Yohimbine & Berberine Protect From Death Due to LPS Intoxication; BCAAs Inhibit Serotonin Metabolism & Cause Anxiety, Tryptophan but not SSRIs Help; Sweet Tea Leaves Are PPAR-G Antagonists & Battle High Lipid + Leptin Levels

Skip the fireworks invest the money in some quality ingredients for a fondue or whatever you like and invest the (often non-negligible) rest of the money in a gym membership for the next year.

Actually my figure of the week is 115,000,000 EUR (~152,000,000 US Dollar), which is the sum my fellow country men and women are about to waste on pyrotechnics this year. And a scientifically unconfirmed addition based on my personal observation: 90% of the worst offenders as far as spending money for fireworks goes are at least overweight. Would be interesting to see, if the use of pyrotechnics on New Years Eve is directly associated with fat mass...

I mean, it could be that they spent so much money on their fireworks that they feel they can only afford the junkfood of which everybody and his/her mama still tend to believe that it would be cheaper than buying fresh products and preparing your own food from those.

Ah, I am ranting. That's usually Carl Lanore's task, so I will better go on with the items I have compiled for the today's last installment of On Short Notice in the year 2012:
 

• Berberine + yohimbine - a synergistic duo to prevent LPS toxicity (Li. 2012) -- With all the recent hoopla about the gut microbiome, I suppose that I don't have to tell you what the acronym LPS stands for, right? Hmm... just to make sure it stands for lipopolysaccharide endotoxins which are produced by gram negative bacteria in your gut and are so "toxic" (in fact they cause profound inflammation) that they can be lethal at higher doses.
  
  

  

  Figure 1: Survival rates (%) after ALB/c mice LPS injection (Li. 2012)

  A group of Chinese scientists have now found that aside from berberine the anti-inflammatory effects of which have been known for quite some time now, yohimbine administered in a daily dose of 2mg/kg (human equivalent 0.16mg/kg) does add to the survival rate of berberine treated rodents (human equivalent 4mg/kg) that were injected intragastrically (so not directly into the blood) with a potentially lethal dosage of 20mg/kg LPS. What's more, taken on its own yohimbine is even more potent than the alkaloid that's found in such plants as Berberis aquifolium, Oregon grape, Berberis vulgaris, Berberis aristata, Hydrastis canadensis (goldenseal), Phellodendron amurense, Coptis chinensis and Tinospora cordifolia.
  
  The mechanism is mediated by the prevention of liver injury, an upregulating of IL-10 production (an anti-inflammatory cytokine), and related anti-inflammatory effects resulting from the suppression of phosphorylation of IkBa, JNK, ERK and IRF3 in macrophages.

• Chronic 9-week high BCAA diet impairs brain tryptophan levels and causes anxiety (Coppola. 2012) -- Scientists from the Duke University took another look at the BCAA-tryptophan depression connection, you may have read about in the context of my "Sugar Addicted or Just Stressed Out?" post from January 3, 2012.
  
  According to the results Anna Coppola and her colleagues are about to publish in the American Journal of Physiology  - Endocrinololgy and Metabolism the provision of a BCAA-enriched diet for 9 weeks leads to both reductions in brain tryptophan levels and an increased turnover of serotonin (5-HT) in rodent brains:
  

  

  Figure 2: Composition of low fat  (LF) and high fat (HF) diets with or without added BCAAs (left); effects on the ratio of tryptophan  to the molar sum of large neutral amino acids with and without supplemental  tryptophan in the drinking water and 5HT turnover in the brain (no supplemental trp, right; Coppola. 2012)

  Both groups (BCAA and non-BCAA) consumed about identical amounts of food as the rodents in the complementary (LF or HF) groups, which confirms that the BCAA content did not modify the taste of the chow or rendered it unpalatable (cannot have been cheap bulk powder then ;-). The reduction in both the availability of tryptophan as well as the increase in serotonin (5-HT) turnover in the brain must in fact have been a consequence of the added BCAAs and are most likely the root of the disrupted transport of tryptophan across the BBB in rats, leading to reduced exploratory behavior of rats in EPM testing, a sign of increased anxiety.

  "Recent studies demonstrating a strong  association between BCAA levels, obesity, and obesity-related metabolic disorders, when linked to the findings reported here, may help to explain the strong association between obesity and behavioral abnormalities, including depression and anxiety." (Coppola. 2012)

  

  As the slight differences between the high an low carb diets show, other nutrients can influence serotonin as well (read more)

  In this regard it is important to point out that these negative side effects were mostly reversible by the provision of 15 mg/100 ml tryptophan in the drinking water of the rodents, but were not alleviated by  the administration of the common serotonine reuptake inhibitor fluoxetine (at 10 mg/kg/day for four weeks).
  
  Bottom line: Isolation is not what you want if what your body has been build for is complex food. And while the single serving of BCAAs you may gulp down during or right before a workout, on the other hand, probably isn't going to harm you. The "I need BCAAs every 30min" approach to gaining muscle mass, may well turn you into a psychotic wrack if you follow it day in and day out for months or years - at least without chronically adding some l-tryptophan to the equation.

• Sweet tea leaves protect against obesity: Once more via PPAR-gamma blockade (Zhou. 2012) -- Actually this is probably not news to anyone out there with a degree in Traditional Chinese medicine. After all, Lithocarpus polystachyus Rehd.(Sweet Tea) is Chinese folkloric medicine that has always been used to treat obesity, diabetes, and hypertension in South China:
  

  "Previous experiments revealed that it contains plentiful bioactive flavonoids and polyphenolic compounds, e.g. phlorizin, trilobatin, 3-hydroxy-phlorizin, etc. These components have extensive pharmacological activities, such as anti-diabetes, memory improvement, anti-aging, inhibition of lipid peroxidation and the growth of human colon cancer cells, and so on." (Zhang. 2012)

  From a "scientific" perspective, however, the efficacy of this herbal medicine as an obesity treatment had still to be elucidated.

  

  Figure 4: Effects of oral gavage of 75 mg, 150 mg and 300 mg/kg of body weight/day of sweet tea extract or placebo (DIO) in conjunction with the 8 weeks on a obesogenic diet (Zhang. 2012)

  In this context it is yet worth mentioning that this study demonstrated for the first time that the aqueous dry leaves extract of Lithocarpus polystachyus Rehd. can potently reduce the worst metabolic side effects of obesity, such as the hypolipidemia, hypoleptinaemia and the degree of insulin resistance (FINS, HOMA-IR, cf. figure 3) what it does not answer, however, is whether the decline in PPAR-gamma is tissue specific, what exactly is behind the profound decline in leptin levels and whether or not lean rodents, let alone humans, who don't consume an obesogenic diet will see anywhere similar benefits.
  
  In other words, this is research in progress, but I suppose something you are going to hear more about at the Supppversity in 2013.

* * * * * *

Apropos hearing or rather reading more, I guess you will realize that you have reached the end of today's installment of On Short Notice which means that you will have to progress to the SuppVersity Facebook Wall if you want a second serving of news on...

• The history of vitamin A as a light sensor and beyond - actually a free full-text I guess those of you who like to "think paleo" may enjoy (read more)
• A paper on "good" and "bad" inflammation, where the author points out that soothing inflammation too much can lead to a reduction in energy expenditure and may therefore not be the king's road to getting rid of the last blubber (read more)
• The food-hitlist of young Americans - Featuring sugar, sugary drinks, sugary bakery, sugary ... as their main energy and carbohydrate sources... (read more)
• Problems with synthroid and generics that have surfaced in a recent study on their efficacy in the treatment of congenital hypothyrodism (read more)

as well as a handful of other news, which are already there or are going to be posted within the next hours. Have a great weekend, everyone! 

References

• Coppola A, Wenner BR, Ilkayeva O, Stevens RD, Maggioni M, Slotkin TA, Levin ED, Newgard CB. Branched-chain amino acids alter neurobehavioral function in rats. Am J Physiol Endocrinol Metab. 2012 Dec 18.
• Li H, Wang Y, Zhang H, Jia B, Wang D, et al. Yohimbine Enhances Protection of Berberine against LPS-Induced Mouse Lethality through Multiple Mechanisms. PLoS ONE. 2012; 7(12): e52863. 
• Zhou CJ, Huang S, Liu JQ, Qiu SQ, Xie FY, Song HP, Li YS, Hou SZ, Lai XP. Sweet tea leaves extract improves leptin resistance in diet-induced obese rats. J Ethnopharmacol. 2013 Jan 9;145(1):386-92.

Taurine + BCAAs - Scientists Identify Unkown Synergy of Branch-Chained and Sulfur-Amino Acids: Redutions in DOMS, Faster Recovery and Reduced DNA Damage

If this is true and sore is the new sexy, the combination of taurine + BCAA's may turn you into an ugly worm.

You know them and I would bet that >75% of you have already taken them: Branch-Chained Amino Acids (BCAAs) and the sulfur-amino acid taurine. Maybe you have taken the former for their beneficial effects on skeletal muscle protein synthesis and the latter for its anti-oxidant effects and the cascade of beneficial downstream effects I have written about quite extensively, here at the SuppVersity.

If I am asking you whether you have taken both in conjunction as a means to reduce post-workout delayed-onset muscle soreness and the expression of purported markers of muscle damage, on the other hand, I'd expect only few people to raise their hands... right?

Sometimes it's worth taking another look

The currently available literature on the beneficial effects of BCAAs on DOMS is pretty inconclusive. If you restrict your review of the literature to studies using resistance training as a trigger for muscular damage (Jackman et al. (2010), Howatson et al. 2012; etc.), it does yet appear warranted to say that the chronic ingestion of a high dose of BCAAs can ameliorate the peak in delayed muscle soreness after 24-48h.

You can learn more about taurine & BCAAs at the SuppVersity

Taurine Pumps Up Strength & Recovery?

Taurine Improves Insulin + Glucose Metabolism

Taurine = 180% Testosterone Increase

Leucine Only Tops Ergogenic Effects of BCAAs

43% Reduced Performance W/ BCAAs

BCAA Neurotransmitter Depletion

In the previously referenced 2012 study by Howatson et al. we are talking about a ~50% reduction after 24h and a 25% reduction after 48h. Both statistically and physiologically relevant, but if the decrease in maximal voluntary contraction had not been blunted, as well, it'd been another instance of much ado about nothing. Similar beneficial effects have been observed with taurine, as well:

• 

  Taurine & caffeine make another super-stack; but only at the right ratios | learn more

  Zhang et al. report that taurine can "attenuate exercise-induced DNA damage and enhance the capacity of exercise due to its cellular protective properties" in the musculature of healthy young men (Zhang. 2004)
• Silva et al. observed in a rodent model of skeletal muscle damage in response to eccentric exercise that taurine decreases the oxidative stress, in association with decreased superoxide radical production (Silva. 2011)
• learn more in previous SuppVersity articles about taurine

Against that background it was to be expected that the previously sedentary subjects of a very recent study from the University of Tsukuba in Japan was attenuated, irrespective of whether they were taking 3x3.5g of BCAAs or 3x2.0g of Taurine for 2 weeks before they performed a standardized eccentric exercise test:

"For the ECC protocol, subjects were seated on a bench with their arm positioned in front of their body and resting on a padded support, such that their shoulder was secured at a flexion angle of 0.79 rad (45°) and their forearm was maintained in the supinated position throughout the exercise. Subjects were repeatedly weight-loaded upon
dumbbell lowering to achieve a 90% MVC (34.3 ± 1.3 Nm). Subjects performed six sets of five repetitions of elbow extension from the flexed position at 90° to the fully extended position slowly over 5 s, while maintaining a constant speed of movement by following a verbal metronome provided by the investigator." (Ra. 2013)

What we could not necessarily be sure of is whether these effects would also add up in those 12 untrained male subjects (22.5 ± 3.8 years) who were assigned to the taurine + BCAA group.

Figure 1: Post workout muscle soreness, left; post workout arm circumference in response to cell swelling, right (area under the curve for the 96h after the eccentric exercise test; based on Ra. 2013)

Now that you've taken a glimpse at the data in Figure 1, it's probably pointless to ask you to make an educated guess. It's too obvious that the individual DOMS reducing effects of taurine and BCAAs add up. What's yet even more obvious is that only the combination of both leads to a rapid reduction in muscle swelling, the effect size of which goes far beyond what additive effects could achieve - this indirect marker of muscle damage would thus suggest that there is a special synergy between taurine and BCAAs, a synergy due to which a reduction of 35mm/96h + 45mm/96h (the individual changes for BCAAs and taurine) does not translate into a -80mm/96h, but into a 465mm/96h reduction of this commonly used indirect measure of skeletal muscle damage.

"Synergy" is the name of the game

If you take a parting look at the data in Figure 2, you should actually be able to understand why the combination of branch-chained amino acids and the sulfur amino acid taurine works so well: One excels where the other has only minor effects.

Figure 2: CK, left, and 8-hydroxydeoxyguanosine (8-OHdG), right (area under the curve for the 96h after the eccentric exercise test; based on Ra. 2013)

While the branch-chained amino acids have a more pronounced effect on the expression of CK and LDH (not shown in Figure 2), they do very little to protect the muscle from oxidative damage (as indicated by the quasi non-existent effect on the levels of 8-hydroxydeoxyguanosine (8-OHdG), a marker of DNA damage.

Suggested read: "Rats 'On' Taurine Can't Ever Get Enough... Exercise of Course! What Were You Thinking About? Mice Cover 50% More Distance W/ HED of 3-4G of Taurine Post Workout " | more

Bottom line: Actually there is very little I have to add to the researchers conclusion that "his study confirmed that a combination of 3.2 g BCAA and 2.0 g taurine, three times a day, two weeks prior to
and three days after exercise attenuates some subjective and objective markers of DOMS and muscle damage induced by high-intensity ECC, which could not have been influenced by BCAA or taurine supplementation alone." (Ra. 2013)

I am not 100% sure if they are also correct in their assessment that this supplement is particularly useful for beginners who would be more motivate to continue an exercise program, if it doesn't hurt so much, though. That it could help competitive athletes to train at higher intensities on the other hand, is something I would fully subscribe - whether that's necessarily going to be more productive, on the other hand, is question I would not want to answer without a follow up study ;-)

References:

• Jackman, S. R., Witard, O. C., Jeukendrup, A. E., & Tipton, K. D. (2010). Branched-chain amino acid ingestion can ameliorate soreness from eccentric exercise. Med Sci Sports Exerc, 42(5), 962-970.
• Ra, S. G., Miyazaki, T., Ishikura, K., Nagayama, H., Komine, S., Nakata, Y., ... & Ohmori, H. (2013). Combined effect of branched-chain amino acids and taurine supplementation on delayed onset muscle soreness and muscle damage in high-intensity eccentric exercise. Journal of the International Society of Sports Nutrition, 10(1), 51.
• Silva, L. A., Silveira, P. C., Ronsani, M. M., Souza, P. S., Scheffer, D., Vieira, L. C., ... & Pinho, R. A. (2011). Taurine supplementation decreases oxidative stress in skeletal muscle after eccentric exercise. Cell biochemistry and function, 29(1), 43-49.
• Zhang, M., Izumi, I., Kagamimori, S., Sokejima, S., Yamagami, T., Liu, Z., & Qi, B. (2004). Role of taurine supplementation to prevent exercise-induced oxidative stress in healthy young men. Amino acids, 26(2), 203-207.

Branched Chain Amino Magic: Study Takes Another Step Towards a Better Understanding of the Anabolic & Anticatabolic Effects of BCAAs and Their Essential Cousins

Image 1: Without the other essential amino acids (EAAs), the branched chain amino acids, leucine, isoleucine and valine (BCAAs) have nothing to "build" your muscle from ;-)

Usually, I do not get very excited, when I hit upon another study into the "protein-synthetic response" that is triggered by the ingestion of branched chain amino acids (BCAAs). I mean, let's be honest... we all know that their ingestion will trigger the phosphorylation of the mammalian target of rapamycin and thusly increase protein synthesis, so why would we need another study where instead of a 17.5% increase in protein synthesis, we would see a 18.3% increase? Actually, we don't... the data Marcus Borgenvik, William Apró and Eva Blomstrand from the  Åstrand Laboratory, Swedish School of Sport and Health Sciences and the Karolinska Institutet, in Stockholm, Sweden (Borgenvik. 2011), collected goes yet well beyond what we have seen in most of the previous studies and is thus well worth an individual blogpost here at the SuppVersity.

BCCAs work! How? Little do we know...

If we are honest, we must concede that our (=the scientific) understanding of the complex processes that are triggered when "large" amounts of BCAAs hit our bloodstream, is very limited. What we know is that we can measure increases in mTOR-expression that correlate with likewise measurable increases in protein synthesis. What we do not really know is how exactly one leads to the other and where the influences of amino acid supplementation and exercise training overlap. This is even more true for the complementary side of the protein synthetic equation of which Borgenvik et al. state that

[w]hereas extensive evidence for the stimulatory effect of amino acids, either alone or in combination with exercise, on protein synthesis has been reported, their effect on protein breakdown is elusive.

In that, it is particularly confusing that "previous investigations involving ingestion of essential amino acids (EAA) in connection with resistance exercise have revealed no attenuating effect on protein breakdown", while studies which investigated the effect of BCAA or leucine in isolation, report reduction in protein degradation in subjects at rest or performing eccentric endurance exercise (MacLean. 1994). Reason enough for the Swedish scientists to recruit a group of seven healthy, recreationally active participants (5 men, 2 women; 27 (± 2) years; height 175 (± 5) cm; weight 67 (± 7) kg), put them on a standardized diet (17% protein; 25% fat; 57% carbs; ~2100kcal for women, ~2700kcal for men) for two days and, on the subsequent morning (subjects reported to the lab fasted) and after a thorough warm-up, have them perform

• 4 sets of 10 repetitions at 80% of their predetermined 1 RM, followed by another 
• 4 sets of 15 repetitions at 65% of their 1 RM of single-legged leg presses.

The subjects used the same leg on all exercises and rested ~5min after each set. Before the warm-up, during and immediately after and 15 and 45min after the exercise regimen the subjects consumed either

• 150 mL of BCAAs (2:1:1 ratio) in flavored water, or
• 150 mL flavored water alone.

The total amount of BCAAs was 85mg/kg or 5.695g for the "average" study participant. After four weeks the experiment was repeated with each participant receiving the opposite treatment.

Figure 1: Complete analysis of serum amino acid levels in the trained and untrained leg of subjects receiving BCAA or Placebo supplement before, during and after the completion of a standardized single-legged leg press exercise (data adapted from Borgenvik. 2011)

As far as the study protocol goes this is thus certainly not an extraordinary study. If you take a look at figure 1, where I deliberately plotted all the data the scientists gathered as far as serum amino acid concentrations are concerned, you will yet realize that what makes this study stand out is the sheer amount of parameters Borgenik et al. have analyzed. Similar data is also available for the amino acid concentrations in the exercised muscle and though, the scientists, who set out to investigate the effects of BCAA supplementation on protein breakdown, would probably disagree with me, here, I feel that this data actually has the most real world significance for physical culturists, like you and me.

Figure 2: Relative increase / decrease in intra-muscular BCAAs and other EAAs in BCAA supplemented subjects vs. placebo control at different time-points before, during and after single-legged leg presses (data adapted from Borgenvik. 2011)

After all, a brief glance at the effects the ingestion of ~6g of BCAA had on the respective tissue levels of leucine, isoleucine and valine (figure 2, BCAA) and the other, "missing" essental amino acids (figure 2, EAA - BCAA) should suffice to understand that though BCAAs may be the necessary to trigger protein synthesis, they are yet obviously not sufficient to "build muscle" - or how else would you explain the

pronounced reduction in the concentration of the aromatic amino acids, tyrosine and phenylalanine, in both plasma and muscle as well as muscle EAA (BCAA excluded) during the recovery period

Borgenik et al. observed in their study? The scientists at least conclude that

[s]ince tyrosine and phenylalanine are neither synthesized nor degraded in skeletal muscle, reduction in the levels of these amino acid could be indicative of an improved net muscle protein balance, i.e. an enhanced rate of synthesis and/or decreased rate of breakdown [and] could  be explained by incorporation into protein.

The accrual of muscle mass (whatever that may eventually mean, cf. yesterday's installment of the Intermittent Thoughts) thusly obviously relies on the presence of all essential amino acids and not just the "branched chained holy grail" of protein synthesis, of which the current study revealed that they (BCAA ingestion) reduced the expression of MAFbx, which regulates the protein transcription factor MyoD and the eukaryotic initiation factor-3f (eIF-3f), which, in turn is of importance in the mTOR-p70S6k signaling pathway, by 30% and 50% in the resting and exercising legs, respectively.

Figure 3: Relative (compared to placebo) mTOR and p70S6K phosphorylation in response to BCAA supplementation in exercised (EX) and non-exercised (Rest) leg at different time-points before, and after single-legged leg presses (data adapted from Borgenvik. 2011)

As figure 3 finally goes to show, we see the "usual" increases in mTOR and p70S6K phosphorylation that are commonly held responsible for the downstream increases in protein synthesis, and which were obviously more pronounced in the exercised compared to the non-exercised leg. The latter may be ascribed to what the scientists cautiously label a ...

[...] tendency for BCAA supplementation to attenuate the elevation in the level of Rheb mRNA in both resting (1.7-fold under the placebo versus 1.2-fold in the BCAA condition) and exercising muscle (2.4-fold versus 1.5-fold).

This ameliorative effect on Rheb, the low-molecular weight GTPase located immediately up-stream of mTOR, in combination with the exercise induced reductions in REDD2 expression (another negative regulator of mTOR) the scientists observed in the exercised leg are actually where we are currently at, as far as our understanding of the complex protein synthetic machinery goes. It is here at the gene-level where amino acid supplementation and its effect on Rheb and exercise and its effect on REDD synergize and facilitate those muscle gains trainees have been making for years often without any understanding of the biological underpinnings.

And though we may eventually be able to squeeze out another 5-10% more muscle mass, when we eventually get the "whole picture", I seriously doubt that even the most thorough understanding of the underlying biomolecular processes will change such basic recommendations as "take your 25g of fast digesting whey as a bolus immediately post workout" (cf. "Never Sip Your Whey!") - or what would you say?

Adelfo Cerame - Road to The Wheelchair Nationals '12: My Five Simple Tricks for Guilt Free Thanksgiving Celebrations. Plus: Four Dietary Supplements You Should not Miss!

Image 1: Adelfo on Wednesday before the feast began. We will see how he looks next week ;-)

I guess, this blogpost should actually begin with the words "Happy Thanksgiving" in big, bold letters, but for us Europeans - and me as a German, in particular - today is a regular working day. Nevertheless, it is a day to celebrate, because THIS is actually post #600 here at the SuppVersity and I am in fact grateful (and thusly giving thanks) that you, my dear students, readers, followers or however else you want to refer to yourselves, are still interested in my perspective(s) on what is going on in the world of fitness, bodybuilding, sports and exercise science.

That being said, I do still hope that all of you (who actually have a holiday) are having a good time with your friends and family, just like my friend Adelfo, who has sent me his weekly contest-prep update and his best wishes to all of you last night, already ....

Turkey, pie, ... pie, turkey, ... turkey ... and some supplements!

Ok guys, I’m going to try and make this real brief because I actually started my Thanksgiving Day Feast a day early, so that I’ve been falling in and out of sleep from all the turkey and pie and still have to prepare for another day of furious eating again tomorrow, in order to "peak" (just to stick to the bodybuilding terminology) right on Thanksgiving!

Image 2: I like protein blends and Lean Supreme is one of my favorites. I gets it for $32 for 4.2lbs at my local Nutrition Zone.

As you know from the previous installments of this series, I have planned these 2 cheat / refeed days long ahead. In fact, being able to go somewhat overboard on Thanksgiving, Christmas, New Years Eve and my birthday was part of the reasoning behind starting my contest preparation so early.  But before I give you guys some pointers on how to approach these holiday feasts, I thought it was about time to address the subject of supplements. I know I've hinted at being on a budget and cutting back on my usage of protein powder in a previous episode; luckily I have recently been able to stock back up on my four staples, i.e. those supplements, I think are really worth spending your money on.

For me the "basic four" are

• a high quality protein powder, 
• a bag of bulk BCAAs, 
• a quality multi-vitamin, and 
• the good old creatine monohydrate. 

Protein powder, BCAAs, a mulit and creatine are what I would call the "foundation" of my supplementation regimen. If I cannot afford all of these, I am not going to buy any other  supplement - "prioritizing" is the name of the game. If my budget allows that I am yet planning on experimenting with a few other products in the weeks to come. What I currently have on my mind are a decent fat burner, some bulk D-Aspartic Acid and a test booster if I can find one that actually works...

Image 3: Protein powder, multi-vitamins, creatine monohydrate, and BCAA’s, combined with good training + nutrition, and that’s all you need to build your physique… well at least for me it is ;-)

Remember, all four, the protein powder, the BCAAs, the multi-vitamin and the creatine are add-ons; they are not part of what I call my "nutritional regimen". They do not replace any nutrients (as Dr. Andro would say: "They are called supplements not replacements" ;-) and I am well aware that I won't get anything out of them, if my diet and training are not 100% in check. Never fool yourself into the false security that you are taking supplements, X,Y and Z and could thusly let your nutritional protocol slide. No supplement gives you the excuse to eat like shit or skip a training session! You can do without quality supplements but you can’t do without quality foods - although the colorful adverts in some of the muscle mags, try to make you believe otherwise.

Image 4: Huge thanks to Jack Gurlekian and Dr. Andro for helping me out on the supplement side.

Before I get started on Holiday feasting, I’d like to give a shout out to Jack Gurlekian and thank him for hooking me up with some of the supplements that he distributes and makes himself… I’d also like to thank Dr. Andro himself for helping me out with finances to help me purchase some supplements that should hold me off for a couple months : Creapure(TM), glutamine peptides & ZMA from True Protein. Inner Armour supplements and Jack’s very own amino acid blends! [A note by Dr. Andro: Additional sponsors are always welcome ;-]

My holiday season feasts 101!

The holiday season is approaching and you all know what that means? Epic holiday feasts and it starts with Thanksgiving, continues on Christmas and ends with New Years Eve. And when it’s all said and done, all you’re left with is a GUT full of regret! In order to help you to keep both the gut, as well as the regret at bay, I want to share with you how I like to prepare myself for the holiday food gauntlet…

Image 5: This thanks giving plate looks way better than the government's MyPlate

Actually it all begins with keeping myself in good physical shape and keeping my physique as lean as I can year round. If you read the last installment of Dr. Andro's Intermittent Thoughts Series on "Why to cut before bulking" you will be aware of the hormonal and metabolic advantages which come with being lean. That being said, there is actually little reason, why I could not enjoy occasions like this without any regret, if it's not just a few days out on the next contest, which obviously isn't yet the case... and if you have been following the whole series since September, you will also be aware that I am exactly on schedule - a schedule that allows for a long consistent cut, precisely because I knew that Thanksgiving, Christmas and New Years Eve were coming and being in decent shape by then would allow me do some serious and more importantly guilt free eating ;-)

5 simple rules to survive the feast without too much damage

Being on an intermittent fasting protocol does help here as well, I have already touched on that in a previous installment - when you pack your complete caloric intake into a small feeding window "overeating" (no, not eating like shit!) is part of the game, anyway. Ok, the foods are probably different on Thanksgiving, but the caloric intake may in fact not vary too much. Moreover, the muscle specific improvements in insulin sensitivity that come with training fasting make it less likely that you store all the good and not so good nutrients you are about to eat today in the form of glycogen or even new muscle tissue, rather than as triglyceride droplets in your love handles.

Personally I do adhere to the the following 5 principles for intermittently fasted holiday celebrations in order to keep any potential damage to my physique at a minimum:

1. I give myself a caloric buffer throughout the week before the big day. I don’t starve myself but I don’t eat as much. I try to be like around 200-300 calories shy of my daily caloric intake (1800 kcal). And being on IF, I do not even notice this 200-300 calorie reduction as far as my appetite is concerned.
2. I stick to proteins and moderate fats throughout the week before the big feast. Just sticking to proteins, moderate fats, veggies and fruits, will help you with making that caloric buffer, simply because they are satiating. I still had my PWO chocolate milk, and PWO carbs with my meal.
3. I fast before the big feast. You may want to even throw in an intense and heavy weight training session before, so you can get more bang for your buck when you eat! (Though I wont be doing that because it’s my rest day)
4. Once I get to the party and begin my feast, I focus first on filling up on protein like turkey, chicken, roasts, fish and BBQ meats. This way I’m sure to hit my protein macros and to fill up with some good protein, then I’ll have my desserts like cakes and pies … and a couple shots of liquor and a couple glasses of wine ;-) But once I’m full… That’s it! I wont force myself to eat more just because there’s a lot of food. I’ll wait a couple of hours like I would normally do (on regular days), until I’m hungry again before I’ll eat again. I try to get in the same amounts of meals that I usually would during my feeding hours (3-4 meals) but obviously the meals will be much bigger... yeah, and with a little bit more junk ;-)
5. I push away the remote thoughts of guilt and enjoy the day! I mean, you train hard and eat smart year round to keep yourself lean & mean. One day is not going to reverse all your efforts, maybe some bloat and a little water retention, but that will subsides in a couple days. Other than that… Enjoy your Thanksgiving, because I know I will!

Happy Thanksgiving Everyone! Eat responsibly and don’t eat & drive full ;-)

Chronic High Dose BCAA Supplementation Reduces Endurance Performance by 43% Plus: How Ammonia, Glutamine, Arginine & Low Carb Could be Involved

Tired, exhausted, had to cut your workout short today? Is it the flu, or just too much BCAAs?

When some is good and more is better, even more is not necessarily going to be 'betterer' - and that's not simply due to the fact that there is no comparative to an adjective that's already in the comparative. Therefore it is actually not surprising that a team of researchers from the Department of Food and Experimental Nutrition at the Faculty of Pharmaceutical Sciences, the Department of Nutrition at the School of Public Health and the Department of Physiology and Biophysics at the Institute of Biomedical Sciences of the University of Sã o Paulo in Brazil has just published the results of a study (Falavigna. 2012) which demonstrates that there is an upper limit to the benefits of BCAA supplementation. What I guess will be surprising at least for some not so regular SuppVersity visitors, is that there is more than just a saturation effect: Too much BCAAs can actually have ergolytic (= anti-ergogenic) effects - at least under certain circumstances.

Another chapter in the book of good things that turn against you, when taken in excess

In their latest paper that has just been published in nutrients, Gina Falavigna and her colleagues analyzed effects of chronic BCAA supplementation on exercise performance in male Wistar rats. Based on previous animal and human data and the still widely supported, though actually experimentally non-validated (cf. Meeusen. 2007) theory that BCAAs would work their non-hypertrophy specific, endurance enhancing magic via the blockade of exercise induced 5-HT (serotonin) accumulation in the brain, the researchers speculated that ...

"[...] chronic BCAA supplementation (through the diet, using different BCAA  concentrations) would increase performance in rats subjected to a swimming exhaustion  test." (Falavigna. 2012)

To verify this hypothesis, Flavigna et al. randomized their rats to three different groups receiving either the standard AIN-93M diet for the maintenance of adult rodents (control group) or the same diet with additional additional 3.57% (group S1) and 4.76% (group S2) BCAAs at a ~2:1:1 ratio of lecine : valine : isoleucine (the BCAAs were manufactured by the Brazilian branch of Ajinomoto). The rodents in the S1 and S2 groups did thus receive 50% and 100% more branched-chain amino acids than the rodents in the control group which had to contend themselves with the BCAAs in the casein fraction of their diets (see figure 1, right). In order to assure that the diets would be isocaloric, an amount of starch equivalent to the amoung of BCCAs that had been added to the chow was removed from the supplemented diets.

Overall, the study lasted for six weeks. During this time the rodents were subjected to a 1h/day weight bearing swimming protocol five times a week. In the first two weeks, the rats were ...

"[...] adapted to the water medium and exercised with increasing overloads attached to the tail until an overload corresponding to 5% of total body weight was reached. This final overload was used until the end of the training protocol [...] The overloads were corrected weekly according to the variations in animal weight.  The efficiency of the training protocol was assessed on the basis of maximum activity of the enzyme citrate synthase in the soleus muscle, with a group of sedentary animals being used as the control for this parameter." (Falavigna. 2012)

Neither the overall amount of food nor the body weight gain of the rodents in the control, and the two exercise groups showed any statistically significant difference. The latter cannot be said about the exercise performance, as well as the accumulation of ammonia, though (see figure 1):

Figure 1: Exercise duration and plasma ammonia levels during / after swmming test (left) and macronutrient composition of the experimental diets (right; based on Falavigna.. 2012)

While the rodents in the +50% BCAA group (S1) do show the expected increase in endurance (+37%) their peers in the high dose (+100%) BCAA group (S2) experienced an even more pronounced drop in endurance performance (-43% vs. control), which went hand in hand with a profound increase in blood ammonia (+34%).

"Ammonia is a ubiquitous metabolic product producing multiple effects on physiological and biochemical systems. Its concentration in several body compartments is elevated during exercise, predominantly by the increased activity of the purine nucleotide cycle in skeletal muscle. Depending on the intensity and duration of exercise, muscle ammonia may be elevated to the extent that it leaks (diffuses) from muscle to blood, and thereby can be carried to other organs. The direction of movement of ammonia or the ammonium ion is dependent on concentration and pH gradients between tissues. As such, ammonia can also cross the blood-brain barrier, although the rate of diffusion of ammonia from blood to brain during exercise is unknown. It seems reasonable to assume that exhaustive exercise may induce a state of acute ammonia toxicity which, although transient and reversible relative to disease states, may be severe enough in critical regions of the central nervous system (CNS) to affect continuing coordinated activity. Regional differences in brain ammonia content, detoxification capacity, and specific sensitivity may account for the variability of precipitating factors and latency of response in CNS-mediated dysfunction arising from an exercise" stimulus, e.g., motor incoordination, ataxia and stupor. There have been numerous suggestions that elevated ammonia is associated with, or perhaps is responsible for, exercise fatigue, although evidence for this relies extensively on temporal relationships." (Falvigna. 2012; my emphasis)

Mark the last words of the previously cited paragraph: "[E]vidence for [the role of ammonia] in exercise fatigue relies extensively on temporal relationships". It is thus - as for now - a solely corollary, not yet a causative association, of which I do however feel that it would be very likely to turn into a causal one if someone actually measured the influx of ammonia into the brain during a workout.

Wait, ammonia? But ain't it more likely that the BCAAs block the uptake of tryptophan?

What's for sure is that another hypothesis, which relates to the blockade of tryptophan uptake can be ruled out as an underlying reason of the differences. After all the scientists who argue that ...

"[t]he increased synthesis of serotonin during exercise may be related to the development of central fatigue, because this neurotransmitter has several physiological functions, since it operates by  mood, lethargy, individual behavior, regulation of sleep, body temperature and blood  pressure, appetite suppression and changes in perceived exertion." (Falavigna. 2012)

...actually measured the 5-HT levels and observed no differences between the dietary groups. Overall, the study results to thus clearly indicate that both, medium nor high dose "chronic BCAA supplementation was not effective in improving the main parameters indicative of central fatigue" (Falavigna. 2012) - well, at least as long as we still stick to the hypothesis that the latter is induced by the accumulation of 5-HT in the brain.

Forget about tryptophan and serotonin, focus on ammonia

The fact that neither the high, nor medium dose of BCAAs did exert any effects on the serotonin levels in the brain does yet not explain why the medium dose supplementation regimen produced ergogenic, while the high dose regimen induced ergolytic effects.

The occurrence of direct toxic effects due to (too) high amounts of branched-chain amino acids can be ruled out based on previous studies in which the administration of more than 10g/kg body weight of BCAAs (the human equivalent would be 130g+ per day), as well as dosages of 2.5g/kg body weight chronically did not entail any toxic side effects (Shimomura.  2004). The same is true for other confounding variables, such as the citrate synthase activity, a measure of the general efficiency of the training protocol, bood glucose, insulin,free fatty acids, and lactate levels, as well as liver and muscle glycogen content, which were virtually identical in both groups. This leaves us with the increase in plasma ammonia as our 'last resort' to explain the -58% shorter swimming time in the high (S2) vs. medium (S1) dose BCAA group (-43% lower vs. non-supplemented control).

Figure 2: The reduced performance of the high BCAA group could well be related to peripheral and/or central ammonia build-up as a results of increased BCAA oxidation, camparably low glutamine intakes and the rate-limited enzymantic conversion and recycling of gluatmine (illustration originally from Earrante. 2003). Studies by Snow (2000) and Carvalho-Peixoto (2007) suggest: Both carbohydrate & glutamine supplements could help.

Based on what we know about the mammalian body, the increased build-up of ammonia in the high BCAA group could be a result of the unfortunate combination of temporary energy shortage and learned wastefulness' in a situation, where the otherwise sparse BCAAs are available in abundance. Furthermore, with a glutamine content of only 9-13% in the casein fraction of their diets (Swails. 1992), the rodents in the high BCAA group did ingest more than 2.6-3.8 times more BCAAs than glutamine; a fact which may have contributed to a temporary glutamine deficiency as a result of its increased use in the detoxification of the ammonia that's generated when the BCAAs are oxidized. The resulting peripheral and possibly central ammonia build-up (see figure 2) could then have begun to intoxicate liver and brains of the rodents and thus hampered gluconeogensis (normal levels stimulate, high levels of ammonia hamper gluconeogensis; cf. Fritz. 1988) and induced central fatigue (Wagenmakers. 1990; Nybo. 2004) -- and that not despite, but rather due to the chronic "high dose" BCAA supplementation (HED ~50g/day).

So do I have to drop my BCAAs now or what? Whether these results are relevant for you will probably depend on a whole host of parameters, which include

• the type, intensity and duration of exercise you do, 
• the ratio of BCAAs to glutamine in your diet,
• the amount of arginine, which acts as a substrate for the urea cycle and is therefore necessary to for the excretion of ammonia by the kindeys (Schaefer. 2002),
• the amount of carbohydrates in your diet (with more = less amino acid oxidation = lower ammonia and very low carb = you are in trouble; e.g. Czarnowski. 1995; Snow. 2000; Carvalho-Peixoto. 2007), 

... and those factors I will probably have forgotten to mention now. Unless you don't forget that you can neither lifve from BCCAs and protein alone, but accept the neflglected truth that too much protein is about as bad a too little protein, you can file this post under "show your stupid friends" and get back out, when they complain about feeling sick, bloated and fat "despite" eating a BCAA supplemented high protein, low carb (and often even low fat) diets.

References:

• Carvalho-Peixoto J, Alves RC, Cameron LC. Glutamine and carbohydrate supplements reduce ammonemia increase during endurance field exercise. Appl Physiol Nutr Metab. 2007 Dec;32(6):1186-90.
• Errante LD, Petroff OA. Acute effects of gabapentin and pregabalin on rat forebrain cellular GABA, glutamate, and glutamine concentrations. Seizure. 2003 Jul;12(5):300-6.
• Falavigna G, de Araú jo Junior JA, Rogero MM, de Oliveira Pires IS, rio Graç a Pedrosa R, Martins Junior E, Alves de Castro I, Tirapegui J. Effects of Diets Supplemented with Branched-Chain Amino Acids on the Performance and Fatigue Mechanisms of Rats Submitted to Prolonged Physical Exercise. Nutrients 2012. 4; 1767-1780.
• Fritz S, Bohnensack R. Stimulation of alanine metabolism in rat liver by ammonia. Biomed Biochim Acta. 1988;47(12):923-32.
• Meeusen R, Watson P. Amino acids and the brain: do they play a role in "central fatigue"? Int J Sport Nutr Exerc Metab. 2007 Aug;17 Suppl:S37-46.
• Nybo L, Dalsgaard MK, Steensberg A, Møller K, Secher NH. Cerebral ammonia uptake and accumulation during prolonged exercise in humans. J Physiol. 2005 Feb 15;563(Pt 1):285-90. Epub 2004 Dec 20. 
• Schaefer A, Piquard F, Geny B, Doutreleau S, Lampert E, Mettauer B, Lonsdorfer J. L-arginine reduces exercise-induced increase in plasma lactate and ammonia. Int J Sports Med. 2002 Aug;23(6):403-7.
• Shimomura, Y.; Murakami, T.; Nakai, N.; Nagasaki, M.; Harris, R.A. Exercise promotes BCAA catabolism:  Effects  of BCAA supplementation on skeletal muscle during exercise.  J. Nutr.  2004, 134, 1583S–1587S.
• Snow RJ, Carey MF, Stathis CG, Febbraio MA, Hargreaves M. Effect of carbohydrate ingestion on ammonia metabolism during exercise in humans. J Appl Physiol. 2000 May;88(5):1576-80.
• Swails WS, Bell SJ, Borlase BC, Forse RA, Blackburn GL. Glutamine content of whole proteins: implications for enteral formulas. Nutr Clin Pract. 1992 Apr;7(2):77-80.
• Wagenmakers AJ, Coakley JH, Edwards RH. Metabolism of branched-chain amino acids and ammonia during exercise: clues from McArdle's disease. Int J Sports Med. 1990 May;11 Suppl 2:S101-13.

Leucine Only Tops Ergogenic Effects of BCAAs: Increased Alanine Cycle Activity Spares Muscle Glycogen, Boosts Endurance Performance - BCAAs Have Opposite Effect

Alanine is the liver's favorite gluconeogenic amino acid and leucine appears to increase its usage.

Being among the first to learn about the "Glucose-Repartitioning Effect of Iso-Leucine" in February 2013 (read up on it), you, as SuppVersity reader, belong to the selected few who know that valine and isoleucine may be more than unnecessary props in the leucine-powered BCAA show. With the recent publication of a rodent study from the University of Sao Paulo in Brazil (Campos-Ferraz. 2013), however, it looks as if you had to revise your perspective on the purportedly auxiliary BCAAs - at least, with respect to their ability to reduce fatigue, and muscle and liver-glycogen degradation, in trained rats and possibly (!) humans.

So what did the Brazilian researchers do?

Basically, the idea Campos-Ferraz et al. had in mind, when they came up with their 8 week exercise + 2 week supplementation protocol (see Table 1) was to ...

Table 1: Exercise progression; suppl. was initiated in w7 after lactate test

"evaluate effects of the use of supplementation with leucine or a mixture of BCAAs in trained rats submitted to an exercise-induced protocol of glycogen depletion.

Furthermore, we attempted to investigate muscle and liver biochemical parameters that were not performed in the previous study in order to elucidate the role of BCAAs in glycogen depletion. " (Campos-Ferraz. 2013)

In other words: The researchers wanted to find out whether or not leucine would exert identical, less or more pronounced effects on muscle glycogen use and endurance performance in rodents that the full spectrum of branch-chained amino acids, i.e. leucine, valine and isoleucine.

Contrary to what bro-science and the shiny ads of the supplement industry are suggesting, the scientists' fundamental hypothesis was that the BCAAs supplementation would impair the rodents endurance capacity, because the branched-chain amino acids would be used in muscle to yield acetyl-CoA. This, in turn could reduce the activity of the glucose-alanine cycle, by which the muscles are supplied with alanine-derived glucose from the liver and (once the BCAAs got burne) result in an earlier onset of fatigue.

BCAAs are "glycogen depleters"?!

If you take a look at the data Campos Ferraz et al. gathered in the testing sessions at the end of the supplementation period, in the course of which the rats received an oral gavage of 166mg/kg per day (in human terms this would be ca. 3-3.5g per day) of BCAAs or leucine, it is quite obvious that the  the leucine group had a significantly lower muscle and liver glycogen degradation ratios than the BCAA group.

Figure 1: Liver & mucle glycogen degradation and time to exhaustion (expressed relative to placebo); muscle TCA intermediate content and enzyme activity / concentration (Campos-Ferraz. 2013)

Compared to the placebo group, only the ratios were different.  While the placebo group had the lowest liver glycogen use and a high muscle glycogen use, the supplemental leucine induced a shifted from muscle to liver glycogen and did thus exert muscle specific glycogen sparing effects.

As the researchers point out, these observations stand in line with their original hypothesis: Leucine can spare a significant amount of muscle and liver glycogen and thus produce a highly significant increase in resistance to exhaustion compared to the mixture of BCAAs (P<0.001).

This is not the first study to cast a bad light on BCAA supplementation. As a SuppVersity veteran, you will remember my November 2012 article "Chronic High Dose BCAA Supplementation Reduces Endurance Performance by 43%" | read more, as well as the more recent investigation into the  "Neurotransmitter Depleting Effects of Branched Chain Amino Acids (BCAAs) and Their Potential Ergolytic, Anxiogenic & Depressive Downstream Effects" | read more.

If we compare the endurance performance of the leucine rodents to that of the placebo group, this does yet cast a slight shadow on the overall image of the glorious ergogenic, and, even more so, the purported performance enhancing effects of BCAAs. Despite measurable differences in the time to exhaustion, the actual endurance increase in response to the leucine supplement is relatively small.
 
If you take another look at the data in Figure 1 you will probably notice the significant increase in TCA cycle intermediates (citrate and malate) in the BCAA group. These changes provide further evidence that the provision of all three branch-chain amino acid emphasized the use of glucose as a main substrate to sustain the endurance activity.

"Mouse vs. man": Can we ignore the differences in BCAA metabolism?


At this point, it may however be about time to point out that the activity of the BCAA catabolizing enzyme branched-chain keto acids dehydrogenase complex (BCKD) in humans is quite different from that in rats.

"In the latter [the rat], liver BCKD is almost completely unphosphorylated (activated) in basal state, making it possible to metabolize more rapidly BCKA from the portal blood; in humans, BCKD in liver is normally phosphorylated (inactivated) in order to spare BCAAs for protein synthesis." (Campos-Ferraz. 2013)

In other words: While rodents use BCAAs mostly as an energy source, the human body spares them as a potential protein anabolic.

In view of the fact that the BCAAs are not used to the same degree as an alternative substrate in the human vs. the rodent liver, it is actually not very surprising that the results of the study at hand appear to conflict with data from a previous study by the same laboratory (Gualano. 2011). In the corresponding experiment, Gualano et al observed measurable increases in exercise capacity and lipid oxidation in human subjects during endurance exercise after muscle glycogen depletion in response to the provision of 300mg/kg BCAAs per day.

So, the study is totally irrelevant, right? Not really, no. The fact that we are not able to use BCAAs as a readily available energy source like rodents does after all not mean that they must necessarily have the opposite effects on us. In fact, you all know that the vast majority of studies investigating the beneficial effects of BCAAs on endurance performance in humans yielded a null-result (!) - despite the fact fact that generations of researchers have been convinced that the inhibition of tryptophan uptake must blunt the exercise induced onset of fatigue (learn more in the articles cited in the red box).

Don't forget the endurance reducing increase in glucose usage that appears to be caused by isoleucine (and maybe valine) can also be beneficial: "The Glucose Repartioning Effects of Isoleucine" | read more.

The actual new information this study brings to the table is thus not that BCAAs are not ergogenic. It's rather the previously overlooked leucine induced acceleration of the glucose alanine cycle in liver. It is the activation of this (catabolic!) powerhouse by the means of which leucine "might have an interesting use in physical performance in prolonged or submaximal exercise, where muscle glycogen stores are more likely to be depleted" (Campos-Ferraz. 2013). It should be noted, though, that these effects are probably only observed after the glycogen levels are fully depleted - after an intense workout, towards the end of a race or after an fasted training - in those situations, the performance benefits may even be more more significant than in the study at hand.

Reference:

• Campos-Ferraz PL, Bozza T, Nicastro H, Lancha AH Jr. Distinct effects of leucine or a mixture of the branched-chain amino acids (leucine, isoleucine, and valine) supplementation on resistance to fatigue, and muscle and liver-glycogen degradation, in trained rats. Nutrition. 2013 Nov-Dec;29(11-12):1388-94.
• Gualano AB, Bozza T, Lopes De Campos P, Roschel H, Dos Santos Costa A, Luiz Marquezi M, et al. Branched-chain amino acids supplementation enhances exercise capacity and lipid oxidation during endurance exercise after muscle glycogen depletion. J Sports Med Phys Fitness 2011;51:82–8

VPX Pre- & Post-Workout Nutrition Gets "Sponsored" Scientific Approval: +4% Lean Mass, -6% Body Fat, +13% Upper and +21% Lower Body Strength in 29 Days

Image 1: Supplemental double-whammy. VPX' now
"scientifically proven" pre- & postworkout products

This is one of those cases, where I cannot decide whether I should applaud VPX or just shake my head... the scientist in me says: "Hey, you know how that is - with a research grant from the government cutting edge science is impossible, especially if you want to investigate something as 'profane' as building muscle". The cynic skeptic, on the other hand, whispers: "Come on, what results would you expect, if the study was financed by the producer of the supplement under scrutiny?" I guess I will applaud skeptically and exercise special caution in my analysis of the latest study from the Department of Health and Performance at Baylor University (Willoughby. 2011).

As in previous studies (Willoughby. 2007; Willoughby. 2009), Darryn S. Willoughby and his colleagues availed themselves of a buckload of VPX supplements and recruited 19 previously recreationally active, yet untrained (*) men with an average age of 22.8 +/-4.67 years, a height of 179.5 +/-6.38 cm and a total body mass of 79.1 +/-16.13 kg for another study into the effects of two supplements, which are supposed to "advance you to the next level of fitness" (VPX. 2011). Strength and body composition (body fat measured reliably by DEXA, not body-impedance), venous blood sampling and muscle biopsies were performed on day 0 and day 29 of the 4-week study period, in the course of which the participants underwent a standardized resistance training protocol (upper-/lower-body split, 4x à week), which mirrored the one that had been used in Willoughby. 2009 already (*).

Figure 1: Illustration of the training regimen (based on Willoughby. 2011)

The bodybuilding-type beginner 2x split training regimen is unquestionably a huge plus of the study (cf. figure 1). Performed twice a weak, this is what real world training would look like and so that it stands out of question that the results of the study will translate into practice - at least for everyone who has not touched a dumbbell more than thrice a week within the last 12 and abstained from all sorts of performance enhancing supplements and drugs within the last 3 months (*).

The NO Shotgun approach to protein NO SyntheSize??? 

Figure 2: Ingredient profiles of
No Shotgun and No SyntheSize

More important than the identical training regimen was yet obviously the supplementation protocol, to which the participants were assigned in a double-blind randomization process (on a side note: "double-blind" means that not only the subjects, but the scientists, as well, did not know which participants received the placebo and which ones the VPX products). While half of the subjects consumed a maltodextrose placebo (27g pre, 27g post workout), the subjects in the "NOSS" group consumed the same amount of NO Shotgun and NO SyntheSize as their pre- and posworkout supplement, respectively. Now, as the names imply, both supplements are intended to increase nitric oxide production and protein synthesis, yet with a focus on the former in NO Shotgun that is loaden with arginine and a heap of stimulants and a focus on the latter in NO SyntheSize, the composition of which is pretty similar (cf. figure 2), yet without the "Redline Energy & Meltdown Fat Burning Technology" ;-)

Although there were no specifically dietary guidelines, the research did at least collect some nutritional data based on a 4-day questionnaire all participants had to answer at the beginning and end of the study. While there was a slight reduction in the total caloric intake in the carb group (interestingly mainly from carbohydates), neither the intra-group changes, nor the inter-group differences reached statistical significance.

More muscle, less fat! Trainee, what more can you ask for?

That there were no differences is yet something you cannot say of the changes in body composition the study participants underwent in the course of this 28-day intervention.

Figure 3: Relative changes (compared to baseline) in body composition after 16 strength training sessions in 28 days with either 54g of maltodextrin or 27g of NO Shotgun and 27g NO Synthesize pre- and postworkout (Willoughby. 2011)

As a passing view of the relative changes (compared to baseline) in figure 3 show, the NOSS group (receiving NO Shotgun prior and NO SyntheSize post workout) registered significantly more pronounced elevations in fat free mass (p<.023 indicates that the chances that this was sheer coincidence are 23%) and - contrary to the carbohydrate group - lost -6% of their body fat, while the carb eaters added another 2% of adipose tissue to their love-handles.

Figure 4: Changes in upper and lower body strength (in kg/kg body weight during bench press and leg press at 1RM) after 16 strength training sessions in 28 days with either 54g of maltodextrin or 27g of NO Shotgun and 27g of NO Synthesize pre- and postworkout (Willoughby. 2011)

Interestingly, the lean mass increase went hand in hand with likewise (statistically) significantly greater (p-values see figure 4) increases in both upper (+13% vs. +1%) and lower (+21% vs. +11%) strength in the subjects in the NO Shotgun + NO SyntheSize groups.

* you may have wondered what all the asterisks in the previous paragraphs meant... well, they indicate specificities in the study design detractors may call "precautions that ensure that the VPX supplements are sitting pretty" ... I mean the exact same supplementation protocol performed on a bunch of veteran bodybuilders would probably not have elicited any measurable effects on body composition - keep that in mind when you interpret the results.

Now, it obviously should not surprise you that the protein (and leucine) loaden and creatine, beta-alanine spiked workout supplements outperform simple sugar water. It is thus more interesting to take another look at the data from the 2009 "NO Shotgun only"-study, Willoughby et al. have done (Willoughby. 2009). On the exact same training protocol, yet with only 27g of NO Shotgun or placebo 30min preworkout, the participants lost less body fat (-1.21%), but gained the exact same ~4% of lean mass and comparable increases in bench press and leg press 1RM (+8.82% and +18.4%, respectively).

Scientifically proven ingredients make scientifically proven products

I leave it up to you whether or not the results of this study will influence your next supplement purchase - after all, even the VPX guys will be aware that their supplements are not so unique that intelligent people like you would not be able to identify the key ingredients in their products (EAAs, hydrolized protein, creatine, beta alanine, some workout-boosting stimulants, etc.) and realize that there are way more than those two products which would probably have produced identically (within statistical margins) results, if, and here we've come full circle, if their respective manufacturers had the money and the balls to do scientific studies on their products.

Intermittent Thoughts On Intermittent Fasting - AMPK II/III: Leucine, HMB and a Glimpse on Other AMPK Modulators

Image 1: You pick a health, diet or diabetes supplement and I find the study that shows that in one way or another its effect is related to AMPK ;-)

I ended yesterday's installment of the Intermittent Thoughts on Intermittent Fasting Series on a pretty bold statement about the benefits of preworkout BCAA supplementation that would, at first sight, contradict common sense, or rather what common sense would dictate based on all you have read about the beneficial effects of BCAA supplementation on mTOR-related muscle protein synthesis (MPS) and the complementarity of mTOR and AMPK as regulators of anabolic (e.g. MPS, adipogensis ,etc.) and non-anabolic "scraping, rebuilding, recycling and repairing" processes. Since, after all, Bomb Jack, who posted a comment on last weeks installment of this series, is right: It would be logical that supplementation with BCAAs (he mentions HMB specifically) during the fast should result in dephosphorylation (~deactivation) of AMPK and thus negate its desirable effect on (metabolic) health.

And in fact, in the Wilson study I wrote about on Saturday the postprandial increase in AMPK phosphorylation, was blunted by the provision of carbohydrates, leucine or a combination of both (cf. yesterday's news) and you would assume that HMB supplementation would do the same, but the latter is - at least for chronic supplementation with low amounts (320mg/kg in rats ~ 52mg/kg in humans) of HMB - not the case (Pimentel. 2011), as the data I plotted in figure 1 clearly shows:

Figure 1: Effect one month of saline (control) or 80mg/day HMB on mTOR and AMPK phosphorylation and GLUT-4 expression in extensor digitorum longus (EDL) muscle of rats (Pimentel. 2011).

In the Pimentel study, there was, if anything, a non-significant increase in the AMPK and its purported downstream effect on GLUT-4 mediated glucose uptake  - both of which common sense would have told us to be compromised by HMB supplementation. While the lack of information on the "timing" or, more specifically, the interval between the last feeding and the intragastric administration (gavage) of 320 mg/kg body weight of HMB is a drawback in view of the significance of these results in an intermittent fasting context, rats usually eat at night and thus the administration of the 80mg of HMB (the rats weighed only 250g) "daily at the same time (during the light period)" will probably have coincided with a "fasting" period.

How can we explain that mTOR expression increased, while AMPK remained constant?

Are the different result a consequence of the metabolic magic of HMB? Well, before we analyze that in detail, there is another significant difference, we have to account for - in fact, a much more obvious one, which the amount of amino acids the rats were given in the Wilson and the Pimentel study, respectively (cf. figure 2).

Figure 2: Dosage, not type of supplement would be the most probable explanation for the different effects of leucine and HMB supplementation on AMPK phosphorylation in the Wilson vs. the Pimentol study.

I hope you did not already forget that, the main function of AMPK is to prevent that your cells run out of fuel or, to be precise, to avoid the ratio of "used" energy ADP and AMP (adenosine di- and monophospate) to ATP (adenosine triphospate) to continue to rise beyond a tolerable level. I further assume that you will be familiar with the fact that branched-chain amino acids bypass oxidation in the liver and thus become readily available energy sources for skeletal muscle (Renny. 2011). Now, if you put one and one together the answer seems pretty obvious: If the dosage of amino acids is sufficient (remember that those 270mg leucine are 4x more leucine than the the rats in the Wilson study got for "breakfast") to restore ATP levels to "appropriate" levels, the decrease in the ADP/ATP ratio will allow part of the AMP-activated protein kinase to be dephosphorylated.

According to our current understanding, BCAAs in general and leucine in particular trigger the ATP related decrease in AMPK and the complementary increase in mTOR by two distinct pathways, of which Tokunaga et al. write (Tokunaga. 2004)

[...]leucine stimulates p70α phosphorylation via mTOR pathway, in part, by serving both as a mitochondrial fuel through oxidative carboxylation and an allosteric activation of glutamate dehydrogenase. This hypothesis may support an idea in which leucine modulates mTOR function, in part by regulating mitochondrial function and AMPK.

In plain English: Leucine increases ATP when it is "burned" as fuel and it docks directly to the the non-active site of glutamate dihydrogenase enzyme and thusly increases the conversion of glutamate to alpha-ketoglutarate which in turn can be fed into the citric cycle to ultimately produce ATP.

Is it all about (cellular) energy ...

Figure 3: AMPK phosphorylation in Escherichia coli at different ADP/ATP ratios (data adapted from Xiao. 2011)

In April 2011 Xiao et al. published a study in Nature with some interesting quantitative data on the ADP/ATP ratio, on the one hand, the phosphorylation status of AMPK, on the other (Xiao. 2011). As my plot of the data in figure 3 shows, with increasing ATP levels (at constant ADP levels of 30µM) the phosphorylation of AMP-activated protein kinase in Escherichia coli BL21 cells declines by roughly -20% from 44% at a 30/0 ADP/ATP ratio to 22% at a 30/800 ADP/ATP ratio.

Yet, although these results would confirm the hypothesis that the main reason for the discrepancy is dose, or rather, energy related, and each and every nutrient that could potentially raise ATP levels, would eventually decrease AMPK, this still does not explain the increase in mTOR Pimentel et al. observed, despite (statistically non-significant) increases in AMPK.

... or is there a place for the "magic" of HMB?

As you probably know, beta-hydroxy-beta-methylbutyrat (HMB) is an oxidation product of leucine and / or its keto-acid alpha-ketoisocaproate (KIC) (Koevering. 1992). In 1998 Lembert et al. found that even KIC is not a direct substrate for ATP production, instead "KIC must transaminate with glutamate or glutamine to yield alpha-ketoglutarate and leucine" (Lembert. 1998). We may thus assume that similarly HMB cannot be used (directly) to restore cellular ATP pools. Moreover, HMB is thought to be the second (non-energetic) pathway by which leucine acts on protein synthesis / breakdown. According to a 2011 review of the literature by Zanchi et al. (Zanchi. 2011)

Nissen et al. (1996) suggested that HMB or some other metabolite (since there is no specific inhibitor to BCAT) is the main component responsible for the anti-catabolic effects of HMB because when adopting inhibitors of BCAA transamination, the only BCAA capable of anti-proteolytic effects is leucine, which undergoes a process capable of generating HMB (Slater and Jenkins 2000). Such effects were not observed when other BCAAs were tested (isoleucine and valine), suggesting that HMB or some metabolite may be the key element in promoting the [anticatabolic] effects.

When usually 5% of the dietary leucine is metabolized into HMB (Wilson. 2008), and these 5% are responsible for the non-ATP dependent effects on phosphorylation of mTOR, p70^S6k, and 4E-BP1 of leucine (Eley. 2007), it is no wonder that chronic intake of 80mg of HMB did stimulate mTOR in the absence of increased ATP levels (which would obviously have led to a decrease in AMPK expression that was not present in the Pimentol study), while 270mg leucine, yielding only 13.5mg HMB, did not stimulate mTOR, but was (ab-)used as a substrate to increase cellular ATP levels, thusly reduced AMPK levels and increased protein anabolism - different pathways, similar results: an increase in net protein synthesis.

Figure 4: Simplified illustration of the two distinct pathways by which leucine can work its muscle protein synthetic (MPS) magic and a hint on the compensatory (/) / amplifying (+) effects of exercise.

There is however, a third major pathway to the metabolic effects that are brought about by common intermittent fasting programs and this third player makes things even more complicated (cf. figure 4) - it's exercise! You probably remember from yesterday's installment that

1. during exercise in the fasted state temporarily AMPK increases and the energetically costly muscle protein synthesis (MPS) is reduced, while
2. after exercise (regardless of whether it was performed fasted or not, cf. "Glycogen-Free Growth") muscle protein synthesis increases due to an exercise-induced stimulation of the mTOR protein synthetic cascade

Before we dig deeper into this modulatory effects of different modes of exercise in the next installment of the Intermittent Thoughts on Intermittent Fasting, however, I want to conclude today's thoughts with a preliminary list of supplements / medications that have been shown to modulate the phosphorylation state of 5' AMP-activated protein kinase.

Image 2: If you insist on trying HMB, don't be stupid and buy a capped products, the prices for bulk HMB powder have lately been crushed - a major European carrier, for example, sells 250g at <13€ atm; HMB is thus cheaper than BCAAs, which cost 16Euros in the small 250g pack - did you hear me say that even 13€ is too much, no - you must be mistaken ;-)

"Should you prefer HMB over leucine as a dietary supplement to promote lean mass gains and prevent muscle loss during the fast?" I assume this is a question many of you will now be pondering about. My answer to this question would be "NO!" Firstly, if you are no construction worker or pursue a similar physically demanding profession, the fear of losing muscle (which is different from "feeling flat", my bodybuilding friends ;-) during a ~16h fast is hilarious, which means that BCAA, Leucine or HMB supplementation, while you sitting fasted at your desk in the office is simply unwarranted. Secondly, when you are exercising the increased energy demand will negate / compensate the negative effect the increase in ATP has on AMPK activity. And thus, thirdly, a large bolus of leucine (or a complete BCAA or EAA product) taken pre-workout will not only ward off proteolysis (as HMB would do) it will also provide the necessary energy to train harder and thus help to increase the exercise induced stimulus on protein synthesis.

All that and the absence of conclusive scientific evidence that would demonstrate the superiority of HMB supplementation over the provision of adequately dosed BCAA or EAA mixtures (it stands to reason that you cannot compare 3g of HMB to 3g of BCAA) are arguments against the use of β-Hydroxy β-methylbutyric acid. If you wanted to try it, anyway (and have no problem swallowing a powder that tastes like poison), the prices for bulk-powders have gone through the floor, lately ;-)

How to modulate AMPK "artificially" -  supplements, medications, hormones and more

In view of the fact, that the discussion of the effects of leucine (BCAAs and HMB) alone took much longer than I had expected and this whole episode took a different turn than I would have expected, the following list is more a preliminary overview than a comprehensive explanation of the effects of various supplements, medications, hormones and hormone-like substances on the AMPK. The latter will follow, as promised, but for today, you will have to content yourselves with what I would like to call a sneak peak on the AMPK-mTOR modulation handbook of which I hope that it will be one of the outcomes of all the past and future work that is going into this series ;-)

AMPK promoters:

• Adiponectin (Yamauchi. 2002; Handy. 2010; Iwabu. 2010)
• Alpha lipoic acid (Lee. 2005; Wang. 2010; Packer. 2011)
• Artemisia sacrorum Ledeb (Yuan. 2010)
• Aspirin (Sung. 2011)
• Astragalus (Zhou. 2009)
• Berberine (Ma. 2011)
• Brain-derived neurotrophic factor (Matthews. 2009)
• Caffeine (Jensen. 2007; Spirydopulos. 2008; Egava. 2009) 
• Capsaicin (Kim. 2010)
• Catechins / green tea (Huang. 2009; Murase. 2009; Reiter. 2010)
• Chromium (Wang. 2009)
• Ciliary neurotrophic factor (CNTF) (Watt. 2006)
• Cinnamon (Huang. 2011)
• Conjugated Linolic Acid (CLA) (HSU. 2011)
• Coptidis Rhizoma (CR), the dried rhizomes of Asian herbs (Egawa. 2011) 
• Cordiceps sinensis (Wong. 2010)
• Creatine (Ceddia. 2004)
• Curcumin (Kim. 2010)
• DHA (Jing. 2011)
• Ecklonia Cava (Kim. 2010)
• Garlic (Lee. 2011)
• Ginseng (Juang. 2010)
• Glucosamine (Kong. 2009)
• Grape seed extract (Meeprom. 2011)
• Hibiscus sabdariffa extract (Yang. 2010)
• Interleukin-6 (Kelly. 2009)
• Kidney bean extract (Lee. 2009)
• Malva verticallata seeds (Jeong. 2011)
• Metformin (Zhou. 2001; Freyer. 2002; Ohira. 2009)
• Mulberry extract (Tsuduki. 2009)
• Nicotine (Cheng. 2007)
• Prostaglandin E2 (Zhu. 2011) 
• Pu-erh tea (Way. 2009) 
• Quercitin (Jung. 2010)
• Raishi Mushroom (Lane. 2011)
• Resveratrol (Fullerton. 2010; Lin. 2010; Breen. 2008)
• Tiozolidinediones (diabetes meds. like rosiglitazone) (Freyer. 2002; LeBrasseur. 2005; Yang. 2011) 
• Vanadium (Hwang. 2011)

I still have two things to add to this list, firstly, this list is the result of a VERY cursory and 100% random search and is not even intended to be complete (at this time ;-). The intention (at least for in this installment) is to show you that an overwhelmingly large percentage of purported health supplements, diabetes and obesity treatments work via the AMPK pathway. And, secondly, I decided to limit the references to 1-3 per compound, even if in cases such as Metformin, ALA & Co the number of relevant studies is probably >500. Therefore you better consider the given references as evidence that I did not make up any associations between compound X and AMPK phosphorylation - and, if you want to know more before the release of the next installment, I suggest you go to PubMed and enter the respective keywords and do some digging on your own (your SuppVersity homework of the day - so to say ;-)

I hope you do not mind that I did not manage to tackle the effects of sleep and exercise in this installment, as I had originally intended. It is, after all, the central characteristic of this series that I sit down in front of the computer and start thinking at point "A", then I dig, here, get distracted there and follow up on "A1" to "A743", so that the output is by no means as structured and straight forward as my lectures and seminars or my SuppVersity blogposts on isolated topics... so, I can only hope that you enjoyed the turn this installment took (at best, because you learned something new) and in the unfortunate case that you did not enjoy what you have just read, you can at least look forward to the next episode of the Intermittent Thoughts on Intermittent Fasting Series ;-)