Natural Sildenafil & Testosterone Alternatives: Pedalium Murex & Paederia Foetida +150% Testosterone and +200% Erectile Function. Plus: Icariin, Aromatase & Stronger Bones. Probiotics, Phytosterols & Thyroid Activity.

If the Aliens have seen these Bugarash apocalypse pilgrims, they have probably turned tail and fled ;-)

I guess the SuppVersity figure of the week was a date "12/21/2012", I mean this is a once in a life-time event! Or do you really believe there will be another chance for mankind to be snatched from the jaws of extinction?

Ok, enough of the sarcasm, the world is still there and the SuppVersity is still operating, so let's get to the not so short On Short Notice items for today.

If there had been more room in the headline I would probably have labeled it "Endocrine Special" and I guess once you have gone through the posts, you will agree that this would certainly have made sense.

• Traditional treatment of osteopenia with Epimedium brevicornum works, 'cause it's a potent aromatase promoter (Yang. 2012) -- When a group of researchers from the Chengdu Institute of Biology, Chinese Academy of Sciences and the School of Chinese Pharmacy, Chengdu University of Traditional Chinese Medicine in Chengdu, China, set out to investigate the mechanism behind the anti-osteoporotic effects of the dried leave extract from Epimedium brevicornum, they already suspected that it would probably be related to some sort of endocrine modulation.
  
  

  

  Figure 1: In vitro effect of Forskolin and Icariin on aromatase expression in KGN cells (Yang. 2012)

  With estrogen deficiency being the major cause of osteoporosis, a disease which does by the way affect over 200 million people worldwide (Riggs. 1998), being their the most likely mechanism behind the bone building and bone mineral density (BMD) protective effects, it was obvious to study, whether there was some sort of interaction between the extract and the data in figure 1 shows that Yang et al. were right to do so (note: the cell line used in the study was a KGN granulosa cell that's closely associated with the developing female oocyte, so effects could vary from one tissue to the other)
  
  Now all that certainly sounds as if this was bad stuff no sane male human being should be taking. Luckily, for many of you, my fellow men, who may have been exposed to Epimidium / Icariin from various supplements, this is not the case. Why? Simply because estrogen is not the enemy and Epimidium has not only been found to have anti-aging effects (whole extracts; Yan. 2009), it also prevents neurotoxicity from beta-amyloid plague the key feature of Alzheimer's and similar diseases (Zeng. 2010; listen to Thursday's Science Round-Up for other amyloid beta inhibitors / protectant's) and the neurotoxic effects of excess corticosteroids (Liu. 2011). In addition, Epimidium pubescen flavenoids have been shown to reverse the negative effects even passive exposure to cigarette smoke may have on the bone mineral density of male rats (Gao. 2012) and Icariin the common denominator in all of the members of the Epimidium family exerts direct proliferative and thus pro-fertility effects on the sertoli cells in male rodents (Nan. 2012).
  
  

  

  Table 1: Analysis of Chinese Epimidium species (MDida. 2010)

  Ah, and last but not least the beneficial effect Horny Goat Weed (likewise one of the members of the Epimidium family) has on male libido are likely partly a result of the increased aromatase activity, as well. If it works for you, your estrogen levels are probably pretty low to begin with (this reasoning is based on the profound beneficial effects of estrogen on male libido in men expressing little to no aromatase enzyme; Carani. 1999).

• Pedalium murex Linn. fruits may not be as potent acute libido and erectile performance enhancers as sildenafil, but the effects accumulate and persist just as its unique testosterone boosting effects (Sharma. 2012) -- In case taking a potential proestrogenic compound like Horny Goat is nothing you feel would do anything good for your libido, you may want to use some Pedalium murex Linn. in traditional Indian medicinea herb that has been used in traditional Indian medicine for centuries to treat male sexual dysfunction and impotency.
  

  

  Figure 2: Effects of different doses of an ethanolic extract from Pedalium murex (P.m.) and Sildenafil citrate (5mgkg) on penile erection scores and post ejaculatory interval (time it takes to be able to have sex again) exrpessed relative to saline treated control (based on Sharma. 2012)

  As the data in figure 2 shows, its effects on erectile performance are slightly less pronounced than those of sildenafil citrate, but at an appropriate dosage and after some time, it will probably do its job sufficiently for most men with respective problems. What's really exiting though, is that the data in figure 3 clearly shows that it does so not by simple nitric oxide effects (P. murex at 10 mg/
  ml exhibited a relative nitric oxide release of 15.3 mM as compared to a nitric oxide release of 39.3472.7 mM with sildenafil citrate), but probably (also) by boosting testosterone levels. 
  
  

  

  Figure 3: Serum testosterone levels during and after Pedalium murex or Sildenafil citrate administration expressed relative to saline control (Sharma. 2012)

  Since the testosterone levels remained elevated for 14 days after the treatment was seized these observations make P. murex a potential candidate for the next best natty test booster.to be powered by advertisement lines like "up to 125% increase in testosterone that last for more than 2 weeks" even when you take some time off. I guess, the only serious downside here is the close relation of P. murex to the notoriously useless Tribulus, which does also contain furostanol glycosides. Luckily Magnle and Jolley found another potentially active ingredient in P. murex fruit. The compound goes by the name diosgenin and appears to be a direct precursor for the synthesis of sex hormones including testosterone (Mangle. 1998).
  
  Since I don't think you will have to wait very long until the first major players in the supplement business will jump on this bandwagon - you know that a single rodent study is enough to make fortune until people realize that this stuff does not work, at all - you will probably soon be able to test it on yourself. In case you intend to do that, you should certainly look for a standardized ethanolic extract with ~20%+ of diosgenin (that was the content of the extract Sharma et al. used) and a serving size of 16.2mg/kg per body weight, the HED of the 100mg/kg group (that would be 1,300mg /day of the extract for an 80kg adult).

• Paederia foetida Linn. (P. foetida) yet another Indian libido and testosterone boosting herb (Soni. 2012) -- If you like the appeal of exclusiveness a climbing plant found in the Central and Eastern Himalayas, at elevations of up to 5000 ft, you may prefer the an ethanol extract of Paederia foetida Linn. (P. foetida) leaves over the previously discussed Pedalium murex fruit extract.
  
  

  

  Figure 4: Penile erection index and testosterone levels of rats after 15 and 28 days on P. foetida, data expressed relative to saline control (Soni. 2012)

  According to a recent study from the same group of Indian scientists (yet a different lead author), the effects of P. foetida and P. murex are in fact virtually identical. While the general protocol was very similar (in this done on rat, not mice) the positive control was not Sildenafil citrate, as in the previously mentioned experiment, but an intramuscular injection of 0.5 mg/kg body weight of testosterone suspension in arachis oil twice a week. It's actually a pity the scientists didn't measure the actual testosterone levels in the animals in the testosterone group. I would be curious how the 2.5x increase in the high dose group (200mg/kg, human equivalent ~2,600mg/day) would compare. When it comes to the erection quality, it did at least easily top the results of injectable testosterone (see figure 4), which is yet allegedly not the best erection booster, anyway. And if we went by the data on body weight and the weight of the testes, seminal vesicles, prostate glands and epididymis, it even appears to be more anabolic than testosterone; after all the body weight gain in the PF groups were 15%. 23% and 34% higher than in the control group and comparable if not higher to those that were induced by testosterone (+29%).
  
  And for all the supplement producers and fans of stacks it may be important to know that the leaves of P. foetida, which is also used as carminative, antiinflammatory, astringent, spasmolytic, antidiarrhoeal, diuretic and antilithic do contain a whole host of well- and lesser known compounds, e.g. iridoid glycosides, sitosterol, stigmasterol, campesterol, ursolic acid, hentriacontane, hentriacontanol, ceryl alcohol, palmitic acid and methyl mercaptan, but no diosgenin. So Ayurvedabol (TM) or whatever name ending on -bol, -drol or -ripp-off has not been used by the creepy competition, already, could benefit from having both Paederia foetida and Pedalium murex in it ;-)

• Combination of probiotics and phytosterols ramps up thyroid function and (re-)establishes a healthy lipid profile (Awaisheh. 2012) -- While I could offer you a minimum of two additional libido enhancing, potentially pro-anabolic testosterone booster, I know that not all of you are into these products (am I right ladies?), so I thought it prudent to close this installment of On Short Notice with a study on thyroid metabolism, which could be is of interested for all of you.
  
  
  Which phytosterols are there and where can I find them? The major phytosterols in the human diet are sitosterol (high in nuts, amaranth, avocados, grape leaves), stigmasterol (high in coriander, chocolate and soy), campesterol (high in various vegetable oils, spec. conola / rapeseed) and brassicasterol (high in cabbage, broccoli, other brassica, but also coriander, seafood and rapeseed oil).
  Contrary to some previous studies and anecdotal reports on the Internet, a recent paper by scientists from the Department of Food Science at the Al-Balqa Applied University, in Salt, Jordan, suggest that phytosterols don't have thyroid inhibiting, but promoting effects.
  
  In their latest study Awaisheh et al. investigated the effects of a probiotic stack containing two strains of each of Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus gasseri, and Lactobacillus reuteri and a phytosterol supplement on the lipid metabolism in rodents. Their results show that the addition of the phytosterol supplement promoted the already pronounced effects the probiotics had on the hypercholesterolemia of the rodents who were fed with a high-fat-high-cholesterol basal diet for 8 weeks. In addition, the phytosterols, yet not the probiotics alone, elevated the levels of serum total thyroxine (TT4), total triiodothyronine (TT3), and free triiodothyronin (fT3), which could have exerted additional benefits on the non-measured triglyceride levels.
  
  These results do actually come as a surprise and I would like to see more research - specifically in human trials - before I would buy products from producers who seize the scientists suggestion that this renders probiotics and phytosterols potential candidates for "functional foods" - I mean, let's be honest ever since the first "functional foods" hit the market people have been getting sicker: Do you really believe that's because they don't eat the "good" cholesterol lowering margarine instead of their beloved Kerrygold butter? I don't think so.

That's it for today.. well, aside from the promise to include the other testosterone and libido boosting herb studies in another SuppVersity post and a selection of the latest  SuppVersity Facebook News, of course:

• Iron deficiency makes H Pylori go on a rampage - Iron deficiency enhances H. pylori virulence and increases risk of gastric cancer (read more)
• Danish folk medicine for depression? Not really, but there are a couple of promising natural MAO-A inhibitors the Danes have been using for centuries (read more)
• Strength training equally heart healthy as aerobics - 6-weeks of strength training show particular beneficial effects in African American men (read more)

Since I it's still pretty early and I am certainly not going to the city, before the shops close and all the people who believed they wouldn't need Christmas presents this year, since the world would *put whatever apocalyptic catastrophe you like here*, have gone home, I may be adding some more news later. Until then, I hope you have some fun with what's already there and are looking forward for the 2nd installment of the "Making HIIT a Hit" series that will be published tomorrow.

References:

• Awaisheh SS, Khalifeh MS, Al-Ruwaili MA, Khalil OM, Al-Ameri OH, Al-Groom R. Effect of supplementation of probiotics and phytosterols alone or in combination on serum and hepatic lipid profiles and thyroid hormones of hypercholesterolemic rats. J Dairy Sci. 2012 Nov 22.
• Carani C, Rochira V, Faustini-Fustini M, Balestrieri A, Granata AR. Role of oestrogen in male sexual behaviour: insights from the natural model of aromatase deficiency. Clin Endocrinol (Oxf). 1999 Oct;51(4):517-24.
• Gao SG, Liu H, Li KH, Liu WH, Xu M, Jiang W, Wei LC, Zhang FJ, Tian J, Xiao WF, Yang Y, Song Y, Lei GH. Effect of Epimedium pubescen flavonoid on bone mineral density and biomechanical properties of femoral distal end and femoral diaphysis of passively smoking male rats. J Orthop Sci. 2012 May;17(3):281-8.
• Liu B, Zhang H, Xu C, Yang G, Tao J, Huang J, Wu J, Duan X, Cao Y, Dong J. Neuroprotective effects of icariin on corticosterone-induced apoptosis in primary cultured rat hippocampal neurons. Brain Res. 2011 Feb 23;1375:59-67. 
• Mangle MS, Jolley CI. HPTLC studies on Tribulus terrestris (Chota ghokru) and Pedalium murex (Bada ghokru). Indian Drugs. 1998; 35:189–194.
• MDidea Extracts Professional. Horny Goat Weed or Epimedium Herb: Botanical Origin, Archeology, Traditional and Pharmacological findings of Epimedium species, fractions and isolated components. 08th, Oct. 2010. < http://www.mdidea.net/products/herbextract/icariin/data10.html > retrieved on 12/21/2012. 
• Nan Y, Zhang X, Yang G, Xie J, Lu Z, Wang W, Ni X, Cao X, Ma J, Wang Z. Icariin stimulates the proliferation of rat Sertoli cells in an ERK1/2-dependent manner in vitro. Andrologia. 2012 Nov 7.
• Riggs BL, Khosla S, Melton LJ 3rd. A unitary model for involutional osteoporosis: estrogen deficiency causes both type I and type II osteoporosis in postmenopausal women and contributes to bone loss in aging men. J Bone Miner Res. 1998 May;13(5):763-73. 
• Sharma V, Thakur M, Dixit VK. A comparative study of ethanolic extracts of Pedalium murex Linn. fruits and sildenafil citrate on sexual behaviors and serum testosterone level in male rats during and after treatment. J Ethnopharmacol. 2012 Aug 30;143(1):201-6.
• Soni DK, Sharma V, Chauhan NS, Dixit VK. Effect of ethanolic extract of Paederia foetida Linn. leaves on sexual behavior and spermatogenesis in male rats
• Yan S, Wu B, Lin Z, Jin H, Huang J, Yang Y, Zhang X, Shen Z, Zhang W. Metabonomic characterization of aging and investigation on the anti-aging effects of total flavones of Epimedium. Mol Biosyst. 2009 Oct;5(10):1204-13.
• Zeng KW, Ko H, Yang HO, Wang XM. Icariin attenuates β-amyloid-induced neurotoxicity by inhibition of tau protein hyperphosphorylation in PC12 cells. Neuropharmacology. 2010 Nov;59(6):542-50.

Weight Loss Supplements Exposed: Green Tea & Probiotics. Fat Loss, Energy Expenditure, Fat Oxidation, Sex & More

Yesterday at Starbucks: "I just ordered a bottle of probiotics!"

In view of the fact that all the feedback I got in response to the re-installment of the Short News was positive, I guess you won't mind if I use the chance and bundle the two soon-to-be-published weight loss studies from the British Journal of Nutrition into a Weight Loss Supplement Mini-Special of the SuppVersity Short News.

If you were actually sitting next to you, I would probably ask you, whether you'd prefer the good, or the bad news, first!? Well, I guess I'll start with the bad one, then: Green tea sucked - again!

ZERO effect of EGCG supplementation in overweight women

To examine the effects of green tea epigallocatechin-3-gallate (EGCG) on the changes in body composition (! not just weight), energy and substrate metabolism, cardiometabolic risk factors and liver function enzymes after an energy-restricted diet intervention in obese women, a group of researchers from the University of the Basque Country in Spain recruited a group of 83(!) obese (BMI 30-40 kg/m2) pre-menopausal women (Mielgo-Ayuso. 2013).

The women were randomly assigned to consume either 3x100 mg/d of EGCG or placebo (lactose) with each of their three main meals for 12 whole weeks. During those twelve weeks, all women followed a specifically designed low-energy mixed (55 % carbohydrates, 30 % lipids and 15 % proteins) diet that provided ca. 600 kcal/day energy less than the women would need to maintain their body weight. The energy content and macronutrient composition of diets were designed to achieve a weight loss of 0.5 to 1 kg per week, as it was observed by Davis et al. (2006) and Bantle et al. (2008) on very similar regimen. As the scientists point out, the "dietary instructions were reinforced weekly by a dietitian" (Mielgo-Ayuso), to optimise compliance.

Figure 1: Changes in body composition, energy expenditure and fat oxidation, left; changes in glucose, cholesterol metabolism and inflammation, right (Mielgo-Ayuso. 2013)

I am not sure how compliant the participants actually were, but in view of the fact that the women were advised not to change their physical activity habits during the energy restriction program, the relatively meager and statistically non-significant changes in body weight (-0·3 kg, p > 0.05) and fat mass (-0·7 kg, p > 0.05) are probably not really surprising. It is nice to see, though, that the women lost more fat than total mass - muscle loss was thus not an issue for the ladies.

What was not to be expected, though, - at least if you believe a single word of the hype about green tea supplements - were the non-existent effects of the purported weight loss supplement on  energy expenditure, fat metabolism, HOMA-IR (insulin sensitivity), total cholesterol, LDL-cholesterol, or triglycerides. In fact, the only good thing about the whole EGCG intervention was that the recently observed negative effects on the liver did not occur, either.

SIGNIFICANT Effect W/ 16 Million CFU of Nestlé's Lactobacillus rhamnosus strain

Want to check out the patent?

Despite the fact that the overall results are much more exciting than those in the previously discussed green tea study, I'd advise you to keep calm. We are after all dealing with another Nestlé study on a patented strain of Lactobacillus rhamnosus (LPR), i.e. "CGMCC1.3724" (date patented: 2012-05-10; #20120114622), and cannot tell how many never published negative study results the Nestlé guys had to dispose of, before Marina Sanchez et al. finally produced study results that pleased the marketing division of this multinational corporation.

What? Ok, ok... let's get back to the facts: The scientists from the Laval University and the Nestlé Research Center randomized a group of one-hundred fifty-three 18 to 55 year-old obese men and women to receive either a placebo or the said LPR formulation with 1·6 × 10⁸ colony-forming units of LPR and additional oligofructose and inulin per cap for a total of 24 weeks.

In the course of the first 12 weeks (phase 1), each participant received a personalised diet plan that would have him or her consume 500 kcal/d less than he or she'd need for weight maintenance (just as an aside, that's 100kcal more than for the subjects in the green teas study). During phase 2, each participant received a personalised diet plan without energy restriction. The good thing, the resting energy expenditure (REE) was actually measured: after a 12 h overnight fast in subjects having had rested for at least 15 min in a standardised supine position. This procedure was repeated thrice: (1) At baseline, (2) after the weight-loss and (3) after the second phase weight-maintenance periods using indirect calorimetry.

Figure 2: Changes in body composition (all data in kg) in men (left, blue) and women (right, orange) after weight loss (ΔW12) and weight maintenance (ΔW24) phase (Sanchez. 2013)

The data in figure 2 confirms what the abstract says: "The intention-to-treat analysis showed that after the first 12 weeks and after 24 weeks, mean weight loss was not significantly different between the LPR and placebo groups when all the subjects were considered."

Figure 3: Changes in metabolic parameters, i.e. energy intake (kcal/day), resting energy expenditure (REE, kcal/day) and respiratory quotient (RQ, remember: low RQ = high fat, low carb oxidation) after 12 and 24 weeks (Sanchez. 2013)

It does yet also confirm - and that there was a significant treatment × sex interaction, observed with the women in the treatment group losing significantly more weight than those in the placebo group (P= 0·02). More importantly, though...

"[...w]omen in the LPR group continued to lose body weight and fat mass during the weight-maintenance period, whereas opposite changes were observed in the placebo group."

For the unlucky men, on the other hand, the (unquestionably expensive) supplement didn't do sh*t: Their "changes in body weight and fat mass during the weight-maintenance period were similar" irrespective of whether they received the placebo or the active treatment.

Whether this was the reason or a consequence of the fact that the the men didn't show similar significant reductions in circulating leptin, as the women is questionable. Based on the fact that the relative abundance of bacteria of the Lachnospiraceae family in faeces increase only in women, we do yet have to assume that the missing reduction in leptin, as well as the absence of the significant body fat reductions, the researchers observed in their female subjects was simply a results of ...

• under-dosing - the same the 1·6 × 10⁸ colony-forming units of LPR that was sufficient for the average woman (body weight ~89kg) could have been too low for the guys (body weight ~104.3kg) 
• dietary interference - there could have been something in the diets of the guys that ruined the effects of the supplementation (lactobacilli are not exactly friends of meats and we all know that men love their meat ;-)
• different baseline gut microbiome - it goes without saying that you cannot place a group of rabbits in forest full of predators and expect them to survive; similarly the LPR spores may have come off second in the guts of the men, because they have a less "LPR-friendly" baseline colinization
• fundamental sex differences - at the moment I am not sure what the underlying reasons could be, but it's not impossible that hormonal difference could have played a role as well

I am pretty sure that I could come up with a whole host of additional, increasingly bizarre ad-hoc explanations for the null-effect Marina Sanchez and her colleagues from the Laval University and the  Nestlé Research Center in Lausanne observed in their male study but would rather conclude this news-item with the scientists own funky, but not unlikely explanation: Men are simply too good at dieting!

True: Women have a harder time losing weight even with high protein | more

As the authors point out, we know from previous trials (and corresponding SuppVersity posts, read more) that men are generally more prone to respond to a negative-energy balance intervention than women - and that's true irrespective of whether it is an exercise-training programme (Tremblay. 1984), a diet– exercise programme (Doucet. 1999), or a session of exercise and of mental work (Pérusse-Lachance. 2013). Plus, if you look at the data in figure 2, you'll see that this is actualy "concordant with the results of the present study that shows higher weight loss in men in the placebo group than in the women". Sanchez et al. do now believe that the high baseline success "abolished this difference" (Sanchez. 2013).

In view of the fact that there was a difference in a single low-abundance taxonomic group  (Prevotellaceae) between the baseline gut microbiome of the male and female study participants, I would still not exclude that the different baseline gut microbiomes could at least have added to the 'effect abolishing effect' of the sex-specific ease of weight loss in men. I mean, why wouldn't the feces of the men show an increase in lactobacillus spores, if the supplement worked?

Bottom line: Today's installment of the short news is very characteristic of the dilemma with weight loss supplements. We are just realizing that the classic thermogenic 'rodent fat burner' don't really work in humans. Against that background the rise of supplements that target the gut microbiome and exert much more complex body recompositioning effects comes in the nick of time.  Unfortunately, our understanding of the complex interactions between the gut microbiome and our immune system in the context of the emerging science of immunonometabolism is so incomplete (Mathis. 2011) that we are more or less groping in the dark, whenever we supplement subjects, patients or even ourselves with allegedly healthful bacteria.

All alleged benefits aside,  "specificity", the 2nd Principle of Sensible Supplementation, should keep you away from the next best GNC or online supplement store. The two studies at hand do after all not warrant the use of either green tea or lactobacillus supplements as weight loss aids in lean, healthy and active  men or women.

Accordingly, the observation that green tea supplements won't help sedentary over-weight women to lose weight appears to be much more reliable than the allegedly impressive weight loss effects of the probiotic during the "maintenance phase" of the Sanchez study.

We must however not forget the respective constraints of the research design and irresponsibly over-interpret the results of the EGCG study to (a) the potential benefits of regular 'whole' tea consumption in the average, non-obese individual (Wu. 2003) or (b) visceral fat loss in diet + exercise interventions in obese individuals (cf. Maki. 2009). Similarly, the fact that obese women will lose weight on a LPR supplemented maintenance diet is very unlikely going to translate to lean, athletic folks like you and me. According to the 2nd Principle of Sensible Supplementation, which is "specificity" (learn them all), I don't see you or me heading over to the next best online shop to buy LPR or EGCG supplements - irrespective of the promising results of the Sanchez trial.

References: 

• Bantle JP, Wylie-Rosett J, Albright AL,et al.(2008) Nutrition recommendations and interventions for diabetes: a position statement of the American Diabetes Association. Diabetes Care31, Suppl. 1, S61– S78.
• Davis NJ, Emerenini A & Wylie-Rosett J (2006) Obesity management: physician practice patterns and patient preference. Diabetes Educ32, 557 – 561. 
• Maki, K. C., Reeves, M. S., Farmer, M., Yasunaga, K., Matsuo, N., Katsuragi, Y., ... & Cartwright, Y. (2009). Green tea catechin consumption enhances exercise-induced abdominal fat loss in overweight and obese adults. The Journal of nutrition, 139(2), 264-270.
• Mathis, D., & Shoelson, S. E. (2011). Immunometabolism: an emerging frontier. Nature Reviews Immunology, 11(2), 81-83.
• Mielgo-Ayuso J, Barrenechea L, Alcorta P, Larrarte E, Margareto J & Labayen I (2013). Effects of dietary supplementation with epigallocatechin-3-gallate on weight loss, energy homeostasis, cardiometabolic risk factors and liver function in obese women: randomised, double-blind, placebo-controlled clinical trial. British Journal of Nutrition, available on CJO2013. 
• Tremblay, A., Despres, J. P., Leblanc, C., & Bouchard, C. (1984). Sex dimorphism in fat loss in response to exercise-training. Journal of obesity and weight regulation.
• Wu, C.-H., Lu, F.-H., Chang, C.-S., Chang, T.-C., Wang, R.-H. and Chang, C.-J. (2003), Relationship among Habitual Tea Consumption, Percent Body Fat, and Body Fat Distribution. Obesity Research, 11: 1088–1095.

Stevia Kills Good Gut Bacteria - One Study Enough to Stop Using the Natural Sweetener? Probably Not in View of its Anti-Diabetes, Anti-LDL, Anti-Viral & Anti-Cancer Effects

Study indicates stevia kills healthy gut bacteria. So, how bad is it? Are the effects significant, will they have an impact on your overall health and does this mean you must not use stevia any longer?

A recent study from the Institute of Microbiology and Biotechnology at the University of Latvia in Riga shows the impossible: Stevia, the "natural" sweetener that's everybody's darling, could mess up your gut microbiome by killing large numbers of the beneficial Lactobacillus Reuteri bacteria in your tummy - exactly those bacteria of which several studies have shown that supplementing will help cure acute diarrhea in young children (Shornikova. 1997), is capable of reducing frequency and intensity of antibiotic-associated side-effects during eradication therapy for H. pylori. (Lionetti. 2006), confers broad-spectrum protection against disease in humans and animals (Casas. 2000), has cholesterol lowering effects (Jones. 2012) and much much more.

You can learn more about the gut & your health at the SuppVersity

Bugs Dictate What You Crave

Sweeteners & Your Gut

Foods, Not Ma- cros for the Gut

Lactulose For Gut & Health

Probiotics Don't Cut Body Fat

The Macrobiotic MaPi2.0 Diet

In view of the fact that it would appear as id Lactobacillus reuteri was clearly one of the "good guys" it seems that the results I. Denin a, P. Semjonovs, A. Fomina, R. Treimane and R. Linde report on their latest study in Letters in Applied Microbiology (Denin. 2014) were really bad news:

Figure 1: Influence of stevioside (a) and rebaudioside A (b) on biomass formation in Lactobacillus reuteri strains (24 h | Denin. 2014).

"In samples supplemented with stevia glycosides, the growth of all Lact. reuteri strains was slightly inhibited – however, a statistically significant concentration-dependent inhibitory effect was not observed for all strains (Fig. 1).

Comparing both the glycosides, the inhibitory effect of stevioside was more pronounced for strains 44 and 16, while the effect of rebaudioside A was more pronounced for strains 16 and 19. Statistically significant concentration-dependent inhib itory effect was observed for lactic acid and acetic acid synthesis. The decrease in lactic acid and acetic acid production was observed for both stevioside and rebaudioside A. [...] Although the inhibitory effect of stevioside on pH was observed at different stevioside concentrations, the effect was evident for all strains. Rebaudioside A had a more pronounced inhibitory effect on pH values of certain strains including Lact. reuteri 12, 16, 43 and 44" (Denin. 2014 | my emphasis).

The good news, however, is in the details: The inhibitory effect was "slight" (see quotation above) and the design of the study leaves it open, whether similar effects would occur in vivo and thus outside of a glycoside, stevioside and rebaudioside laden Petri dish.

Previous studies seem to refute significant effects of stevia on the human microbiome! In 2003, Gardana et al. found no effect of stevia on the make-up of human fecal cultures when they were incubated with either stevioside or rebaudioside A. Only the fact that bacteroides, i.e. the "enemies" of lactobacilli, were the most efficient in hydrolyzing Stevia sweeteners to steviol would suggest that there may be an overall effect on the human microbiome form stevia (ab-)use.

And while we have little in vivo evidence that stevia is bad for you, a brief review of the contemporary scientific literature on Stevia yields the following "proven" (mostly only in a handful, if not just a single study) benefits:

• Stevia has been implicated in diabetes and hyperlipidemia treatment and its effects on blood glucose levels are not a mere result of the corresponding reduction in sugar intake.

  

  Figure 2: Effects of stevia vs. diabetes drug Glibenclamide on blood glucose and lipid levels in diabetic rodents; data expressed relative to healthy control (Singh. 2014)

  In a recent rodent study that compared the effects of stevia against those of the often-prescribed diabetes-drug Glibenclamide, the natural sweetener outperformed the drug in many in its ability to reduce LDL and blood sugar and was not far off of what the Glibencamide did for the diabetic lab animals in terms of its effects on HDL and VLDL (see Figure 2).
  
  Previous human studies indicate that stevia extracts will also increase the increased 16 healthy human volunteers whose plasma glucose levels during an oral glucose tolerance tests were significantly lower after having consumed 5 grams of aqueous leave extract at regular 6-h intervals for 3 days (Curi. 1985).

  

  Figure 3: Effects of stevia and aspartame replacement of sucrose in test meals that were fed to obese and normal-weight volunteers on postprandial blood glucose levels (Anton. 2010)

  Moreover, in a more recent study by Anton et al. where stevia was compared to aspartame, it had the same beneficial effects on total energy intake and let to statistically significant reductions in postprandial glucose levels of both obese and lean study subjects (see Figure 3) that did not reach significance when the sucrose content of the test meal was replaced by aspartame.
• In-vitro stevia appears to have anti-cancer effects, as well. That's at least what studies by  Jayaraman et al. (2008) observed with stevia extracts. An effect that may be related to both it's anti-microbial, as well as its potent anti-oxidant activity (Tadhani. 2007) of the whole leaves and leave extracts of which Tahani et al. found that they contain significant effects of folic acid (52.18 mg/100 g) and vitamin C, as well as 130.76 μg catechin and 15.64 μg quercetin for leaves and 43.99 μg catechin and 1.57 μg quercetin for cellus at mg of water extracts, respectively.
  
  Furthermore, Tadhani et al.'s results showed that the leaf extracts contained higher amounts of free radicals, hydroxyl radicals and superoxide anion radical scavenging activities than those of the callus extracts or the anti-mutagenic effects Cariño-Cortés et al. report in their 2007 study. Whether anything similar can be observed with the white "stevia" powder that is used by most people to sweeten their foods is yet questionable - it's after all pure steviosid and thus devoid of all of the previously mentioned compounds.

  

  Figure 1: Several natural constituents of the stevia plant, including steviosides, which are the naturally sweet agents in stevia have potent anti-viral activity against Epstein-Barr virus; values in brackets
  represent % of untreated control dish (Konoshima. 2002)

  Another possible anti-cancer mechanism may be related to stevia's ability to kill viruses like the Epstein-Barr virus that has been implicated in the pathogenesis of Burkitt’s lymphoma, Hodgkin’s disease, non-Hodgkin’s lymphoma, nasopharyngeal carcinoma, and lymphomas, as well as leiomyosarcomas arising in immunocompromised individuals.in humans (Thompson. 2014).

Against that background it seems questionable that the new evidence of negative effects on allegedly healthy gut bacteria (just want to remind everyone that we have no clue what the optimal gut microbiome would look like) is significant enough to have us all reconsider our use of tiny amounts of stevia as a sweetener in our foods.

Read more about the effects artificial sweeteners have on the microbiome in a prevoius article | go ahead!

Interim conclusion: While I am not all too scared that stevia will mess with my gut microbiome in a way that makes me sick, fat and what not, I truly believe that the effects of artificial sweeteners on the make-up and density of the human gut microbiome is still massively under-researched - and that in spite of the fact that it could have a significant effect on the health of us all.

As s SuppVersity reader you will also be aware that this is not a stevia-specific effects. Only recently I have written about similar effects for a bunch of artificial sweeteners - an article I can only recommend to anyone who hasn't read it yet | Comment on Facebook.

References:

• Anton, Stephen D., et al. "Effects of stevia, aspartame, and sucrose on food intake, satiety, and postprandial glucose and insulin levels." Appetite 55.1 (2010): 37-43.
• Casas, Ivan A., and Walter J. Dobrogosz. "Validation of the probiotic concept: Lactobacillus reuteri confers broad-spectrum protection against disease in humans and animals." Microbial ecology in health and disease 12.4 (2000): 247-285. 
• Curi, R., et al. "Effect of Stevia rebaudiana on glucose tolerance in normal adult humans." Brazilian journal of medical and biological research= Revista brasileira de pesquisas médicas e biológicas/Sociedade Brasileira de Biofísica 19.6 (1985): 771-774.
• Deniņa, Ilze, et al. "The influence of stevia glycosides on the growth of Lactobacillus reuteri strains." Letters in applied microbiology 58.3 (2014): 278-284. 
• Gardana, Claudio, et al. "Metabolism of stevioside and rebaudioside A from Stevia rebaudiana extracts by human microflora." Journal of agricultural and food chemistry 51.22 (2003): 6618-6622. 
• Jayaraman, Sathishkumar, Muthu Saravanan Manoharan, and Seethalakshmi Illanchezian. "In-vitro antimicrobial and antitumor activities of Stevia rebaudiana (Asteraceae) leaf extracts." Tropical Journal of Pharmaceutical Research 7.4 (2008): 1143-1149.
• Jones, M. L., C. J. Martoni, and S. Prakash. "Cholesterol lowering and inhibition of sterol absorption by Lactobacillus reuteri NCIMB 30242: a randomized controlled trial." European journal of clinical nutrition 66.11 (2012): 1234-1241.
• Konoshima, Takao, and Midori Takasaki. "Cancer-chemopreventive effects of natural sweeteners and related compounds." Pure and applied chemistry 74.7 (2002): 1309-1316.
• Lionetti, E., et al. "Lactobacillus reuteri therapy to reduce side‐effects during anti‐Helicobacter pylori treatment in children: a randomized placebo controlled trial." Alimentary pharmacology & therapeutics 24.10 (2006): 1461-1468.
• Shornikova, Aino-Vieno, et al. "Lactobacillus reuteri as a therapeutic agent in acute diarrhea in young children." Journal of pediatric gastroenterology and nutrition 24.4 (1997): 399-404.
• Singh, Sunanda. "Antidiabetic, Antidyslipidymic and Antioxidative potential of methanolic root extract of Stevia rebaudiana (Bertoni) on Alloxan induced Diabetic Mice Sunanda Singh and Veena Garg Department of Bioscience and Biotechnology, Banasthali Vidyapeeth, Banasthali, Rajasthan, India." (2014). 
• Tadhani, M. B., V. H. Patel, and Rema Subhash. "In vitro antioxidant activities of Stevia rebaudiana leaves and callus." Journal of Food Composition and Analysis 20.3 (2007): 323-329. 
• Thompson, Matthew P., and Razelle Kurzrock. "Epstein-Barr virus and cancer." Clinical Cancer Research 10.3 (2004): 803-821.

Vitamin A Educates T-Cells, Joins Forces With Vitamin D Against Liver Cancer. Milk Better Than Sugary Electrolyte Solutions for Rehydration? Helicobactor Pylori: Probiotics from Breast Milk & Feces Better Than Amoxicillin!

Lactobacilli are hip, vitamin A is not - at the SuppVersity you still get news on both

1kg! That's the amount of weight you could probably lose if you rid yourself of all the microbes in your gut - from the weight of the bacteria alone, of course. Whether this would be a good idea or not, is however very questionable. On the one hand, we do have the still not fully understood studies on obesity-resistant germ free mice and an accumulating amount of evidence that having the "wrong" bacteria in the gut is at least associated with an increased obesity risk (Blaut. 2012). On the other hand, however, we are seeing new studies on the various benefits of having the "right" gut microbiome being published on an almost daily basis. So what?

Before we take a closer look at a definite benefit of having the "right" gut bacteria, though, let's start out with another likewise gut-related news item on the role of retinoic acid in T-cell education. In a way it's funny, it starts right where the bacteria reside, could have immune-modulatory effects that are way more pronounced and far reaching than probiotics and is still hardly discussed.

Vitamin A is of critical importance to (intestinal) T-cell education

If you have ever asked yourself how the immune cells in your body know what they are supposed to do, Catharine Ross' latest paper that was published in the American Journal of Clinical Nutrition and is based on a short talk the researcher from the Department of Nutritional Sciences at the Pennsylvania State University held at a conference earlier this year may provide at least some additional insides into the role a still way underrated molecule plays in this "T cell education" (Ross. 2012): Vitamin A!

Figure 1: Model of T cell differentiation, from uncommitted naive T cells into different T cell subsets that produce different cytokines and thus promote different functional activities (adapted from Ross. 2012)

As you can see in figure 1, retinoic acid does not simply promote the differentiation of regulatory T cells, which help to suppress inflammatory reactions, it also plays a significant role in normal mucosal immunity (in the gut, the airways and elsewhere) by modulating T cell activation and regulating cell trafficking. Moreover, vitamin A promotes antibody responses to T cell–dependent antigens. Needless to say that

"[...] in a state of vitamin A deficiency, inflammatory T cell reactions may be inadequately opposed and therefore become dominant [...] Although data from human studies are still needed, the framework now developed from studies in mice and rat models suggests that adequate vitamin A status, [...] is  important for maintaining a proper balance of well-regulated T cell functions and for preventing excessive or prolonged inflammatory reactions." (Ross. 2012).

Discovery a beta carotene derived vitamin A receptor blocker is only one of a couple of intriguing findings wrt to vitamin A.

One thing that sticks out from the complex interactions (see figure 1), really is the way by which the interaction of vitamin A with the T-cells in the gut crucially determine the efficiency of the 'fist line defenses' and their downstream effects on the whole organism. It is by no means co-incidental that diarrhea is rampant in areas of the "third world", where a large amount of the population is vitamin A deficient (Beaton. 1994). And in fact studies have shown consitently that

"RA is essential for 'imprinting' gut-homing specificity on T cells activated by intestinal DCs [dendritic cells] and suggested that MLN DCs are a source of RA that drives T cell differentiation toward the gut-homing phenotype" (Ross. 2012)

Moreover, oral tolerance to foreign antigens and thus an allergy free live requires a form of immune suppression, which can be proffered or hampered by sufficient and insufficient vitamin A intakes. In that, the exact effects of vitamin A will depend on the cytokine milieu the T-cells are exposed to. Examples are...

• an exaggerated IL-17 response with vitamin A deficiency, on the one hand, and
  
• an increase of the inflammatory response due to high vitamin A in an IL-15 environment 

Based on these observations, Ross rightly points out that "when RA is used for therapeutic purposes, it should be used cautiously in subjects with various inflammatory bowel conditions and sensitivities to dietary antigens." (Ross. 2012) People with gluten intolerance, celiac and other allergic reactions, for example would probably be better off avoiding the consumption of any form of supplemental vitamin A (on top of what's in their regular diet). Someone with high IL-17 and IL-6 levels as they have been observed in non-celiac inflammatory bowel disease, type 1 diabetes, multiple sclerosis and rheumatoid arthritis, on the other hand, could actually benefit from vitamin A's (especially ATRA) presence during activation of CD4⁺ T cells, because it will - even in the presence of IL-6 - "favor the development of the a Treg lineage at the expense of T cells secreting IL-17" and could thus help reduce chronic inflammation and keep autoimmune reactions at bay (Schambach. 2007; also Ramgolam. 2010).

More news

• 

  Figure 2: Who cares about cell viability, the survival time (in days) matters

  Combination therapy with vitamin A and a vitamin D (not D3, but calcitriol) analog EB1089 kills liver cancer cells. And it does so more effectively than any of the two molecules alone. That's the actually unsurprising result of a study that has been conducted at the Beijing Army General Hospital in China. The researchers injected nude mice with molecules that made them develop hepatocellular cancer. Afterwards, the rodents received either 10 μmol/L retinoic acid (vitamin A), 10 nmol/L EB1089 or both as a combination treatment.
  
  Compared to vitamin A or the calcitriol analog alone, the combination treatmend resulted in a significanlty higher reduction of the viability of hepatocellular cancer cells. Based on TUNEL analysis, Zhang et al. did also establish that individual cancer cells had a higher apoptotic ratio in the combined drug group than in the groups for which the drugs were used separately. Most importantly, however, the tumor weight was decreased and the mice on the combination treatment lived significantly longer (see figure 2; Zhang. 2012)

• 

  In the same publication, Pritchett and Pritchett recommend 1.0-1.5ml / kg body weight per hour of chocolate milk as the optimal post-workout drink to be consumed in the 2 h after a workout.

  Skimmed milk, the ideal post-workout rehydration formula? According to L James' paper in Lamprecht's compendium Acute Topics in Sport Nutrition, milk is a way better choice then the standard sugar + electrolyte rehydration formulas. Interestingly this is not due to the minerals in the milk, or the sugar, but, as James argues, a direct consequence of the milk proteins, which help restore "fluid balance after exercise-induced dehydration to a greater extent than a carbohydrate-electrolyte sports drink." As James points out it will yet have to be elucidated, whether the simple addition of whey protein to a standard sugar + electrolyte formula would exert similar effects (James. 2013).
• Probiotics to kill Helicobacter Pylori? While not every bacteria stands a chance against the nasty gut bug H. Pylori, certain Lactobacillus spp. strains obviously do. At least, if the results of a recent in-vitro + in vivo rodent study by Pei-Shan Hsieh can be replicated in human studies.

  

  Figure 3: Urease activity in H. pylpori after co-incubation with the specific probiotic and resulting bacteriostatic ratio (100% = bacteria free; data adapted from Hsieh. 2012)

  Lactobacillus acidophilus TYCA08, L. acidophilus TYCA15, L. johnsonii MH-68, and L. salivarius subsp. salicinius AP-32 were the most effective strains the researchers from National Chung Hsing University in Taichung, Taiwan, analyzed. And believe it or not, the latter of these, i.e. L. johnsonii MH-68, and L. salivarius subsp. salicinius AP-32, both of which are  by the way found in feces, were even minimally more potent effective than Amoxicillin, a moderate-spectrum, bacteriolytic, β-lactam antibiotic used to treat bacterial infections. L. acidophilus TYCA15, however, steals the show. This probiotic that occurs naturally in breast milk reduced the urease activity of H. Pylori by -97.1% (see figure 3).
  
  In the consecutive rodent study, Hseieh et al. did yet still use 10⁹ CFU/mL of either AP-32 alone, MH-68 alone, or an equal mix of cultures of the two strains and both, "either alone or as a mixture in powder form were effective in reducing H. pylori load in gastric mucosa and help in reducing gastric inflammation and in regulation of gastric acid production." (Hsieh. 2012)

Thats it for today and for this weekend. As mentioned yesterday, there was simply not enough time to do the necessary research for the follow up to the Athlete Triad Series, so that this will have to wait. So don't dig an even deeper whole in the mean time. Maybe you want to do some of the psychomotor tests mentioned in yesterday's news, and check whether you are already overtrained!? How steady are your hands, for example? And whatever the result may be, don't forget to enjoy the rest of the weekend!

References:

• Beaton GH, Martorell R, Aronson KA, Edmonston B. McCabe, G, Ross, AC, Harvey, B. Vitamin A supplementation and child morbidity and mortality in developing countries. Food Nutr Bull 1994;15(4): 282–9.
• Blaut M, Klaus S. Intestinal microbiota and obesity. Handb Exp Pharmacol. 2012;(209):251-73.
• Hsieh PS, Tsai YC, Chen YC, Teh SF, Ou CM, King VA. Eradication of Helicobacter pylori Infection by the Probiotic Strains Lactobacillus johnsonii MH-68 and L. salivarius ssp. salicinius AP-32. Helicobacter. 2012 Dec;17(6):466-77.
• James L. Milk Protein and the Restoration of Fluid Balance after Exercise. In Lamprecht M (ed): Acute Topics in Sport Nutrition. Med Sport Sci. Basel, Karger, 2013, vol 59, pp 120–126. 
• Pritchett K, Pritchett R. Chocolate Milk: A Post-Exercise Recovery Beverage for Endurance Sports. In Lamprecht M (ed): Acute Topics in Sport Nutrition. Med Sport Sci. Basel, Karger, 2013, vol 59, pp 127–134.
• Ramgolam VS, Markovic-Plese S. Interferon-beta inhibits Th17 cell differentiation in patients with multiple sclerosis. Endocr Metab Immune Disord Drug Targets. 2010 Jun;10(2):161-7.
• Ross AC. Vitamin A and retinoic acid in T cell-related immunity. Am J Clin Nutr. 2012 Oct 10.  
• Schambach F, Schupp M, Lazar MA, Reiner SL. Activation of retinoic acid receptor-alpha favours regulatory T cell induction at the expense of IL-17-secreting T helper cell differentiation. Eur J Immunol. 2007 Sep;37(9):2396-9. 
• Zhang J, Zhang H, Zhang X, Yu Z. Synergistic effect of retinoic acid and vitamin D analog EB1089-induced apoptosis of hepatocellular cancer cells. Cytotechnology. 2012 Oct 16.

Fructose Toxicity or Bacteria Deficiency? 10 Billion CFU of Probiotics Ameliorate, But Don't Solve the Fat Problems That Are Brought About by a Crappy Diet + Fructose Water

The mere fact that probiotics can ameliorate some of the damage that's done by the way people eat this day, does not imply that they are the origin of the obesity epidemic; and the study at hand shows: They are not the solution either.

A recently published study from the Key Laboratory of Dairy Biotechnology and Engineering at the Inner Mongolia Agricultural University could hold a "solution" - or rather a way to ameliorate the damage that comes with the overconsumption of fructose.

Cause or solution!? I don't care if it works!

Based on previous observations the researchers speculated that the provision of  10,000,000,000 CFU/day Lactobacilli Casei Zhang bacteria  to male Sprague Dawley rats who were guzzling "rodent coke", i.e. 25 % fructose water, 24/7 for either

• 9 weeks after the supplementation was initiated, or
• 4 weeks after 9 weeks on fructose water

would ameliorate the weight negative side effects of the addition of fructose to an already 100% unhealthy diet comprising 50 % corn and wheat starch, 25 % wheat bran, 20 % soybean pieces, 2 % bone powder, 2 % fish powder, 0.9 % table salt and a 0.1 % vitamin mixture would have on the impaired glucose tolerance and the incidence and severity of the other typical side effects that occur, when you are chronically poisoning rodents with this type of dieting.

So, what did the scientists find?

While it appears to be self-evident, the first trivial, yet extremely important result of the study at hand is the observation that the bacteria did not just make it to the gut of the rodents, but actually settled and survived there. This lead to a significant increased numbers of Lactobacillus and Bifidobacterium (what we currently think are the "good guys") and a concomitant decrease in the number Clostridium bacteria in the intestine.

Figure 1: Liver gylcogen, serum insulin, osteocalcin, and MDA levels expressed relative to rodents on control diet after 9 and 13 weeks on different diet + supplementation regimen (Zhang. 2013)

These microbial changes went hand in hand with normalization of liver glycogen in all groups. Only the animals who had been pretreated with probiotics did yet show lower insulin and GLP-2 levels.
The improved response to the glucose tolerance test, on the other hand was observed in both the pre-treated group, in which the rodents received fructose and probiotic bacteria together for 9 weeks, and the "therapeutic" group, in which the probiotics were gavaged to already obese rodents for only four weeks.

Figure 2: Glucose response (AUC) in tolerance test and body weight development (Zhang. 2013)

And while there were no significant differences in body weight, the administration of L. casei Zhang could at least improve the compromised glucose tolerance, increase the dropping osteocalcin levels and restore the bile acid secretion in the "therapeutic group".

---

Bottom line: In conjunction with the decreased serum MDA and the upregulation of LXR-a, PPAR-gamme and AdipoR2 gene expression the overall picture that emerges is somewhat similar to that you would see with some of the "better" diabetic drugs. Yet while this may sound great, at first, it is in the end a total bummer.

Aside from the people whose businesses flourish, when people are sick enough to need medication, but not so sick that they would die away anytime soon, "solutions" like these don't help anyone. And let's face it - we all know that the root cause of the problem is not a messed up gut microbiome. That may be a consequence, yes. The root cause, on the other hand, is the way millions of people "eat".

References:

• Zhang Y, Wang L, Zhang J, Li Y, He Q, Li H, Guo X, Guo J, Zhang H. Probiotic Lactobacillus casei Zhang ameliorates high-fructose-induced impaired glucose tolerance in hyperinsulinemia rats. Eur J Nutr. 2013 Jun 25.

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".

Do You Have the Gut(s) to Lose 8% Belly Fat in 12 Weeks? Lactobacillus gasseri (LG2055) Can Fix Your Gut Problems

As a diligent student of the  SuppVersity, you'll obviously know that beer is not the main cause of the eponymous belly (Bobak. 2003). Although..., when when you come to think about it: For every glass of beer you drink, you're probably drinking a glass of probiotic yogurt less ;-)

You've done everything you could and still feel like a whale? You've been low-carbing, even have made sure you are in ketosis by consuming an only 10% protein diet and still the fat won't disappear? Well, I guess it sounds unfair, if I do now tell you that you YOU JUST DON'T HAVE THE GUTS it takes to get rid of that pouch that blocking the view on your toes, when you are standing.

Now calm down, I am not talking about not being able to curb your insatiable appetite or skipping 90% of your workouts, here. I am talking about your gut(s), literally! Your digestive tract and it's inhabitants. Those critters which are also at the heart of the soon-to-be-published paper by scientists from the Milk Science Research Institute in Fukoaka, Japan (Kadooka. 2013) - a paper, the results of which I am about to present in the next few paragraphs.

Probiotic in fermented milk shred visceral fat

I am not telling you something new, when I say (or write) that scientists have long identified the modulation of the gut microbiome, i.e. the bacterial ecosystem in your intestines, could be the long-sought-for lever to get rid or at least ameliorate the negative effects of the way we live and eat promotes the expansion of the highly inflammatory "stress fat" that fills the room between the organs in your trunk. Most of the hitherto published peer-reviewed research has yet been conducted in rodents. The existing evidence from well-controlled human studies, on the other hand is scarce.

Suggested read: "Probiotics for Athletes: The Supplemental 10 Billion CFS Leaky Gut Solution for the Fermented Food Refusinek?" (read more)

One of these studies has been conducted by Kadooka et al. in 2010 (Kadooka. 2010). In the pertinent paper the researchers from the Milk Science Research Institute in Japan reported that the provision of 200g/day of fermented milk containing 10^8 colony-forming units (cfu)/g of Lactobacillus gasseri strain SBT2055 (LG2055) led to significant reductions in visceral fat in male and female subjects with " higher body mass index" (BMI: 24.2-30.7 kg/m²) who had been randomized to the active arm of the study. Now, roughly three years later, are about to publish a follow-up paper with great practical relevance, as it summarizes the results of what Kadooka et al. say is the first human trial that examines the effect of different doses of probiotics on abdominal adiposity.

"We know that it works, but we don't know how much we need to elicit the desired effects"

The design of the study at hand is essentially identical with the one in the 2010 trial. It is based ona multi-centre, double-blind, parallel-group RCT with 210 healthy Japanese adults with large visceral fat areas (80.2–187.8 cm²), who were now randomly assigned to three instead of two groups who consumed 200g of fermented milk with either 1,000,000 CFU (1m), 10,000,000 CFU (10m) or just the normal yogurt probiotics (control) per gram for 12 weeks.

The intention was to determine whether the effect of LG2055, which is by the way a probiotic lactic acid bacterium that originates from the human intestine, would be observed with lower / more realistic dosage regimen as well and whether possible reductions in the effect size would be dose-dependent.

Figure 1: Reduction in visceral and subcutaneous adipose tissue, as well as waist circumference after 8 and 12 weeks on 1,000,000 CFU/g or 10,000,000 CFU/g yogurt drink; data expressed relative to group baseline (Kadooka. 2013)

If you take a look at the results, the first thing that strikes the eye in figure 1 is that the lactobacilli did their job and that in both the low and high dose group:

• Even in the 10x and 100x lower dosages there was a significant reduction in the amount of visceral fat the  33/36 respectively 36/35 men and women in the in the 1 million and 10 million CFU/day groups, respectively.
• The effect size of both dosages of LG2055 was identical, so that it would not actually matter if you consume just 100g or 1kg of an imaginary commercially available milk drink with 1,000,000 CFU of LG2055 per 100g.

In view of the >8% reduction in visceral fat mass, it should not come as a surprise that the reduction in waist size was likewise statistically significant - both on its own, as well as in comparison to the control group, What is yet not so obvious and probably somewhat disappoint for you is the fact that the reduction in the ugly, but benign subcutaneous fat was statistically and practically non-significant.

"Visceral fat loss, only? But what about my thigh fat?"

Not all probiotics are created equal: As the scientists emphasize in the discussion, the "it is also worth noting that the test FM containing both LG2055 and yogurt cultures reduced abdominal adiposity when compared with the control FM containing yogurt cultures alone, even though the bacterial counts of LG2055 (106 and 107 cfu/g) were lower than those in the yogurt cultures (109 cfu/g)." (Kadooka. 2013) The observed superiority of this particular strain of bacteria stands in line with previous rodent studies and is imho one of the reasons we are seeing so much research in this area. How's that? Well, easy: If you came up with a particularly potent strain and patented that, this would be the literary cash machine.

If you are no newbie to the SuppVersity you will yet be aware that it is by no means extra-ordinary that "healthy fat burners", which work their weight loss magic mostly by modulating local and/or whole body inflammation, have little or no effects on the amount of subcutaneous body fat.

If you discarded any statistical shenanigan and simply went by the height of the small bars in the middle of figure 1, you could yet probably argue that it appears as if there was a trend towards greater subcutaneous fat loss in the high(er) dose LG2055 group. If you went a step further and made the unwarranted assumption that you could simply compare the results from the study at hand to those of the aforementioned 2010 study b Kadooka et al. you could support this argument by stating that there was a clear trend here with 1.2%, 2.6% and 3.3% reductions in subcutaneous fat with LG2055 at concentrations of 1, 10 and 100 million CFU per gram of the fermented milk drink.

In the overall context it is yet important to point out that it is as of yet unknown whether the composition of colonic microbiota has a direct influence on abdominal and/or subcutaneous adipose tissue (Hildebrandt. 2009; Conterno. 2011). Moreover, ...

"[...] environmental factors, including genetics and age, makes it difficult to discuss the causal relationship between adiposity and intestinal microbiota." (Kadooka. 2013)

The latter are particularly difficult to access in a study like this, where the subjects maintain their habitual mode of living, including diet, and no strict dietary control is applied. Kadook et al. are thus right to point out that "the relation ship between adiposity and intestinal microbiota" has to be investigated more closely "under a strict diet, together with the latest methodology" (Kadooka. 2013) to finally make some progress in our understandings of the intricate interaction between the human gut microbiome and our metabolism.

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Even the nicest subtenants can become a real problem, when they come over without being asked. Unfortunately, all sorts of gut bacteria (even the "good" ones) have a similarly nasty habit of translocating through a leaky gut wall into parts of your body, where you don't wont them (learn more)!

Bottom line: In view of the fact that the subjects shed the superfluous body fat in the absence of a reduced energy intake and/or exercise / increases in physical activity, the results are unquestionably impressive. To speak of "probiotic fat burner" would yet still not be warranted. After all, the subjects were all overweight and had an average body fat content of 32%, the baseline diet does - at least by the available data - not look really optimal (protein intakes of 0.8g/kg; macro-composition of 15/57/28% from protein, carbs and fats) and the non-disclosed insulin and HbA1c levels as well as the markers of inflammation will probably have left more than enough room for the "get healthy lose weight" effect.

Against that background I have my doubts that you can expect anywhere similar effects in lean, let alone athletic individuals. This does yet not negate the general usefulness of probiotics, after all, health is something you take for granted only, when you've never been sick - if you want to avoid that, the incorporation of fermented foods makes sense for everyone from the sedentary slob to the elite level athletes*...

* Note: I am writing the above although it gives me tummy aches (pun intended). Specifically with respect to the use of high dose probiotic supplements, I am asking myself, whether we really know enough about the potential downstream effects of long-term / high dose supplementation. In that, I am particularly concerned about shifting the balance from an "A"-dominant and unhealthy state in which the attributes "good" and "bad" are all of the sudden reversed and the previously dominant "bad" strain X gets totally replaced by the allegedly "good" strain Y that will then turn out to be just as nasty as X, once its natural arch enemy X is no longer present.

References:

• Bobak M, Skodova Z, Marmot M. Beer and obesity: a cross-sectional study. Eur J Clin Nutr. 2003 Oct;57(10):1250-3.
• Conterno L, Fava F, Viola R, et al. Obesity and the gut microbiota: does up-regulating colonic fermentation pro- tect against obesity and metabolic disease? Genes Nutr. 2011; 6:241–260.
• Hildebrandt MA, Hoffmann C, Sherrill-Mix SA, et al. High-fat diet determines the composition of the murine gut microbiome independently of obesity. Gastroenterology. 2009; 137:1716–1724.
• Kadooka Y, Sato M, Imaizumi K, Ogawa A, Ikuyama K, Akai Y, Okano M, Kagoshima M, Tsuchida T. Regulation of abdominal adiposity by probiotics (Lactobacillus gasseri SBT2055) in adults with obese tendencies in a randomized controlled trial. Eur J Clin Nutr. 2010 Jun;64(6):636-43. 
• Kadooka Y, Sato M, Ogawa A, Miyoshi M, Uenishi H, Ogawa H, Ikuyama K, Kagoshima M, Tsuchida T. Effect of Lactobacillus gasseri SBT2055 in fermented milk on abdominal adiposity in adults in a randomised controlled trial. Br J Nutr. 2013 Apr 25:1-8. [Epub ahead of print]