Leucine + B6: 82% More Weight + Twice As Much Body Fat Loss in First 12 of 24 Weeks on -500kcal/day Diet W/ 2.25g Leucine + 30mg Pyridoxine - To Good to Be True?

Check your nutrition labels, I bet you get 2.25g+ leucine and 30mg of B6 from one of your supps already

I am sorry to say that and actually this is usually something that goes into the bottom line, but I do have serious doubts whether the results reported in this study can be replicated in any of you. Yes, it is a human study, but (a) the subjects, 12 men and 12 women, were obese (34.76 ± 2.57) and (b) if the mere addition of 2.25 g leucine and 30 mg vitamin B6 would have such profound effects, everyone who has ever taken a commercial BCAA product would have noticed a doubling in fat loss, already, as almost all of them contain 2.25g+ of leucine and tons of pyridoxine (B6).

So what are you saying here? Is this study a fraud?

I have no evidence that the study was a fraud and I am certainly not implying this, but you must wonder about the magnitude of the weight loss and fat loss difference between the regular and the NuShape supplemented dieters (-500kcal from baseline energy demands calculated simplistically by  1.3× result of the WHO formula for basal metabolic rate), who ingested 2x 1.125 g leucine and 15 mg pyridoxine per day (whether that's on empty is not disclosed in the paper).

Figure 1: Changes in insulin (μU/ml), HOMA-IR, waist circumference (cm), body weight (kg), fat weight (kg) after 12 (blue) and 24 weeks (orange), relative change from 12-24 weeks is indicated in % below the bars (Zemel. 2013)

According to the researchers, who have published corresponding data for rodents and an analysis of the effects of the same combination of leucine + pyridoxine on total fat oxidation (>30g increase per day in a randomized controlled study; Zemel. 2012), the specific effects on visceral obesity could be a result of the

"reductions of oxidative and inflammatory stress biomarkers, such as plasma malondialdehyde (MDA), 8- isoprostane F-2α, tumor necrosis factor-α, and C-reactive protein, as well as an increase in the anti-inflammatory marker adiponectin" (Zemel. 2013)

they observed in a previous trial after 4 weeks, even in the absence of weight loss (Zemel. 2012). This would certainly be beneficial for both the lean and the fat individual, but with already low levels of CRP, MDA and TNF-alpha as you, as a healthy follower of physical culture should have them, these changes will be so small that they are unlikely to have any visible effects on you.

There decreased weight / waist / fat loss within the 2nd two weeks in the supplement group (see figure 1, yellow arrow for the discussion of waist circumference as an example) does actually provide evidence that the effect size will decrease the learner you are... even "worse", but rather hypothetical: The effect could also be diminishing, because the human body get's used to the new steady influx of leucine and B6 and their beneficial effects on the mitochondria, Sirt-1 and AMPK expression (learn more; note: as stated in this older article of mine, it works also without resveratrol)

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Learn more about leucine, HMB, B6, mitochondrial health and weight / fat loss in a previous post (read more)

Bottom line: You are probably already doing what 50% of the subjects in this (sponsored) study have been randomly assigned to, i.e. consuming an extra 2.25g of leucine and 30mg of vitamin B6 (this is ~3800% of the RDA) per day if you consume either a "sports multi", a "high potency B-vitamin" or simply the next best BCAA or other amino acid / protein product with added B6. So, let me ask you a question, then. Did you get ripped to the shreds from this practice? No? Well, in that case you will probably share my doubts about the relevance of the data and the usefulness of the said supplement.

References:

• Zemel MB, Bruckbauer A. Effects of a leucine and pyridoxine- containing nutraceutical on fat oxidation, and oxidative and inflammatory stress in overweight and obese subjects. Nutrients. 2012;4:529–541
• Zemel MB, Bruckbauer A. Effects of a leucine and pyridoxine-containing nutraceutical on body weight and composition in obese subjects. Diabetes, Metabolic Syndrome and Obesity: Targets and Therapy. 22 August 2013.

Mitochondrial Super Food: R-ALA, Acetyl-L-Carnitine, Biotin, Nicotinamide (B3), Riboflavin (B2), Pyridoxine (B6), Creatine, CoQ10, Resveratrol & Taurine Optimize Mitochondrial Function.

From China, the biggest (and cheapest) producer of raw materials for dietary supplements comes a study (Sun. 2011) on the effectiveness of a mitochondrial nutrient combination on performance and mitochondrial biogenesis in exhaustively exercised rats, which may well have consequences on the number of items on your next supplement shopping list.

For 4 weeks, Sun et al. supplemented exhaustively exercising rats with a combination of R-a-lipoic acid, acetyl-L-carnitine, biotin, nicotinamide, riboflavin, pyridoxine, creatine, CoQ10, resveratrol and taurine (cf. table 1)

Table 1: Ingredients of the "mitochondrial nutrient supplement";dosage used in rat study (data adapted from Sun. 2011); and calculated human equivalent doses 

This nutrient combination had beneficial effects on standard markers of exercise induced oxidative stress and muscular breakdown. Specifically, it "significantly inhibited the increase in activities of alanine transaminase, lactate dehydrogenase and creatine kinase". The supplementation protocol also had beneficial effects on antioxidant status reversing increases in malondialdehyde and inhibiting the decrease in glutathione S-transferase and total antioxidant capacity in plasma. It also suppressed the elevation of reactive oxygen species in the spleen and thus protected splenic lymphocytes from apoptosis [cell death].

These effects were accompanied / mediated by significant increases mitochondrial biogenesis, evidenced by increases in

[...] the protein expression of mitochondrial complexes I, II and III, mtDNA number and transcription factors involved in mitochondrial biogenesis and fusion in skeletal muscle.

Taken together these results underline that proper nutrition, not only on a macroscopic, but also on a microscopic level, is of paramount importance to exercise performance and metabolic health. Interestingly, the amount of the supplemental "mitochondrial nutrients" used in this study is not even exorbitantly high. In fact, the human equivalent doses (cf. table 1) would be easily attainable by a nutrient-rich diet and some cheap and readily available supplements.

Sunlight, L-Tryptophan & Vitamin B6 With Breakfast Increase Serotonin and Wakefulness During the Day and Melatonin and Restful Sleep at Night

Image 1: For these kids the high tryptophan + B6 breakfast would be useless unless they already got their 10min+ of direct sun exposure this morning

A recently published study that was conducted by a team of international researchers led by Miyo Nakade from the Gakuen University in Nagoya, Japan (Nakade. 2012), concludes that sunlight exposure and vitamin B6 and l-tryptophan intake with breakfast could be profound modulators of circadian rhytmicity in young children (N=816, age = 2-6 years). The results the researchers published in the latest issue of the Journal of Physiological Anthropology clearly suggest that the increased production of serotonin from l-tryptophan and the vitamin B6 metabolite pyridoxal 5’-phosphate (also known as PLP or p5p) exerts beneficial effects on the morningness-eveningness (M-E) score - a measure for the natural and highly desirable difference in wakefulness in the morning and sleepiness in the evening, the disturbance of which is among the emerging correlates of metabolic diseases, such as diabetes, obesity and abnormal lipid metabolism.

[C]hildren showed a tendency to be more morning-typed if they ate breakfast with
a high estimated Trp content. This study also confirmed a similar trend with estimated Vi-B6
content. Among essential amino acids, the Trp content in food is quite small, and thus it is
necessary to make a special effort to consume a sufficient amount in one’s diet.

That those foods with specifically high tryptophan content are, as so often, eggs, meats, and fish is something I probably don't have to tell you - just as I don't have to tell you that soybeans, and other soy products, which also contain significant amounts of tryptophan should not constitute a major part of your diet; regardless of whether you are a man or a woman.

Sunlight must not be hidden behind curtains or seen just through the windows your car

Interestingly, all the vitamin B6 and tryptophan in the world appears to be of little use, if you get too little sun-exposure (Rosenthal. 1997). And, now listen up!, they can become disturbed when you sleep with black out curtains and use an alarm clock to wake you up (Harada. 2003) - in how far these effects are mitigated, or even reversed (i.e. using black-out curtains = beneficial) in people who would otherwise be constantly exposed to unnatural light-exposure (e.g. a street lantern right before your window) would require further investigation, though.

Figure 1: Distribution of early birds (low M-E score on the x-axes) and late risers (high M-E score) among the study population (data calculated based on Nakade. 2012).

Anyways, as you can see in figure 1 the effect of the curtains was statistically significant, but not as pronounced as the scientists' emphasis in their report would have it. There are tendencies for more morning types (lower M-E scores) in the children sleeping without blackout curtains, yes. And there are no real evening types in the children sleeping without blackout curtains, but the broad majority falls into the same 18-23 M-E score range on the 7-28 early bird to long sleeper scale Nakade et al. employed.

Vitamin B6 and inflammation: Although this is only indirectly related to the topic of circadian rythmicity and wake-sleep patterns, it is quite intriguing that vitamin B6 is still touted as an anti-inflammatory agent. And though even very recent population based studies confirm that there is in fact an inverse relation of low plasma vitamin B6 and its active metabolite pyridoxal-5-phosphate (PLP) are inversely associated with the "overall inflammation" score in the US population (Sakakeeny. 2012), another recent study by Ulvik et al. clearly suggests that these associations are corollary, at best (Ulvik. 2012). In their trial, the results of which have been published in the May issue of the American Journal of Clinical Nutrition, Ulvik et al. administered 40 mg pyridoxine hydrochloride per day to patients with stable angina pectoris. Contrary to their expectations, this practice did neither replete the low vitamin B6 levels, nor sooth the inflammation. On the contrary, the rapidly metabolized vitamin B6 appeared to preserve or even increased the expression of inflammatory markers.

What's good in the morning won't hurt at night

With tryptophan being a serotonine precursor and vitamin B6 a necessary co-factor in its synthesis, it may come as a surprise that both, and the sunlight induced serotonin synthesis boost can boost "morningness" and make you more alert. After all, you will probably have read on the label of various dietary supplements that tryptophan is supposed to make you sleepy, right? What those labels don't tell you is that the exposure to sunlight and not the shear availability of precursors and co-factors, let alone the position of the hands on your clock modulate its destiny. And determine whether the tryptophan will get you going, as it obviously does in the children with the highest breakfast intake of tryptophan and vitamin B6 (cf. figure 2), or helps you fall and stay asleep at night.

Figure 2: M-E index relative to group mean in high / low tryptophan & vitamin B6 breakfast intake groups (data calculated based on Nakade. 2012)

That both is possible and obviously works quite fine, is thus the main message of this study, the results of which Nakade et al. interpret as direct evidence for the potential usefulness of

[...] a higher Trp and Vi-B6 intake [to] promote the synthesis of serotonin via light stimulation in the morning and have a natural sleep-inducing effect when converted to melatonin at night [and thus] help prevent a phase delay in young children’s circadian clocks and promote their morningness against the effects of the 24-h commercialization of society.

Against the background, that similar improvements have been observed in 35 middle-aged/elderly (aged 55-75 year) volunteers who consumed a tryptophan enriched  (+30mg per serving) cereal at breakfast and dinner for three weeks by Bravo et al. (Bravo. 2012). And since there is no effect without "side effects", I guess I better mention that the Spanish researchers also observed significant increases in

Table 1: Tryptophan content of various foods (in g per 100g, 2nd column), ordered by tryptophan per protein content (3rd column; data based on Wikipedia)

• sleeping time,
• sleep efficiency, and
• immobile time

as well as concomitant decreases in

• sleep latency (almost 25%!),
• wake bouts,
• total activity, and
• sleep fragmentation index

If you take into consideration that their subjects' total antioxidant capacity levels and mood improved, as well and that the treatment elicited a -50% decrease in state anxiety and a significant drop on Beck's Depression Inventory Index, both of which returned to their initial value, when the treatment was seized, it should be obvious that you better make get out in the sun, immediately after you made the right food choices (and no, I am not talking about tryptophan-enriched breakfast cerals, here) at breakfast and watch the sunset after dinner ;-)

References:

1. Bravo R, Matito S, Cubero J, Paredes SD, Franco L, Rivero M, Rodríguez AB, Barriga C. Tryptophan-enriched cereal intake improves nocturnal sleep, melatonin, serotonin, and total antioxidant capacity levels and mood in elderly humans. Age (Dordr). 2012 May 24.
2. Harada T, Matsumura A, Takeuchi H: Effects of the usage of a blacked-out curtain on the sleep-wake rhythm of Japanese University students. Sleep Biol Rhythms 2003, 1:179–181
3. Nakade M, Takeuchi H, Taniwaki N, Noji T, Harada T. An integrated effect of protein intake at breakfast and morning exposure to sunlight on the circadian typology in Japanese infants aged 2-6 years. J Physiol Anthropol. 2009 Sep;28(5):239-45. 
4. Nakade M, et al. Can breakfast Tryptophan and Vitamin B6 intake and morning exposure to
   sunlight promote morning-typology in young children aged 2-6 years? Journal of Physiological Anthropology 2012, 31:11
5. Sakakeeny L, Roubenoff R, Obin M, Fontes JD, Benjamin EJ, Bujanover Y, Jacques  PF, Selhub J. Plasma Pyridoxal-5-Phosphate Is Inversely Associated with Systemic Markers of Inflammation in a Population of U.S. Adults. J Nutr. 2012 May 23.
6. Rosenthal N, Schwartz P, Tumer E, Nalm S, Matthews J, Hardin T, Barnett R, Wehr T:
   The psychobiology of SAD and the mechanism of action of light therapy. Biol Psychiatry
   1997, 42:57S.
7. Ulvik A, Midttun O, Ringdal Pedersen E, Nygård O, Ueland PM. Association of plasma B-6 vitamers with systemic markers of inflammation before and after pyridoxine treatment in patients with stable angina pectoris. Am J Clin Nutr. 2012 May;95(5):1072-8. 
8. Wikipedia contributors. Tryptophan. Wikipedia, The Free Encyclopedia; 2012 May 26, 00:42 UTC [cited 2012 May 27]

Vitamins B1, B2, B5 & B6 & Glucose Management | Part VII of the "There is More To Glucose Control Than Low Carb"- Series. Any Real Benefit From Supplementing With "Bs"

Funny or obscene? A woman w/ low vitamin B and thus fortified cornflakes is among the "top images" Google will show you, when you search for B-vits

There is an often overlooked reason I am addressing thiamin (B1), riboflavin (B2), panthotenic acid (B5) and pyridoxine (B6) in one installment of the "There is More to Glucose Control Than Carbohydrates"-Series (read previous installments): They are all necessary to store glycogen in the liver (Supplee. 1942).

In general, a whole foods diet, as recommended in previous SuppVersity articles will easily cover the B-vitamin needs of the average sedentary and physically active individual - in spite of minimally increased requirements for B2 & B6, in particular (Manore. 2000; Woolf. 2008).

You can learn more about this topic at the SuppVersity

Proteins, Peptides & Blood Glucose

SFA, MUFA, PUFA & Blood Glucose

Vitamin D & Diabetes

Glucose Manager Calcium?

Flush & No-Flush Niacin & Diabesity

Vitamin C & Glucose Control

As a SuppVersity reader you do yet know that "adequate" and optimal intakes can differ significantly and the fact that the provision of additional B-vitamins does not have ergogenic effects does not exclude the possibility that it may have beneficial effects on blood glucose management.

The initially mentioned inability to convert glucose to glycogen and to store the latter in the liver, for example, would already set you up to increases in blood glucose levels. The latter will in turn increase the urinary loos of the water-soluble vitamins, so that a deficiency in one of the initially named B-vitamins could trigger a whole "pro-diabetic" cascade that leaves the by then (pre-)diabetic individual deficient even in those of the B-vitamins of which he or she is actually getting enough from his or her diet (+ supplements).

Annual spending Alzheimer patients >65y in the US from 2010 to 2050 (projection, in billion U.S. dollars;  Alzheimer's Association. 2010)

This article is exclusively about the beneficial effects of b-vitamins on glucose control: The conclusions I draw based on the evidence presented in this article do not affect potential cognitive benefits from "optimal" (=within the RDA) intakes of B-vitamins (in particularly folate, and B-12, which are not part of this overview, anyway) in the young (Herbison. 2012) and old , where they are furthermore "confined to participants with high homocysteine (above the median, 11 µmol/L) and that, in these participants, a causal Bayesian network analysis indicates the following chain of events: B vitamins lower homocysteine, which directly leads to a decrease in GM atrophy, thereby slowing cognitive decline" (Douaud. 2013).

Conclusive evidence for anti-diabetic or insulin-sensitizing effects of B-vitamin supplements is yet still scarce. Even the notion that (pre-)diabetics suffer from low levels of the said B-vitamins is still controversial. This does not mean, though, that there were no promising study results I could report. For thiamine, for example, ...

• 

  Figure 1: Effects of lipophilic thiamine on HbA1c (top) and insulin requirements (bottom) of type I diabetics (Valerio. 1999(

  Valerio et al. report that the provision of a lipophilic form of thiamine (benzoyloxymethyl-thiamin) at 50mg/day lead to improvements in HbA1c and reduced insulin requirements in children with type I diabetes (Valerio. 1999) - the difference between the active and the placebo arm of the study did yet not reach statistical significance
• Obrenovich et al. report in a 2003 that thiamine, or rather benfothiamine bocks the oxidative damage due to the presence of excessive amounts of glucose in the blood of a rodent model of diabetes - their results have been replicated in human studies by Stirban et al. an other researchers several times over the past decade (Stirban. 2006)

Corresponding evidence for riboflavin is hard to find. While there are studies that suggest the presence of reduced levels of this b-vitamin in both type I and type II diabetics, direct beneficial effects of vitamin B2 supplementation on glucose management have not been reported.

A very similar picture, i.e. reduced levels in type II diabetics, but no reports of direct metabolic benefits from the provision of supplemental vitamin B5 from randomized controlled human trials, emerges if you do a database search for panthotenic acid.

Figure 1: 2h glucose and insulin response to oral glucose tolerance test before (white) and after 25 days of B5 depletion (red), as well as during B5 refeed (violet) in a healthy male subjects (Bean. 1995)

The results of a study from the mid 1950s, when scientists still put healthy individuals on nutritionally deficient diets still indicate. After 25 days without significant amounts of panthotenic acid in the diet, the subjects' insulin sensitivity was notably compromised (Figure 1, red) and was not normalized within only 10 days on a diet with 133x the normal amount of panthotenic acid (Figure 1, violet).

Mind the vitamin <> vitamin interactions: Even if there is no reason for high dose pantothenic acid supplementation to inhibit the cellular uptake of glucose directly, it's well possible that it messes with glucose metabolism via interactions with other water solube vitamins like vitamin B6 aka pyridoxin, the excretion of which is increasing, whenever the intake of panthothenic acid exceeds an (in humans undetermined) sane threshold.

In fact, the extreme elevation of the insulin levels in the "reload phase" would rather suggest that extreme amount of vitamin B5 will compromise, not improve your insulin sensitivity - contrary to edema, severe fatigue, joint pains, reduced protein metabolism, reduced phosphorus, raised VLDL triglycerides, calcification (from calcium pantothenate), dehydration, gastrointestinal symptoms, and depression, a decreased insulin sensitivity is yet not on the "official list of side effects"* of high panthotenic acid intakes (*by "official" I refer to the lists everyone copies ad pastes from the major health information outlets on the Internet).

And what about B6? It's in all my supplements, so it must be good!

If I had to write the bottom line to today's installment of the "There is More to Glucose Control Than Carbohydrates" series now, it would probably be very short and certainly very disappointing for the various supplement junkies out there. Luckily (?) there is still one of the B-vitamins missing: Pyridoxine or vitamin B6 - and you should expect the only B-vitamin that can produce severe toxic effects when it is consumed in very high amounts chronically (peripheral nerve damage) should be able to bring about at least minimal increases in insulin sensitivity / cellular glucose uptake, as well, right?

Well, unfortunately, that's not the case. In 1980, already, a group of scientists from the Gandhi Medical College Hospital in India were able to show that the provision of 40mg of pyrodixine per day had "did not bring about any significant alterations in either the oral glucose tolerance or the insulin response to glucose" in thirteen adult maturity-onset diabetics - and that in spite of the fact that 7 of them were actually vitamin B6 deficient!

Mind the "hidden" B-sources: If you are still concerned that you may not be getting your Bs in, you are probably an OTC supplement junkie. In that case I suggest you briefly take a look at the pre-workout, post- workout and whatever other products in your stack... what? Oh, they all contain 10x the RDA and more of these B-vitamins - that's surprising, right?

A major disappointment? Although this article focused exclusively on the benefits of the water-soluble B-vitamins on glucose control, the results are still paradigmatic for the overall "potency" of vitamin-B-supplements. They are all the rage, but the benefits are overblown, in many cases simply non-existent.

If we discard the well established beneficial effects of benfothiamine on the side-effects of elevated blood glucose levels, and the highly disputed benefits of pyridoxine in diabetic peripheral neuropathies (alleviation of sympthoms, no change in nerve damage; Bernstein. 1988 & 1990), there is actually no reason to even consider taking extra amounts of any or all of these vitamins if you are (a) no diabetic and (b) no junk food eater - and let's be honest, if either (a) or (b) applies you have got more important issues to deal with than potentially suboptimal B-vitamin intakes and their effects on glucose tolerance.

Reference:

• Bean, William B., et al. "Pantothenic acid deficiency induced in human subjects." Journal of Clinical Investigation 34.7 Pt 1 (1955): 1073. 
• Bernstein, A. L., and C. S. Lobitz. "A clinical and electrophysiologic study of the treatment of painful diabetic neuropathies with pyridoxine." Current topics in nutrition and disease (USA) (1988).
• Bernstein, Allan L. "Vitamin B6 in clinical neurology." Annals of the New York Academy of Sciences 585.1 (1990): 250-260.
• Herbison, Carly E., et al. "Low intake of B-vitamins is associated with poor adolescent mental health and behaviour." Preventive medicine 55.6 (2012): 634-638.
• Manore, Melinda M. "Effect of physical activity on thiamine, riboflavin, and vitamin B-6 requirements." The American journal of clinical nutrition 72.2 (2000): 598s-606s.
• Supplee, G. C., R. C. Bender, and Z. M. Hanford. "Interrelated vitamin requirements. The influence of thiamin, riboflavin, pantothenic acid and vitamin B6 on liver glycogen reserves." Journal of the American Pharmaceutical Association 31.7 (1942): 194-198.
• Valerio, G., et al. "Lipophilic thiamine treatment in long-standing insulin-dependent diabetes mellitus." Acta diabetologica 36.1-2 (1999): 73-76.