Intermittent Thoughts on Building Muscle: IGF-1 and its Splice Variants MGF, IGF-IEa & Co - Master Regulators or a Bunch of Cogs in the Wheel of Muscle Hypertrophy?

Image 1: With regard to IGF-1 and its splice-variants like MGF, there is probably 10x-100x more bro- than pro-scientific data out there - this does not help us, though, since you never know which of the bro-reports is bogus and which is not.

In view of the fact that we have not covered much ground with the last installment (we did build a pretty solid foundation, though ;-), I will try my very best to steer a middle course between presenting impressive amounts of facts and explaining the complex and in part not even completely elucidated physiological underpinnings of skeletal muscle hypertrophy, or, as the bros would say, getting big and buffed! A pros pros Bro, you will unquestionably have read on one of the myriads of bodybuilding-related bulletin boards how the injection of X amounts of IGF-1 right into the muscle made BigGuns, or whatever the poster's pseudonym may have been, grow "3 inches in 2 weeks"... ok, his profile picture looks impressive, but is that credible? Does IGF-1 really have such profound effects on muscle growth? And about what type of growth are we talking here? The myostatin-negative "ballooning up" of the muscle, which leaves you with overblown myogenic domains and dysfunctional muscles?

IGF-1: Insulin, growth hormone, or what?

To be able to answer these and related question we will first have to understand what exactly this "insulin-like growth factor 1" actually is. From a (bio-)chemical perspective it is nothing but a bond of 70 amino acids which are entangled into a specific peptide structure that is characteristic for somatomedin C, as IGF-1 is also called. Both the "growth" in IGF-1, as well as the "somato" in its old-fashioned appellation already suggest that what we are dealing with, here, is a "growth hormone related" polypeptide. And in fact, the synthesis of IFG-1, which, in the case of the systemically available fraction, takes place primarily in the liver, and is triggered by systemic growth hormone (somatotropin) levels.

Figure 1: Changes in systemic IGF-1 levels after 5-weeks on either a "normal" (=55:15:30 carbs:protein:fats) or a low carb "high protein" (=20:30:50) diet in 8 men with untreated type II diabetes (data adapted from Nuttal. 2006)

The "insulin" in its name, however, is pretty misleading... or I should say people mislead themselves, by not reading  the name correctly: It's not "insulin-growth factor", but "insulin-like growth factor" and the "like" refers to the structure of the molecule and does not imply that it is released in response to insulin spikes, as you may have read it on one of the aforementioned bulletin boards. If you do take a look at the growth hormone and IGF-1 levels of eight male subjects in a 2006 study on the metabolic of 5-weeks on what the scientists call a "high protein, low carbohydrate diet" (Nuttall. 2006), you will see that an increase in protein and fat from 15% to 30% and 30% to 50%, respectively elicited an 34% increase in serum IGF-1 levels over the treatment period, a finding that is corroborated by the recently published results of Matthew B. Cooke and his colleages from the Department of Health, Human Recreation and Performance at Baylor University.

Figure 2: Serum IGF-1 levels in response to whey vs. maltodextrin supplementation and subsequent lower body resistance training (data adapted from Cooke. 2011)

In their randomized double-blinded cross-over study, Cooke et al. had a group of 10 recreationally active men (2-3 non-resistance training exercise sessions per week) perform a lower body exercise program (leg presses and knee extensions, 4 sets, 8-10 reps at 80% of the individual 1RM) with either 10g of maltodextrose or 10g of whey 30 minutes before the exercise bout (Cooke. 2011). The results of the study (equal IGF-1 response regardless of whey or carbohydrate supplementation) imply that even in the short term, in healthy subjects and in conjunction with exercise the ingestion of carbohydrates is not superior to the provision of fast acting protein sources as a means to either increase or maintain systemic IGF-1 levels.

On a side note: The insulin-mediated induction of Akt, which subsequently triggers the phosphorylation of the mammalian target of rapamycin (mTOR) and thusly does its bit to elevate protein synthesis, has no direct relation to IGF-1, which - I cannot emphasize that enough - has a structure resemblance to insulin, nothing more, nothing less. And what's more, the insulin response in the aforementioned study by Cooke et al. was identical in the whey vs. maltodextrin arm of the study.

Systemic vs. local IGF-1 expression: A crucial distinction

If you have been following the daily research updates here at the SuppVersity over the last months, you may now be wondering why I am even caring about those growth hormones (after all you should, after reading the first paragraph, realize that IGF-1 is something like the active incarnation of somatotropin), when Stuart Phillips lab has quite conclusively shown that even the exercise induced elevation of testosterone does not correlate with subsequent increases in muscle protein synthesis. Certainly a good question, but nevertheless not difficult to answer:

1. The previous installments of the Hypertrophy 101 (Part 1, Part 2) should have made it quite clear that protein synthesis alone is not sufficient to grow. Without intra-muscular restructuring / reorganization and the recruitement of new myonuclei from satellite cells, you would sooner or later grow beyond the maximally allowed myonuclear domain sizes (assuming that by whatever means you block the healthy upregulation of mystatin that will prevent that) and end up as an over-muscled but completely dysfunctional wrack.
2. In a very recently published study, the results of which I have actually been holding back, because I thought I would get to them much earlier in this series, the very same Stuart Phillips whose studies are "responsible" (in fact it is the way they are discussed by the lay-press and abused by the supp-companies that is actually "responsible") for the current over-emphasis on acute increases in the protein synthetic response to exercise and/or supplements, reports that there actually was a statistically significant correlation between exercise induced growth hormone release and increases in mean type I fiber (p<0.06) and type II (p<0.04) cross-sectional area (CSA) in 56 healthy previously non-resistance trained healthy young men in response to a 12-week, 5-day per week resistance training regimen (West & Phillips. 2011).
3. While we have hitherto been talking about systemic IGF-1, it has become evident in the course of the last decade that the hepatic IGF-1 output, which is the main determinant of circulating IGF-1 levels, has little to no impact on the IGF-1 induced increases in skeletal muscle mass and remodeling of muscle tissue that has been previously studies in Petri dishes. In fact, recent research suggests that, just like the liver produces IGF-1 for "the whole body", muscles produce their own IGF-1, or I should say, their own IGFs-1, whenever they are challenged to grow and/or repair (Velloso. 2010), and that the decline of muscle mass with age is at least in parts attributable to a defect / reduction in the expression of local IGF-1 splice variants (for an explanation of what this is, see red box below).

If we now count 2. and 3. together the result is not 5. but rather that it is the growth hormone mediated, exercised-induced local expression of IGF-1 splice variants, which drives the repair and restructuring process that allows for continuous (healthy) muscle growth.

Did you know that the intra-muscular (=autocrine, meaning directly in the tissue where it is supposed to work) "construction process" of the mature 70 amino acid polypeptide IGF-1 gives rise to three different splice variants of insulin-like growth factor (note: the structure of IGF-1 gene does theoretically allow for 6 variants)? And though we are just beginning to understand the physiological roles of IGF-IEa, IGF-IEb and IGF-IEc, also known as MGF (mechano-growth factor), their distinctly timed expression in response to physical overload appears to constitute one of the major driving forces of myocellular hypertophy.

In order to fully understand the role "the" insulin-like growth factor 1 plays in the physiology of muscle growth, it is thusly important to realize that the common perception of IGF-1 as a systemic hormone is, at best, incomplete - I would even venture to say that it is totally flawed.

MGF?! Yeah, I have heard of that one!

Figure 3: Stained myocyte migration (top) and infiltration (bottom) essays for IGF-1 and MGF; more stains = greater effect (taken from Mills. 2007).

Of the three primary splice variants that are expressed in skeletal muscle, IGF-IEc, or MGF (Mechano-Growth Factor) has probably received the greatest attention - so much attention that even the aforementioned bros, will probably have grasped the notion that this is somewhat of a local isoform of IGF-1 which is expressed in response to exercise induced muscle damage and could potentially be the magic bullet to grow beyond what we have hitherto believed to be possible... and, guess what, in essence this appears to be correct.

In one of the earlier studies on the cellular effect of MGF, Yang et al. were able to show that MGF stops the IGF-1 mediated cell differentiation process (in practice this means that it stops the satellite cells from differentiating = specializing and becoming muscle cells) and increases their proliferation. Or put more simply: While in vitro exposition to IGF would suffice to build muscle, as long as there are enough progenitor cells (satellite cells) available, MGF is necessary to replenishes the satellite cell pool of which you have learned in the previous installments that it is necessary to a) repair damaged muscle tissue and b) increase the number of myonuclei in order to grow beyond the physiological growth limit that arises due to the muscle-type-specific upper limit to the myonuclear domain size (cf. previous installments).

Figure 4: Cell proliferation data in response to MGF treatment after blocking the IGF-I receptor.

As the data in figure 4 goes to show the effects of the complete polypeptide IGF-1 and its splice variant MGF appear to be mediated, at least partly via distinct receptors. And while recent research suggest that MGF also exerts similar effects on tendon (Olesen. 2006), brain (Dluzniewska. 2005) and nervous tissue (Aperghis. 2004), our primary concern here, is its pivotal role in muscle repair, which involves the activation of satellite cells, their proliferation (Yang. 2002) and migration (Mills. 2007).

A series of studies by Hammad et al., which was originally intended to investigate the effects of age on the expression of the different IGF splice variants, goes to show that the "muscle (re-)building effects" of MGF are not restricted to the test tube. In their 2002 study (Hamed. 2002), the researchers were able to show profound increases in the MGF expression in the quadriceps muscles of 8 healthy young men (age 29.5 ± 1.5 years, body mass 81.1 ± 2.4 kg, height 179.3 ± 1.8 cm) 2.5h after a single muscle-damaging leg-extension exercise (10 sets of 6 repetitions at 80% 1-RM, 2 min rest between sets):

Figure 5: MGF (ng mRNA / 10^8 µg RNA) and IGF-IEa ng mRNA / 10^5 µg RNA) expresion in quadriceps muscle of young subjects before and 2.5h after 10 sets of 6 repetitions at 80% 1-RM on a leg-extension machine with 2min rest between sets (data adapted from Hamed. 2002)

If you take a closer look at the data in figure, you will probably notice that there was one subject with an extreme MGF response, the scientists explain by a particularly high type-IIx fiber content of the quadriceps of this individual. If you remember the mouse studies and the analysis of the muscle composition of bodybuilders from the previous installments, you will be aware that the shift from type IIb to type IIx muscle fibers is one of the main characteristics of "getting real big". The extreme MGF response (>10x higher than the mean MGF expression across the other subjects) in this subject thusly suggests the increased growth capacity of type IIx muscle fibers is in part due to their ability to release MGF in response to strenuous exercise and thusly multiply / replenish their satellite cell pool to prepare for future growth.

Figure 6: MGF (ng mRNA / 10^8 µg RNA) and IGF-IEa ng mRNA / 10^5 µg RNA) expresion in quadriceps muscle of young subjects after eccentric HIIT exercise on cycle ergometer (data adapted from Hamed. 2008)

Interestingly, a 2008 follow up study (this time involving nine healthy young men aged 20–27 years, cf. Hamed. 2008) with a completely different training protocol that consisted of

60min of opposing the rotation of the pedals down to 60 r.p.m. Subjects performed the following program of six working intervals: six working intervals: 0–6min at 50%, 6–12min at 75%, 12–20min at 100%, 20–25min at 130%, 25–40min at 100% and 40–60min at 75% of the load  eliciting concentric VO2max

illicited surprisingly similar results (cf. figure 6). And in both cases, it appears to be the MGF splice variant not the IGF-IEa variety that drives the short term (hours to days) response to strenuous exercise.

HIIT and resistance training a dynamic duo for MGF expression

Assuming that you are following each and every post here at the SuppVersity (you know you should be ;-), this should remind you of a previous blogpost of mine (cf. "HIT Your Satellite Cells to Increase Your Gains!"), in which I explained that one of the many advantages of high intensity training (not even interval) over classic "cardio" training is that it can increase satellite cell proliferation. Now, with this installment of the Intermittent Thoughts you finally understand, why this is the case.

Image 2: This is not the kind of muscle damage you should be aiming for in the gym.

Now, while protein synthesis and increases in domain size are partly mediated via nutrition, the intra-muscular expression of the IGF-IE splice variants appears (at least based on the current research) to depend solely on exercise, or I should say the wear and tear that goes hand in hand with heavy exercise. In that it seems to be less important, whether you are "pumping away" or "cycling like maniac", as long as its "hard" - to put that into perspective, in the 2008 study by Hamed et al. the subjects underwent ~3600 eccentric muscle contractions in only 1 h, their creatine kinase (CK) levels (marker of muscle damage) increased by +183% and all subjects reported profound muscle soreness.

This controlled amount of muscle damage ties in nicely with the topic of next week's installment which will center around the the intricate relation of the inflammatory response to exercise, the expression of the well-known and less known inflammatory cytokines, TNF-alpha, IL-6 and IL-15 (sorry, Trevor, I have already gone overtime, so your question will have to wait till next week ;-) and the muscle (re-)building effects of IGF-1 and its intra-muscular children.

Alanyl-Glutamine or Alanine + Glutamine? Dipeptide or Free Form Aminos? What Offer Maximal Muscle Protection?

"Wouldn't have happened if she'd used alanyl-glutamine instead of regular that cheap alanine + glutamine combo!" - True or False? Recent study says: False!

If you combine your liver's favorite gluconeogenic amino acids, i.e. alanine and glutamine, into a single peptide the result is called alanyl-glutamine and marketed as the ueber-potent alternative to regular l-glutamine supplements. It goes without saying that a comparison like this is about as stupid as comparing french fries with mayo to regular french fries and saying that the former are worse because they contain more fat, or whatever. Even if we didn't care about the physiological significance of the effects of alanyl-glutamine, we would obviously have to compare the purported cryogenic effects of this "innovative" dipeptide to those of a simple combination of free form amino acids to deserve the bragging rights for having created an advanced form of glutamine.

Alanine + glutamine vs. alanyl-glutamine - fight!

By now you are probably asking yourselves why I am bothering you with things like this. Right? Well, the reason is that Éder Ricardo Petry and his colleagues from the University of Sao Paulo must recently have been pondering the same question. To answer it, they conducted an experiment that would allow them to verify if the oral supplementation with l-glutamine and l-alanine as dipeptide has more pronounced muscle protective effects than a simple mixture of l-glutamine and l-alanine (GLN+ALA, both in their free forms) in a group of Wistar rats that are subjected to intense aerobic training (treadmill).

I know what you are thinking now: "Not another rodent study...", but think about it: How many people are willing to pay $50 and more on supplements without any in vivo evidence of their efficacy let alone long-term safety? Against that background Petry's rodent is a major advancement - isn't it?

True or False: You can (ab-)use glutamine to replenish your glycogen stores!? True! It sounds strange, but according to a study from the late 20th century glutamine is a pretty effective glycogen replenisher, even in the absence of your bodies favorite nitrous glucose precursor alanine | learn more

Don't get me wrong, there are a few alanyl-glutamine studies in humans, but there is not a single one that would compare the dipeptide to a reasonable placebo in an exercise scenario. I mean, who tells me that the basketball players in the 2012 study by Hoffman et al. wouldn't have experience the same beneficial effects on basketball skill performance and visual reaction time if their rehydration solution had contained alanine and glutamine or even glutamine alone? Yes, I know... the increased absorption: Well, let's just look at a fair comparison, i.e. the study at hand, and see what happens when the dreams of supplement formulators and reality meet ;-)

Ok, back to the facts - the exercise & supplementation protocol

The male Wistar rats, the researchers used in their experiment were exercised 5x per week - at increasing intensities: Starting with 30 and 45 min of treadmill running (incline 3°) at 20 and 22.5 m/min in the first three weeks, the speed and duration of their treadmill runs increased to 60 min at a speed of 25 m/min in week four and remained like that for the rest of the 8-week study period.

The supplements were administered via oral gavage in the course of the last 3 weeks, only. The daily doses for the animals in the dipeptide (DIP) and free form amino acid groups (GLN+ALA) were...

• 1.5g/kg alanyl-glutamine in the DIP group,
• 0.67g/kg l-alanine + 1.0g/kg l-glutamine in the GLN+ALA group, and
• plain water in the control group

The amount of of alanyl-glutamine the scientists used was calculated in such a way that the total amount of l-glutamine was the same as that of l-glutamine administered in its free form.

Changes? YES! Dipeptide benefits? Not really...

The gavage was provided 1 h after the end of each session of exercise, after which the animals had with free access to water and chow. To make sure that the results of the examinations on the last day of exercise would not reflect the acute effects of a single dose of the supplements, the animals were killed 10 h after the last exercise session.

Figure 1: Plasma glutamine, glutamate, ammonium, malondialdehyde, myoglobin, and creatine kinase activity in Wistar rats supplemented with alanyl-glutamine (DIP) or regular glutamine + alanine; data expressed rel. to control (Petry. 2013)

The virtually identical increases in l-glutamine and l-glutamate, you see in Figure 1 should thus represent the baseline and not the 'immediately post supplementation level' of these amino acids. For the exercise-induced accumulation of ammonium, malondialdehyde (MDA; indicates lower lipid oxidation), myoglobin and creatine kinase (both indicate lower muscle damage) the timing is not that important, anyway. What is important, however, is the fact that there were no physiologically relevant advantages for the "super glutamine".

DHEA & estrogen are alternative muscle protectors. Despite the fact that estrogen has repeatedly been shown to have muscle protective-effects, I would not suggest you steel your granny's HRT medication. DHEA on the other hand, may be something to consider - specifically if you are about to overreach, like the male subjects in a 2012 study by Liao et al. (learn more)

If we take a closer look at the p-values and the statistical significance of these changes, it turns out that, the minor increase in glutamate aside, all of the difference to the placebo group were statistically significant. The DIP vs. GLN+ALA differences, on the other hand, were marginal and reached statistical significance only in the case of the marker of myoglobin. Where the dipeptide has a physiologically probably irrelevant edge of 9% over the GLN + ALA combination.

Figure 2: Glutathione (GSH) and glutathione disulfide (GSSG = used glutathione) levels in soleus and gastrocnemius skeletal muscles of the rodents; data expressed relaitve to control (Petry. 2013)

For the muscular GSH levels, it does not look much different. In this case, there is however not even a statistical difference between alanyl-glutamine and the simple l-alanine + l-glutamine mix - neither for the universal anti-oxidant glutathione (GSH), nor for its "used form" glutathione disulfide (GSSG).

Does that mean that alanyl-glutamine is another supplemental rip-off?I would say that it's too early to use such harsh words. There was after all one statistically, and maybe even physiologically relevant difference between the two groups I didn't mention, yet: The dipeptide group presented with a different heat-shock protein response: They had higher HSP-70 and lower HSF-1 levels in the soleus and lower HSP-70 and lower HSF-1 levels in the gastrocnemius.

"Will training your biceps, heal your heart & protect your brain!?" - a study on the effects of exercise induced HSP increases suggests so | more

In view of the fact that the subsequent "deficit in HSP70 expression" is supposed to "impair recovery from these injuries" Petry et al. are probably right to point out that

"one cannot discard the possibility that part of the beneficial effects of high-intensity exercise training may be due to the enhancement of HSP70 expression which is exacerbated by glutamine supplementation."

In view of the fact that the total amount of proteins from the HSP70 and HSF1 family was increased in both groups, and the differences appear random, it is impossible to tell, whether the slight differences in HSP expression actually matter and whether this is an advantage for alanyl-glutamine or rather for the cheap free form amino acids.

Before future studies provide additional data based on which we can decide whether these differences are relevant and why they differ between slow- (soleus) and fast-twitch (gastrocnemius) skeletal muscle fibers, I'd say that the study at hand would suggest that alanine and glutamine have muscle protective effects irrespective of whether they are bound or not, when you ingest them.

References:

• Cruzat VF, Rogero MM, Tirapegui J. Effects of supplementation with free glutamine and the dipeptide alanyl-glutamine on parameters of muscle damage and inflammation in rats submitted to prolonged exercise. Cell Biochem Funct. 2010 Jan;28(1):24-30. 
• Cruzat VF, Tirapegui J. Effects of oral supplementation with glutamine and alanyl-glutamine on glutamine, glutamate, and glutathione status in trained rats and subjected to long-duration exercise. Nutrition. 2009 Apr;25(4):428-35.
• Hoffman JR, Williams DR, Emerson NS, Hoffman MW, Wells AJ, McVeigh DM, McCormack WP, Mangine GT, Gonzalez AM, Fragala MS. L-alanyl-L-glutamine ingestion maintains performance during a competitive basketball game. J Int Soc Sports Nutr. 2012 Mar 7;9(1):4.
• Petry ER, Cruzat VF, Heck TG, et al. Alanyl-glutamine and glutamine plus alanine supplements improve skeletal redox status in trained rats: Involvement of heat shock protein pathways. Life Sciences. 20 November 2013 [ahead of print]
• Rogero MM, Tirapegui J, Pedrosa RG, Castro IA, Pires IS. Effect of alanyl-glutamine supplementation on plasma and tissue glutamine concentrations in rats submitted to exhaustive exercise. Nutrition. 2006 May;22(5):564-71.

"Just One More Set" (1/2): Metabolic Response to 10,000kg vs. 20,000kg Regimen. EPOC: Do Reps and Loads Both Figure? And What About Elite Athletes Do They Need More?

"Ah come on, just another set!" ... "I don't know man, we've already pumped away 100,000kg today... do you really believe that's going to be productive, I mean, yeah, we are cuttin', but still... I mean I don't dig this epic!", "EPOC man, it's called EPOC!" *shakes his head* "Call it whatever you want, bro, I am out!"

If you want, you can think of today's SuppVersity post as an extension to yesterday's "Bigger, Stronger, Faster" special of the On Short Notice series; to be more precise: As a practically more relevant version of the rodent study on hypertrophy vs. strength training that was part of the aforementioned post. Yep, we are "talking volume" today. How much is too much?  And though this is never-ending debate, it appears that at least as far as research goes, a little more debating certainly would not hurt. Therefore I am happy to have not one, but two studies for you which don't just address this issue, but have also been conducted with human subjects!

In view of the fact that these are no "short notices", I will discuss one today and the other tomorrow - yep, that means that you can already make a mental note to come back tomorrow ;-)

"Just one more set, ..." - how productive can that be?

Today's study comes from the Human Performance Laboratory at the Florida State University and deals with the energetic side of things - specifically the often-cited EPOC (excess post-exercise oxygen consumption), which is often touted as one of the most important aspects why strength training in general and higher volume / intensity strength training, in particularly, would have the edge over cardio training. The reasoning is easy: You don't burn so much energy while you work out, but in the time after, your body will (a) still expend more energy per minute / hour and (b) has the advantage of emptied glycogen stores, which will force it to tap into its body fat stores the source for the required energy.

All of you who have read the complete Athletes' Triad series, will by now already know that at least argument (b) is pretty idiotic, because it wouldn't allow you to replenish your muscle and liver glycogen after workouts and thus pave the way into the dreaded vicious circle of the athlete's triad. The former argument, on the other hand has - on a way more general level - only been confirmed a couple of days ago (see "Scientists resolve the paradox of stable muscle metabolism but greater mitochondrial respiration in muscle of inactive vs. active subjects", read more), the question still remains: How much weight do you have to lift to set the 'afterburner' into full gear? 

10 metric tonne or 20 metric tonnes? What do you say?

I see, you are laughing, but basically the above question is what the George J. Abboud and his colleagues tried to find out, when they recruited 8 healthy men aged19-29 yrs who had 

"at least 12 months of RT experience with no more than 2 wks rest at a time, less than a total of 4 wks off within the last 6 months, or 9 wks off within the last 12 months [and] reported no prior or current use of illegal performance enhancing substances." (Abboud. 2012)

Suggested read "Three is More Than One: Higher Volume Increases Strength Gains in Legs, and Satellite Cell Recruitment and Fiber Size in Legs & Traps."

As usual the subjects had to fill food logs for the three days before the testing and were instructed to replicate the same eating pattern on the second occasion in which they were randomly assigned to perform a standardized resistance training (RT) regimen consisting of 4 exercises performed on a non-counterbalanced smith machine so that the range of motion during

• bench press, 
• squat, 
• bent-over row and 
• Romanian deadlift 

could be controlled for easily. Other than the equipment and the exercises, which were identical on both occasions, the volume of the training sessions varied and if you express this volme in kg or metric tons, it was a competition of 10,000 kg (10 metric tonnes) vs. 20,000 kg (20 metric tonnes) of weight. 

"The loads were divided between the 4 exercises as follows: 35% to squats, 30% to bench press, 20% to bent-over rows and 15% to Romanian deadlift. For each set, subjects lifted approximately 85% of their 1RM for 6-8 repetitions. If 6 repetitions could not be completed at any point, the load was reduced by 10% for the subsequent set." (Abboud. 2012)

Both sessions were supervised by three testers : One monitored the metabolic cart, one made sure the proper range of motion was used and one monitored the proper lifting form. The subjects had to perform the concentric portion of each lift with maximal speed and ensure a controlled eccentric descent. A specific time interval was not dictated. Sets were stopped if "subjects broke form" (Abboud. 2012) and 2 minutes of rest were given between sets. In this fashion the subjects simply kept lifting set after set until the volume prescription for the respective trial was reached.

Figure 1: Resting metabolic rate (RMR) per kg body weight, 30min energy expenditure and respiratory exchange ratio (RER; lower values = higher fat, lower glucose  oxidation)  after low and high volume trial (based on Abboud. 2012)

As you can see in figure 1 there were differences as far as the effects of the high (20,000) vs. low (10,000kg) regimen on the resting metabolic rate, 30 min energy expenditure, and the respiratory quotient (lower values = higher fat, lower glucose oxidation), but in view of the fact that the high volume group moved 2x more weight and should thus (at least theoretically) have expended twice the energy (assuming they performed all reps with perfect form and identical speed), those differences are more than disappointing. 

The minuscle effect size is yet not the most "disappointing" (or "surprising" ?) result

In fact, contrary to the low volume workout the 20,000kg workout did not produce any increases in resting metabolic rate and 30min energy expenditure, at all - put simply: There was no EPOC after the high volume training. And this did not change over the whole 48h period (and I know you guys, you won't rest longer anyway ;-).

Now you may say that this was a crazy protocol, but let's do the math, let's assume the guys did squat 100kg, benched and rowed their own body weight of ~80kg and deadlifted 125kg. With 10 reps per set thats 1,000kg + 2x800kg + 1,250kg per set respectively. If they did three sets per exercise they would thus already be up to 9,950kg! If you still think that's crazy, let's hear what the scientists have to say:

"As subjects in the present study were well adapted to RT, the training stimulus needed to elicit increases in EPOC arguably needed to be much higher compared to that used in previous research  Two studies using intensities of 70% 1RM report significant increases in RMR. Melby et al. had subjects perform 6 setsof 10 different exercises for a total of 60 sets. The repetition range for this protocol was 8-12 repetitions per set. This amounts to approximately 600 repetitions performed during the course of the exercise bout. The range of load-volume lifted by these subjects was 15,000-38,000 kg. [...]" (Abboud. 2012)

The list goes on and you just have to go to your gym and I guarantee you, no matter how few people are on the floor you will see a guy who (often without noticing is) will be pounding away much more word within a single workout. Moreover, the the subjects in the present study completed their trials
with a drastically lower number of repetitions -  a mean of 199. Had they performed the crazy rep-volume of the Melby study, they would probably have come close to 50,000 kg. This raises an interesting question is "volume" correctly defined by giving the total amount of weight you lift? Or is the number of reps maybe more important as far as the after-burner EPOC is concerned?

Too much of a good thing? But what if you are a highly trained athlete?

A previous study, by Hackney et al. would support the notion that heavy lifting is an obligatory part of the EPOC equation. In the latter study, EPOC trained individuals who used a lower load-volume than the trainees in the study at hand  had increased resting metabolic rates for up to 72h (Hackney. 2008). Since the Hackney study also put an emphasis on eccentric contractions and will thus probably have lead to even greater muscle damage than the protocol of the study at hand (CK(10,000kg) = 729U/L vs.CK(20,000kg)  = 1,159IU/L), Abboud et al. speculate that ...

"[a]s protein synthesis required for repair is energetically expensive, it is logical that untrained subjects will show greater and longer alterations in EPOC post-RT. Judging by training history, strength levels and CK responses, subjects in the present study had most likely reached a higher level of adaptation than ones in previous studies, and therefore were less sensitive to the metabolic effects of recovery from RT." (Abboud. 2012)

In other words, for you, probably a seasoned strength trainee, the 'more is more' principle is not going to yield better results - even if your goal is to shed body fat. And ...

"Although RT is an important component in any weight loss program to attenuate the loss of fat free mass and therefore better preserve RMR, it is unlikely that the total energetic cost (during and post-exercise) of a typical duration workout will be adequate for significant weight reduction in highly trained recreational lifters without caloric restriction and/or additional aerobic or high intensity interval training." (Abboud. 2012)

And since I rarely encounter a conclusion that's so to the point, I'll leave you with that for today and remind you to come back tomorrow to learn, when and for which body parts doing somewhat more may still be beneficial - read me tomorrow ;-)

References:

• Abboud GJ, Greer BK, Campbell SC, Panton LB. Effects of Load-Volume on EPOC after Acute Bouts of Resistance Training in Resistance Trained Males. J Strength Cond Res. 2012 Oct 18.
• Hackney KJ, Engels HJ, and Gretebeck RJ. Resting energy expenditure and delayed-onset muscle soreness after full-body resistance trainingwith an eccentric concentration. J Strength Cond Res. 2008; 22: 1602-1609.
• Melby C, Scholl C, Edwards G, and Bullough R. Effect of acute resistance exercise on postexercise energy expenditure and resting metabolic rate. J Appl Physiol. 1993; 75: 1847-1853.

HMB Supplementation: Pre- or Pre- and Post-Workout? Anti- or Pro-Inflammatory? MA Thesis Offers Food for Thought

Supplement facts: "Is as potent as the weak androgen Oxmethalone aka Anavar!" If that's how you advertise an expensive dietary supplement that tastes like poison, you better make sure your product delivers. For HMB the supplement companies must have overlooked that their clientele is in no way similar to the elderly subjects from their references with their protein deficient diets... the result? An epic fail for both the consumers who felt ripped off and the producers who probably expected this to be a long-term investment!

HMB was once hailed to be as effective as a "weak" androgen such as Anavar. No wonder that dozens of consumers were pretty  disappointed, when they added it on top of their already protein and, at that times more or less coincidentally, leucine-laden diets and saw... nothing. Well, at least no gains that would even remotely remind anyone of Oxymetholone. With no costumer being interest to pay the extra bucks for the (at that time still) very expensive product, it was no wonder that the leucine metabolite β-Hydroxy β-methylbutyric acid (HMB) disappeared from the market relatively quickly.

I bet, the fact that it tastes like poison did not really help either. after all it is downright impossible to add an effective amount of HMB into a powdered supplement, if you do not want to totally ruin the taste of the product.

As a SuppVersity reader you will yet be aware that HMB is still no epic fail (read all older posts on HMB). Its marginal utility, however, is exactly that: Marginal -- at least when someone is taking it on top of tons of leucine rich protein powders, BCAAs and whatever else.

HMB is not useless and there appears to be much we have to learn about it

Despite the fact that it is very unlikely that taking HMB will turn you into a second Phil Health within weeks, its hitherto not fully understood beneficial effects on body fat, its effects on GH and IGF-1 as well as open questions that are related to our own bodies ability to produce HMB from leucine and whether this conversion mediated some of the benefits of the #1 among the BCAAs (=leucine) clearly indicate that there remains a lot to learn about Dr. Steven L. Nissen's 1996 discovery (Nissen. 1996).

One of those things we still have... or I should say had to learn pertains the effect of HMB on the exercise induced expression of inflammatory cytokines, which turned out to be totally different from what Paul Raymond Vulcan, a student who has recently submitted his Master Thesis on the "Role of β-hydroxy-β-methylbutyrate (HMB) on inflammation after eccentric exercise" at the Iowa State University probably expected, when he recruited the 16 female and 16 male, untrained volunteers (mean age 3±0.30y, mean weight 67.5±0.9kg, and mean hight 172.2±0.7cm) for his study.

'Extend your legs' till they burn ;-)

The study consisted of a single exercise + supplementation trial in the course of which the participants underwent the following supplementation regimen:

A note on the supplementation protocol: The way Volcan describes the protocol is at least "suboptimal". If I am getting it correctly (mostly by looking at a graphical outline) the participants in the pre/post trial received HMB not just on day 0, but also on day 1, day 2, day 3 and day 4 - always with lunch and dinner.

"Experimental groups received supplement in one of the following manners: placebo pre- & post-exercise (CON), HMB pre-exercise (PRE) in either a calcium salt form or a free acid gel, HMB pre- & post-exercise (PRE/POST) in either a calcium salt form or a free acid gel.Supplements were given in a double-blind protocol so that investigators were also blind to the contents of the supplements throughout the study.  Originally, the study called for 5 treatment groups with a separation of calcium salt groups from the free acid gel groups. Due to sample sizes, groups receiving the same quantity of HMB were consolidated for statistical reasons." (Volcan. 2012)

While it is certainly somewhat disappointing that we don't have a comparison of the salt and the gel variant of HMB (you will see the first gels hit the market, very soon, believe me),  this is understandable given the small sample size. What is yet a clear greenhorn mistake, however, is that Volcan does not disclose the amount of HMB salt the participants received. He states that the gel syringes contained 3ml but since I have no clue what else (besides HMB) is in the gel, I cannot tell you how much HMB the groups actually received (let alone calculate the equivalent in term of calcium-hmb).

Standard protocol, surprising results

Aside from this lapse the general protocol of the study looks pretty solid. The original challenge on day 0, which consisted of 3 sets of 50 eccentric leg extensions (both legs, 0° to 90°, 60°/sec; 3s per rep, 2 min rest between sets), was preceded and followed by a combination of

• urine collection, 
• muscle soreness test, 
• leg circumference measurements, 
• blood collection and a 
• strength test

which took place on day 0 (before the exercise protocol), 24h, 48h, 72h and 96h post. And I guess you could say "fortunately", the analysis of the respective data yielded not exactly what Volcan had expected. Firstly many expected effects did not occur (at least not if you consider only statistical significant inter-group effects), e.g. there were no differences for markers of muscle damage:

• creatine kinase (CK) 
• lactate dehydrogenase (LDH) and 
• 3-methyl histidine (3-MH)

Some did however show a trend (which did not reach statistical significance due to the small sample size) and / or did reach statistical significance at a certain time point only:

• 

  Figure 1: Peak performance force (in N) on the days after the leg extensions (based on Volcan. 2012). As you can see it's not like there had not been any effects, but with the small sample size few made it over the p < 0.05 hurdle, or put simply were "statistically significant".

  creatine kinase (CK) showed a non-statistically significant reduction in PRE/POST, while it was identical in PRE and CON (1593 IU compared to 3514 IU and 4068 IU; this is a candidate that would almost certainly have reached statistical significance with a larger number of participants, because the CK response shows high inter-individual variability)
• the decrease in right leg peak force was about 16% in the CON group compared to 9% and 7% in the PRE and PRE/POST groups, respectively
• the muscle soreness was also not significantly different between groups
• the difference in the loss of peak force was only significant on day two when the peak performance in the control group dropped significantly (see figure 1)

So far the somewhat disheartening but not actually novel part of the study. With the data in figure 2, however we are actually approaching the real news part of today's post:

Figure 2: Makers of inflammation (IL-1 and TNF-alpha) after 3x50 leg extension on the test day, as well as 24h, 48h, 72h and 96h post (based on Vulcan. 2012)

Take a close look and don't be fooled like I was, when I initially looked at the results and almost automatically assumed that there was a reduction in TNF-alpha and IL-1 in the group that recevived the most HMB (I must say I had only the poor black and white graphs from the original study and not the full-service pack you get, here at the SuppVersity, though ;-)

"I mean, good things reduce inflammation, right?"

Wrong, at least in the case of HMB, which certainly is a good thing, this is not the case. HMB does not reduce inflammation and that despite the fact that it works like a charm for those people of whom we are told that inflammation was the last thing they would need, namely the elderly. And still it appears as if the increased inflammatory response to the muscle damaging exercise on day 0 could actually be part of how HMB works. Volcan realizes that and states:

Just in case you don't want to believe that inflammation could play a beneficial role in the regeneration and even the subsequent super-compensation process at the end of which both your muscle strength and muscle size will go up, I suggest you read the respective installment of the Intermittent Thoughts

"The results of TNF-α and IL-1ra support the theory that inflammation is affected by HMB. In  both cases the CON group experienced a decline in serum concentration and the dip was reduced or limited by supplementation of HMB.  These results could be interpreted as an increase in the inflammatory response following HMB supplementation [...] This suggests that HMB creates a greater inflammatory response which may improve recovery of damaged tissue. When exactly this occurs and how, whether direct or indirect, is not evident from this study. We already kno that proteolysis is affected by HMB and it is also possible that inflammatory cytokines are mediating an optimal recovery." (my emphases in Volcan. 2012)

Edited: While this sounds like an excellent hypothesis there is one thing Volcan has overlooked (and me too, at least initially, thus the update). The current scientific evidence suggest that the IL-1 receptor is like a multifaceted chimera. Or put simply, in its conventional form it will accept IL-1 and thus have exert pro-inflammatory downstream effects. IL-1-RA, which is the variant Volcan measured here, actually does the opposite, though: IL-1-RA blocks the inflammatory effects of IL-1 (Arend. 1997).

In the end, this does not really change my previous assertion that "You need to go beyond the 'all inflammation is bad' paradigm and see 'inflammation', or what we usually refer to as inflammation as what it really is, namely a per se physiological reaction of our bodies that can be good, appropriate and highly desirable or bad, misplaced and highly detrimental depending on the circumstances." It just adds another level of complexity to that statement. And this added dimension can actually explain why HMB works in the elderly, but does not work (at least not to the same degree in young people). Old people have high IL-1 (the pro-inflammatory varieties), young and healthy people don't, so blocking IL-1 signaling will have more pronounced effect in the older ones of you than in the young chaps, who will probably fare quite well with nothing but a leucine rich protein shake.

References:

• Arend WP. Interleukin 1 receptor antagonist. A new member of the interleukin 1 family. J Clin Invest. 1991 Nov;88(5):1445-51.
• Nissen SR, Sharp M, Ray JA, Rathmacher D, Rice JC, Fuller Jr, Connelly AS, Abumrad N: Effect of leucine metabolite beta -hydroxy-beta -methylbutyrate on muscle metabolism during resistance-exercise training. J Appl Physiol 1996, 81:2095-2104.
• Volcan PR. Role of β-Hydroxy-β-methylbutyrate (HMB) on inflammation after eccentric exercise. Graduate Theses and Dissertations. 2012; paper 12501.

SuppVersity Science Round Up Seconds: Wheat Gluten Hydrolysates Fail, Exposure to Air Pollutants During Workout Reduces Brain Benefits, Homocysteine, B-Vitamins, Cognitive Impairment and Mortality

Before the profound weight loss (A) you don't see any of the glucose sucking and fad burning brown fat depots (black spots in B) on the neck of the in (B) 'foermerly obese', now only 'overweight' subject (also take a look at how the visceral fat in the abdominal region in (A) is actually pushing the organs upwards; img Vijgen. 2012)

Those of you who have listened to yesterday's show will have noticed that despite its flow the number of things you can discuss in a 1h podcast is simply very limited, to say the least. This is also why these Friday posts are probably never going to be simple summaries of the SuppVersity Science Round Up of the day before. The same is true for today and still I decided not to use the allegedly lame logo I did for the first two installments, but provide you with some 'real science' evidence of the absence of brown adipose tissue on the obese and it's magical reappearance after shedding 100lbs+ subsequent to a gastric bypass operation, instead (see image on the right).

Assuming that you have no idea what this "evidence" is for, I would suspect that you missed the live show yesterday and also did not find the time to download and listen to the podcast, yet -- right? Well, you should either download and listen to the show now and digest the Seconds later, or you read the following paragraphs first and download the podcast later.

What is not an option, however, is to miss one or another - I mean you can hardly want to eat the seconds if you have not had the main dish yet... and after listening to the podcast, I cannot imagine you don't want at least some seconds. Apropos seconds, here are today's seconds...

• Wheat gluten hydrolysate is not the new goto protein supplement - certainly not for female distance runners and probably not for anyone else, either! These are the kinds of studies that really annoy me. Studies that start out with blatant statements like "WGH [Wheat gluten hydrolysate] has been reported to suppress post-exercise rises in serum creatine kinase in male distance runners" (Hirao. 2012).
  
  

  

  Figure 1: CK, AST, ALT response in the "success trial" with men. In women even the miniscule beneficial effect on CK was not there. No reason to even think about buying a gluten hydrolysate as you new go-to protein supplement with only 5.6g of leucine/100g (whey has 50% more) and almost no GSH replenishing cysteine in it (0.9g vs. 3g+ in whey, which is more than +200% more).

  Sentences like that make the null-results of the study they precede look like the exception to the rule and are still nothing but a concession to a bias (let's hope not due to the grant from Nisshin Pharma Inc. which was the manufacturer of the wheat gluten hydrolysate used in this study). A bias, due to which an isolated observation as the slightly blunted increase in CK is blown up as if a slightly lower CK level was what could turn a sedentary pencil pusher into the next Hussein Bolt (Aoki. 2012).
  
  So, even if you are not afraid of the evil in gluten (which I believe not everyone has to), I strongly caution against making the switch from a high EAA protein with ton's of GSH boosting cysteine in it like whey to a mediocre grain protein, which is a potential allergen and contains tons of glutamine your body will readily turn into glucose, once it passes through the portal veign into the liver (I bet a large part won't even make it into systemic circulation).
  
  And as far as the purported "gender difference" goes the study at hand tries to blame the null result on (Hiriao. 2012), I suspect that it is rather the indisputable difference between the long-distance running at a continuous pace the women in the study at hand did, versus the totally different strains the guys in the previous study were exposed to during a soccer training + mini-match, which made the difference.
• Working out next to a street takes away some of the beneficial cognitive effects due to ultrafine particulate matter (UFPM) exposure. "Working out in the fresh air will promote weight loss more than working out inside." You heard me state that in one of the previous installments of the SuppVersity Science Round Up on Super Human Radio. Now this is still correct and based on sientific evidence, but at least as far as the cognitive benefits are concerned, working out outside does also have its downsides - at least for those of you who live in the inner city area.
  
  

  

  You better watch what you breath while you run.

  During a 12-week program the researchers from the Universiteit Brussel, the Hasselt University and the Royal Military Academy measured the improvements in physical performance, changes in serum markers and corresponding ultrafine particulate matter (UFPM) concentrations in the enviromnent in which their 15 previously untrained subjects conducted their aerobic training program thrice a week (Bos. 2012). What Bos et al. found was that the UFPM levels were signfificantly higher in the urban compared to the rural environment and that the higher UFPM exposures correlated with increases in leukocyte counts (p = 0.02), neutrophil counts (p = 0.04), and eNO levels (p = 0.002) that were exclusively observed in the group that trained in the urban environment.
  
  With the latter being markers of inflammation which exert their effects systemically, i.e. not just in the lung or musculature of which you may be thinking now, but also in the brain, it is no wonder that
  

  "reaction times on the Stroop task improved in the rural group (p = 0.001), but not in the urban group" (Bos. 2012). 

  What's comforting, though, is that the physical fitness did increase to a similar extend in both arms of the study.
• Homocysteine levels, mortality, cognitive impairment and which nutrients can offer some protection. I am not sure about what your impression is, but for whatever reason homocystein seems to be 'out of vogue' -- probably no room for it on the research agenda with all the hype surrounding vitamin D. It used to be all the rage in CVD risk research and today's news item is actually ain't about cardiovascular health, either.
  
  What the researchers from China and Taiwan actually were interested in was the correlation of high and low homocysteine levels with cognitive impairment and the corresponding nutrient intakes. In that Xiu et al. paid particular attention to the "B-vitamins" and found the following correlations between the mortality, cognitive status, homocystein levels and nutrient intake of their 1412 study participants (Xiu. 2012):
  

  • 

    Figure 2: Unadjusted mortality in the four quartiles of homocysteine levels (top); mortality according to homocysteine levels in subjects with different degrees of cognitive impairment (based on Xiu. 2012)

    if you go by the unadjusted data in figure 2, it's plain obvious that the all-cause mortality increases linearly from one quartile to the other 
  • this relation between plasma homocysteine levels and all mortality remained statistically significant after adjustments for age, sex, smoking status, BMI, physical function and general health were made
  • of the general foods, the scientists assessed, only regular fish intake had a statistically significant effect on homocysteine levels, with higher intakes being associated with lower homocysteine levels
  • of the b-vitamins choline was the only one with a significant association with plasma homocysteine levels (suggested read "Old School Supplement Choline Could Save Your Live and Liver!") 
  • neither betaine, nor vitamin B1, B2, B3 or B6 intakes did show statistically significant correlations with plasma homocysteine (not even "borderline significant; p > 0.15 for all, most way hither)
  • of the plasma markers, folate showed a highly significant correlation with homocysteine (14.4 nmol/L in the lowest HCY and 8.70 nnmol/L in the highest HCY group)
  • PLP, the active form of vitamin B6, came in close second with 70.3 nmol/l in the lowest HCY quantile and only 44.4 nmol / l in the highest quantile.
  Now, if you consider the fact that higher intakes of B-vitamins are probably not doing much to lower homocysteine levels int he elderly (at least not dose-dependently, when they are already getting enough) oddity #1, another look at the data in figure 2 will reveal oddity #2: The surprisingly high mortality in the lowest homocysteine quartiles in the patients with severe cognitive decline - how come? I mean, with low homocysteine they should not be at risk of having severe cognitive decline, anyway - right?
  
  Actually if you follow this rationale you can almost answer the question yourself. If you have low homocysteine and severe cognitive decline, the severe cognitive decline can hardly be from high homocysteine levels, so it must have another obviously pathological reason, or as the scientists have it
  

  "The joint effects of the 2 variables [homocysteine and cognitive decline] were most pronounced with severe cognitive impairment where mortality HRs ranged from 5- to 18-fold across a wide range of homocysteine concentrations. The findings with hypohomocysteinemia provide some insight into what might be an optimal range for this analyte in peripheral blood and tissues. The low concentrations may be seen with severe illness and malnutrition, and our study population comprises the health-vulnerable aged. For these reasons, we adjusted these associations for BMI (using the World Health Organization chronic energy deficiency category of, 18.5 kg/m 2 ), and we excluded those who died in the first year of follow-up. The findings were unchanged. Because mortality among the very old may have skewed the joint effects, these are presented for those ≤75 years and over, but again with similar findings." (

  A sarcastic person would now probably say: "We all have to go some time!" and just wave his hands at these results. True! And I am the last to advice you to become over-anxious. Yet in the mean time it would appear prudent to make sure to get your homocysteine levels checked from time to time, not to forget that choline is a b-vitamin as well and not to fall for the idea that you cannot overdose on B-vitamins - I don't have to remind you of the negative effects, specifically folic acid supplementation can have on all sorts of cancer (e.g. breast cancer, where a high folic acid intake from foods and supplements is associated with a +30% risk of cancerous growth; cf. Kim. 2006).

In case you are looking for the post on "ammonia accumulation brain-fog, toxicity, liver 'pathologies' and workout performance", yeah it was on the list, but I decided it would be a shame to tackle that within a short two paragraph seconds items. Don't worry I am not going to forget about it, after all its in my humble opinion one of the main reasons the diets and workout regimen of the many ambitious physical culturists fail. If you are still looking for more and have not listened to the podcast, yet, this would be the right moment to download the file from the Super Human Radio Network server (click here to download), otherwise the latest short news on the SuppVersity Facebook Wall may offer some diversion ;-)

References:

• Aoki K, Kohmura Y, Suzuki Y, Koikawa N, Yoshimura M, Aoba Y, Fukushi N, Sakuraba K, Nagaoka I, Sawaki K. Post-training consumption of wheat gluten hydrolysate suppresses the delayed onset of muscle injury in soccer players. Exp Ther Med. 2012 Jun;3(6):969-972. Epub 2012 Apr 3.
• Bos I, De Boever P, Vanparijs J, Pattyn N, Panis LI, Meeusen R. Subclinical Effects of Aerobic Training in Urban Environment. Med Sci Sports Exerc. 2012 Oct 15.
• Cankurtaran M, Yesil Y, Kuyumcu ME, Oztürk ZA, Yavuz BB, Halil M, Ulger Z, Cankurtaran ES, Arıoğul S. Altered Levels of Homocysteine and Serum Natural Antioxidants Links Oxidative Damage to Alzheimer's Disease. J Alzheimers Dis. 2012 Oct 29.
• Guest PC, Urday S, Ma D, Stelzhammer V, Harris LW, Amess B, Pietsch S, Oheim C, Ozanne SE, Bahn S. Proteomic analysis of the maternal protein restriction rat model for schizophrenia: Identification of translational changes in hormonal signalling pathways and glutamate neurotransmission. Proteomics. 2012 Oct 16.
• Hirao T, Koikawa N, Aoki K, Sakuraba K, Shimmura Y, Suzuki Y, Sawaki K. Female distance runners show a different response to post-workout consumption of wheat gluten hydrolysate compared to their male counterparts. Exp Ther Med. 2012 Apr;3(4):641-644.
• Kim YI. Does a high folate intake increase the risk of breast cancer? Nutr Rev. 2006 Oct;64(10 Pt 1):468-75.
• Vijgen GH, Bouvy ND, Teule GJ, Brans B, Hoeks J, Schrauwen P, van Marken Lichtenbelt WD. Increase in brown adipose tissue activity after weight loss in morbidly obese subjects. J Clin Endocrinol Metab. 2012 Jul;97(7):E1229-33. Epub 2012 Apr 24.
• Xiu LL, Lee MS, Wahlqvist ML, Chia-Yu Chen R, Huang YC, Chen KJ, Li D. Low and high homocysteine are associated with mortality independent of B group vitamins but interactive with cognitive status in a free-living elderly cohort. Nutr Res. 2012. Ahead of print.

High and Low Dose BCAA Supplementation Have Minimal, Non-Significant Effects on Markers of Muscle Damage 24h and 48h Post Heavy Resistance Training

Image 1: Cover of the September issue of the International Journal of Wrestling Science - don't tell me you don't have a subscription, yet!

I don't know about you, but I feel that it's quite interesting to look at the highly heterogeneous dosage suggestions on the labels of the ever-increasing number of BCAA supplements on the market. Interestingly, almost every producer claims in his "non FDA-approved" statements that his supplement contains "scientifically supported" or "clinically validated" amounts of branch-chained amino acids in the "optimal" (whatever that may be) ratio of 2:1:1, 3:1:1, 4:1:1, 8:1:1, ... and all the other variations that appear to be limited only by the patent applications and lawyers of the financially more potent players in the business. From a scientific perspective, however, this "optimal" amount has still to be elucidated - at least to my knowledge, no respectable scientist has yet claimed to have found the "optimal" amount and composition of free form amino acids for a given subgroup of athletes, let alone strength athletes, bodybuilders or figure competitors, in general.

At this point I would like to add that no respectable scientist would ever dare to make the claim that he or she has found the "optimal free form amino acid supplement" for all, or even a significantly large group of athletes, unless he or she would be interested in losing his reputation as a "respectable scientist" ;-)

In a recently published study scientists from the Department of Physical education and Sports Science University of Tabriz in Tabriz, Iran, set out to establish whether there is at least a significant difference between the effects of ~15g (210mg/kg) or 33g (450mg/kg) of branched chain-amino acids taken before and after the completion of an intense resistance training regimen comprised of 7 exercises à 3 sets of 10 repetitions (Amirsasan. 2011). Yet, despite the fact, that even the "low dose" of 15g of BCAAs (in the customary 2:1:1 ratio, i.e. 7.5g of leucine + 3.75g of iso-leucine + 3.7g of valine) was about 1.5x higher than what I have seen as "suggested dosing" or "serving size" on very high-dosed commercial supplements, the effects of this amino acid overkill were "sobering", to say the least.

Figure 1: Effects of "low" (210mg/kg) and "high" (450mg/kg) dose BCAA supplement on enzymatic markers of muscle damage relative to pre-values in the placebo group (data calculated based on Amirsasan. 2011)

As the data in figure 1 goes to show both the "low" as well as the "high" (or should I say "overkill" ;-) dose of pre- and post-workout BCAAs had only marginal, and certainly statistically non-significant effects on creatine kinase (overall - CK; muscle specific - CK MB) and lactate dehydrogenase activity, both established indicators of (exercise-induced) muscle damage.

Comparison of results between groups in mean and amplitude changes of serum indexes of cell damage (CK-LDH-CKMB), 24 and 48 hours after the exercise performance showed no significant difference between the 3 groups. In other words, different amounts of BCAA did not significantly affect the serum cell injury indexes (CK-LDH-CKMB), 24 and 48 hours after the heavy resistance activity.

These results are interesting, because they contradict previous findings by Sharp et al. who reported "significantly reduced" creatine kinase levels with BCAA supplementation in likewise previously strength-trained athletes on a similarly intense (8 exercises; 3x 6-8 repetitions) resistance training protocol (Sharp. 2010), as well as the results of studies in endurance athletes and previously untrained subjects, where the provision of BCAAs decreased creatine kinase and lactate dehydrogenase enzyme expression, across-the-board (Greer. 2007; Koba. 2007; Matsumoto. 2009).

"To supplement or not?" This question may arise if you have a look at the data from this study. Thor, in a comment to this posts poses the question whether his "personal experience" that "having the fast digesting aminos seemed to increase [his] ability to have a more successful work out" is, after all "only in [his] head" and while I cannot say for sure how much of it may be the result a placebo-effect in his case, I can provide you with the results of a 2011 study by Greer et al. who found no increases in exercise performance despite reduced perceived rates of exertion with BCAA supplementation after a 90-minute cycling bout (Greer. 2011). These results seem to confirm the "central fatigue hypothesis" according to which BCAAs exert their beneficial effects agains (perceived) fatigue via modulation of the availability of the serotonin precursor tryptophan. In a 2007 review of the literature, Meeusen and Watson do yet conclude that the "nutritional manipulation of these systems [neurotransmitter] through the provision of amino acids has proven largely unsuccessful" (Meeusen. 2007)... All that does not take away from the established beneficial effects of chronic low-dose BCAA (in particular, leucine) supplementation on endurance performance and strength adaptations to exercise (e.g. Crowe. 2006; Matsumoto. 2009). In the respective studies, dosages in the 1.5-3.0g/day range have yet been sufficient, to elicit these beneficial effects - and that in subject groups that are not particularly well-known for their exorbitantly high protein intakes ;-)

Image 2: When bought in bulk and without the addition of a ton of fancy extras BCAAs have become reasonably priced - whether they are a "necessary" part of your supplement regimen may yet depend on your dietary protein intake.

Probably - this would at least be my first guess - the outcome of these studies was not so much affected by the actual study protocol, but rather by the habitual dietary protein intake of their subjects. With endurance athletes (Koba. 2007; Matsumoto. 2009),  recreationally active (Sharp. 2010) and untrained (Greer. 2007) we usually see much lower dietary protein intake than with professional wrestlers, which prompts me to repeat my previously stated skepticism towards the usefulness of large boluses of additional free form amino acids in a group of athletes whose habitual dietary protein intake is way beyond the 1.5g/kg level, anyway... but hey, that's just the opinion of a brainy physicist; so if your brawny guru says you need those 150g of BCAAs on top of your 5x50g whey protein shakes and your 3 pound of lean meat - go for it!

Is Hydrolized Whey, the New Way to Go? 12 Week Human Study Suggests: Yes, If Your Goal is to Ward Off Oxidative Damage. No, If You Want to Build Muscle & Lose Fat

The typical soccer player is no longer a stick on muscular legs, these days. The sport has changed and so have the physiques of the players.

I guess ever since I published the article "The Glucose Repartioning Effects of Isoleucine: Falsely Underappreciated BCAA and Its Dipeptides Maximize GLUT-4 Expression and Ramp Up Muscular Glucose Uptake" (read it) that discussed the beneficial effect of the small isoleucine peptides in hydrolized whey on glucose metabolism, some of you may have been wondering, whether theh previously sneered at even more insulinogenic fast-digesting, bad-tasting, highly-processed whey protein hydrolysates (WPHs) may not be an alternative, if not the better alternative to whey concentrates or isolates.

Personally, I have always favored the "whole" over its individual parts, but the evidence that there is something special about WPH is accumulating.

"So what kind of new evidence is accumulating here?"

That being said, the latest evidence that would support this notion comes from the Universidade Estadual de Campinas in São Paulo, Brazil (Lollo. 2013). Where Pablo Christiano B. Lollo et al. investigated the effects the provision of whey protein (WP), hydrolysed whey protein (WPH), or a non-protein placebo (maltodextrin, MALTO) would exert on selected biochemical, anthropometric and performance parameters in 24 soccer player over the course of 12 weeks.

The iso-caloric supplements which contained 0.5g of protein (or placebo) per kg of body mass had to be before and after each training session, as well as on rest days (i.e. on Monday). The overall protein intake was relatively low (typical of a sport with a clear endurance focus) and was designed so that the protein of the diet plus that of the supplement would represent 15% of the total daily caloric intake. Both, the diets, as well as the sleep and training schedule were standardized. The intensity and training volume were identical for all individuals having plaing the same position (e.g. striker, defender, etc.). And the amino acid content of the two whey supplements was identical, so that the only difference between the WP and the WPH group was the chain-length of the proteins and peptides in the drinks they ingested.

Figure 1: Changes in markers of exercise induced damage (left) and body composition changes (in %) over the course of the 12-week study period (Lollo. 2013)

As you can see in figure 1 this minute difference was yet enough to result in significantly different responses to the protein supplement in the WPH and WP groups.Only, in the former, i.e. the whey protein hydeolysate group did the scientists observe significant decreases in the muscle damage indicators, creatine kinase (-42%) and lactate dehydrogenase (-30%). The minimal changes in the whey protein group, on the other hand, were not superior to the maltodextrin control.

"The foremost features of this investigation were greater than 40% and 30% decrease in CK and LDH, respectively, obtained after 12 weeks of supplementation with the hydrolysed whey protein. This outcome contrasted even more with the increase of CK, which was approximately  +35% obtained by supplementing with maltodextrin alone." (Lollo. 2013)

As you would expect there were no apparent adverse effects observed in either of the groups whose total total protein intake remained below 2.3 g/kg per day. The renal function and protein metabolism parameters, uric acid and creatinine remained within the physiological limits of normality in all groups. 

WPH the whey for elite athletes? 

In their discussion of the results, Lollo et al. make an interesting point, when they compare their observations to those in previously conducted studies and state:

Right from the archives: "Looking at Fast, Slow & Total Protein Intake. More Than 2g/kg Protein = Madness?" (read more)

"Since studies of the effects of protein supplementation have normally been carried out with non-athlete volunteers in acute experiments (Bolster et al., 2005; Pennings et al., 2011; Tipton & Ferrando, 2008), it was interesting to note that the data collected from elite athletes engaged in a real championship pointed to hydrolysed whey protein being the only form of supplement that effectively diminished the levels of muscle damage biomarkers.

Those results could be understood in light of the high antioxidant capacity of the hydrolysate." (Lollo. 2013)

If that could be confirmed for other elite athletes and highly trained gymrats, as well, the question I raised in the headline of this article may well be answered affirmatively: "Yes, for elite athletes with a high training workload, hydrolized whey proteins may in fact be the better way to go."

If you take a closer look at the data in figure 1 (right hand side), you will have to concede that this could well be a question of the kind of athlete we are looking at. A bodybuilder for example would fare better with regular whey. While soccer training is an allegedly bad model for bodybuilding, the way in which regular whey has the most favorable effects on body composition, i.e. a statistically significant 3.4% increase in muscle mass and non-significant 6% decrease in fat mass, is something I personally would not ignore (on a side note, you did see that the fat loss maxed out and reached statistical significance with the pure maltodextrose, right?). 

---

Metabolic ward study shows: Higher than RDA protein intake turns weight loss into a fat loss diet, yet still it's not the more the better (learn more)

Bottom line: Despite or maybe even due to its high anti-oxidant prowess whey hydrolysate may not be the ideal protein for the average gymrat trying to build muscle, lose fat and improve his overall body composition. Allegedly, a study in soccer players and above all one with only 24 subjects is certainly not a reliable gauge, but why would you quit using what has worked for you before, if the evidence that the alternative may be good, but not exactly conducive to your primary goals?

Moreover, let's not forget, if you are simply trying to pack in some additional protein into your diet, price and taste are certainly things you want to consider, as well. And let's face it the taste and mouth-feel of the few true hydrolysates out there (most supplement producers mix various forms of whey and will still sneak a "hydro" into the name) are not exactly what you would expect from something you'd consume as a "liquid snack", right?

References: 

• Bolster DR, Pikosky MA, Gaine PC, Martin W, Wolfe RR, Tipton KD, Maclean D, Maresh CM, Rodriguez NR. Dietary protein intake impacts human skeletal muscle protein fractional synthetic rates after endurance exercise. Am J Physiol Endocrinol Metab. 2005 Oct;289(4):E678-83.
• Lollo PBC, et al. Hydrolysed whey protein reduces muscle damage markers in Brazilian elite soccer players compared with whey protein and maltodextrin. A twelve-week inchampionship intervention. International Dairy Journal. August 2013 [epub ahead of print]
• Tipton KD, Ferrando AA. Improving muscle mass: response of muscle metabolism to exercise, nutrition and anabolic agents. Essays Biochem. 2008;44:85-98.
• Pennings B, Koopman R, Beelen M, Senden JM, Saris WH, van Loon LJ. Exercising before protein intake allows for greater use of dietary protein-derived amino acids for de novo muscle protein synthesis in both young and elderly men. Am J Clin Nutr. 2011 Feb;93(2):322-31.