smaller visceral fat pads, the rats from the experimental groups (X1, X2) were just as overweight (X1: 525.1 +/- 9.6g; X2: 540.4 +/- 9,9g) as their mates (HFD: 543.2 +/- 10.2) who were crammed with what I would like to call a "high-fat corn diet". Furthermore, there were no significant differences in the amount of energie the rats from the different groups consumed and the lean mass of the soleus muscle, the only muscle variable measure in the study, was almost identical. Against that background the improvements in glucose management (cf. figure 2), the scientists observed, could simply be related to the decreased carbohydrate intake. 

Figure 1: Serum markers of glucose and lipid metabolism in high-fat (HFD) and high-fat xylitol-substituted (X1, X2) fed rats after 8 weeks of treatment (data adapted from Ama. 2011)

So, if it were not for the statistically significant increases in adipose gene expression of PPAR-gamma (key regulator of adipocyte differentiation) and the insulin sensitizing hormone adiponectin, as well as the increased lipolytic enzyme-activity (hormone sensitive lipase, HSL; adipose triglyceride lipase, ATGL) within the adipose tissue of the xylitol fed animals, all of you, who have already gotten off the westernized fast-food diet, the scientists were emulating in this study, could easily discard their results as meaningless. Until now, however you will have to wait for future studies on the effects of xylitol on metabolic health and body composition in animals (or humans) who are not fed a diet the detrimental health effects of which cannot be fought off by any supplement or drug on the market. And remember: As always, the SuppVersity is where you will hear about those studies first ;-)

Coumarins from the Stem Barks of Fraxinus rhynchophylla Inhibit Adipocyte Differentiation. A Novel Candidate for Weight Loss Supplement?

A recent study by Shin et al. (Shin. 2010) identified another potential weight loss drug: The Coumarins from the stem barks of Fraxinus rhynchophylla inhibited adipocyte differentiation and thus the basic mechanism of irreversible fat gain.

Figure 1: Effects of Coumarins from F. rhynchophylla on Adipocyte Differentiation
in 3T3-L1 Cells on days 1-6 (Shin. 2010)

As figure 1 shows, Esculetin a Coumarin from Fraxinus rhynchophylla is highly effective in inhibiting early adipocyte differentiation (day 1). With regard to the underlying mechanism of action, the scientists speculate:

Esculetin (1) significantly blocked induction of PPARg expression in differentiated adipocytes, as measured by Western blot analysis (Fig. 4). In addition, esculetin (1) significantly inhibited adipocyte differentiation induced by troglitazone, a PPARg agonist (Fig. 5). These results suggest that esculetin (1) inhibited adipogenic differentiation, in part, via inhibition of PPARg -dependent pathway.

If the results can be reproduced in vivo, supplementing with Fraxinus rhynchophylla bark may prove a preventive measure against adiposity. If, however, you are already fat and dieting, inhibition of adipocyte differentiation won't be your major concern, anyway.