Inulin & Beta Glucan Reduce Body Fat Gain By -50% & -33%! Both Have Similar Effects on the Gut Microbiome, But Only Inulin Appears to Be More Than An Appetite Suppressant

What do these Jerusalem artichokes, agave, bananas, burdock, camas, chicory, coneflower, costus, dandelion, elecampane, garlic,jicama, Leopard's-bane, mugwort, onion, wild yams, yacon and a whole host of other foods have in common? Right! They contain inulin. Whether you will be able to get a whopping amount of 10% inulin in your diet w/out the use of supplements or "enriched" foods, is yet as questionable as how beneficial this actually is for friends of physical culture.

The gut microbiome is not just one of the hottest topics in the (health-)blogosphere, it is also a subject of ongoing research. Research, however, that is, if we are honest, still very much in its infancy. As impressive as the results from the latest studies into the metabolic downstream effects of the administration of fermentable fiber to rodents may be and as obvious as their relation to certain changes in the gut microbiome of the animals may appear - in the end, our understanding of the underlying mechanisms does not allow any reliable prognoses like "double the amount of lactobacilli and you will eventually be able to lose that pouch of body fat you've been carrying around for years now". And yet, if the results from the latest rodent experiments at the Imperial College in London, could be reproduced in humans, I can already foresee that both, the consumption and use of the foods I listed in the caption of the image to the right, as well as related products, extracts and supplements, which contain more or less significant amounts of the naturally occurring polysaccharides, we usually refer to as inulin, will increase in the months and years to come.

Fermentable fiber and the gut-brain-axes: The key to lifelong leanness?

If this is not your first visit to the SuppVersity, you will certainly be aware that the idea of a magic pill (or fiber) that will allow you to eat whatever, whenever and in whichever amounts without having to cope with the metabolic consequences is illusive. When the addition of 10% inulin (or beta glucan) to the diets of 36 male C57BL/6 mice had an "anti-obesogenic" effect, this does not mean that the poor critters who were kept on a hypercaloric high fat (41.8%) diet for 8 weeks did not get obese. What it does mean, though, is that the addition of 10% fermentable (=being food for certain gut bacteria) fiber in the form of

*the producers of these products did not fund or support the study (at least the scientists don't mention that in the respective disclosure ;-)

• inulin from Synergy(TM)*, a fructan based preparation containing both long and short chain
  fructooligosaccharides, or
• beta-glucan from Glucagel(TM)* a highly rich (,80%) barley derived b-glucan preparation

to their otherwise iso-caloric diet (the HFD control contained cellulose) was not without helped to mitigate the negative effects of this diet - a fact the majority of you, of whom I would expect that they are not on a fast-food diet should keep in mind, before they head over to their favorite online supplement vendor and type "Synergy inulin" into the search box.

Figure 1: Effect of addition of 10% fermentable fiber as inulin or beta glucan to the high fat diet of male mice on cumulative weight gain (left), body composition and fatness (middle) and food intake (right) over the course of 8 weeks (data adapted from Arora. 2012)

In spite of that, the results are simply too impressive not to think about their implications in otherwise healthy and even more so previously obese individuals. This is particularly true, because the same microbial changes about which the authors write in a previously published paper from May 2012 that the ...

"[...] increases in both Bifidobacteria and Lactobacillius and a significant increase in short chain fatty acids (SCFA) [went hand in hand with] increase in neuronal activation within the arcuate nucleus (ARC) of animals that received In [inulin] supplementation" (Anastasovska. 2012)

do not (and this is a result of the researchers very latest experiments) simply blunt the rodents appetite. If that was the case, the rodents that received the beta glucan supplemented chow and consumed 12% less energy should have had the most favorable body composition. A cursory glance at figure 1 will yet tell you that this was not the case, though.

Inulin beats beta glucan when it comes to body fat reduction / repression

If we take a closer look a the differential effects of inulin and beta glucan, there yet only one figure that really sticks out and that's the accumulation of fat within the musculature of the animals. The "beautiful marbling" people are looking for in their steaks, however, usually is a harbinger of impeding or even existing skeletal muscle insulin resistance. A muscle fat content above the high fat control (it's certainly a weakness that we don't have a "real" control group on standard rodent chow, here) as Arora et al. observed it in the tissue samples of the beta glucan group, does thus tell you something about its potential usefulness, or rather uselessness of this specific type of fermentable fiber.

Figure 2: Effects of the different types of fermentable fiber on cecal microflora groups (figures are in scientific notation, this means "1E+6" equals 1mio, "1E+9" would be 1 billion etc.; data based on Arora. 2012)

In conjunction with the information about the corresponding changes in the gut microbiome (see figure 2), which appear virtually identical in both groups (specifically the extreme increasesin in both Bifidobacteria (BIF) and Lactobacillius (LAB) really stick out), this does however suggest that the modulatory effect on the composition of the gut flora, or at least the part of it the scientists evaluated in the study at hand, cannot be the only driving force behind the beneficial metabolic effects of inulin.

Inulin or beta glucan? This is not a question... 

While the latter, i.e. inulin, which has by the way been found to directly suppress lipogenesis in a 2011 study by Belgian scientists in a similar HFD rodent model (Dewulf. 2011), appears to be promising for everyone, regardless of whether he or she is poisoning him- or herself with the standard American diet (which is, with its high fat and high carbohydrate content de facto an identical twin of the so-called "high fat diet" in rodent studies), the ingestion of larger amounts of the former, i.e. beta glucan, does at least appear questionable.

If you want to use inulin to your metabolic advantage, you better make sure you get your self a more comfortable place to answer the call of nature - it could call thrice as often! Moreover, large amounts of inulin and other fermentable fiber can induce gastrointestinal distress-

The question is therefore not so much whether it's worth supplementing (it's certainly worth to incorporate some of the initially mentioned foods into your diet, as most of them contain a whole list of other advantageous micronutrients) with inulin or beta glucan - the answer would obviously be inulin - but rather whether it's worth adding larger amounts of inulin to an already healthy diet. And while we cannot answer this question based on the results of the previously cited rodent studies, we could argue that Marwa Zenhom and her colleagues from the Christian Albrecht University in Kiel have already supplied relevant evidence that this would be the case (Zenhom. 2011). After all, the German researchers have been able to show that the PPAR-gamma related anti-inflammatory effects (significant reductions IL-12 secretion in Caco-2 cells and gene expression of IL-12p35, IL-8, and TNFa as well as NF-kB) of oligosaccharides are not (exclusively) brought about by their effects on the gut microbiome, because bacteria simply were not present in their in-vitro study with human Caco-2 cells (cells from the gut lining). Bassaganya-Riera et al. even argue that this effect could be beneficial for IBS patients (Bassaganya-Riera. 2011).

Whether having 10% of your diet in form of inulin, or to make this more conceivable, having 1 tablespoon of plain inulin for every 9 tablespoons of whatever else you eat is either feasible or reasonable, is a whole different story (to put that into perspective: The average inulin intake of Westerners is 1-10g per day (van Loo. 1995). Even 10g would yet only be enough if you ate only 100g of food within 24h!)... and I must forewarn you, if you go by the fecal volume of the mice in the Arora study, it is possible that you will spend >3x more time on the toilette than usual ;-)

References:

• Arora T, Loo RL, Anastasovska J, Gibson GR, Tuohy KM, Sharma RK, Swann JR, Deaville ER, Sleeth ML, Thomas EL, Holmes E, Bell JD, Frost G. Differential effects of two fermentable carbohydrates on central appetite regulation and body composition. PLoS One. 2012;7(8):e43263.
• Anastasovska J, Arora T, Sanchez Canon GJ, Parkinson JR, Touhy K, Gibson GR, Nadkarni NA, So PW, Goldstone AP, Thomas EL, Hankir MK, Van Loo J, Modi N, Bell JD, Frost G. Fermentable carbohydrate alters hypothalamic neuronal activity and protects against the obesogenic environment. Obesity (Silver Spring). 2012 May;20(5):1016-23.
• Astegiano M, Pellicano R, Terzi E, Simondi D, Rizzetto M. Treatment of irritable bowel syndrome. A case control experience. Minerva Gastroenterol Dietol. 2006 Dec;52(4):359-63.
• Bassaganya-Riera J, DiGuardo M, Viladomiu M, de Horna A, Sanchez S, Einerhand AW, Sanders L, Hontecillas R. Soluble fibers and resistant starch ameliorate disease activity in interleukin-10-deficient mice with inflammatory bowel disease. J Nutr. 2011 Jul;141(7):1318-25.
• Dewulf EM, Cani PD, Neyrinck AM, Possemiers S, Van Holle A, Muccioli GG, Deldicque L, Bindels LB, Pachikian BD, Sohet FM, Mignolet E, Francaux M, Larondelle Y, Delzenne NM. Inulin-type fructans with prebiotic properties counteract GPR43 overexpression and PPARγ-related adipogenesis in the white adipose tissue of high-fat diet-fed mice. J Nutr Biochem. 2011 Aug;22(8):712-22.  
• van Loo J, Coussement P, de Leenheer L, Hoebregs H, Smits G. On the presence of inulin and oligofructose as natural ingredients in the western diet. Crit Rev Food Sci Nutr. 1995 Nov;35(6):525-52.
• Zenhom M, Hyder A, de Vrese M, Heller KJ, Roeder T, Schrezenmeir J. Prebiotic oligosaccharides reduce proinflammatory cytokines in intestinal Caco-2 cells via activation of PPARγ and peptidoglycan recognition protein 3. J Nutr. 2011 May;141(5):971-7.

Leucine + Resveratrol - Synergistic Sirtuin Boosters: +118% Fatty Acid Oxidation, 60% Increase In Muscular Glucose Uptake, -30% Visceral Fat & More - To Good to be True?

Image 1: Can you really team up leucine (or HMB) and resveratrol to make tired mitochondria get a move on? NuSirt Sciences says "YES!" And in the dish and rodents it's actually already working.

What happens if you marry a well-known AMPK promoter and exercise mimetic, with an even more prominent exercise adjuvant and nutritional mTOR booster? Will they neutralize each other? Think about it.... ok, now gimme your answer: What happens if you put resveratrol and leucine together? At first it does not really make sense, does it? Right, it doesn't, at least not unless you follow the same train of thought, the researchers from NuSirt Sciences. NuSirt? That rings a bell, hah? Yeah those were the guys who did a study on their 250mg leucine + 30mg vitamin B6 proprietary blend NuFit (see "Testosterone - 12% Drop /W 75g Glucose? Fat Loss - Adzuki, Leucine + B6 or HiMaize & More") and actually, the leucine + resveratrol combination is sort of a spin-off of this initial research.

If you put Sirt1 & Sirt1 together, it suddenly makes sense!

In their latest study (and you bet a future product!) Bruckbauer et al. build on their previous research on the agonistic effects HMB, alpha-KIC or leucine have on skeletal muscle Sirt-1 activity (Bruckbauer. 2011) and rationalize that it seems legit to combine one Sirtuin portein promoter with another one in order to achieve an even more pronounced effect - makes sense, right? Resveratrol the proven AMPK-promoter and igniter of the longevity, gene transcription, cell survival and apoptosis regulating Sir2 proteins (=sirtuins) and leucine the mTOR promoting and, as of late, proven Sirt1 agonist, they could actually form a synergistic duo for fat oxidation, glucose management, the reduction of oxidative stress and inflammation and even longevity!

Figure 1: Effects on sirtuin & AMPK expression in muscle and fat cells upon incubation with leucine, HMB and resveratrol and the respective combinations (left) and effects fatty acid oxidation in isolated rat skeletal muscle upon incubation in low and high glucose conditions (data based on Bruckbauer. 2012)

Now, aside from Sirt1, which is mainly expressed in the nucleus of a cell, another one of the Sir2 proteins, Sirt3, which is expressed predominantly in the mitochondria has as of late gathered quite some attention, as mitochondrial dys- or malfunction is one, if not the common denominator of many of the pathological features of the metabolic and neuro-endocrine ailments the Western diabesity society is suffering from: insulin resistance, type II diabetes, Alzheimer's , you name them! No wonder the NuSirt guys (and girls) are striving to find a marketable way to set them both in full gear and if you take a closer look at the data in figure 1 their initially counter-intuitive approach to bath muscle and fat cells in resveratrol  + HMB / leucine solutions yields impressive results:

• resveratrol, leucine and HMB, alone, exerted only weak independent effects on Sirt1, Sirt 3 and AMPK
• resveratrol and leucine or HMB, combined, yielded Sirt1 and Sirt3 activity increases in the ~50% range (p < 0.05) and AMPK increases of +42% and +55% (p < 0.03); particularly noteworthy are the ~125-175% increases (p < 0.02) muscle cells (remember: Sirt3 is expressed in the mitochondria!)
• the ensuing increases in fatty acid oxidation in incubated muscle cells reached statistical significance in the presence of low (5 mM) glucose levels, only, when and 5 µM HMB or  0.5 mM leucine were co-incubated with 200 nM (~18%; p < 0.05), in the high glucose condition, however, all treatments broad about significant increases in fatty acid oxidation, of which those in the leucine- and HMB-resveratrol combination treatments were the most pronounced (118% and 91% stimulation, respectively; p < 0.005)

Especially the last finding, i.e. the increase in fatty acid oxidation in an in-vitro condition that resembles the hyperglycemic state the average type II diabetic who is not popping tons of metformin and/or injecting insulin is constantly in, makes these results particularly interesting, as it appears as if a "non-pharmacological" (what by the way is "pharmacological" and what isn't?) solution to the diabesity problem could already be hidden on the shelves of your GNC right next door (I assume they carry leucine and resveratrol products ;-)!

Outside of the box... ahh, I mean, ... the petri dish!

In view of the fact that 75% of the in-vitro high performers suck in the rodent model already and of those another 75% don't work in human trials you will be pleased to hear that NuScirt Sciences' resveratrol + leucine / HMB combination has already overcome the first of these hurdles: At least in DIO (diet-induced-obese) rodents who on a 6-week high fat diet regimen, the combination works.

Figure 2: Weight gain, visceral adipose volume, PET measured palmitate uptake, respiratory rate (lower levels = higher relative fat oxidation), heat production relative to body weight, food intake; all value expressed relative to DIO mice who were maintained on an unsupplemented control diet (data based on Bruckbauer. 2012)

Now, it's not as if the rodents would have made it to the Mr Olympia stage, but if you take a closer look at the pattern that's emerging here, it's quite clear that the sirtuin booster does its job in this rodent model. Aside from its ameliorative effect on weight gain, the combination of resveratrol and leucine, led to statistically significant improvements in glucose management and improvements in inflammatory markers (including the anti-inflammatory adipokine adiponectin, see figure 2).

Figure 2: Glucose, insulin and HOMA IR levels, muscular glucose uptake (left), C-reactive protein , IL-6, MCP-1 and adiponectin (right) ; all value expressed relative to DIO mice who were maintained on an unsupplemented control diet (data based on Bruckbauer. 2012)

Most importantly, however it effectively cut through the exuberant amount of visceral adipose tissue (>30% reduction), ramped up the palmitate (fatty acid) uptake, oxidation and heat production (=thermogenesis). Despite all these metabolic improvements which took place in the absence of a simple reduction in food intake, there are still a couple of things left to be desired:

What are the human equivalent doses, here? Since I know you would be asking I did the math for you and you will be pleasantly surprised (HED for 80kg humans)

• 12.5mg resv. = 9mg
• 225mg resv. = 136mg
• 2g HMB = 1.1-1.4g
• 10g HMB = 7.2g
• 24g leucine = 14.3g

I am well aware that it must look as if I had the typical poor arithmetic abilities of the average physicist who has totally forgotten how to calculate using figures instead of letters, but the reason for the discrepancies is that I calculated the exact HEDs based on body weight and food intake for each of the groups.

1. Supplementation with the respective human equivalent doses should yield the same astonishing results in humans as it did in the diet-induced obese mice.
2. The protocol should have effects not just in morbidly obese diabetic human beings, but also in overweight and ideally even lean individuals.
3. The supp must work if you don't put it into the chow, but pop it in separate doses (e.g. 3x/day) as a capsule or tablet.

The good news however is that if 1-3 apply, you could start benefiting from this "super supplement" right now! After all, the resveratrol dose of 12.5mg per kilogram of chow (the mice in the study did not consume more than max. 4g(!) per day) is so low that the 10g package I just saw for 20$ over at the webshop of a major bulk supplier would last you literally forever ...

Unfortunately, this is exactly why I don't believe that LeuResSirt, or whatever other stupid name the final product will be given, is going to work - I mean, come on, you can't tell me that there are not already people out there who get 15-20g of leucine everyday and pop resveratrol in 100x the necessary dose of 8-9mg everyday!? And did they turn into a beast, become fast-food resistant or lose fat magically? What? Yeah... that must be Phil Heath secret, right... how come I did not realize that before? ;-)

Bottom line: Regardless of the probably justified skepticism, I will still keep you posted on whether or not NuSirt knocks out another incredible (in the literal sense) human study like the one on NuFit (see "Testosterone - 12% Drop /W 75g Glucose? Fat Loss - Adzuki, Leucine + B6 or HiMaize & More"). So stay tuned, you all know that no supplement will ever more ergogenic than your daily dose of SuppVersity news!

References:

• Bruckbauer A, Zemel MB. Effects of dairy consumption on SIRT1 and mitochondrial biogenesis in adipocytes and muscle cells. Nutr Metab (Lond). 2011 Dec 20;8:91.
• Bruckbauer A, Zemel MB, Thorpe T, Akula MR, Stuckey AC, Osborne D, Martin EB, Kennel S, Wall JS. Synergistic effects of leucine and resveratrol on insulin sensitivity and fat metabolism in adipocytes and mice. Nutr Metab (Lond). 2012 Aug 22;9(1):77.

High Fish, Soy, Lard & Low Fat Diets: How Do They Affect Body Composition, Lipid and Glucose Metabolism? Results From Study on Rats That Don't Get Obese on High Fat Chow

(Un!)Surprising results: The "healthy" soy oil is the only fat in the study at hand that causes NAFLD.

I know, it is not certain whether or not the results of rodent studies will transfer to human beings and believe me, I would rather have seen this study conducted on humans or at least pigs. And while the latter are simply to expensive, the former are unreliable and don't like to be caged in metabolic wards... well, unless they receive a monetary compensation that costs about as much as a whole pigsty ;-)

That being said, let's stick with what we have and take a look at the results, Yoko Hashimoto and her colleagues published in a recent issue of the medical journal Lipids (Hashimoto. 2013).

The "obesity resistant" wistar rat and it's reaction to different dietary fats

I promise, the results are interesting and probably highly relevant, because the strain of Wistar rats (Slc:Wistar/ST) used in this study does not become obese simply because there is some fat in their chow. In fact, this is exactly the reason why the Japanese researchers picked the Wistar/STs. They wanted to examine the effects of various high-fat diets on plasma and hepatic lipid parameters and lipid metabolism in an obesity "resistant" rodent strain (everyone and every rat can become obese, but these don't get obese just by feeding them fatty chow) who were kept on either standard low fat chow or 3 different high-fat diets for 4 weeks (45% of the energy from fat) .

Figure 1: Fatty acid composition of the different diets (Hashimoto. 2013)

The primary outcomes of the study were the body and tissue weights, total food consumption, fatty acid composition, and energy metabolism, as well as the plasma and liver lipid profiles of the rodents.

Figure 2: Food intake, organ/body weight (Hashimoto. 2013)

If you take a look at the first set of results, namely the energy intake and body/organ weights, and focus on the distribution of white and brown fat you will realize that all rats on the high fat chow were significantly fatter than their low fat fed peers. For the fish oil group, however, the fat gain was almost exclusively from metabolically active brown fat.

Figure 3: Serum an liver lipid profiles (Hashimoto. 2013)

Against that background it is actually not surprising that the "fish oil rats" were the ones with the most favorable blood lipid profile. The rats in the soy group, on the other hand carried the highest amount of white fat and that also shows up in their messed up liver lipids.

The changes in the blood lipid levels the researchers observed went hand in hand with a few, but statistically significant differences in the local expression of important epigenetic regulators of fatty acid oxidation Acat1 / Acox1, in particular in the "fish oil rats":

Suggested read on the effects of fish oil on liver fat: "TTA + Fish Oil - Fat Burning Superfats or Hepatoxic Pro-Oxidants?" (read more)

"Nineteen genes involved in inflammation response genes as well as lipid metabolism-related genes were selected and their mRNA expression levels were measured by qRT-PCR. No significant differences were detected between the groups in the expression levels of genes encoding microsomal triglyceride transfer protein (MTP), ACAT2, 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMG- CoA R, EC 1.1.1.34), and prostaglandin-endoperoxide synthase 1 (PTGS1, COX-1). In contrast, the expression levels of Acat1 and Acox1 mRNAs were the most abundant (p < 0.01) in the livers of the [fish] group." (Hashimoto. 2013)

Interestingly enough, the fish oil group was yet not the only group with metabolically relevant changes in the genetic landscape of the liver:

"Lipogenesis was not significantly increased in the [soy] group [...] Moreover, the level of VLDL secretion in the [soy] group was lower than that in the [lard] group group, because the mRNA expression levels of Apob and Mttp were not up-regulated in the [soy] group. The normal levels of VLDL secretion contributed to hepatic lipid accumulation in the [soy] group"(Hashimoto. 2013)

Overall, the results of the study at hand do not simply highlight the differential effects of various forms of fatty acids on the development of metabolic derangements, they are also testimony to the fact that their effects on the metabolism of obesity-resistant Slc:Wistar/ST rats are much different from those of the obesity-susceptible animals that are usually used in studies like that. The latter become hyperphagic (ravenously hungry => overeating) and acquire hepatic lipid accumulation, almost irrespective of the source of dietary fat. The results of Hashimoto et. al. on the other hand demonstrate that "obesity-resistant Slc:Wistar/ST rats are isocaloric and do not exhibit hepatic lipid accumulation even when consuming high-fat diets, except one that includes soybean oil." (Hashimoto. 2013)

---

These results remind me of another SuppVersity article with the telling title "If You Go 'High Carb', You Better Go Really High! Seven Meals/Day, More than 800g of Carbs, Less Than 50g of Fat & 1000kcal Over Maintenance and Still Lean Gains!" (read more)

Bottom line: For me there are three messages to take away from this study. (1) The Slc:Wistar/ST rat could be a much better model to study the effects of high fat diets on human metabolism, than the regular rodents that are in fact actually selected for their high susceptibility to become obese on diets with 45% vs. just 11% of the energy in form of fat. (2) In a high fat diet scenario fish oil is the most effective way to keep the liver clean; the increase in body fat on the other hand is identical and the higher brown (=metabolically active) vs. white fat accumulation is probably irrelevant for humans. And what's more, if you take into consideration that all animals weighed the same at the end of the study, you just have to do the math to know that the "low fat rodents" were not just leaner, but also more muscular than their peers. (3) Lastly, there is the issue of omega-3 vs. omega-6 fatty acids where the comparison between the effects of lard and soybean oil do not contradict the importance of ratios, but still underline that the ratio interacts with the total PUFA (and omega-6) intake and that nothing compares to high soybean oil, when it comes to ruining your health deliberately.


References:

• Hashimoto Y, Yamada K, Tsushima H, Miyazawa D, Mori M, Nishio K, Ohkubo T, Hibino H, Ohara N, Okuyama H. Three Dissimilar High Fat Diets Differentially Regulate Lipid and Glucose Metabolism in Obesity-Resistant Slc:Wistar/ST Rats. Lipids. 2013 Aug;48(8):803-15

Saturated Fat Kills Gut Bacteria & Modifies Genes in the Distal Small Intestine - Another Reason Why We Get Fat? Plus: Bacteria, Fiber, SCFA, GLP-1 & PYY Revisited

Image 1: Bacteria, there are >100 trillion of them right inside of your digestive track, you can hardly know them all and scientists do neither - the only thing we are beginning to understand, though, is that it may be a good idea to get them to know at least somewhat better ;-)

I guess some of you have already noticed that I was (and probably am now, again) somewhat behind, as far as answering your questions, comments an wise remarks are concerned. Actually it is still more of a coincidence that today's SuppVersity news, which, as you see is not an Adelfo Cerame post (don't forget to keep the fingers crossed for him! This is his weekend!), could actually be interpreted as my somewhat lengthy response to a comment from Vincente on the effects of GLP-1 on chocolate preference in rats and an interesting hypothesis of his, on how this could all relate to my previous post on the fat burning effects of GLP-1 ("Eat More, Burn More and Lose Fat Like on Crack with GLP-1!?"). What, that was Vincente's reasoning, what, if those obese individuals had just messed up their gut bacteria an would lack those beneficial bacteria, which convert the fiber and resistant starch that makes it through your small intestine, right down into your long one to short chain fatty acids?

Does obesity come from within?

I guess by now some of you may already be asking themselves, where all that relates to GLP-1 and eating more, burning more and losing fat like on crack. Well, the missing link if you will is actually not a link, but rather a receptor - the free fatty acid receptor, FFR, which "sniffs" the presence of the short chain fatty acids and triggers the release of GLP-1 and PYY. Those two incretin hormones, of which researchers have found within the past 10 years or so that they are way more than mere "satiety signals. Several research studies in rodents have shown that the anti-obesogenic effects of GLP-1 and PYY are if at all, only partly mediated by reductions in food intake, yet mostly via complex downstream effects on total energy expenditure, glucose and fatty acid oxidation.

Contrary to exogenously administered GLP-1, which is actually being used in the treatment of diabetes an the metabolic syndrome, the in-vivo data from rodent studies, which suggests that high fiber diets protects those little critters from diet induced obesity (Aziz. 2008; Shen. 2008; Zhou. 2008) have, as Robertson et al. pointed out only recently, not yet been confirmed in humans trials (Robertson. 2012). Moreover, the latest results from the Merck Reserach Lab show, contrary to previous evidence from the Cambridge Institute for Medical Research (Tolhorst. 2012), that even our current assumption with respect to the underlying mechanism, could at least be incomplete (Lin. 2012). This does not mean that the short chain fatty acids would not produce the desired increase in GLP-1 nad PYY, but rather that their effects are not solely mediated by  the aforementioned free fatty acid receptor in the gut.

Let's make things even more complicated and bring some long chain fatty acids to the table!

What is yet self-evident though is that the way GLP-1 and PYY modulate energy utilization punches yet another huge hole in the prostrated "calories in vs. calories out hypothesis", one that has little to nothing to o with insulin and one that acquires yet another shade of gray, when we look at the long-chain counterpart of the "bacterial excrements" the dreaded or beloved (depending on the standpoint of the individual) saturated fatty acids (SFA) and a recently published study by scientists from the Wageningen University in the Netherlands (De Wit. 2012), who investigated the long-term effects (8 week, study conducted on mice) of high fat diets with fats from different fat sources

• palm oil - representing the saturated fatty acids,
• olive oil - representing the mono-unsaturated fatty acids, and
• safflower oil - representing the polyunsaturated fatty acids

on body weight gain, liver triglycerides and the whole other standard parameters and their relation changes in the gut microbiome and the amount of fat that "left" the animals undigested.

Figure 1: Fecal fat and energy loss, total energy intake and relative (to control on normal chow) liver triglycerides, oral glucose tolerance and weight gain over the 8 week study period (de Wit. 2012)

A casual look at the data in figure 1 should suffice to see that there is a profound mismatch between almost all classic features of the metabolic syndrome of which we would usually expect that they would be closely associated:

• the rodents in the palm oil group ate the least amount of energy, excreted the greatest amount of fat and total energy in their feces and still gained the greatest amount of body weight and had the highest amount of liver triglycerides (beginning non-alcoholic fatty liver disease)
• the rodents in the olive oil group did not consume significantly more amount of energy or excrete significantly more amount of fat / energy in their feces and still gained ~40% less body weight and did not exhibit similarly high triglyceride storage in the liver as the rodents on the saturate fat (palm oil)
• the rodents in the safflower oil group were comparably ravenous (+20% energy intake), but although they did not excrete more energy and fat than their peers, their bosy weight gain was profoundly reduced and their liver triglycerides were better than in the "non high fat control group" and yet their glucose tolerance was not the best, but the worst of all the three groups

All that does only make sense, when a second parameter, or I should say another 100 trillion bacterial parameters come into play and the SFA induced reduction in microbial diversity and
composition (increased the firmicutes/bacteroidetes ratio) are accounted for, as well. those, this is at least what de Wit et al. believe are namely responsible for the complex changes in genes that regulate the fatty acid metabolism and expression of inflammatory markers, the scientists observed

Chicken or egg, cause of correlation? Or just gut optimization?

Even tde Wit et al. do yet point out that their observations do not provide significant evidence to establish a causal relationship between the bacterial changes, which are a direct result of an overflow of (selectively) antimicrobial saturated fats into the distal part of the intestine, the subsequent disturbances in the bacterial balance and (human!) gene expression in the gut and the  particularly pronounced obesogenic effects of saturated fatty acids.

You could, at least in my humble opinion, even argue that these are simply adaptive effects that ensure that the "host", in this case the rodents, "gets the most" out of his diet - after all, this is exactly what we are seeing here: A modulation of genes related to the conservation and storage of energy, such as the downregulation of the Bcmo 1 gene that predisposes to the development of obesity and non-alcoholic fatty liver disease (Hessel. 2008),  which allows for maximal energy efficiency despite greater fecal energy loss.

Conclusion? Drink safflower oil?

That these results should not be taken as an incentive to guzzle safflower oil (or drop your coconut oil for the latter) should be obvious. Just as obvious, by the way, as the realization that despite all the hoopla and my own excitement about the newly discovered importance of the gut microbiome as one of the possible contributers to the global obesity epidemic. We are understanding way too little about its interactions with its host, i.e. us, to exclude that we are not - yet again - confusing cause and effect, causation and correlation and take our gut microbiome, which is eventually nothing else than a mirror of our healthy or unhealthy lifestyle for the real deal, and try to modulate and fix the mirror image with anti-, pro- or prebiotics without working on what stands right before the mirror: The sedentary, convenience food consumer, who works to jobs and rather watches TV till late at night instead of getting his 7-8h of sleep....

References:

1. Aziz AA, Kenney LS, Goulet B, Abdel-Aal el-S. Dietary starch type affects body weight and glycemic control in freely fed but not energy-restricted obese rats. J Nutr. 2009 Oct;139(10):1881-9. Epub 2009 Aug 19. 
2. Hessel S, Eichinger A, Isken A, Amengual J, Hunzelmann S, Hoeller U, Elste V,  Hunziker W, Goralczyk R, Oberhauser V, von Lintig J, Wyss A. CMO1 deficiency abolishes vitamin A production from beta-carotene and alters lipid metabolism in mice. J Biol Chem. 2007 Nov 16;282(46):33553-61.
3. Lin HV, Frassetto A, Kowalik EJ Jr, Nawrocki AR, Lu MM, Kosinski JR, Hubert JA, Szeto D, Yao X, Forrest G, Marsh DJ. Butyrate and propionate protect against  diet-induced obesity and regulate gut hormones via free fatty acid receptor 3-independent mechanisms. PLoS One. 2012;7(4):e35240.
4. Robertson MD. Dietary-resistant starch and glucose metabolism. Curr Opin Clin Nutr Metab Care. 2012 Jul;15(4):362-7. 
5. Shen L, Keenan MJ, Martin RJ, Tulley RT, Raggio AM, McCutcheon KL, Zhou J. Dietary resistant starch increases hypothalamic POMC expression in rats. Obesity  (Silver Spring). 2009 Jan;17(1):40-5. Epub 2008 Oct 23.
6. Tolhurst G, Heffron H, Lam YS, Parker HE, Habib AM, Diakogiannaki E, Cameron J, Grosse J, Reimann F, Gribble FM. Short-chain fatty acids stimulate glucagon-like peptide-1 secretion via the G-protein-coupled receptor FFAR2. Diabetes. 2012 Feb;61(2):364-71.
7. Zhou J, Martin RJ, Tulley RT, Raggio AM, McCutcheon KL, Shen L, Danna SC, Tripathy S, Hegsted M, Keenan MJ. Dietary resistant starch upregulates total GLP-1 and PYY in a sustained day-long manner through fermentation in rodents. Am J Physiol Endocrinol Metab. 2008 Nov;295(5):E1160-6.
8. de Wit NJ, Derrien M, Bosch-Vermeulen H, Oosterink E, Keshtkar S, Duval C, de Vogel-van den Bosch J, Kleerebezem M, Müller M, van der Meer R. Saturated fat stimulates obesity and hepatic steatosis and affects gut microbiota composition by an enhanced overflow of dietary fat to the distal intestine. Am J Physiol Gastrointest Liver Physiol. 2012 Jun 14.
9. Zhou J, Martin RJ, Tulley RT, Raggio AM, McCutcheon KL, Shen L, Danna SC, Tripathy S, Hegsted M, Keenan MJ. Dietary resistant starch upregulates total GLP-1 and PYY in a sustained day-long manner through fermentation in rodents. Am J Physiol Endocrinol Metab. 2008 Nov;295(5):E1160-6. 

Produce Your Own No-Bullshit-Bulletproof Frying Oil W/ the Right Herbals: Sage & Rosemary Work Best! + Rosmarinic Acid As Potent as Metformin - Anti-Diabetes & Add. Benefits

Rosemary and Garlic Steak - This is only one out of thousand recipes you will find all over the Internet that inform you how to use rosemary and reap (some) of the health benefits (thing about dosage for taste vs. for medical effects) I am about to outline in todoay's SuppVersity article.

I know that I still owe you the 2nd par of the "Perfect Frying Oil" series, and I promise I will write it as soon as I have found convincing evidence to argue that there actually is a "perfect frying oil". In the mean time, I'd suggest you focus on the "tricks" I am about to reveal in today's SuppVersity article and produce your own no-bullshit-bulletproof frying oil by adding... no, not butter and coconut oil, but sage, thyme, and rosemary.

If you belong to the extremely studious 10% of the SuppVersity students, you will probably remember that you can easily produce an anti-bacterial marinade based on green tea, lemon and turmeric (read more). If you put this "no-bullshit-bulletproof" marinade on your meats and fry them in oil you've made "no-bullshit-bulletproof" with sage, thyme, or rosemary that would be truly bulletproof ;-)

You can learn more about potential negative sides of too many / the wrong antioxidants:

NAC = GSH ￪, Anabolism ￬

Too Much "Vit C" For Gains?

Protein requ. of athletes

Block inflamma- tion, choke fire

C + E Get Avg. Joes Ripped

ROS Management Not Eradication

But enough of the sarcasm for today. A recent study from the Faculty of Agriculture at the South Valley University does in fact confirm that adding sage, thyme or rosemary to a commercially (canola based) frying oil can more than double its induction period, i.e. the time it takes until the oil is literally inedible ... and profoundly unhealthy.

Figure 1: Oxidation stability (y-axis) of refined rapeseed oil treated w/ different herbs during frying for 0-32h (Taha. 2014).

As you can see in Figure 1 thyme and rosemary, or "Rosmarinus officinalis", as the science geeks would say, are the most potent of the three additives. With an increase of approx. 63% over that of pure rapeseed oil (the North American SuppVersity readers know "rapeseed" as "Canola oil", which is actually a special, mostly GMO variety of rapeseed oil - of which you can expect that it will react similar - unless it's the "high MUFA" version, in that case I would expect increases but by no means as pronounced increases in oxidative stability).

Accordingly, both, the rosemary and sage enhanced oils retained a significant albeit highly reduced alpha- and gamma-tocopherol content even after 32h of frying, while the control oil and the thyme-enhanced oil lost all its vitamin E after only ~14h and ~29h of frying, respectively.

There's one "on the other hand": You will probably know by now, that there no SuppVersity article with a simplified bottom line that does not at least mention the "other hands", i.e. things you should keep in mind, when reading the bottom line. For the study at hand this is the increase in free fatty acids with frying time. This is another, albeit very unspecific marker of the deterioration of a given oil during the frying process. In view of the superior outcome for both the oxidative stability, as well as the reduced depletion of tocopherols (vitamins E) I will still whole-heartedly conclude that adding sage and especially rosemary to your oils is a good idea.

Although both will work, there are a couple of things that speak in favor of rosemary, which are not directly related to the results Taha et al. (2014) present in their latest study. If you look at the results of a recent study from the University of Madras, for example you will see that rosmarinic acid, one of the major active ingredients in rosemary will also have protective effect on your glucose and lipid metabolism (Jayanthy. 2014).

In said study, the provision of 100mg/kg of rosmarinic acid (human equivalent: 8mg/kg) reverted almost all of the detrimental changes in liver and glucose metabolism in a rodent model of diet-induced diabetes. And as the authors point out, the obtained results were almost comprable to those of the anti-diabetes drug, metformin (see Figure 2).

Rosemarinic acid is on par with metformin!

Noot bad, given the fact that the same compound from the fragrant, evergreen, needle-like leaves and white, pink, purple, or blue flowers of the perennial shrub from the Mediterranean has a whole host of additional benefits. It...

Figure 2: Effect of Rosmarinic acid (100mg/kg | human equivalent: 16mg/kg) and metformin (200mg/kg | human equ.: 32mg/kg) on the insulin sensitivity in a rodent model of experimental diabetes (Jayanthy. 2014)

• inhibits seasonal allergic rhinoconjunctivitis in humans (Takano. 2004) and the immune over-reaction to mite allergens (Sanbongi. 2004).
• has potent (in-vitro) anti-Parkinson's effects (Wang. 2012)
• ameliorates diabetic nephropathy in a rodent model of diabetes (Tavafi. 2011).
• appears to help with arthritis and rheumatoid joint degeneration (Youn. 2003; Khanna. 2007)
• induces apoptosis (cell death) in various cancers, including human colorectal cells (Xavier. 2009)
• has anti-hypertensive and pro-metabolic (glucose + fat metabolism) effects in rodents on high fructose diets (Karthik. 2001)

And that's only the tip of an iceberg, which involves potential beneficial effects on all types of auto-immune diseases and a whole host of other metabolic benefits. In spite of the fact that the rosmarinic acid content of regular rosemary is limited, rosemary is thus one of the herbs you should consider adding to your kitchen cabinet... and frying oils ;-)

Table 1: Total polyphenol (gallic acid equ.) and ORAC (trolox equ.) values of common cullinary herbs (Zheng 2001).

Bottom line: It's funny how many of the common herbs humans all across the world have been using for centuries for culinary reasons turn out to have profound health effects. Rendering frying oils bulletproof with sage, thyme and obviously rosemary is after all only one of the many examples. Making your meats bacteria resistant with green tea, lemon and turmeric is another one, and in the end, even the citric acid + vitamin C people have drizzling on their fish filet for centuries will have beneficial health effects. Fascinating, isn't it? And certainly healthier than butter in a pot of hot coffee ;-)

How to you do it: If you want to do it exactly the way the scientists did it, add 50g of plant material (best freshly grounded!) to 1l of the oil of your choice, stir it for 24h and vacuumfilter the particles afterwards. Then you put it in the fridge and store it there for ~1 week. If you can't filter the oil, use less oil and prepare a fresh concoction every week (e.g. 20g + 400ml oil).

Reference:

• Jayanthy, G., and S. Subramanian. "Rosmarinic acid, a polyphenol, ameliorates hyperglycemia by regulating the key enzymes of carbohydrate metabolism in high fat diet–STZ induced experimental diabetes mellitus." Biomedicine & Preventive Nutrition (2014).
• Khanna, Dinesh, et al. "Natural products as a gold mine for arthritis treatment." Current Opinion in Pharmacology 7.3 (2007): 344-351.
• Sanbongi, C., et al. "Rosmarinic acid in perilla extract inhibits allergic inflammation induced by mite allergen, in a mouse model." Clinical & Experimental Allergy 34.6 (2004): 971-977.
• Taha, Eman, et al. "Stabilization of refined rapeseed oil during deep‐fat frying by selected herbs." European Journal of Lipid Science and Technology (2014).
• Takano, Hirohisa, et al. "Extract of Perilla frutescens enriched for rosmarinic acid, a polyphenolic phytochemical, inhibits seasonal allergic rhinoconjunctivitis in humans." Experimental Biology and Medicine 229.3 (2004): 247-254.
• Tavafi, Majid, et al. "Rosmarinic Acid Ameliorates Diabetic Nephropathy in Uninephrectomized Diabetic Rats." Iranian Journal of Basic Medical Sciences 14.3 (2011).
• Wang, Jieyu, et al. "Neurorescue effect of rosmarinic acid on 6-hydroxydopamine-lesioned nigral dopamine neurons in rat model of Parkinson's disease." Journal of molecular Neuroscience 47.1 (2012): 113-119.
• Youn, Jeehee, et al. "Beneficial effects of rosmarinic acid on suppression of collagen induced arthritis." The Journal of rheumatology 30.6 (2003): 1203-1207.
• Xavier, Cristina PR, et al. "Salvia fruticosa, Salvia officinalis, and rosmarinic acid induce apoptosis and inhibit proliferation of human colorectal cell lines: the role in MAPK/ERK pathway." Nutrition and cancer 61.4 (2009): 564-571.
• Zheng, Wei, and Shiow Y. Wang. "Antioxidant activity and phenolic compounds in selected herbs." Journal of Agricultural and Food chemistry 49.11 (2001): 5165-5170.

Science Round-Up Seconds: Breast Cancer, GH Induced Insulin Resistance, Stretch + Contraction Increase Molecular Hypertrophy Signals and Probiotics & the Obesity Pandemic

When we are talking about the extinction of endangered species such as the Siberian tiger, we are adopting a perspective that will also help us to understand such things as the pitfalls of probiotic supplementations as a "solution" to the diabesity epidemic

It's Friday the day after the SuppVersity Science Round-Up (download the podcast) and you all know what that means: Right! Time to summarize the stuff that did not make it into the show and provide you with a couple of thoughts, as well as additional information on the topics, Carl Lanore and I did already cover (this does also imply that you have to listen to the show if you want the info on the genetic and non-genetic underpinnings of breast cancer).

If I had to come up with a motto, something that connects the topics in the show and thus obviously also the ones that will be discussed in this article, I guess it would be "ecosystems". I have repeatedly pointed out in the past that I am not a believer in the "back in the good old days everything was better" interpretation of "Paleo". What I do believe in, however, is the notion that exposition entails adaptation.

You can't control the adaptation, but you can control the exposition

Now, these adaptation processes whether they be negative such as the growth of cancerous tissue in the breast or ovaries of a woman like Angelina Jolie are beyond our reach: The environment that triggers them, on the other hand, can be manipulated - whether that's by getting your breasts and ovaries removed to rid yourself of the 50-80% (you heard me right! the "real" chance carriers of a defect BRCA1/2 gene to develop cancer ranges from 50% to 80%, by the age of 70) or - and this is obviously a pretty drastic change of subjects, by emphasizing peak contractions in a semi-stretched position to benefit from the upregulation of the proteins p-Akt, p70S6K, p38 MAPK and ERK 1/2, i.e. the driving forces of protein synthesis, it's always your manipulation of the environment that will bring about "adaptation" (used in the broadest sense).

Figure 1: Graphical summary of the main results, i.e. the stretch initiated activation (+, ++, +++) of the signaling molecules on different conditions with either short, long (at maximal peak contraction) or slightly longer (+25%, but still way from lockout) positions and the corresponding tension on the soleus muscle of the rodents (Van Dyke. 2013)

If I was successful, the above introduction should have set the scene and thus provided an environment for your perspective on things to change / align with mine. I've manipulated the "cognitive environment" that determines or at least influences the way you are about to understand what follows, i.e. the missing side-kick on the recently observed negative effects of growth hormone on insulin sensitivity and glucose uptake in healthy men and the comprehensive discussion of the (imho) misunderstood role of the gut microbiome as both, a trigger and solution to the obesity epidemic.

Can exogenous growth hormone trigger insulin resistance?

Within our new cognitive framework you will probably have transformed the above question as follows, by now: "How does the exogenous administration of growth hormone change the the endocrine environment and why does this entail an adaptive response of which most people would say that it's highly detrimental?" From this "mini-evolutionary perspective", as you may call it, it's surprisingly easy to understand what exactly has happened in a recent study from the University of Aarhus in Denmark (Vestergaard. 2013).

In their effort to probe the hypothesis that there was a connection between ghrelin, growth hormone and the increases in retinol-binding hormone (RBP4, to be specific) that are observed in patients with developing and/or full-blown metabolic syndrome, Vestergaard et al. conducted two studies of which only the second one, which involved nine totally healthy young men in their twenties (23y; BMI 23kg/m²), will - without major qualifications - transfer to you me or other healthy physical culturists.

The men received a course of daily 2mg GH injections while consuming a previously standardized diet containing 50–60 % carbohydrates, <30 % fat, and roughly 10–15 % of protein (in % of total energy intake). Before and after the intervention period, the participants had to report to the lab at fasted and thus optimally prepared for the hyperinsulinemic euglycemic clamp test.

Figure 2: Basal fasting glucose and insulin concentrations, as well as M-value (=marker of insulin resistance) in healthy young men after eight days of placebo and GH administration, respectively (left; p-values over bars); glucose infusion rates (GIR) indicated by during hyperinsulinemia after placebo / GH administration (right, Vestergaard. 2013)

The selected results I plotted in figure 2 are quite unambiguous. As expected there was a huge increase in the IGF-I concentrations in response to the 8-day course of 2mg GH per day.

The 8% increase in fasting plasma glucose and even more so, the doubling of the insulin levels that were required to keep these elevated glucose levels stable, certainly come as a surprise, if you're thinking about it from an broscientifically influenced mechanistic point of view. I mean, doesn't broscience tell us that GH is the good guy that's going to make you lean and ripped?

How come we are seeing a -34% reduction in glucose sensitivity (this is actually what's measured as the so-called "M value" in the euglycemic clamp studies).

Within our previously established cognitive framework, the answer to this question is actually quite straight forward. To get to the bottom of the counter-intuitive effects of GH we just have to think about the "natural" environment that will trigger the release of GH from the somatotropic cells within the lateral wings of the anterior pituitary gland.

Did you know that the normal GH response to hypoglycemia is blunted in the obese and the reduced obese (Ball. 1972)? Without GH to get their blood sugar back up, overweight individuals and formerly obese will thus have to resort exclusively to corticosteroids (cortisol) to regulate their blood sugar levels.

In this context, it's also worth mentioning that insulin induced hypoglycemia is still considered a valid - yet maybe not optimal - in children who are suffering from retarded growth. With the injection of exogenous insulin and the kids becoming hypoglycemic the body should compensate for that by secreting growth hormone. If that's not the case, this is a good indicator of a general malfunction of the pituitary gland.

Hah? Right! Hypoglycemia or borderline hypoglycemia is among the primary triggers of somatotropin aka GH release. Now, think about it - would it make sense that a hormone that's supposed to increase fatty acid oxidation to supply your body with glucose would at the same time increase insulin sensitivity and thus have your muscles suck up all the precious glucose your brain is longing for?

On the contrary, it is only logical that GH - just like it's falsely vilified potent cousin cortisol (yeah, you heard me right, you won't see both at the scene on the same time, because they serve a similar purpose at least wrt to glucose metabolism) - will decrease the insulin sensitivity of your muscles in order not to "waste" the precious glucose which should be scarce once GH goes up.

In the study at hand, the environment in which the high growth hormone levels occur are totally different: Once you plug people to a euglycemic clamp, there is a mismatch between the actual environment, which is normal / high glucose, and the natural (if you will "evolutionary) expected environment for elevated GH levels - and what you see in figure 2 is the inevitable consequence of this mismatch: a significantly lowered glucose sensitivity... quite cool, how a small change in the way we are looking at things allows us to understand phenomena that do initially appear totally inexplicable, right?

Lifestlye changes, not probiotic supplements are the solution to the diabesity epidemic

Now that we have first-hand evidence for the explanatory power of our new perspective let's stick to it and apply it to the false expectations studies such as the recently released investigation into the "anti-obesity" effects of the common gut bug Akkermansia muciniphila in a rodent model of diet-induced obesity (Everard. 2013).

Figure 3: The AM count is reduced in response to the dietary enviroment, the environment - not the lack of bacteria - is the root cause of the negative effects on fasting glucose, fat mass gain and thickness of the intestinal mucus lining all of which can be partly (the latter even fully) restored to normal with supplementation (Everard. 2013)

If we take a look at the data in figure 3 it is undebatable that the provision of exogenous live (this does not work at all with dead bacteria) Akkermansia muciniphila (AM) can significantly reduce the weight and fat gain in the rodents receiving a species inappropriate high fat diet. As the data on the AM content of the gut microbiome of the obese mice on the right hand side of figure 3 reveals this effect is yet a simple result of the restoration of the "original" microbial composition in the intestinal tract of the rodents.

Siberian tigers on corn fields!? Does this really make sense?

Since the term "original" refers only to the amount of the bacteria Dr Antoon Akkermans identified as one of the many bacteria the count of which is decreased / changed in obese rodents (and humans), it is yet not surprising that we are seeing nothing but ameliorative effects.

Glutamine may offer another way to accelerate gut healing and improve amino acid absorption or rather avoid the "abuse" of BCAAs, arginine etc. for other metabolic purposes (learn more)

Within our cognitive framework of "ecosystems" and "adaptations", the chronic administration of living bacteria to counter a specific aspect of the diet induced detrimental effects on the gut microbiome has may be compared to the laughable efforts of biologists to save certain animals from extinction by breeding them in a Zoo and releasing them into an environment, where deforestation, pollution and all the other nasty things, we, the crown of evolution, enjoy about as much as the average Westerner likes to wash down his super-sized fast food menu with a "refreshing" *rofl* 2L XXL pot of coke (drinking plenty is healthy for you, ain't it? ;-), will have them die before their time unable to reproduce in time to contribute to the species' natural survival.

Once you understand that, you will have to realize that you are wasting your time and money on false promises if you don't change the environment, i.e. the way you live and the foods you eat first before you even think of using probiotic supplements as an adjunct to accelerate the normalization process, which should - just like the detoriation - take place irrespective of the supplementation as a mere results of the dietary modulation of your gut microbiome, anyways.

With these insights, I am going to release you into the weekend... and who knows, maybe you'll notice the effects the environment you expose yourselves to in the coming days has all sorts of beneficial or detrimental influences on your physical and mental health. Listening to even more non-science-based Angelina Jolie news on the television and radio, for example, is probably not going to have beneficial effects on your psyche ;-)

References:

• Ball MF, el-Khodary AZ, Canary JJ. Growth hormone response in the thinned obese. J Clin Endocrinol Metab. 1972 Mar;34(3):498-511.
• Everard A, Belzer C, Geurts L, Ouwerkerk JP, Druart C, Bindels LB, Guiot Y, Derrien M, Muccioli GG, Delzenne NM, de Vos WM, Cani PD. Cross-talk between Akkermansia muciniphila and intestinal epithelium controls diet-induced obesity. Proc Natl Acad Sci U S A. 2013 May 13.
• Van Dyke JM, Bain JL, Riley DA. Stretch activated signaling is modulated by stretch magnitude and contraction. Muscle Nerve. 2013 Apr 26.
• Vestergaard ET, Krag MB, Poulsen MM, Pedersen SB, Moller N, Jørgensen JOL, Jessen N. Ghrelin and growth hormone induced insulin resistance: no association with retinol-binding protein-4 Endocr Connect EC-13-0019; published ahead of print May 7, 2013, doi:10.1530/EC-13-0019

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.

Lactobacillus vs. Oscillibacter! Does (Saturated) Fat Tip The Scale Towards Leaky Gut, Obesity & Visceral Inflammation?

Image 1: Location of the different fat depots of the human body (sorry Evilyn, had to borrow this from your Carbsane blog ;-)

Having tons of subcutaneous fat is certainly unaesthetic, but as science would have it, probably more healthy than a mediocre amount of superfluous visceral fat. But why is that? I mean, what makes the difference? It cannot be the location, can it? Well, a recently published study on metabolic dysfunction in diet induced diabetic mice suggests that it could be as simple as that (Lam. 2012).

You cannot spot reduce fat, but can you "spot inflame" it?

In order to assess the effects of normal vs. high fat (60%) diets on gut Permeability and microbiota the Yan Y Lam and his (or her?) colleagues from the University of Sidney kept a group of 16-week old Female C57BL/6J mice (those are the "normal" lab mice used in these experiments) on a regular chow diet (control; 10% fat) or a high saturated fat (34%) high fat diet (HFD; 60% of total energy from fat). After 12 weeks, the rodents in the HFD group were obese and diabetic, had elevated insulin and amyloid A3 serum levels and reduced adiponectin.

What are amyloid A3 and adiponectin? Not to long ago, I would probably have had to explain what insulin is, but in these days of carbophobia, I better stick to the aliens, amyloid A3 and adiponectin. While the former is an acute phase protein elevations of which are indicative of adipocyte (=fat cell) inflammation and have also been observed in Creutzfeldt Jacob and Alzheimer patients (Baker. 2003), adiponectin is a negative control factor for body fat and energy expenditure, with increasing adiponectin expression from adipose tissue being linked to increases in fatty acid oxidation, improvements in glucose metabolism and reduced triglyceride storage within existing body fat stores.

While these are results we have seen in countless of studies before, the interesting part of the study began with the collection of stool samples in the last three days and the "termination" of the animals at the end of the 12-week study period.

Figure 1: Gut microbiota composition in control and high saturated fat diet (HFD) fed mice after 12-weeks (Lam. 2012).

As you can see, there was a telling association between body weight and the Lactobacilli (fig. 1, middle) and Oscillobacter (fig. 1, right) content of the stool samples; with p-values (chance that this is coincidence) way below the significance criterion, i.e. p < 0.05 = 5%:

At the phylum level, HFD mice had more Firmicutes (73 6 1.5% vs 68 6 2.3% for controls; P = 0.041) and fewer Bacteroidetes (156 1.7% vs 196 1.3% for controls; P = 0.026) and thus a significantly higher Firmicutes: Bacteroidetes ratio ( P = 0.019). HFD also induced significant shifts in fecal microbiota composition at the genus level of taxonomic resolution. Within members of the Firmicutes, notably there was a 75% decrease in Lactobacillus ( P ,0.001) and a 279% increase in Oscillibacter( P = 0.004) as compared to controls and these changes in abundance were closely associated with weight gain.

These gut microbiotal changes, in turn, were closely associated with differences in gut permeability, as measured according to Wang et al. 2001 (Wang. 2001), showing positive correlations (r = 0.52; P = 0.013) with the abundance of Lactobacillus and negative correlations with Oscillibacter (r = - 0.55;P = 0.007) in the proximal colon.

Notably, increased Oscillibacter abundance was also associated with a reduction in the mRNA expression of ZO-1 (r = - 0.37; P = 0.039) and, although not statistically significant, a similar trend was observed with proglucagon (r = - 0.34; P = 0.061).

While the reduced expression of the macromolcule ZO-1 is symptomatic, maybe even causative for the "leakiness" of the gut (Assimakopoulus. 2011), the reduction in proglucagon, a pro-peptide to GLP-1 the fat-burning effects of which I have discussed in a previous blogpost (cf. "Eat more, burn more and lose fat like on crack"), could partly explain the diet induced hyperphagia (unsatiable hunger) and pronounced weight gain, which is yet perpetuated by what could eventually turn out to be a life-threatening down-stream effect of the leaky gut: Inflammation in the adjacent visceral adipose tissue:

Figure 2: Macrophage infiltration (per 100 adipocytes) and adipocyte area in the different fat depots of the mice on normal (control) and high fat diet (HFD) at the end of the 12-week study period (data adapted from Lam. 2012).

As the data in figure 2 shows, the 12 weeks on the high fat diet and the subsequent changes in the gut microbiom and permeability lead to an increased macrophage infiltration in mesenteric (58%; P = 0.020) and epididymal (71%; P = 0.006) fat, but was without effect in perirenal and subcutaneous fat. This interesting as neither the peri-renal = next to the kidneys, nor the subcutaneous fat (obviously right beyond the skin) is in close proximity to the (now leaky) gut.

Causal, corollary, chicken or egg?

Image 2: Until we know it's not the saturated fat that tips the scale in favor of Oscillibater you better have Sauerkraut with your Weißwurst ;-)

There are still way to o many uncertainties to state that a "leaky gut", let alone a high fat diet induced increase in Oscillobacter and decrease in Lactobacillus is the root of visceral obesity, or even the obesity epidemic. It is however quite telling how nicely these pieces of the puzzle are falling in place and that initial studies on the effects of lactobacillus supplementation on the onset of diet-induced obesity in HFD fed rodents (e.g. Takemura. 2010) suggest that there may be merit to rebalancing diet induced changes in the microbiota composition of the gut. In fact, so compelling that it will be hard to argue with the researchers' conclusion that "the gut is a central player in the aetiology of diet-induced metabolic diseases".

So happy to be a "Kraut" ;-)

Whether it is a or even the central player and whether the saturated fat content and not the lack of other nutrients cause the undesirable shift in the microbiota composition of the gut still has to be determined. I guess, in the meantime you better make sure to have Bavarian Weißwurst with Sauerkraut ;-)

Update: If you want to know more about the correlation between different food-types and nutrients and the gut microbiom, check out my previous post on the "Gut Type Diet".