80g Glycerol + 2L Water Decreases Body Weight in Athletes & Increases Overall Performance in Sedentary Subjects

If it does not make you as swole as the colorful ad promised it must not be working, right? The jury was not even any longer "out there" for glycerol, but a recent study makes you rethink, whether you just have to look in the right place to see the benefits.

Another of the "odd" Thursdays without an update from "Your's Truly" Adelfo Cerame. And since there is holiday today, over here, I even thought there would not be a SuppVersity Science Round Up today. But hey, you are lucky you (and Carl) got to work, so you can tune in live at 1PM EST, or even better, start listening live at 1PM in order not to miss the Strength and Hypertrophy Round Table!

As far the  topics for today's installment of the SuppVersity Science Round Up are concerned, you are actually only a couple of lines away from reading about one that's on the list:  The effects of glycerol on exercise performance. I don't have to tell you though that this is not everything. Other things I believe you may be interested in are...

• childhood obesity, physical education and attention at school
• wheat gluten hydrolysate and how they don't come up to the expectations early trials have raised
• ammonia accumulation brain-fog, toxicity, liver 'pathologies' and workout performance
• running next to a street entails 'particular downsides' ("particular" is to be taken literally, here ;-)
• homocysteine levels, mortality, cognitive impairment and more
• epigenetic programming by nicotine and different protein contents before birth

These topics alone obviously won't fit into a single episode, but by now you should be aware that the SuppVersity Science Round Up Seconds, which are always published one day after the show aired, will provide you with the things we have missed and additional information, suggested reads and graphs to the topics we covered...  apropos "won't fit in", since the above is not even everything I have up my sleeve, I thought it would be wise to take the glycerol news from the compilation and tackle it on its own, today.

Can the backbone of bad triglycerides really be good for you?

Glycerol, a 3-carbon sugar alcohol that provides the backbone of triglycerides and is naturally found in foods as a component of dietary fats (Burke. 2011), is one of those supplements that have been all the rage for some time, didn't produce the expected instant results everybody was looking for (in this particlar case mostly "skinbursting pumps" and have eventually, in the course of one or two cycles of the regular yearly reformulations of the pre-workout supplements, completely disappeared from the market. I was therefore surprised, when I hit onto a recent study by researchers from the Physical Education and Sport High School in Konya (Turkey) that was published in one of the latest issues of the Journal of Human Kinetics (Patlar. 2013).

Is glycerol save? There have not been any reported toxicity effects up to doses of 5g/kg body weight. Glycerol does accumulates in body fluids, with the exception of the brain and the eyes and increases osmotic pressure (which was the reason why people used it in "pump supplements"), as well as the total volume of water in the body. If anything was 'dangerous', or I should probably rather say 'detrimental' to it, it would probably be its energy content (it is subject to gluconeogensis in the liver), which puts you at 'danger' of adding one or another pound of body fat you would probably want to avoid. The results of the study at hand to yet suggest that this is not an issue as long as you are active.

In essence the study protocol is nothing extraordinary: Take a couple of guys, 40 in this case (age 22.82 ± 1.49 years), and feed and water them using...

• 1.2 g/kg body weight) followed by water (26 ml/kg body weight) to 10 sedentary individuals (GS) and 10 soccer players from the University team
• just plain water to the another 10 sedentary men and 10 soccer players

Conduct a baseline test in the course of which all subjects are familiarized with the exercise equipment, a cycle ergometer (Monark 814-E) and required to perform an...

• Astrand Cycle Ergometer Test and an
  
• Wingate Anaerobic Power Test
  *the videos are don't show the study participants

in a room that is kept at 30°C and a barometrical pressure of 668 mm-Hg. For the next twenty days, have half of the guys (athletic groups E and GE) perform a 20-m shuttle run test every day. And finally perform a second follow up to see and evaluate the individual and joint effects of exercise and glycerol supplementation.

What's a shuttle run? I guess those of you who play soccer or basketball will know similar drills (at least I have been tormented by my trainers with them before in both sports and could imagine they are also among the standard repertoire of football coaches - though I have never played that myself):

Video 1: The shuttle run is every trainer's darling and would actually make a nice conditioning workout to be implemented into your own routine - whatever it may you are training for (note: the video is a random pick from YouTube and has no relation to the study at hand).

"The subjects warmed up for several minutes by jogging followed by stretching. The test program was installed on the computer and  initiated. A single beep was emitted at regular intervals. The subjects had to complete a lap or shuttle (foot on or over the line) with each beep. If the subjects completed a lap early they had to wait for the beep before starting the next lap. A triple beep indicated the start of a new level with a slightly faster speed required to complete each  lap.

The subjects were encouraged to complete as many levels as possible. An observer monitored the progress of a given subject, recording each completed lap on the recorder form. The subjects were instructed to turn by pivoting and not to run in a wide arc. The test was terminated when a subject was two or more steps from the line, for two consecutive laps. The observer alerted the subject at this time." (Patlar. 2013)

The shuttle run was followed by a couple of minutes of walking to cool down and a stretching exercise. The data was collected, logged and archived for evaluation.

Now if we take a look at the results of this undertaking they are unquestionably somewhat surprising - at least at first sight (see figure 1). In absolute terms it looks as if we had an across the board, almost identical increase in performance due to the daily shuttle runs in the exercise groups and a surprisingly large beneficial effect of glycerol only in the sedentary subjects (which would by the way be in line with many of the more or less disappointing trials on the benefits of glycerol supplementation in athletes; cd. Burke. 2011):

Figure 1: Changes in anaerobic and aerobic performance - relative values on the left, absolute before (white) and after (black) on the right (Patlar. 2013)

If you take a look at the relative pre-post changes in figure 1 (left), instead of the absolute changes a more distinct picture emerges:

• the benefit the sedentary subjects derived from the supplementation looks even more pronounced,
• the aerobic performance of the soccer players in the exercise group did likewise benefit, albeit less than the performance of the sedentary group, and
• shockingly the increase in anaerobic performance which looks pretty much identical is not statistically significant, yet still reduced in the glycerol supplemented athletes in the exercise + glycerol group (note: there is an increase, it's only relatively smaller)

Now, we all know that hyperhydration goes hand in hand with an increase in body water. In figure 1 I did even plaster a huge red sticker with "hyperhydration" onto the graph to give you an idea of a possible mechanism of action. So, if we wanted to be fair, we would have to take that into account... what? Yeah and you want to know if it will make you blow up like a wale, right, ... so let's see:

Figure 2: Changes in body weight and relative power (watts per body weight) in the course of the trial (data based on Patlar. 2013)

If we assume you are a sedentary slob at 80kg you could in fact gain 1.6kg... whether that's only water or if there is some fat there, as well, I cannot tell. Notwithstanding, I mentioned in the red box on safety issues, already that you can hardly expect to down 80g of glycerol with an energetic value of 4.32kcal/g (i.e. 350kcal per day) extra everyday without gaining at least some weight (assuming all other parameters are constant; plus, this could be muscle as well - well, not if you don't work out, though ;-).

You cannot expect to lose weight, but surprisingly it may still happen that you do if you consume those 350kcal of glycerol with 2l water right before your daily shuttle run. 

At least this is what happened to the soccer players in the supplementation group: They lost 2.66lbs of body weight on average. "Weight" is the unfortunate key word here, because we have no way of telling whether that was muscle, water or fat weight, as the scientists did not measure that separately. But let's be honest, it appears more than unlikely that it is (a) water or (b) muscle. After all the relative anaerobic power increased equally in both groups and why on earth would you lose water when you hyperhydrate? Ok, it could be one of those counter-regulatory reactions our bodies love. That again should however lead to performance decrements we did not see... you see, it's like the idiomatic dog that's chasing his tail. Why don't you play ginea pig and let us know what happens ;-)

WADA Warning for competitive athletes: If you are a WADA controlled athlete, you better avoid glycerol. It may sound hilarious, but it is on the WADA list of prohibited supplements since 2013. Why? Well, the increase in blood volume could mask the use of testosterone and co. because the /dl count would be lower if the total blood volume is higher -- this is something the WADA officials consider call a "masking agent" (Wada. 2013).

How much do you need? Don't forget, for glycerol to work its hyperhydrating magic, you must consume it with similarly hilarious amounts of water as the subjects in the study at hand. According to van Rosendal et al. an effective protocol comprises 1-1.5 g/kg glycerol + 25–35 ml/kg of fluid. Assuming you weigh 80 kg you can't get way with anything below 80g of glycerol + 2l water! Obviously way more than what any of the hitherto no longer available 'pump' or pre-workout supplements contained (at least I have not come across one that has a 80g scoop and says "consume with at least 2l of water on the label" - have you?)

Bottom line: I want to be honest, I still have to make up my mind about the usefulness of this supplement. I guess what actually does the trick is the combination of hyperhydration + energy availability. I have been preaching more than enough about the importance of energy availability over the last couple weeks, so I don't think I have to go into any more details here.

What I do think, however, is that few of you will be aware of the 2008 paper by Judelson et al. in which they report that hydration status is a fundamental determinant of the endocrine response to exercise, with dehydration leading to inappropriately high cortisol and norepinephrine levels that go hand in hand with an attenuation of the testosterone response to exercise, and negative effects on carbohydrate and lipid metabolism (Judelson. 2008).

Since you should by now have gotten the notion that insufficient energy does exactly the same, glycerol could well provide a means to counter this ergolytic double whammy. Against that background it is however strange that the athletes could not derive any athletic benefit from it... and weight loss without dieting (at least they were advised to stick to their habitual diets)?

If there is one definitive message you can take home from this study, though, it would be related to the dosage advice in the blueish info-box on the top right of this last paragraph: You better know how to use a supplement correctly! And this goes for the manufacturers of supps, as well as for the consumers: While the formers should finally stop putting ingredients into their supps to have them on the label, consumers should learn to identify hilariously underdosed and thus useless 'kitchen sink supplements' that 'have it all', but in doses where 'all' does not produce 'any' effect... how you can do that? Easy: Just make sure you get your daily dose of educative SuppVersity posts every day!

References:

• Burke LM, Stear SJ, Lobb A, Ellison M, Castell LM. A-Z of nutritional supplements: dietary supplements, sports nutrition foods and ergogenic aids for health and performance--Part 19. Br J Sports Med. 2011 Apr;45(5):456-8. 
• Judelson DA, Maresh CM, Yamamoto LM, Farrell MJ, Armstrong LE, Kraemer WJ, Volek JS, Spiering BA, Casa DJ, Anderson JM. Effect of hydration state on resistance exercise-induced endocrine markers of anabolism, catabolism, and metabolism. J Appl Physiol. 2008 Sep;105(3):816-24
• Patlar S, Yalcin H, Boyali E. The Effect of Glycerol Supplements on Aerobic and Anaerobic Performance of Athletes and Sedentary Subjects. Journal of Human Kinetics. 2013; 34: 69-70. 
• Van Rosendal SP, Osborne MA, Fassett RG, Coombes JS. Guidelines for glycerol use in hyperhydration and rehydration associated with exercise. Sports Med. 2010 Feb 1;40(2):113-29.
• World Anti Doping Association. WADA Prohibited List 2010. < http://www.wada-ama.org/Documents/World_Anti-Doping_Program/WADP-Prohibited-list/2013/WADA_Prohibited_List_2013_EN.pdf > retrieved Nov. 01, 2013. 

Trimethylglycine aka Betaine Sets the Anabolic Stage for Increased Muscle Growth: Higher IGF-1 & Lower Cortisol - Statistically Significant, but Physiologically (Ir-)Relevant?

Figure 1: Betaine content (in mg/100g) in some common food items (data based on Craig. 2004). Makes me wonder if Popeye ate wheat germ as well or whether he was celiac and stuck to spinach to get his daily dose of pro-anabolic betaine?

Trimethylglycine (TMG) the sciency name for a molecule most of you probably know by the name "betaine" is actually no longer a new-comer to the supplement scene (please note that this is not betaine HCL(!), the stuff you will find in digestive aids). I have already written about its purported ergogenic effects several times and there are actually quite a handful of proprietary blends with mostly undisclosed, but judged based on the total serving size and amount of ingredients in them, hilariously underdosed amounts of the zwitterionic compound and a methyl derivative of glycine in it on the market.

Friends and followers of the SuppVersity will also be aware that betaine is also found naturally in a variety of food sources such as sugar beets, wheat bran, spinach, shrimp, and many others (see figure 1) and that it can be synthesized from choline in your body, when dietary intake exceeds your current metabolic demands (Ueland 2011).

What you probably don't know, however, is...

... that the latest study from the Human Performance Laboratory at the Department of Kinesiology,
of the University of Connecticut, shows that "betaine (vs. placebo) supplementation enhanced
both the anabolic endocrine profile and the corresponding anabolic signaling environment, suggesting increased protein synthesis" (Apicella. 2013).

Figure 2: Effects of a standardized full-body workout (see text for details) on growth hormone (µmol/L), IGF-1 (nmol/L) and cortisol (µmol/L) levels in 12 recreationally trained young men after 2 weeks supplementation with betaine (2x 1.5g/day) or placebo (data based on Apicella. 2013)

And if we temporarily lose sight of the fact that the devil is in the detail, the data in figure 2 certainly looks as if you should run to the next best store with fishing equipment and get yourself a huge pot of trimethylglycine, of which the shop assistant will probably tell you that "This is a good choice Sir! The carps love the sweet taste!" But I am digressing, so let's get back to what's really sweet, namely ...

• stable growth hormone levels (vs. -17% in the placebo group)
• an 18% increase in IGF-1 (vs. a -10% decrease in the placebo group), and
• a -5% reduction in cortisol (vs. a 6% increase in the placebo group)

- they all sound pretty sweet, as well. Especially in conjunction with the stable p-AKT levels the scientists observed, when they analyzed the tissue samples. Unfortunately (but earnestly), Apicella et al.'s conclusion, still contains one word, too many people who read the abstract, are probably going to ignore:

"Betaine (vs. placebo) supplementation enhanced both the anabolic endocrine profile and the corresponding anabolic signaling environment, suggesting increased protein synthesis." (Apicella. 2013; my emphases)

Which one is it? A tip: It is none of the words I emphasized in bold. After all, that would make it way too easy for you... ha? Yeah! I see you've done your homework. Suggest(-ing) is in fact the most important word in this and the conclusions of many objectively written scientific papers.

So, the study "suggests increased protein synthesis"...

... and this means it does not even prove that the protein synthetic response in the immediate vicinity of the workout was increased in response to the to the two weeks of BID (=twice daily) supplementation with 1.25 g of betaine. In other words, all we know is that the funky gene essays for p-AKT and serum tests for growth hormone, IGF-1 and cortisol "suggest" that it could be the case, if we assume that marginally higher IGF-1 levels, stable growth hormone levels and lower cortisol levels (rememeber we are not talking about increasing any of them into the supraphysiological range, here) would

1. result in increased protein synthesis and ultimately
2. greater lean mass accrual,

because, if we are honest, no one is interested in a number you can measure, when you infuse a marked amino acid into the circulation and check how much of it goes into the muscle, but doesn't come out of it, afterwards.

Something to remember: What I find remarkable - and this is by no means something you will see only in this study, neither is it "fraud" or whatever, is how by simply adding a break into the Y-axes of the graph and thus omitting the lower 80% of the bar Apicella et al. give the impression that the effects on IGF-1 were more than twice as large than they actually are... remember that, because you will encounter that in many studies, and reproductions of graphs from scientific papers, especially if they are used to market certain products.

What we want is to get bigger, stronger and all that faster, and whether the 12 recreationally trained men (age 19.7±1.2 years; lean body mass 65.2±8.8 kg; fat mass 15.6±8.5 kg; body fat percentage 18.7±7.0 %; BMI 28.2±4.0) would have gained even a single inch of muscle more on whatever body part, if they had performed a real workout instead of the funky "AES" (=acute exercise session) that consisted of

• 10x maximal vertical jumps without pause, 
• 1x 10-s isometric squat, 
• 1x 10-s isometric bench press on a smith machine, and
• 1x 10 min of repeated box lifting (RBL)

is more or less guesswork. In view of the previously discussed results from the researchers at the McMasters University in Ontario (see "Anabolic Workouts Revisited"; a brief reminder. the systemic hormonal response to an acute exercise bout is irrelevant, if anything higher cortisol levels correlate with greater increases in lean muscle mass) and the "statistically significant", but physiologically probably irrelevant increases and decreases in IGF-1 and cortisol at least highly questionably (please take a look at the additional information in the red box to the right, as well).

... but there are still way too many "ifs" in here!

The sheer number of "suggests", "is touted", "is likely", "also possible", "as we presume", etc. is honest and speaks in favor of the quality of the study, but against the reliability of the statement that followeed the initially cited "suggests" in the conclusion. Moreover, the researchers freely admit that...

"[...] the mechanisms by which betaine may have affected the hormones measured in this study are still unclear and require further research" (Apicella. 2013)

so that even the fact that betaine is an organic osmolyte and could thus help stabilize skeletal muscle protein, promote / maintain optimal hydration and protect against

• hypertonic stress (Alfier. 2006), 
• urea-induced inactivation of muscle myosin ATPases (Ortiz-Costa. 2002), and 
• structural changes in myosin due to urea accumulation (Ortiz-Costa. 2002)

does lend credibility to the hypothesis that betaine could help you build muscle, but it does not prove it. In conjunction with the results of previous trials, like...

Betaine does not increase nitric oxide While I have no idea why everyone is so keen about those nitrate supplements, one thing is for sure: Betaine has no effects on serum nitrate or nitrite levels. The vasolidation effect of beet roots / beet root juice is simply a result of the nitrate that's in there along with the betaine (+ the sugar and the insulin spike, which will also trigger an increased NO-response).

At least this is what a study by Bloomer et al. which consisted of three independent experiments using 1.25 and 5.00g B, acutely, 2.5g per day for 14 days, chronically, and a combination of chronic (6g for 7 days) + acute (6g acutely before the test) betaine supplementation (Bloomer. 2011).

• Hoffman. 2009 - 2.5/day for 14 days; jump squat, squat, bench press; "Two-weeks of betaine supplementation in active, college males appeared to improve muscle endurance of the squat exercise, and increase the quality of repetitions performed." 
• Lee. 2010 - 2x 1.25g/day for 14 days; bench squat and jump tests; "[Betaine] supplementation increased power, force and maintenance of these measures in selected performance measures, and these were more apparent in the smaller upper-body muscle groups."
• Hoffman. 2011 -  2.5g/day for 15 days; 5 training + testing sessions; "15 days of betaine supplementation did not increase peak CON or ECC force outputs during an isokinetic chest press but did appear to reduce subjective measures of fatigue to the exercise protocol"
• Trepanoswki. 2011 - 2.5g/day for 14 days; resistance training; "moderate increase in total repetitions and volume load in the bench press exercise, without favorably impacting other performance measures."
• del Favero. 2013 - 2g/day for 10 days; muscle strength and power, muscle PCr content, and body composition, three "familiarization sessions" preparing the participants only to perform the tests; "we showed that betaine supplementation combined or not with creatine supplementation does not affect strength and power performance in untrained subjects."
• Pryor. 2013 - 2.5g/day for 7 days; cycling performance; "betaine ingestion significantly increased average peak power (3.4%; p = 0.026), maximum peak power max (3.8%; p = 0.007), average mean power (3.3%; p = 0.034), and maximum mean power (3.5%; p = 0.011) in recreationally active males and females"

... there is still room for long-term improvements in muscle gains as a consequence of the general ergogenic effects of betaine (every rep more counts!), but it appears unlikely that the "anabolic" hormonal milieu observed in the study at hand are the fundamental cause of the latter.

Reminder: If you want to try it, you got to get yourself "trimethylglycine" (TMG) not "betaine HCL" and you better don't buy it in capped form if you don't have lots of money to burn. I just checked with the next best bulk supplier - they got 1kg for $33.50. Even if you double dose, i.e. take 2x 2.5g per day (most studies mixed it with Gatorade) this will last you for 200days(!), which is probably the time it will take until you can actually see and not just measure any potential, possible, suggested, etc. anabolic effects ;-)

References:

• Alfieri RR, Bonelli MA, Cavazzoni A et al (2006) Creatine as a compatible osmolyte in muscle cells exposed to hypertonic stress. J Physiol 576:391–401.
• Bloomer RJ, Farney TM, Trepanowski JF, McCarthy CG, Canale RE. Effect of betaine supplementation on plasma nitrate/nitrite in exercise-trained men. J Int Soc Sports Nutr. 2011 Mar 18;8:5.
• Craig SA. Betaine in human nutrition. Am J Clin Nutr. 2004; 80: 539–549.
• del Favero S, Roschel H, Artioli G, Ugrinowitsch C, Tricoli V, Costa A, Barroso R, Negrelli AL, Otaduy MC, da Costa Leite C, Lancha-Junior AH, Gualano B. Creatine but not betaine supplementation increases muscle phosphorylcreatine content and strength performance. Amino Acids. 2013 Jun;42(6):2299-305.
• Hoffman JR, Ratamess NA, Kang J, Rashti SL, Faigenbaum AD. Effect of betaine supplementation on power performance and fatigue. J Int Soc Sports Nutr. 2009 Feb 27;6:7.
• Hoffman JR, Ratamess NA, Kang J, Gonzalez AM, Beller NA, Craig SA. Effect of 15 days of betaine ingestion on concentric and eccentric force outputs during isokinetic exercise. J Strength Cond Res. 2011 Aug;25(8):2235-41.
• Lee EC, Maresh CM, Kraemer WJ, Yamamoto LM, Hatfield DL, Bailey BL, Armstrong LE, Volek JS, McDermott BP, Craig SA. Ergogenic effects of betaine supplementation on strength and power performance. J Int Soc Sports Nutr. 2010 Jul 19;7:27. 
• Ortiz-Costa S, Sorenson MM, Sola-Penna M (2002) Counteracting effects of urea and methylamines in function and structure of skeletal muscle myosin. Arch Biochem Biophys 408:272–278
• Pryor JL, Craig SA, Swensen T. Effect of betaine supplementation on cycling sprint performance. J Int Soc Sports Nutr. 2013 Apr 3;9(1):12.
• Trepanowski JF, Farney TM, McCarthy CG, Schilling BK, Craig SA, Bloomer RJ. The effects of chronic betaine supplementation on exercise performance, skeletal muscle oxygen saturation and associated biochemical parameters in resistance trained men. J Strength Cond Res. 2011 Dec;25(12):3461-71.
• Ueland PM. Choline and betaine in health and disease. J Inherit Metab Dis. 2011;34:3–15.