No Advantage of Bolus Ingestion of EAAs in Young Men!? Cereal Bread Not Better for Weight Control. Saturated Fat & the Heart. Plus: Serine for Your Weekend Alcohol Binge!

The "muscle full effect" indicates you don't have to consume 4 scoops at once.

With the publication of the latest issue of The Journal of Nutrition came a handful of interesting scientific papers I will briefly introduce in today's SuppVersity Nutrition Science Update.

The corresponding studies deal with the link of saturated fat to heart disease (Puaschitz. 2014), the effects of proteinogenic amino acid serine (one of the non-essential amino acids) on homocysteine metabolism in a rodent model of alcoholic fatty liver disease (Sim. 2014).

And when we're through with those, we will take a closer look at the effects of cereal enriched breads on the appetite ratings and postprandial glucose, insulin, and gastrointestinal hormone responses related to hunger and satiety in healthy men and women (Gonzalez-Anton. 2014), and the "muscle full effect", or rather limits to maximal protein synthesis in man (Mitchell. 2014).

Read more short news here at the SuppVersity

Obesity Research Upd. Nov. '14

Exercise Res. Upd. Nov '12(1)

Exercise Res. Upd. Nov '12(2)

Nutrition Res. Update Nov. '14

Weight Loss Tricks & More

Reductive Stress, Iron & the Military

• Saturated fat and your heart - Right from the Haukeland University Hospital in Norway comes a new study that investigated the associations between self-reported dietary SFA intake and risk of subsequent coronary events and mortality in patients with coronary artery disease (CAD).
  
  The study included patients who participated in the Western Norway B-Vitamin Intervention Trial and completed a 169-item semiquantitative food-frequency questionnaire after coronary angiography - 2412 patients, total, 81% men, 19% women with a mean age of 61.7 y.
  After a median follow-up of 4.8 y, a total of 292 (12%) patients experienced at least one major coronary event during follow-up.  And while a gigh intake of SFAs was associated with a number of risk factors at baseline, "there were no significant associations between SFA intake and risk of coronary events [age- and sex-adjusted HR (95% CI) was 0.85 (0.61, 1.18) for the upper vs. lower SFA quartile] or any secondary endpoint. Estimates were not appreciably changed after multivariate adjustments" (Puaschitz. 2014).
  

  

  Figure 1: Hazard ratios according to % saturated fat intake of total energy intake compared to minimal saturated fat intake (HR = 100%) in 2412 subjects (Puaschitz. 2014).

  In other words, if you ask researchers from Northern Europe, their answer to the question, whether our high intake of saturated fats is the reason we are dying prematurely from heart disease is "no". This stand in line with a recent review of the current evidence by O'Keffee et al. who point highlight that the different results (which often depend on the country, where the studies are conducted) may be attributable to the fact that "not all SFA are created equal and the food sources of SFA". Accordingly the researchers from the King's College in London, the Luke’s/Roosevelt Hospital, the New York Nutrition Obesity Research Centre and the Columbia University in New York recommend that "individual characteristics of the SFA, such as chain length, should be considered in dietary recommendations" (O’Keeffe. 2014)... and I would like to add: In every future study, as well.
  
  I mean, this and the foods from which the subjects in the study at hand got the majority of their saturated fat intake may well be the reason that there was a statistically significant correlation between high fat intakes and the occurernce of coronary artery disease (remember: all participants had CAD, already) in the cohort Western Norway B-Vitamin Intervention Trial.
• L-Serine as super-supplement for binge drinkers? At least in rodents the provision of 200mg/kg body weight (for humans this would be ~1.2-1.5g/day) serine in the diet led to an attenuation of alcohol-induced increases in serum homocysteine and hepatic triglyceride (TG) concentrations (>5-fold in the control mice) by 60.0% and 47.5%, respectively.
  

  

  Figure 2: Liver triglyceride levels, serum ALT and serum homocysteine levels in control mice (C) and "binge drinking mice" (EV) with and without 20mg/kg (ES20) and 200mg/kg (ES200) serine in their diets (Sim. 2014)

  Moreover, in the chronic ethanol study, l-serine also decreased hepatic neutral lipid accumulation by 63.3% compared with the ethanol group and ramped up the glutathione and S-adenosylmethionine content of the liver by 94.0% and 30.6%, respectively.
  
  If we assume that serine is only half as powerful, when it is given to humans, I would recommend you drink your Vodka Red Bull with serine in the future ;-)
• Super-satiating cereal enriched breads - I guess "super-satuating" is an exaggeration, but there is no doubt that the addition of variety of cereal flours (wheat, oat, and spelt) and 22% dried fruits (figs, apricots, raisins, and prunes) to regular bread lead to a significant improvement of appetite control by reducing hunger and enhancing satiety in 30 healthy adults (17 men and 13 women) aged 19–32 y with body mass index of 19.2–28.5 who participated in an experiment that was conducted at the University of Granada in Spain (Gonzalez-Anton. 2014).
  

  

  Figure 3: The hormonal changes would indicate increased satiety, the subjects reported increased satiety, but their 4h energy intake was identical in both condition (Gonzalez-Anton. 2014)

  Whether the decrease in prospective consumption and increased satiety is enough to have long-term benefits on weight control is yet questionable, because the subsequent ad libitum energy intake in a 4 h period after the ingestion of the "enriched" bread did not differ from that in the control condition, even though the postprandial blood glucose, insulin, ghrelin, were lower and the pancreatic polypeptide AUC (an indicator of satiety) was higher than with the control bread.
  
  Speaking of insulin: In view of the fact that the latter actually is a satiety hormone and its release is closely related to glucagon-like peptide (GLP) 1 and gastric inhibitory polypeptide (GIP) where the AUC (areas under the curve) were lowered as well, it's eventually maybe not too surprising that the "enriched" bread was not better than the regular one.
• Muscle full? What's limiting protein Synthesis? Scientists from the Clinical, Metabolic, and Molecular Physiology, MRC–Arthritis Research UK Centre of Excellence for Musculoskeletal Ageing Research at the University of Nottingham and the Royal Derby Hospital in the United Kingdom recently determined the effect of Bolus (=all the aminos at once) vs. Spread EAA feeding in young men, hypothesizing that muscle-full is regulated by a dose-, not delivery profile–, dependent mechanism; and what they found was surprising for us - not for the researchers, though:

  

  Figure 4: Even though the study was conducted in young men, the overall dosage of 15g may potentially have had an effect on the outcome. On the other hand: If you "overdose" it would actually be more likely for spread protein ingestion to have superior effects. Against that background the "low" dose of "only" 15g of pure EAAs is not an argument that would falsify the results of the study at hand (Mitchell. 2014)

  "Despite distinct plasma and muscle profiles, Bolus feeding provided no anabolic advantage over Spread feeding (or vice versa); these findings are in keeping with our hypothesis of there being an intrinsic muscle-full state in young men at rest.
  
  Bolus feeding led to rapid aminoacidemia with a brisk upstroke and high peak plasma EAA and leucine concentrations. Spread feeding, by comparison, resulted in lower, later peak concentrations. Despite this, identical MPS responses were observed, even with the same latency (of ~90 min) and amplitude.
  
  Furthermore, with both feeding strategies, basal MPS was observed 180 min after consumption of either Bolus or the initial Spread doses. This preceded the peak Spread plasma EAAs, in keeping with the onset of a muscle-full state.

  As the scientists point out, their results do thus "suggest that, in healthy young men, it is dose dependent mechanisms that regulate the size of the anabolic response to feeding and that this response" and that this dose-dependent anabolic response "is not perturbed by later arriving, lower-amplitude aminoacidemia." The researchers also highlight hat it would seem "vital to have such a mechanism in place"; because of the "stability of muscle mass from year to year in healthy younger populations" (Mitchell. 2014). Eventually, the differences may well be explained by the existence of three distinct phases in the postprandial period, the scientists argue:
  

  

  Figure 5: Absolute changes in FSR from fasted (2120 to 0 min) to fed (0 to 240 min) (A), actual FSRs (B) and plasma EAA and insulin concentrations, phospho- 4EBP1 Thr65/70 and muscle protein synthetic rates, normalized to their own data spans shown on the same axis (C and D) in young men after consumption of 15 g of mixed-EAA meals by Bolus or Spread treatment. The black arrows represent ingestion of 15 g EAAs once, and the gray arrows represent ingestion of 3.75 g EAAs 4 times (Mitchell. 2014)

  "After the onset of essential aminoacidemia, a latent period exists when a significant negative arteriovenous EAA balance is detectable (Mitchell. 2013) but incorporation of EAAs into newly synthesized myofibrillar proteins is not. The existence of a similar latent period in response to Bolus and Spread EAA ingestion suggests that providing time for adequate intracellular EAA accumulation, even with rapid aminoacidemia with Bolus, is crucial before MPS can be ‘‘switched on.’’ After this latent period, a transient stimulation in MPS, lasting ;90 min (Bohé. 2001), occurs before the onset of the muscle-full state restores basal MPS despite sustained, near-peak postprandial EAA availability" (Mitchell. 2014).

  Put simply, it takes long enough for the muscle protein synthesis to gain full speed to incorporate all the amino acids the healthy subjects received in 4x45min boluses.
  
  Practically speaking this does not necessarily mean that you should give up your previous protein feeding strategies. With intact proteins, of which you know that they are more than the sum of their EAA parts (see "Whey Beyond Brawn"), studies by Moore et al. (2012 | learn more) and Burke et al. (2012 | learn more) yielded different results... albeit with less frequent biopsies that were taken across the postprandial period and thus a lower temporal resolution that does not exclude that said studies simply overlooked the dose-dependency of the muscle-full effect Mitchell et al. demonstrate in the study at hand.

10+ Things You Probably Didn't Know Whey Protein | more

So what are the take home messages from today's research update? I guess the one you will be most interested in, is the related to the Mitchell study which indicates that protein timing and / or the importance of bolus ingestions may previously have been overrated - at least in the short run. We should not forget, after all, that this is a result that would stand in line with Alan Aragon's & Brad Schoenfeld's recent review (Aragon. 2014  on nutrient timing which found a significant effect for the amount of protein people consume, but no evidence of the purported importance of protein timing.

This is yet not the only myth that is tumbling. The idea of heart disease triggering saturated fats and the notion that you could make bread a superfood by adding cereals and dried fruits did not get away unscathed either. With the impressive effects of serine in the rodent study by Sim et al. (2014), we do have another myth to bother with - one of which I would like to remind you that it has to remain a myth until the results have been confirmed in human beings | Comment on Facebook!

References:

• Aragon, Alan Albert, and Brad Jon Schoenfeld. "Nutrient timing revisited: is there a post-exercise anabolic window." J Int Soc Sports Nutr 10.1 (2013): 5.
• Bohé, Julien, et al. "Latency and duration of stimulation of human muscle protein synthesis during continuous infusion of amino acids." The Journal of physiology 532.2 (2001): 575-579.
• Burke LM, Hawley JA, Ross ML, Moore DR, Phillips SM, Slater GR, Stellingwerff T, Tipton KD, Garnham AP, Coffey VG. Preexercise aminoacidemia and muscle protein synthesis after resistance exercise. Med Sci Sports Exerc. 2012 Oct;44(10):1968-77.
• O’Keeffe, Majella, and Marie-Pierre St-Onge. "Saturated Fat and Cardiovascular Disease: A Review of Current Evidence." Current Cardiovascular Risk Reports 7.2 (2013): 154-162. 
• Mitchell, William Kyle, et al. "Development of a new Sonovue™ contrast‐enhanced ultrasound approach reveals temporal and age‐related features of muscle microvascular responses to feeding." Physiological reports 1.5 (2013). 
• Mitchell, William Kyle et al. "A Dose- rather than Delivery Profile–Dependent Mechanism Regulates the ‘‘Muscle-Full’’ Effect in Response to Oral Essential Amino Acid Intake in Young Men."J. Nutr. February 1, 2015
• 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.
• Puaschitz et al. "Dietary Intake of Saturated Fat Is Not Associated with Risk of Coronary Events or Mortality in Patients with Established Coronary Artery Disease." J. Nutr. February 1, 2015 jn.114.203505
• Sim, et al. "l-Serine Supplementation Attenuates Alcoholic Fatty Liver by Enhancing Homocysteine Metabolism in Mice and Rats." J. Nutr. February 1, 2015 jn.114.199711.

Protein Wheysting?! No Significant Increase in PWO Protein Synthesis W/ 40g vs. 20g Whey, But 100% Higher Insulin, 340% More Urea & 52x Higher Oxidative Amino Acid "Loss"

No, I don't think the results would have been different, if the subjects had been young women. For older guys and gals, on the other hand, I am not 100% sure.

It has been a while since we've been taking a look at one of the two or three dozen "whey increases muscle protein synthesis" studies and, officially, we would have to wait not just for Santa, but actually until January 2014 to take a glimpse at the results Oliver C Witard, Sarah R Jackman, Leigh Breen, Kenneth Smith, Anna Selby, and Kevin D Tipton present in their soon-to-be-published paper in the journal of the American Society for Nutrition (Witard. 2014).

The intention of the researchers was (yet again) to "characterize the dose-response relation of postabsorptive rates of myofibrillar MPS to increasing amounts of whey protein at rest and after exercise in resistance-trained, young men", (Witard. 2014). This is nothing new, but still right up the average SuppVersity reader's alley, I suppose.

So what about the study design

The design of the study was simple. The 48 healthy volunteers consumed a standardized, high-protein
(0.54 g/kg body mass) breakfast. Three hours later, they all performed a standardized bout of unilateral exercise, consisting of 8x10 leg presses and leg extensions at 80% of their individual, predetermined one-repetition maximum. "Immediately" (max. 10min) after they were done with the leg workout the volunteers consumed

• 0g, 10g, 20g, or 40g whey protein isolate

as a post-workout protein shake, of which I don't have to tell you that it was likewise... standardized, right! The subjects were then hooked up with the necessary instruments and tools to measure their

• postabsorbtive rates of myofibrillar protein synthesis (MPS) , 
• whole-body rates of phenylalanine oxidation and 
• urea production 

over a 4-h period (the stopwatch started ticking the very moment the subjects had ingested the protein shake) in all four arms of this parallel research design, single-blind study with 7 subjects in each of the 0, 10, 20, and 40g whey protein isolate groups.

Change (%) in myofibrillar and sarcoplasmic protein synthesis after ingestion of 25g whey at rest (FED) and resistance exercise (FED-EX) after 3h and 5h (Moore. 2009a)

Just a reminder: You do remember that there is another muscular compartment where we can measure protein synthesis? Right? The sarcoplasma, i.e. the zone around the myofibers, where the satellite cells reside. At least for the exercised leg, in the study at hand, this may not be that important, though, because "in contrast [to protein feeding at rest], resistance exercise rapidly stimulates and sustains the synthesis of only the myofibrillar protein fraction after protein ingestion" (Moore. 2009; my emphasis). The word "only" is slightly misplaced. If you look at the figure on the left, it's obvious that "mainly" or "more significantly", would probably be more accurate.

Now that you know all the important details about the study design, it's almost time to take a look at the results. Before we finally do that, let's just briefly recapitulate the results of (Cuthbertson. 2005) who observed that 10 g EAAs at rest and (Moore. 2009b) who observed that 20 g egg protein after exercise were "optimal for the maximal stimulation of MPS in young adults". This is after all, what the researchers hypothesis that "20 g of whey protein (~10 g EAAs) would be sufficient for the maximal stimulation of myofibrillar-MPS rates at rest and after resistance exercise in trained, young men" (Witard. 2014) was based on.

Figure 1: Post-exercise serum insulin (AUC, µmol/ml x 4h) and leucine peak (mmol/ml), total phenylalanine oxidation (AUC µmol/ml x 4h x 100), urea production (AUC µmol x 4h) and plasma urea (AUC mmol/l x 4h), as well as myofibrillar protein synthesis (MPS) in the 4h after the workout (Witard. 2014).

As you can see, the actual study results confirm the scientists suspicion: The 20g of whey protein did maximized the myofibrillar protein synthetic response to a hypertophy-oriented leg training workout (see bottom line for an explanation of why I chose to underline the word "leg") in rested and exercised muscle of ~80-kg resistance-trained, young men.

We are talking about statistical significance here: I know what you are going to tell me, now. And yes, you are right. The protein synthesis was in fact higher, but that's more of a matter of how sustained the increase was and not a matter of a "faster" protein synthesis. In other words, with 20g of a fast absorbing whey protein and a whole meal with slow absorbing proteins 30-40min after you will achieve the same - if not higher muscle protein synthesis rates in the long(er) run (>2h)... ah, and by the way: The response in the untrained leg confirms: There is not additional MPS stimulus from 40g vs. 20g of whey (much contrary to the insulin spike, by the way ;-).

The side-finding that this in medical terms "high" amount of whey also lead to significant increases in urea production is - at least in my humble opinion not surprising. The increased ammonia production due to higher protein oxidation rates does after all have to be cleared from the body. Against the background that this process is facilitated by the urea cycle, anything but the observed increase in urea production would have been startling.

"Confirmed: All Wheys, Not Just Hydro Whey Boost Glucose Uptake and Liver + Muscle Glycogen Supercompensation. Plus: How Can Taurine help?" | more

The fact that this increase in urea production and plasma concentrations did occur in the first place, on the other hand, is a clear cut sign for the onset of "wastefulness" with higher protein consumption - or as Selby et al. put it:

"Indeed, in the current study, urea production rates , as well as plasma urea concentrations, were markedly raised with the ingestion of 40 g protein.

Thus, instead of incorporation into muscle protein, the metabolic fate of excess exogenous amino acids contained in the 40WP was predominantly the oxidation or excretion as an indication that a state of amino acid excess was reached." (Selby. 2014)

Whether you consider this a "waste" of valuable dietary protein or not is probably a matter of your personal concept of protein nutrition.

If you are on the "protein worshipper" side of the devide, you will probably argue that you better "burn" protein for energy than carbs or fats, because otherwise you would have to eat less protein and  more carbohydrates + fat and would "become fat". It goes without saying that this is bullshit - not to mention that anyone who is interested in performance and the sanity of his doctor. The poor guy would freak out, when he'd see the elevated AST and ALT levels the combination of "protein only" diets + intense physical exercise are going to produce.

Your doctor's mental sanity or the excited calls of his receptionist are probably not really your concern, but I would still not discard the performance and, in the long run, metabolic and psychological detriments from running on protein only. From an (bio-)energetic perspective it's the least effective of the three macronutrients and thus not exactly a suitable fuel source for high performance athletes.

"So what would you put into a post-workout shake, Adel?" Personally, I have ~30g of whey protein and some fruit, like 1-2 bananas, a ton of water melon, or whatever else I have lying around. If no fresh fruit is available, I just grab some instant oats. And while I know that the carbs won't help with protein synthesis (Koopman. 2007), there is hardly any better timepoint to use the massive isulin spike and shuttle the glucose into the muscle than after the workout (van Loon. 2000). For me personally, the addition of carbs also prevents the brainfog, I get due to low blood sugar after an intense leg-workout and a protein shake without carbs. So, if you feel like you're not thinking straight or would have to go to bed after your shake, I would try to fix that by adding some carbs to the equation.

Bottom line: With the study at hand we (will) get further confirmation of the existence of a protein threshold of ~20g of whey protein, beyond which we won't see additional increases in acute myofibrillar protein synthesis after having a high protein breakfast and the completion of a standardized hypertrophy-oriented leg workout in young, healthy, male individuals.

If you wonder about the many underlined words in this conclusion, I may remind you of the fact that all these words describe boundary conditions that won't be fulfilled for everyone: There are more than enough people who don't have a high protein breakfast. There are people who train their whole body in a single session and would thus upregulate the protein synthesis in more than just the leg muscles. Not everyone is still young (and there is albeit inconclusive evidence that older individuals need more protein). For long-term muscle gains the sarcoplasmic protein synthesis may and the long-term (not acute) net protein balance definitely is more important than the acute increase... I could go on, but I guess you see, where this is heading: Theoretically, we'd have to do another 100 studies, but I am not sure whether Glaxosmith Kline who support Tiptons research would want to finance all of these ;-)

Reference:

• Cuthbertson, D., Smith, K., Babraj, J., Leese, G., Waddell, T., Atherton, P., ... & Rennie, M. J. (2005). Anabolic signaling deficits underlie amino acid resistance of wasting, aging muscle. The FASEB journal, 19(3), 422-424.
• Koopman, R., Beelen, M., Stellingwerff, T., Pennings, B., Saris, W. H., Kies, A. K., ... & Van Loon, L. J. (2007). Coingestion of carbohydrate with protein does not further augment postexercise muscle protein synthesis. American Journal of Physiology-Endocrinology And Metabolism, 293(3), E833-E842.
• Moore, D. R., Tang, J. E., Burd, N. A., Rerecich, T., Tarnopolsky, M. A., & Phillips, S. M. (2009a). Differential stimulation of myofibrillar and sarcoplasmic protein synthesis with protein ingestion at rest and after resistance exercise. The Journal of physiology, 587(4), 897-904.
• Moore, D. R., Robinson, M. J., Fry, J. L., Tang, J. E., Glover, E. I., Wilkinson, S. B., ... & Phillips, S. M. (2009b). Ingested protein dose response of muscle and albumin protein synthesis after resistance exercise in young men. The American journal of clinical nutrition, 89(1), 161-168.
• van Loon, L. J., Saris, W. H., Kruijshoop, M., & Wagenmakers, A. J. (2000). Maximizing postexercise muscle glycogen synthesis: carbohydrate supplementation and the application of amino acid or protein hydrolysate mixtures. The American journal of clinical nutrition, 72(1), 106-111.

Maximal Protein Synthesis in the Elderly: How Much Protein Does it Take? Another Study to Suggest More is Better!

Maximal protein synthesis requires protein, but how much exactly you need will depend on your age - the older you are the more PWO protein you'll need.

Scientists from the University of Auckland were fed up with the lack of information about the differential response in protein synthesis in response to the ingestion of various amounts of protein. Accordingly, Randall F. D’Souza et al. conducted a study to characterize the changes in intramuscular levels of EAAs and BCAAs and the expression of the "protein pump" p70S6K at Thr389, a marker of protein synthesis, in response to resistance exercise and graded ingestion of whey protein in older men.

As a regular SuppVersity reader you will probably already think: "Where is the actual measurement of the fractional protein synthesis?" The unfortunate answer: It's not there.

You can learn more about protein intake at the SuppVersity

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Previous research had show that the ingestion of graded amounts of high-quality protein such as whey after resistance will maximize with "only" 20g of egg protein (Moore. 2009) or whey (Witard. 2014) in young men. Multiple studies in older adults (>60 years), on the other hand, suggest that they exhibit a lower anabolic signaling and MPS response to protein feeding, resistance exercise, and the combination of feeding and exercise when compared to young men (Cuthbertson. 2005; Fry. 2011; Burd. 2013). Scientists call this phenomenon age-related "anabolic resistance" (Yang. 2012b).

Figure 1: In contrast to the fractional protein synthesis in the elderly, which increases with increasing amounts of protein, the FSR of young men shows a ceiling effect at 20g+ whey protein (Yang. 2012a; Moore. 2009)

As you can see in Figure 1 from a 2012 study by Yang, the same 20g of extra-whey (total dose 40g) that was useless in young men, lead to a significant increase in protein anabolism in elderly men. Compared to young men, the MPS response to feeding 40 g of protein was yet still slightly lower in older vs. count men (Yang. 2012a; Churchward Venne. 2013b).

What is particularly relevant for the study at hand, and the previously criticized absence of actual MPS measurements is the fact that deficits in feeding induced p70S6K phosphorylation may at least partially underpin anabolic resistance in aged skeletal muscle (Cuthbertson. 2005), which is why measuring the p70S6K phosphorylation in older human subjects (mean age 71 years) in response to the graded ingestion of whey protein after a leg workout consisting of three sets of 8–10 repetitions of bilateral barbell smith rack squat, 45°leg press, and seated knee extensions at 80% of the subjects' predetermined 1R is not as irrelevant at it may initially have seemed.

Workout + supplements, that's the "whey to go" ;-)

The exercises were performed in a circuit manner with 1 min rest between each exercise and 3 min rest between subsequent sets, the exercise protocol took approximately 20 min to complete. Following completion of the exercise protocol, subjects were immediately provided with a fixed-volume (350 mL) beverage, containing a flavored noncaloric placebo, or oneof the four doses of whey protein concentrate (10 g, 20 g, 30 g, or 40 g).

Figure 2: Intramuscular amino acids. This figure is a heat map which shows groups means fold changes from the resting fasted condition. Green represents a decrease in amino acid content, white represents no change, and red represents an increase in amino acid content (D’Souza. 2014)

Subjects were instructed to ingest the beverage within 2 min and were required to ingest the total volume provided. Following consumption of the supplements, subjects rested in a supine position throughout the 4 h of post-exercise recovery with additional muscle biopsy samples collected at 2 and 4 h post exercise.

Figure 3: Higher protein intake = higher increase in p70S6K phosphorylation (left graph). This increase is linearly associated with intramuscular leucine levels (right graph | both from D’Souza. 2014)

As you can see in Figure 3, there was a similar dose-dependent increase in p70S6K as it was observed previously for MPS in skeletal muscle of elderly subjects by Yang et al. (2012b). In fact, the fold change in the phosphorylation of p70S6K (Thr389) at 2 h post exercise was correlated with the dose of whey protein consumed (r =0.51,P<001) and was found to be significantly correlated with intramuscular leucine content (r =0.32,P=0.026).

Moreover, the intramuscular BCAAs, and leucine in particular, appear to be important regulators of anabolic signaling in aged human muscle during post-exercise recovery via reversal of exercise-induced declines in intramuscular BCAAs.

Suggested Read: "Protein Timing Does Matter! Yet Only in Trained Men. More Than 2x Higher Relative Protein Retention W/ Immediate vs. 6h Post Whey Consumption in Bodybuilders vs. Rookies" | read more.

Bottom line: In the absence of a young control group and actual muscle protein synthesis (MPS) measurement, the study at hand cannot finally answer the question, whether older men require higher amounts of protein than young ones to achieve maximal increases in post-workout protein synthesis, but it is at least another piece of evidence that "more helps more" - at least in the elderly.

As mentioned in other recent posts, there are yet still many confounding variables that would have to be controlled and modified as well to answer the important (?) question: "How much protein does it take to achieve maximal post-workout protein synthesis?" Which confounding factors that would be? Well, what about the training experience? The baseline muscle mass? The protein content of the diet? And so on and so forth || Comment on Facebook!

References:

• Burd, N. A., S. H. Gorissen, and L. J. van Loon. 2013.  Anabolic resistance of muscle protein synthesis with aging. Exerc. Sport Sci. Rev. 41:169–173.
• Churchward-Venne, T. A., N. A. Burd, C. J. Mitchell, D. W. West, A. Philp, G. R. Marcotte, et al. 2012. Supplementation of a suboptimal protein dose with leucine or essential amino acids: effects on myofibrillar protein synthesis at rest and following resistance exercise in men. J. Physiol. 590:2751–2765.
• D'Souza, Randall F., et al. 2014. Dose‐dependent increases in p70S6K phosphorylation and intramuscular branched‐chain amino acids in older men following resistance exercise and protein intake. Physiological Reports 2.8: e12112.
• Churchward-Venne, T. A., L. Breen, and S. M. Phillips. 2013a. Alterations in human muscle protein metabolism with aging: protein and exercise as countermeasures to offset sarcopenia. BioFactors 40:199–205.
• Churchward-Venne, T. A., C. H. Murphy, T. M. Longland, and S. M. Phillips. 2013b. Role of protein and amino acids in promoting lean mass accretion with resistance exercise
  and attenuating lean mass loss during energy deficit in humans. Amino Acids 45:231–240.
• Churchward-Venne, T. A., L. Breen, D. M. Di Donato, A. J. Hector, C. J. Mitchell, D. R. Moore, et al. 2014. Leucine supplementation of a low-protein mixed macronutrient beverage enhances myofibrillar protein synthesis in young men: a double-blind, randomized trial.
  Am. J. Clin. Nutr. 99:276–286.
• Cuthbertson, D., K. Smith, J. Babraj, G. Leese, T. Waddell, P. Atherton, et al. 2005. Anabolic signaling deficits underlie amino acid resistance of wasting, aging muscle. FASEB J. 19:422–424.
• Moore, D. R., M. J. Robinson, J. L. Fry, J. E. Tang, E. I. Glover, S. B. Wilkinson, et al. 2009. Ingested protein dose response of muscle and albumin protein synthesis after resistance exercise in young men. Am. J. Clin. Nutr. 89:161–168.
• West, D. W., and K. Baar. 2013. May the Force move you: TSC-ing the mechanical activation of mTOR. J. Physiol. 591:4369–4370.
• West, D. W., N. A. Burd, J. E. Tang, D. R. Moore, A. W. Staples, A. M. Holwerda, et al. 2009a. Elevations in ostensibly anabolic hormones with resistance exercise enhance neither training-induced muscle hypertrophy nor strength of the elbow flexors. J. Appl. Physiol. 108:60–67 .
• West, D. W., G. W. Kujbida, D. R. Moore, P. Atherton, N. A. Burd, J. P. Padzik, et al. 2009b. Resistance exercise-induced increases in putative anabolic hormones do not enhance muscle protein synthesis or intracellular signalling in young men. J. Physiol. 587:5239–5247.
• Witard, O. C., S. R. Jackman, L. Breen, K. Smith, A. Selby, and K. D. Tipton. 2014. Myofibrillar muscle protein synthesis rates subsequent to a meal in response to increasing doses of whey protein at rest and after resistance exercise. Am. J. Clin. Nutr. 99:86–95
• Yang, Y., L. Breen, N. A. Burd, A. J. Hector, T. A. Churchward-Venne, A. R. Josse, et al. 2012a. Resistance exercise enhances myofibrillar protein synthesis with graded intakes of whey protein in older men. Br. J. Nutr. 108:1780–1788.
• Yang, Y., T. A. Churchward-Venne, N. A. Burd, L. Breen, M. A. Tarnopolsky, and S. M. Phillips. 2012b. Myofibrillar protein synthesis following ingestion of soy protein isolate at rest and after resistance exercise in elderly men. Nutr. Metab. 9:57.

45x More Testosterone Yet Identical Increase in Protein Synthesis: MPS Response to Exercise + 25g Whey in Men vs. Women Challenges Common Wisdom About Androgens

Image 1: Is it not testosterone that makes the difference?

It is an open secret that women are having a much harder time building muscle than men, and it is another instance of (bro-)scientific wisdom that the obvious lack of testosterone in female strength athletes would be the underlying reason. Right from Stuart M. Phillips lab at the Department of Kinesiology of the McMaster University in Hamilton, Ontario, Canada, comes a new study (West. 2012) which puts yet another questionmark behind the anabolic prowess of testosterone (if you still believe that a transient increase in testosterone will help you build muscle, I suggest you read up on "The Big T" in the Intermittent Thoughts on Building Muscle).

Women are different, but it's not about protein synthesis

In the recently conducted trial Daniel W.D. West, who has also been the lead author of the "Never Sip Your Whey" study, I covered back in November 2011, undertook another attempt to identify the intricate endo- and paracrine mechanisms of skeletal muscle hypertrophy and its sex-specific variability. To this ends, West et al. recruited 5 male and 5 female subjects, who "who were habitually engaging in two to five sessions of physical activity per week including", yet did not train legs more than twice a week.

Image 1: In this case, testosterone took a backseat, as well. With synthol, protein synthesis is yet unnecessary anyway.

Note: The selection of advanced trainees as study participants is the first huge plus of this study. After all, we all know that the exercise induced hypertrophy response diminishes with training and those of you who read the whole Intermittent Thoughts on Building Muscle series will also be aware that the protein synthetic response is limited by the maximal domain size. Further growth thusly requires restructuring / the recruitment of satellite cells and installment of new myonuclei (cf. "Growing Beyond Temporary Physiological Limits"), a time-consuming and complex process which is probably one of the underlying reason for the "growth difference" between beginners and advanced strength athletes.

On the day of the experiment, the study participants, who had consumed a standardized diet containing 15% fat, 30% protein and 55% carbohydrates (the macronutrient ratio was adapted to their habitual diets) on the previous day, reported to the lab at 6am. After the infusion of the tracer that is necessary to evaluate the protein flux and an initial biopsy, all subjects performed a bout of  intense, high-volume lower body exercise consisting of

• 5 sets of 10 repetitions of leg press at ~90% of their individual 10RM, and
• 3 super-sets of 12 repetitions of leg extension/leg curl at ~90% of 12 RM

The rest intervals between the sets were 60s, so that the whole workout should not have lasted longer than max. 20min. Directly thereafter, the subjects consumed the "obligatory" (for Phillips lab this has in fact become obligatory ;-) 25g of whey protein from the usual New Zelandian source, Phillips et al. used in all their previous study (as ridiculous as this may sound but this is a nice means of standardization ;-) and rested in a supine position for the rest of the trial. Biopsies were taken and the subjects who were sent home with a launch packet consisting of their standardized meals had to report back to the lab on the following morning for another three biopsies 24h, 26h and 28h after the test workout (the subjects remained fasted and received another 25g of whey 26h post, i.e. before the last four blood samples were drawn and the last biopsy at 28h post was performed).

Figure 1: Serum testosterone levels (in nM) and myofibrillar fractional protein synthesis rate (in %/h) before and after the resistance workout, as well as on the morning and at noon of the 2nd day (data adapted from West. 2012)

As the data in figure 1 shows, the (expected) huge difference in both basal as well as exercise induced increases in circulating androgen levels (45-fold in men vs. women) had no (not even a statistically non-significant) beneficial impact on the exercise induced increase in protein synthesis in the 28h window of opportunity (cf. "Opening the 'Anabolic Barn Door' with the Key of Science").

Akt Ser473 phosphorylation increased at 1h ( P < 0.001, main effect for time) and to a greater extent in men (sex × time interaction, P = 0.018). Phosphorylation of mTOR Ser2448 was increased at 1, 3 and 5 h (P < 0.001; Figure 4B); there was a main effect for sex (men > women, P = 0.003). Phosphorylation of mTOR Ser2448 was elevated similarly between sexes after next-day protein feeding, approximately 26 h after the exercise bout (sex  × time interaction,  P = 0.49; main effect for time, 28 > 26 h,  P = 0.006).  Phosphorylation of p70S6K1 Thr389 increased at 1, 3 and 5 h (all  P < 0.001; sex × tim e interaction,  P = 0.13) and there was a significant interaction with next-day feeding (28 > 26 h in women only, sex × time interaction,  P = 0.016; data not shown). Androgen receptor content was greater overall in men (P = 0.049) but there was no significant interaction ( P = 0.47).  

The greater increase in mTOR and Akt (both hitherto regarded as the "gas pedals" of the skeletal muscle protein synthetic machinery) are not only less pronounced, than one would expect if there was a direct interaction with testosterone levels, they also lack real world significance. After all, the area under the myofibrillar protein synthesis curve (a measure for the total protein synthetic response to exercise) was identical in the 1-5h period right after the exercise and - although West et al. did not include the respective data in their article - I would suspect that the data from the subsequent day (cf. figure 1, right) would even suggest that it must have been slightly greater in the female participants.

Testosterone useless and mTOR and Akt unreliable indicators at best?

Now, which conclusions shall be drawn from these results? Is testosterone useless? Does it not contribute to the overall greater muscle mass in men compared to women? It stands to reason that this conclusion would be about as flawed as the notion that testosterone alone would suffice to build muscle. Rather than its "inefficiency" in building muscle, this study only shows that its importance in relation to the exercise-induced increase in protein synthesis is probably way overrated.

A similar point could be made for mTOR and Akt, as well, though. Or as West et al. put it in their discussion of the results and the respective implications for future studies:

In light of this disconnect, it is worth recognizing that the phosphorylation of signalling proteins is a temporal snapshot of the propagated signal for translation initiation. It is also unclear if there is a minimum threshold signal required to initiate and completely activate or  ‘turn on’ translation. If  there is such a threshold then it seems plausible that greater phosphorylation above such a  threshold would be unlikely to further amplify the signal/lead to increased rates of translation.

For a physicist or anybody who knows a thing about "energy levels" the existence of "threshold" levels in processes taking place at a molecular level should not come as a surprise.

I suspect, we are still missing the boat with our focus on protein synthesis alone

Another question, I have been hinting at in many of my previous blogposts on the insightful studies from Stuart Phillips lab at the McMaster University, is yet whether or not the acute increase in protein synthesis (alone) is actually an acceptable predictor of skeletal muscle hypertrophy, a process which, as I have explained in detail in the Intermittent Thoughts on Building Muscle is only partly mediated by the simple accrual of amino acid chains (=proteins) within existing myofibrillar domains.

Figure 2: Graphical illustration of the processes and their respective triggers which contribute to the exercise induced increase in skeletal muscle mass (click here for detailed elaborations).

If you take another look at the complex network of endo- and paracrine signalling cascades and the number of factors which contribute to a process that is generally reffered to as "skeletal muscle hypertrophy" (cf. figure 2) and is, at least in my mind, falsely reduced to the influx of amino acids into the muscle, it should be clear that testosterone does play a central role in the actual exercise induced growth response. That the latter is less pronounced than bro-science would have it (esp. when we are talking about physiological levels, cf. "Quantifying the Big T") and that testosterone itself and its metabolites, DHT and estrogen are probably of greater importance in the "restructuring" process, which in turn facilitate the accrual of even more protein within the muscle, does after all not imply that the huge differences in androgen levels are not the reason for the differential hypertrophy response in men and women - and I guess, I don't have to tell you that you just have to take a glimpse at the female IFBB (International Federation of Bodybuilding and Fitness) competitors to know that androgens can make a difference ;-)

Where Protein Fails, Protein + Resistance Training Succeed: Lifting Corrects Diet-Induced Decrease in Postprandial Protein Synthesis, But Fails to Normalize Net Retention

It takes pains to maintain your gains!

You will certainly remember the shocking revelation that simply eating more protein is not going to prevent the diet induced muscle loss that occurs whenever you consume less energy than you expend (read up on "Protein Intake & Muscle Catabolism: Fasting Gnaws on Your Muscle Tissue and Abundance Causes Wastefulness" | go for it!)...

Don't rejoice, the study at hand does not refute this - protein is still unable to counter the increase in atrogin-1 and other muscle cannibalizing proteins, but there is a "tweak" by the means of which you can at least avoid that its pro-anabolic affects are also impaired.

You can learn more about protein intake at the SuppVersity

Are You Protein Wheysting?

Cod protein for recovery

Protein requ. of athletes

High EAA protein for fat loss

Fast vs. slow protein

Too much ado about protein?

What this "tweak" is? Well, that's easy: Heavy lifting. If you are familiar with the "muscle loss in zero gravity" research that has been conducted by and for the NASA in the past decades (e.g. Ferrando. 2002).this shouldn't surprise you. The NASA studies have after all shown quite conclusively that compared to bed-rest / chronic skeletal muscle unloading, starving yourself is almost "anabolic". No wonder that lifting heavy objects, and not dietary protein is the #1 when it comes to saving your muscular ass from shriveling away on a long and hard diet.

Why does resistance training work, if protein fails?

As discussed in "Protein Intake & Muscle Catabolism" (read it!), it's not a question of the pro-anabolic effects. You, as a suppversity reader know that the p-AKT/mTOR pathway that's activated by protein feeding is sufficient to increase the influx of protein into the musculature. What your beloved protein can't do, though, is to reset a different switch: The "sacrifice muscle to fuel more fundamental metabolic demands switch" which is triggered whenever you are in a long(er) term energy deficit.

"Training For Gains: High Intensity, Low Volume Strength Gains Stick." | more

So what can be done then? Well,... as it is so often the case, the answer lies - once more - open before our eyes: Hit the weights, down the protein and kick your diet's catabolic ass!

I know this sounds too easy, but if you take a peek at the weight loss diets of the average physique athlete and their appearance on stage, it stands out of question that the combination of resistance training and strategic protein supplementation spares muscle mass.

Now the verb "to spare", according to the Oxford English Dictionary, means "to leave (a person) unhurt" (OED.COM), which is - and you probably expected this already, not really accurate. Even the latest data from the School of Medical Sciences at the RMIT University in Melbourne and the Exercise Metabolism Research Group at the Department of Kinesiology of the McMaster University in Hamilton, Ontario, Canada, and the Canadian Sport Institute clearly demonstrates that you cannot switch the diet-induced protein wasting off, completely (Areta. 2014).

Figure 1: The large inter-individual differences make it virtually impossible to tell, whether the MURF-1 levels increased. The similarly catabolic (see overview in the middle) atrogin was yet significantly increased in the early (15g) and late phase (30g) after the workout during ED (Gumucio. 2013; Areta. 2014)

In the corresponding experiment, 16 young, healthy, resistance trained subjects (8 females, 8 males) who had been fed individualized pre-packaged meals delivering 45 kcal/kg FFM (macros: PRO / CHO / FAT 1.4-1.6, 3-3.5 and 0.5-1.5 g·kg BM) per day for five days before they went on a standardized energy 30% energy reduced diet containing approximately

• 1.4-1.6g protein per kg total body mass, 
• 4.0-4.5g carbohydrates per kg total body mass and
• 1.5-2.5g fat per kg total body mass

for another five days. At the end of this "ED" period and five days on rations with only 30kcal/kg fat free mass, all subjects performed a standardized leg press workout (warmup + 6 sets of 8 repetitions at ~80% 1 RM with 3 min rest between set) that was followed by the ingestion of either 15g or 30g of whey protein or an isocaloric placebo.

Figure 2: SLC7A5 AA transporter expression (left) and myofibrillar fractional protein synthesis (% / hour; Areta. 2014)

What a brief glance at the data in Figure 2 does tell you, though, is that resistance training will effectively counter, the diet-induced downregulation of the pro-anabolic response to protein. What it won't do, though is to increase the net protein retention to levels comparable to those on an energy balanced diet!

A high protein intake doesn't normalize the levels of anabolic hormones, either | learn more

Loss ￪, synthesis down ￬ ➲ net protein loss - there is no way out! In conjunction with the concomitant reduction in protein synthesis (-27% in the study at hand), the combination of increased loss and decreased synthesis in a caloric deficit will always entail a net loss of protein (also in view of the endocrine deterioration | learn more). What exercise can do for you, though, is to counter the net-reduction in protein synthesis, i.e. maximize the amount of amino acids that is pumped into the muscle, before it's used for hepatic gluconeogenesis.

Contrary to what Areta et al. may have suspected the restoration of the protein synthetic response in the post-workout period did not restore the expression of the amino acid transporter gene SLC7A5 to normal. It is thus not surprising that...

Highly suggested read: " Evidence From the Metabolic Ward: 1.6-2.4g/kg Protein Turn Short Term Weight Loss Intervention into a Fat Loss Diet" | more

"[...] despite this elevation, exercise merely restored MPS [muscle protein synthesis] to a level that was similar to, but not exceeding, rates measured in EB [energy balance]. Accordingly, it appears the metabolic status of the muscle during short-term (5 days) ED [energy deficit] plus a ~10 h fast may dictate that contractile overload in isolation is not enough to increase MPS to values that otherwise would be observed when subjects are in EB." (Areta. 2014)

The results of this recent study do thus have to regarded as another nail an already boarded up coffin that's loaded with bro-scientific myths about "body recompositioning."

A word on "body recomposition": You cannot build muscle, while you are dieting. You can, however improve your body composition by losing more fat than muscle. In the mirror / on photos, the results will look like "gains" - in spite of the fact that you simply revealed the muscle that has always been hidden beneath the blubber.

Unlike the non-existent changes in amino acid transporter expression, the observation that 30g of protein are more effective than 15g will probably not come as a surprise to you - notwithstanding the fac t that this was "the first [study] to determine the acute muscle anabolic response to resistance exercise with two different doses of protein ingested after exercise during short-term ED", by the way. About as unsurprising as the researchers' (eventually unwarranted - I don't see a 20g protein group, here ;-) conclusion that their ...

"[...]results suggest that the optimal amount of protein to maximize the response to a single bout of resistance training while in ED may be above the level (20 g) found to maximize MPS post-exercise for individuals who are in EB." (Areta. 2014)

And my recommendation, not to worry too much about all the details. There are a couple of simple principles that have been working for generations of athletes thriving to cut weight without having to sacrifice muscle mass; and as you should know if you've read and memorized the "9 Simple Rules Every Dieter Must Follow" (go back) consuming 30g of protein with every meal and lifting heavy objects are both part of a set of rules that's rooted in bro- and supported by pro-science.

"There is Such a Thing As Over- training, Beware! When IGF-1 & Co Plummet and MAFbx Gnaws Away Your Muscles, It'll Be Too Late to Acknowledge" | more

Bottom line: In the end, the results of this study are probably less exciting than the title, i.e. "Reduced resting skeletal muscle protein synthesis is rescued by resistance exercise and protein ingestion following short-term energy deficit" may have suggested.

That's yet not the least owed to the fact that you all know what it takes to maximize lean mass retention. If there wasn't that irrational hope somewhere deep inside your head that there was a hitherto unknown non-pharmacological way to build muscle and lose body fat at the same time, you'd now be hitting the weights or enjoying your post-workout protein shake... ;-)

Reference: 

• Areta, José L., et al. "Reduced resting skeletal muscle protein synthesis is rescued by resistance exercise and protein ingestion following short-term energy deficit." American journal of physiology. Endocrinology and metabolism (2014). Ahead of Print.
• Ferrando, Arny A., Doug Paddon-Jones, and Robert R. Wolfe. "Alterations in protein metabolism during space flight and inactivity." Nutrition 18.10 (2002): 837-841.
• Gumucio, Jonathan P., and Christopher L. Mendias. "Atrogin-1, MuRF-1, and sarcopenia." Endocrine 43.1 (2013): 12-21.

3.2kg of Lean Mass Over Night W/ 40g of Slow Digesting Protein 30min Before Bed!? Over One Year, a Positive Nitrogen Balance and +20% FSR Could Make It Happen!

Image 1: Babies instinctively know how to grow - mother's milk (60% whey, 40% casein, at later stages) + sleep ;-)

Tell me, does the sentence "Where Bro- and Pro-Science Unite in the Spirit of True Wisdom" ring a bell? Anyone? Well, that's what I thought. It's the mantra of the SuppVersity... unfortunately, more often than not, one "science" does not really care about the other, so that studies as the one by Peter T. Res and his colleagues from the University of Maastricht are unfortunately rather the exception than the rule (Res. 2012).

Pre-bed protein intake could be a crucial determinant of 24h protein synthesis

I guess, I won't have to tell you that bro-science has it that the most important thing to do before you go to bed (and for some hardcore "bros" even in the middle of the night) is not to brush your teeth, let alone to shower or at least wash your face, hands, feet and certain other body parts... no! The most important thing to do before you go to bed is to have a huge serving of protein - preferably a "night-time protein", like a slow-digesting casein-based protein shake with some additional fats to further slow the absorption (whether the fat will actually prolong the digestive process beyond what you will see if you ingest intact micelles, which will then be hydrolized in the gut and start clumping is anyone's guess, though). In fact, this is one of those truisms that has been repeated so often on the boards (and the ads) that you may be surprised to hear that Res et al. rightly claim that their study is the first one to investigate, whether this practice does actually povide any benefit for the professional or recreational lifter.

To this ends, the scientists recruited a group of 16 of the usual suspects, ah.. pardon "recreationally active men", which in this case means that they had a weekly physical activity level of 6.3h and 5.2h for the eight men in the protein and the seven in the placebo arm (#8 had a problem with a catheter, so that he had to be excluded), respectively. As you would expect from any study investigating the effect of dietary supplements on exercise performance and/or muscle growth, the subjects received a standardized dinner (0.04kcal/kg; 57% carbs, 13% protein, 30% fat) at the evening before the testing session, as well as "identical" (obviously the energy content was matched to the body weight of the respective individual) meals for breakfast, lunch and dinner on the day of the experiment. The overall protein content of the regular meals was 1.2g/kg body weight and should thusly at the lower end of what the "bros" would prescribe as a baseline protein intake for anyone trying to gain muscle.

Exercise protocol: Leg presses and extensions 8x8 - 45 min total

After a standardized meal at 4:45pm and a whole host of experimental procedures (most importantly to place the catheter for the multiple blood draws during the night), the participants performed 8 sets of 8 reps on a leg press and another 8 sets of 8 reps on a leg extension machine (2 sets at 55% and 65%, 6 sets at 75% of 1RM; "subjects were verbally encouraged during the test to complete the whole protocol"). Rest between sets was 2 min rest between exercises 5 min. At 9pm, ca. 15min after the exercise test, the subjects received a serving of Lucozade Sport Body Fuel and Lucozade Sport Recovery (yes, the study was supported by GlaxoSmithKline ;-), which contained 60g of carbs and 20g of whey and thusly mimics what many non-carbophobic athletes use to replete glycogen stores and ramp up protein synthesis after a workout. After a muscle biopsy at 11:30pm, the subjects received either 40g casein protein or placebo and "remained in a supine position until 0:00am" ... I lover this formulation, because it suggests that with all those catheters every subject fell asleep at exactly 0:00am after "remaining in a supine position" *rofl* - be that as it may, the scientists simply assume that their subjects had slept for 7 hours, when they woke them at 7am for the second muscle biopsy.

Figure 1: Plasma levels of essential amino acids (µmol/L) and overnight mixed muscle fractional protein synthesis rates (measured by phenylalanine tracer) in subjects after receiving 40g of slow acting protein (casein) or placebo 30min before bed (at T=0; data adapted from Res. 2012)

As you can see in figure 1 the EAA levels the scientists measured in the blood of their subjects in the course of the night was profoundly elevated in response to the protein feeding. It is thusly not surprising that the fractional protein synthesis rates the scientists calculated for the 7.5 h of overnight sleep was ~22% higher in the "pre-bed" protein group than in the subjects who received the placebo supplement (cf. figure 1; right). Yet, although this may sound much, we are talking about 0.059% vs. 0.048% fractional muscle protein synthesis per hour and thusly about a 0.011% increase, which was only "borderline significant" (meaning p = 0.05).

Figure 2: Net protein breakdown, synthesis, oxidation (all left) and balance (right) measured over night in previously exercised subjects after receiving 40g of slow acting protein (casein) or placebo 30min before bed (based on Res. 2012)

What is probably more important than the difference in fractional protein synthesis, anyways, is the overall net protein balance, which indicates that contrary to the trainees in the placebo group, the subjects who received a 40g serving of casein 30 min before they went to bed (and hopefully slept 7h ;-) did effectively "gain" muscle, or I should say, muscle protein over the course of their 7.5h nightly "fast", while the subjects in the placebo group ended up losing a minimal amount of skeletal muscle protein.

3.2kg of lean muscle mass in one ear with nothing but a protein shake before bed?!

If we take a look at the abstract numbers the scientists measured, such as an increase in whole body (!) net protein retention of ~50µmol/kg (measured in phenylalanine tracer molecules) over the course of 7.5h and do some math, this tells us that a trainee who weighs ~80kg and followed this practice over the course of one year, where we assume that he trains four times a week (i.e. 208 sessions) this would allow him to store 832mmol or (if I did not miscalculate) ~146g of the phenylalanine tracer in the 208 nights following his training sessions... does that sound much? No, it certainly does not, but we just assume that for each of those phenylalanine molecules another molecule of each of the other EAAs was stored within the muscle (since we are talking about "whole body" protein retention, other organs will get their share as well, though), and further assume that they all weigh about the same (which is obviously bullocks) the 40g of casein every night would result in a net protein gain of 3.2kg! How does that sound?

Image 2: Quark = Natural #1 casein source

Note: Fatfree asked rightly, whether there are not any natural alternatives to protein shakes and as I thought this is relevant for everyone, I decided against answering in the comment area. Personally I would suggest you watch out for either curd/quark (~10g casein per 100g) which has tons of highly bioavailable calcium etc. An alternative with lower protein content is cottage cheese. More fat, but still nice - any other cheese. A huge chunk of steak could work, but I am not sure if that is not problematic in terms of nighttime digestion, which was one of the 2ndary results of the study at hand: Casein is easily digested while we sleep.

Now while this is a pretty optimistic calculation, while we are (again) dealing with "rookies" who obviously gain like crazy, and so on and so fort, the fact that there are still 175 days, where you don't train and your body would still be able to store some protein, goes to show how important a properly timed intake of protein and with it a persistent influx of readily available amino acids is, if you want to gain muscle - and in that it does not matter if that are going to be 500g or 10kg over the course of one year. However, I beg you not to forget that you cannot live on protein alone and that it is highly questionable that by escalating the dose to say 60g or 80g the net gains would increase by 50% let alone 100%, respectively. So keep that in mind before you set up a bathtub full of protein to sleep in ;-)

References:

• Res PT, Groen B, Pennings B, Beelen M, Wallis GA, Gijsen AP, Senden JM, VAN Loon LJ. Protein Ingestion before Sleep Improves Postexercise Overnight Recovery. Med Sci Sports Exerc. 2012 Aug;44(8):1560-9.