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marylin monroe
Showing posts with label protein oxidation. Show all posts
Showing posts with label protein oxidation. Show all posts

Glycerol, An Overlooked Ergogenic Supplement? 10-12g of Glycerol May Increase Lean Mass Without Training. Plus: Effects on Adrenals & AST + Lactate Response to Exercise

Interestingly, the study at hand suggests that glycerol may make you more musclar, even if you don't life. Crazy, but true (photo by awesomebody).
As a SuppVersity reader the potential ergogenic effect of the backbone of triglycerides are no news to you. The number of studies investigating the effects of glycerol on exercise performance and/or the adaptive response to exercise is yet low. Against that background, even a rodent study like the one by Eric Francelino Andrade from the Federal University of Lavras is worth its own SuppVersity article, I guess.

In said study, the Brazilian researchers evaluated the training adaptation and physical performance parameters in rats orally supplemented with glycerol,glucose, or saline, and submitted to moderate aerobic exercise.
Glycerol can be used in conjunction with creatine to hyperhydrate (Easton. 2007).

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Thirty male rats were trained for 6 weeks and administered the supplements during the last 4 weeks of the experiment. Animals were distributed in a completely randomized factorial 2 × 3 design (with or without exercise and 3 substrates) and received 864 mg/kg body of either glucose or glycerol in solution.

For human beings this equals human equivalents of 140mg/kg or ~10-12g of both, glucose and glycerol, for an adult. If this amount of glycerol had the same effects on humans at it did on the rodents in the study at hand, this would mean that it would lead to non-significant increases in lean body mass, even if the men and women who take it, were not working out.
Figure 1: Overview of the study design (top) and changes in protein content (~lean mass) and fat content of the carcass of trained and untrained rodents in the glycerol, glucose and saline groups at the end of the study period (Andrade. 2014)
The data from the rodent study at hand does yet also suggest that glycerol may at the same time non-significantly inhibit the lean mass gains that occur in response to aerobic training.

Glycerol as an adrenal supplement?

In contrast to the previously cited changes in lean body mass, which are not the result of a simple increase in body water (that's the beauty of actually cutting your "hairy subjects" open - you don't have to use BIA or DEXA scans to measure their lean mass), the effects on the weight of the adrenals of the animals was statistically significant in both, the trained and the non-trained rodents.
Figure 2: Organ weights (top row) and lactate (middle row) and aspartate aminotransferase (AST | lower row) levels at the end of the study period and after workouts, respectively (Andrade. 2014)
In the exercise trained rodents, there was also a significant reduction in the protein oxidation marker AST in both, the glucose and glycerol group. An obvious sign that both, pure glucose and the polyol (sugar alcohol) compound were used as alternative fuel during the exhaustive workouts, with the latter providing greater reductions in lactate build-up than the former.
What do previous studies say? Previous studies suggests that both glycerol, as well as combined creatine and glycerol can be used to hyperhydrate before exercise and thus to reduce the thermal and cardiovascular strain (Easton. 2007). This practice can also increase the exercise performance of endurance athletes like cyclists (Montner. 1996). Consumed in large amounts of 80g and in conjunction with 2L water, glycerol has also been shown to decreases body weight in athletes & increase overall performance in sedentary subjects, as previously reported at in a previous article of mine.
What do we make of these results? The increased lean mass in the non-trained rats is good news for every couch potato. The non-significance of the difference in trained rats is good news for athletes. The same goes for the reduced AST and lactate levels which suggest that glycerol is effectively used as alternative fuel and thus a candidate for pre- and intra-workout supplements for low-carbers.

Both the increases in heart and adrenal weight are physiological responses to exercise. In that, it is good news that the heart weight did not increase more in the glycerol group, despite the increased adrenal weight in these rats (otherwise, this may be interpreted as a result of pathological stress). The possible mechanisms for the increased adrenal weight are thus not of sympathetic (stressful) nature. Rather than that, the increased adrenal weight "may be related to an increase in body fluid volume caused by glycerol, decreasing relative sodium concentration (hyponatremia) (Von Duvillard. 2004), and increasing mineralocorticoid (aldosterone) secretion and adrenal gland mass (Decaux. 2003)" (Andrade. 2014). Overall, the results of the study still confirm that the ergogenic effects of glycerol are beyond the well-known hyperhydration properties caused by this substance | Comment on Facebook!
References:
  • Andrade, Eric Francelino, et al. "Adaptation to physical training in rats orally supplemented with glycerol." Canadian journal of physiology and pharmacology 93.999 (2014): 1-7.
  • Decaux, Guy, et al. "Low plasma bicarbonate level in hyponatremia related to adrenocorticotropin deficiency." The Journal of Clinical Endocrinology & Metabolism 88.11 (2003): 5255-5257.
  • Easton, Chris, Stephen Turner, and Yannis P. Pitsiladis. "Creatine and glycerol hyperhydration in trained subjects prior to exercise in the heat." International journal of sport nutrition and exercise metabolism 17.1 (2007): 70-91.
  • Montner, P., et al. "Pre-exercise glycerol hydration improves cycling endurance time." International Journal of Sports Medicine 17.01 (1996): 27-33. 
  • Von Duvillard, Serge P., et al. "Fluids and hydration in prolonged endurance performance." Nutrition 20.7 (2004): 651-656.

2x40g, 4x20g or 8x10g of Whey? Which Feeding Strategy Yields the Greatest Net Protein Retention? Plus: What the Results Can Tell Us About Intermittent Fasting on a "Bulk"

In know, after reading the headline you are probably already urgently waiting for the results of the latest study on protein timing, but before we get to the facts, let me briefly announce that this "bolus vs. intermittent vs. pulse" protein study, which is incidentally the result of an international cooperation between researchers from the Nestlé Research Centre in Lausanne, Switzerland, Canadian researchers from the University of Guelph, the Canadian Sport Centre and (you guessed it) Stuart M Phillips' group at the McMaster University, and their colleagues from the Australian Institute of Sport and the RMIT University in Melbourne, will be one of the topics of today's SuppVersity Science Round Up on Super Human Radio.

Other things I hope Carl Lanore and I will be able to squeeze into today's show, which airs, just as every Thursday live at 1PM EST and will also be available as a podcast later today, either right from the nav-bar on the right ("Physical Culture for Your Ears") or at www.superhumanradio.com, are ...
  • the latest news on natural nitrate supplementation with beet root juice, 
  • how stress and laziness increase breast cancer risk more than hormonal imbalances, and
  • how you can prepare your own powerful stevia-based wound ointment  
There is obviously more to the list, but I have learned from past mistakes and won't announce all I have piled up, when I know that's simply not possible to squeeze all of them into a single 1h show ;-)
A note for those of you who are looking for Adelfo Cerame's weekly contest prep blog: Don't worry it's still alive! You must have over-read that he has switched to a bi-monthly format!
Ok, ok... but NOW tell me hod do I have to spread my protein across the day"

While we know already that more is not necessarily better, when it comes to protein intake and that timing plays a significant role with respect to the returns in protein synthesis, and more importantly net protein retention you get for each gram of additional protein you consume, the question how you best spread your roughly 1.5-2.0g of protein per kg body weight across the day is still a matter of contemporary research and bro-scientific debate.

Suggested read: Protein Synthesis "Beyond the 20g Limit: Study Shows Exercise Facilitates 32% Greater Increases in Fractional Protein Synthesis With 40g vs. of 20g of Whey PWO" (click here to read)
What appears to be widely accepted, though, is the notion that both, the ingestion of a slow digesting protein before, and the intake of a fast digesting protein after a workout can effectively increase protein synthesis and net protein retention. If we assume that the combination of both strategies will yield further benefits (this has to my knowledge not been shown yet and is certainly not necessarly the case!), and regard the peri-workout supplementation as a "stand alone" that's not part of the 1.5g-2.0g /kg body weight baseline protein intake, we still end up with at least 80g of high quality protein (for the real light-weights or ladies ;-) we would have to spread in one way or another across the rest of the day.

8 x 10g, 4 x 20g or 2 x 40g? What's "optimal"?

Now, Moore et al. obviously won't have had my allegedly botchy "real-world" scenario on their minds, when they came up with the exact experimental design of their latest study. Still, if we forget about the 20g+ of protein post-workout, I believe none of you will be willing to abandon, their experimental setup fits the framework pretty nicely. After all, the scientists deliberately picked the 12h period after a workout "to standardise and take advantage of the accentuated protein synthesis in the exercised muscle over this period" and investigate three archetypal means of spreading a total amount of 80g of protein across the day: In 2 x 40g servings, 4x 20g servings or 8x 10g serving (Moore. 2012).

Suggested read: "3.2kg of Lean Mass Over Night W/ 40g of Slow Digesting Protein 30min Before Bed!?" (click here to read more)
The 24 male subjects who were advanced trainees working out 4–6 times per week in what the researchers call a "high intensity resistance training regimen" (note: I don't think this denotes a classic low volume HIT regimen) had to
"[...] follow standardized diet for the 72h prior to the trial that provided an energy availability of 45 kcal/kg fat-free mass with a macronutrient contribution 1.5 g protein/kg/d and 4 g carbohydrate/kg/d, respectively. [Moreover, s]ubjects were instructed to refrain from training and other vigorous physical activity during the 72h period."
When the men reported to the laboratory on the testing day, they had refrained from training or performing any other vigorous activity during the 72h period leading to the intervention and had been fasting 10h (over night). In absence of any other information I assume they remained in the fasted state for the subsequent standardized acute bilateral leg extension exercise session (4x10 sets at 80% 1-RM with 3 min recovery between sets), after which they were randomly allocated to receive their 80g of protein from whey as
  • pulsed feeding (PULSE), 8x10g every 1.5h; 
  • intermediate feeding (INT), 4x20g every 3h ; or 
  • bolus feeding (BOLUS), 2x40g every 6h. 
The supplementation regimen was started right after the workout and the protein synthesis, breakdown and net balance were determined based on previously tested and verified procedures (Hartmann. 2006).
Figure 1: Comparison of effect sizes, p-values (remember only p < 0.05 would be a statistical significant difference) and the scientists qualitative inference's based on the effect of feeding pattern on whole body net protein balance (left) and a detailed breakdown of the feeding specific effects on 12h protein synthesis expressed relative to the bolus group (based on data from Moore. 2012)
As the data in figure 1 goes to show you, the results clearly confirm that the pattern according to which you consume your daily allotment of protein does matter, what it does yet not really tell us is how this will translate into a real-world scenario, in which, as I have pointed out before, not having at least 20g of whey / other protein sources after a workout appears almost negligent. The provision of a 20g whey + 10g casein mix right after a workout could, for example, have undone the minimal (and statistically non-significant) advantage in net protein retention of the intermediate feeding group. And that may still have been the case if the latter had been "upgraded"  to a 4x25g whey pattern.

On the other hand, if we wanted to pick on the study design, the "workout" (leg extension) and the absence of other nutrients (or the lack of information about those in the paper?), which could easily have reduced the amino acid breakdown that nullified the advantage the pulse feeding had with respect to its ability to trigger and sustain (over 12h) protein synthesis, would be more relevant points of critique, anyway. That said the "study" at hand is actually only a "short communication", and I am pretty sure there is more to come in the future (it stands to reason that the SuppVersity is the place to go to read about that, right?)

Note: I still maintain that overnight fasting is healthy, and IF probably one of the best, r at least a very effective way to shed body fat, but that does not mean that it should be the only diet strategy in your "nutritional toolbox", in which other tools are probably better suited to pack on slabs of muscle!
(Preliminary) bottom line: The results Moore et al. present certainly don't provide a definitive answer on "the very best" way to time your protein intake (and even if there was an "optimal" way, no single study will ever be able to elucidate it). They do however make one thing pretty clear: My gut feeling that intermittent fasting and here especially those varieties with very long fasting and very short feeding windows, is probably not the best way of dieting to gain muscle. After all, there is no debating that the bolus regimen (2x40g 6h apart!) is trailing behind.

You can certainly tweak and thus optimize it by (a) adding a third meal in between and (b) cleverly using / combining fast and slow acting proteins (cf. "Whey and Casein Work Hand in Hand for Protein Anabolism"), but if you want level playing fields you would have to apply similar tweaks to the more frequent 4 x 20g and 8x 10g regimen as well... and I that would probably restore, if not magnify the difference.
Update on the real world significance of the advantage: I know that SuppVersity readers are smart and therefore was not suprised that only minutes after I posted this article, Steven Arcera objected that long-term studies don't show this advantage. Now, while Steven is right the implicit assumption that this implies that there is no advantage of spreading your protein across meals is false. If we simply take the exact figures from the study, which would be an added ~0.02g/kg body weight in protein retention over 12h, assume (which is obviously not valid) that the protein retention would be identical over the other 12h of the day in all groups and do the math for the study participants who weighed 80kg, this would be an additional 1.6g of protein retention for the whole body (remember this is whole body protein retention) and therfore even in a long-term study of 12 weeks only 134.4g! This would still be 134.4g more than with bolus feeding but would NEVER make a statistical significant difference in any study. And even the 584g "advantage" you would accumulate over a whole year would make it past the p < 0.05 line! So much about "optimal feeding strategies" and the real world outcomes of the latter :-)

References:
  • Hartman JW, Moore DR, Phillips SM. Resistance training reduces whole-body protein turnover and improves net protein retention in untrained young males. Appl Physiol Nutr Metab. 2006 Oct;31(5):557-64.
  • Moore DR, Areta J, Coffey VG, Stellingwerff T, Phillips SM, Burke LM, Cléroux M, Godin JP, Hawley JA. Daytime pattern of post-exercise protein intake affects whole-body protein turnover in resistance-trained males. Nutr Metab (Lond). 2012 Oct 16;9(1):91.