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marylin monroe
Showing posts with label net protein balance. Show all posts
Showing posts with label net protein balance. Show all posts

5-10% Weight Reduction From Set to Set For Hypertrophy, Heavy Leg Workouts for Cyclists, Garlic For 400% Higher Test/Cortisol Ratios & Max(!) 1g Vitamin C for Muscle Gains

7% increase in breast cancer risk for every 500g above "normal" birthweight for Scandinavian women. Weight is yet not all that counts, mommy's gestational diabetes and even a large body size also precipitate to later disease.
7% per 500g that's the increase in breast cancer risk, the female offspring of Scandinavian women will have, if they are born heavier than normal. This figure is the SuppVersity Figure of the Week and comes from a study I came across a couple of days ago (Troisi. 2012). The statistics are based on birth register data of women from Norway, Sweden or Denmark who were subsequently diagnosed with primary, invasive breast cancer (n=51419) and 10 controls for each case from the birth registries matched by country and year of birth (n = 514,190).

Contrary to what you may think, the birth weight does yet not pose as much of a risk to become obese later in life as being larger than "appropriate" for your gestational age does (Eyzaguirre. 2012). If you also consider that gestational diabetes has been linked with increased risk of metabolic syndrome in the offspring (Davis. 2012) and that obesity in itself is an independent risk factor for breast cancer (Patterson. 2012), these should be more than enough good arguments not to surrender to your occasional food cravings and laziness - pregnant or not.

It's not all in your genes, but most in your hands

Although some people would love, if this was the case, because they could blame their own misery on the mistakes other  may have made, our lives and health are not fully determined by our genes and/or the mistakes our mothers may or may not have made. As Poston and Foreyt wrote in 1999, already: "Obesity is an environmental issue." And we are lucky: It is in our hands to change the environment we are exposing ourselves to and thus influence which of our genetic disposals will become active and are  promoted and which of them won't. Now that's obviously not just the case for obesity, muscular hypertrophy would be another example. Irrespective of your genetic make-up your strength and muscle gains stand and fall with the way you train, eat and supplement... and guess what, all of these points will be addressed in today's installment of On Short Notice.

  • Experimentally validated: 5-10% drop in weights per set is "optimal" for hypertrophy training (Medeiros. 2012) -- Scientists from the Laboratory of Physiology and Biokinetic at the Faculty of Biological Sciences and Health on the UNIG Campus V at Itaperuna in Brazil find: The average resistance trainee - in this case a young man aged 24.0±4.5 years with a body mass of 78.3±10.2 kg and a height of 177±7 cm - can remain in the hypertrophy range (10-12 reps to failure) for most of his sets, when he reduces the weight by 5-10% after each set.

    Whether this will also yield optimal gains was yet not within the scope of this 5-week study. What these results do however tell you is that you are not training hard enough if you perform all your sets with the exact same weight in the exact same rep range - well, unless you don't just like to listen to Super Human Radio, but are actually related to Superman himself ;-)

  • Sir Chris Hoy's legs are not as hilarious as those of the German Robert Forstemann (Robert is the right guy), but I am pretty certain their size and strength played a very important part in becoming the most successful Olympic track cyclist of all times (six gold and one silver Olympic Medal + 11 times world champion)
    Heavy leg training could make the difference between victory or defeat at the end of a cycling race (Hansen. 2012) -- In a soon-to-be-published paper, Ernst A. Hansen et al. report that the addition of 12-weeks of heavy resistance training in the form of 4 lower body exercises (3 × 4–10 repetition maximum) which had to be performed twice a week enhanced the cycling performance of highly trained cyclists by 7% compared to the training outcome of the subjects in a control group who simply followed their regular endurance-only, protocols:
    "Performance was determined as average power output in a 5-min all-out trial performed subsequent to 185 min of submaximal cycling. The performance enhancement, which has been reported previously, was here shown to be accompanied by improved pedaling efficacy during the all-out cycling. Thus, E+S shortened the phase where negative crank torque occurs by ~16°, corresponding to ~14%, which was more than in E (P = .002)" (Hansen. 2012)
    Since the test was conducted at the end of a 3h cycling session, it should be plain obvious that those 15% increases in torque will catapult the strength trained endurance athlete to the forefront on every final sprint.

  • Human dose equivalent of ~0.1g/kg garlic per day could not just boost your testosterone and lower the high protein diet induced increases in cortisol, it could also improve the way your body utilizes dietary protein (Oi. 2012)-- Actually this is not a new study, but since Maxim was not happy with things "so yesterday" as the increases in HDL and LDL the Arabian scientists observed in the garlic study I have been talking about at the end of Thursday's SuppVersity Science Round-Up on SHR, I thought others may be as happy as Maxim will hopefully be to hear that there is more to garlic than "just" its beneficial effects on your heart.

    Figure 1: Higher testosterone levels, an amelioration of the high protein induced increase in corticosteroids and a 40% increase in net protein balance are unquestionably impressive results given the fact that the all those differences were brought about within 28 days and by no more than 0.1g/kg (HED) of "supplemental" garlic in form of heat dried powder that was added to the chow (Oi. 2001)
    In fact, I am almost sure that the >400% increase in the testosterone to cortisol ratio you will see if you take a closer look at the data in figure 1, is probably rather what Maxim would have liked to hear me talk about. Especially in view of the fact that this endocrine effects went hand in hand with a highly significant +60% increase in protein retention (figure 1, top right). Think about it, if only part of he protein that was now no longer excreted in the urine / feces would be used for protein synthesis this would entail exactly those hypertrophy effects you don't see with your average "scientifically proven" herb-based testosterone booster.

    Unfortunately, the scientists did only measure the body weight and visceral fat pads, not the actual muscle mass of the rodents,. But if you go by their ratios it is obvious that the high protein + garlic group were not just the heaviest, but also the leanest.

    With +11 % vs. +5% in both the medium and high protein diets, the animals on the low protein did yet exhibit the most profound benefits as far as the body weight / visceral fat ratio goes. Against the background that their net protein balance remained the same, this observation does actually suggest that the pro-anabolic effects of garlic are not solely a result of a decreased protein excretion (see figure 1).
    Table 1: Principal sulphur compounds of garlic preparations (Hammami. 2012)
    Warning: Don't live on garlic alone! While the provision of 0.8% garlic powder did have beneficial effects on testosterone production in the study at hand, there are a couple of studies which suggest that a diet with 15-30% of crude garlic (Hammami. 2008 & 2009), as well as the administration of Diallyl trisulphide in isolation (Qian. 1986) and raw garlic juice (e.g. 600mg/kg per day for 21 days in Fehri. 1991) can compromise testosterone production and/or testicular function. In view of the difference between 0.8% garlic powder in the diet of the rodents in study at hand and 15-30% of pure garlic in the diet of the animals in the Hamami studies, it is most likely that the effects were dose-depended, but in case you are interested in health benefits of specific sulfor compounds in garlic, the data in table 1 on the left may still come handy to pick "your" preferred form of garlic.
    Rather than that, it appears as if the human equivalent of 0.1g/kg body weight of heat dried garlic powder that contained a total amount of 5.05 mg/g of total diallylsulfide (0.05 mg of monosulfide, 1.0 mg of disulfide, 3.4 mg of trisulfide, 0.6 mg tetrasulfide) had the ability to improve the incorporation of dietary protein into muscles (and other organs).
 
  • Study shows: Vitamin C supplementation does reduce skeletal muscle hypertrophy in response to chronic overload (Makanae. 2012) -- Despite the fact that it has not even been published yet, the paper by Yuhei Makanae et al. actually only confirms what more and more scientists have been speculating about within the last couple of years. The provision of high does of active antioxidants, and as it seems in particular vitamin C, blunts the hypertrophy response to skeletal muscle overload.

    Figure 2: 14-day of 500mg/kg  (HED 0.08g/kg) supplemental vitamin C blunt skeletal muscle hypertrophy in rodents (Makanae. 2012)
    As you can see in figure 2 the effect size was relatively small, but statistically highly significant (p < 0.01) and that despite the fact that the supplementation regimen (500mg/kg body weight; HED: 0.08g/kg body weight) was not even that much higher than what some "vitamin C enthusiasts" are taking on a daily basis in the futile (and useless) effort to boost their serum vitamin C levels to a concentrations your body does - probably not without reason - try to counter by increasing renal vitamin C clearance.

    As the data in figure 2 shows, the same homeostatic mechanism we know from humans worked in the rodents, as well - well, at least with respect to the serum levels. In the plantaris muscle of the supplemented group, on the other hand, there was a significantly higher accumulation of vitamin C than in the placebo group. This increase went hand in hand with an attenuation of the repressive effects the chronic overload of the muscle had on the expression of the catabolic protein atrogin-1 and the increases in the pro-anabolic protein Erk1/2 (p < 0.01) in the non-supplemented animals. Based on this observations and with reference to the results of previous studies and the fact that neither the water content of the muscle, nor a significant reduction in food intake in the vitamin C group could explain the observed differences, Makanae et al. conclude "that oral vitamin C administration attenuates plantaris muscle hypertrophy induced by chronic mechanical load." (Makanae. 2012).

    What the study does not answer, though, is the question whether the effects would be identical in a real-world training scenario, where the temporary, yet more intense wear and tear on the muscle could in fact be sufficient to induce skeletal muscle hypertrophy human despite vitamin C supplementation. But let's be honest in view of the fact that scientific evidence for ergogenic benefits of more than 1g of supplemental vitamin C  per day (in humans) is simply non-existent, the take away message from the study at hand should actually read: Do not escalate your vitamin C beyond the 1g per day, if you don't want to risk compromising the results of all the hard work you are investing into your training.

That's is, another installment of On Short Notice and the first day of the weekend approaching it's peak. If you still have some time before whatever your plan for Saturday night may be and feel like you could use some seconds on today's short news, I suggest you head over to the SuppVersity Facebook Wall and check out the latest news on
  • Ever thought about what green tea, grape seed, curcumin, cranberry, and tons of other Super Food have antimicrobial effects? Considering the LPS-influx from the gut turns out to be a major contributor to all sorts of diseases, I am curious about how much of their effects are actually mediated by the gut microbiome.
    The differential role of intramuscular lipids in trained athletes and sedentary slobs and how the difference between performance enhancement and insulin resistance it all comes back to getting your as off the coach (learn more)
  • Metformin 2.0? Scientists have developed a hypolipidemic, anti-atherosclerotic, anti-obesity, and glucose lowering agent called ETC-1002 (learn more)
  • Confirmed: Grape seed could be the go-to neuroprotector for diabetics - GSE administration was found to be able to ameliorate most of the biochemical altered parameters in diabetic rats (read more)
  • Fermenting your own dairy? Just add some catechin rich teas and the lactobacilli will strive. Makes you wonder about the 'internal' probiotic effects of green and black teas, as well. Doesn't it? (learn more)
There will be more, don't worry - so feel free to check for updates either directly on the SuppVersity Facebook Wall or simply by taking a look at the navigation in the right under "SuppVersity Facebook Wall" from time to time. Obviously, you can also simply "like" the SuppVersity on facebook to make sure you don't miss anything.

    References:
    • Davis JN, Gunderson EP, Gyllenhammer LE, Goran MI. Impact of Gestational Diabetes Mellitus on Pubertal Changes in Adiposity and Metabolic Profiles in Latino Offspring. J Pediatr. 2012 Nov 10.
    • Eyzaguirre F, Bancalari R, Román R, Silva R, Youlton R, Urquidi C, García H, Mericq V. Prevalence of components of the metabolic syndrome according to birthweight among overweight and obese children and adolescents. J Pediatr Endocrinol Metab. 2012;25(1-2):51-6. 
    • Fehri B, Aiache JM, Korbi S, Monkni M, Ben Said M, Memmi A, Hizaoui B, Boukef K (1991) Toxic effects induced by the repeat administration of Allium sativum L. J Pharm Belg 46:363–374.
    • Hammami I, Nahdi A, Mauduit C, Benahmed M, Amri M, Ben Amar A, Zekri S, El May A, El May MV. The inhibitory effects on adult male reproductive functions of crude garlic (Allium sativum) feeding. Asian J Androl. 2008; 10:593–601.
    • Hammami I, Amara S, Benahmed M, El May MV, Mauduit C. Chronic crude garlic-feeding modified adult male rat testicular markers: mechanisms of action. Reprod Biol Endocrinol. 2009; 24:57–65.
    • Hansen EA, Rønnestad BR, Vegge G, Raastad T. Cyclists Improve Pedalling Efficacy and Performance After Heavy Strength Training. Int J Sports Physiol Perform. 2011 Dec 2. 
    • Hammami I, El May MV. Impact of garlic feeding (Allium sativum) on male fertility. Andrologia. 2012 Sep 3.
    • Makanae Y, Kawada S, Sasaki K, Nakazato K, Ishii N. Vitamin C administration attenuates overload-induced skeletal muscle hypertrophy in rats. Acta Physiol (Oxf). 2012 Nov 26.
    • Medeiros Jr HS, Mello RS, Amorim MZ, Koch AJ, Machado M. Planned Intensity Reduction to Maintain Repetitions Within Recommended Hypertrophy Range. Int J Sports Physiol Perform. 2012 Nov 19. 
    • Oi Y, Imafuku M, Shishido C, Kominato Y, Nishimura S, Iwai K. Garlic supplementation increases testicular testosterone and decreases plasma corticosterone in rats fed a high protein diet. J Nutr. 2001 Aug;131(8):2150-6.
    • Patterson RE, Rock CL, Kerr J, Natarajan L, Marshall SJ, Pakiz B, Cadmus-Bertram LA. Metabolism and Breast Cancer Risk: Frontiers in Research and Practice. J Acad Nutr Diet. 2012 Nov 2. doi:pii: S2212-2672(12)01426-8.
    • Qian YX, Shen PJ, Xu RY, Liu GM, Yang HQ, Lu YS, Sun P, Zhang RW, Qi LM, Lu QH.  Spermicidal effect in vitro by the active principle of garlic. Contraception. 1986; 34:295–302.
    • Troisi R, Grotmol T, Jacobsen J, Tretli S, Toft­Sørensen H, Gissler M, Kaaja R,Potischman N, Ekbom A, Hoover RN Stephansson O. Perinatal characteristics and breast cancer risk in daughters: a Scandinavian population­based study. Journal of Developmental Origins of Health and Disease, Available on CJO 2012.

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

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

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

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

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

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

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

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

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

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

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

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

    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

    Drop the weights, grab the shake! Timing matters for advanced trainees.
    I guess you'll all have followed my suggestion to read Brad Schoenfeld's, Alan Aragon's and James Krieger's excellent review of the effects or protein timing on skeletal muscle hypertrophy, last year. In said paper, the two conclude that their review would "refute the commonly held belief that the timing of protein intake in and around a training session is critical to muscular adaptations" (Schoenfeld. 2013)

    Certainly a reasonable conclusion based on the evidence they present. With the recent publication of a study by Hiroyasu Mori from the Department of Nutrition Management at the Hyogo University, future reviews will yet probably have to distinguish according to the training status of the athletes.
    You can learn more about protein intake at the SuppVersity

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    In his latest study, Mori investigated the effect of the timing of protein and carbohydrate intake after resistance exercise on nitrogen balance in trained and untrained young men. By dividing his 20 healthy male subjects between the age of 20 and 29 into two groups, i.e.
    • those Mori calls "bodybuilders" and who regularly performed resistance exercise were assigned to the trained group (n = 10; mean age, 23 ± 4 years; height, 173.8 ± 3.1 cm; weight, 72.3 ± 4.3 kg) and
    • those of which Mori writes that they were "recreationally trained" but without resistance training experience were assigned to the untrained group (n = 10; mean age, 23 ± 1 years; height, 171.8 ± 5.0 cm; w eight, 64.5 ± 5.0  kg),  respectively,
    Mori added an element to the equation that has been overlooked in previous studies. An element that would allow him to answer the question:

    Do the same rules apply for untrained subjects and bodybuilders with 6.2 ± 2.8  years of training experience?

    In view of the fact that the majority of studies that investigate the long(er) term hypertophy effects of resistance training are conducted with training noobs to make sure you can measure meaningful and statistically significant changes after only a few weeks, the previously cited conclusion by Schoenfeld et al. is also based mostly on data from rookies or the infamous "recreationally active" study participants.
    Figure 1: Overview of the experimental design (Mori. 2014)
    As you can see in Figure 1, both groups, i.e. the bodybuilders and recreationally active rookies, were subjected to the same exercise + supplementation protocol in this 4-week randomized crossover trial.
    "In the P0 experimental period, subjects consumed protein and carbohydrate supplements 5 min after resistance exercise, and in the P6 experimental period, subjects consumed the same supplements 6 h after exercise. A washout period >7 days was applied before each experimental period. During each 11-day experimental period, the first 8 days were defined as an adaptation period for muscle to adapt to the energy and nutrients from the experimental food and supplements prepared by the examiner. During the next 3-day period (day 9 to day 11), 24-h urine samples were collected. The two experimental schedules are shown in Figure 1. " (Mori. 2014)
    Because at least 7 days of adaptation and 3 days of urine collection are needed to calculate nitrogen balance (Jordan. 2010), the resistance exercise schedule in the P0 and P6 experiments lasted for 11 days: 8 days of adaptation (two cycles of resistance exercise for 3 days and rest for 1 day) and 3 days of urine collection.
    Are urine collections valid measures of protein retention? I would prefer a 12-week study that measures the net muscle gain in response to immediate vs. 6h post supplementation as well. In the end, the 3+ day urine collection is probably still a better measure of the amount of protein that's actually used to "build lean mass" (remember this way we cannot distinguish where the protein was stored - we only know that is was not metabolized and excreted!) than the acute measures of protein anabolic signalling you see in many other studies - a measure of which Mitchell et al. have shown that it does not correlate with resistance training-induced muscle hypertrophy in young men (Mitchell. 2014), only recently.
    Prior to each experimental period, body composition and one-repetition maximum (1RM) were measured, and questionnaires on daily activity were completed.
     "In daily experimental sessions, subjects performed the following resistance exercises: 4 sets of 8 to 10 repetitions of resistance exercise consisting of leg press, leg extension, and leg curl on experimental days 1, 5, and 9; bench press, shoulder press, and triceps pushdown on experimental days 2, 6, and 10; and lat pulldown, biceps curl, and rowing on experimental days 3, 7, and 11. All exercises were performed at 80% RM, and each set was followed by a 2-min break." (Mori. 2014)
    Before the experimental session each day, subjects used a cycle ergometer (Aerobike 800; Combi Wellness Corporation, Tokyo, Japan) at 100 W for 10 min to warmup. Each exercise session was scheduled to take place between 10:00 and 11:00. The subjects were instructed not to participate in any other sports activities during the experimental period.

    The diets were standardized to 1.5g/kg body weight

    In spite of the fact that the bodybuilder group consumed ~13% more energy per day, both the total (1.5g/kg per day), as well as the supplemental protein intake (0.5g/kg of whey protein + 0.8 g/kg dextrin body weight) were standardized. In that, the amount of protein per meal was defined as follows:
    • P0 experimental period: Subjects had to ingest protein (0.3 g/kg body weight) and carbohydrate (0.8 g/kg body weight) immediately after resistance training.
    • P6 experimental period: Subjects had to ingest protein (0.3 g/kg body weight) and carbohydrate (0.8 g/kg body weight) 6 h after resistance exercise session.
    The total energy intake, total protein intake, and protein intake per  body  weight  (kg) were  calculated  by  a  registered dietitian and the protein balance was assessed by analyzing the nitrogen excretion in the urine.
    Figure 2: Net protein balance per body weight and per lean body mass (LBM) when the post-workout protein + carbohydrate shake was consumed immediately after or 6h after the workout (Mori. 2014)
    And, as you can see in Figure 2, a comparison of the net protein balance in the two groups during the immediate post vs. 6h post consumption periods clearly indicates that protein timing does matter, even if it's just for the experienced resistance trainee, for whom the study at hand shows that his nitrogen balance is less positive than that of the rookie anyway.

    Apropos total protein intake and nitrogen balance, the average total protein intake in the studies reviewed by Schoenfeld et al. was after all slightly higher (1.66 g/kg/day). This, as well as the general believe that bodybuilders should consume tons of protein make me question whether the results would have been different, if total protein intake had been 2.0 or even 2.5g/kg per day instead of just 1.5g/kg. Personally, I don't thinks so, but it would be worth a try, anyway.
    (1) Thou shalt not wait 6h to consume protein after a workout (2) Thou shalt not wait 6h to consume carbs (not necessarily ultra fast digesting, though) after a workout either | learn why
    Bottom line: If you're striving for maximal muscle and performance gains, specificity is key; and the study at hand specifies that you have to specifically make sure to get your post-workout nutrition "immediately" post workout and not 6h later if you (a) want to maximize net protein retention and (b) are already beyond those first happy months in the course of which you just have to look at a dumb- or barbell to grow ;-)

    Ah, and when you're at it, I suggest you also include 5g of creatine monohydrate in your postworkout shake. The latter has after all also been shown to work a tad better, when it's consumed after the workout | learn more.
    Reference:
    • Jordan, Leora Y., et al. "Nitrogen balance in older individuals in energy balance depends on timing of protein intake." The Journals of Gerontology Series A: Biological Sciences and Medical Sciences 65.10 (2010): 1068-1076.
    • Mitchell, Cameron J., et al. "Acute Post-Exercise Myofibrillar Protein Synthesis Is Not Correlated with Resistance Training-Induced Muscle Hypertrophy in Young Men." PloS one 9.2 (2014): e89431.
    • Mori, Hiroyasu. "Effect of timing of protein and carbohydrate intake after resistance exercise on nitrogen balance in trained and untrained young men." Journal of Physiological Anthropology 33 (2014): 24.
    • Schoenfeld, Brad Jon, Alan Albert Aragon, and James W. Krieger. "The effect of protein timing on muscle strength and hypertrophy: a meta-analysis." Journal of the International Society of Sports Nutrition 10.1 (2013): 53.

    Protein Blends, Not Isolates Promote Maximal Skeletal Muscle Protein Retention(!) - It's Not About How Much You Pump into the Muscle, It's About How Much You Retain

    Scientific evidence suggests: There is not one optimal protein to build muscle - it's the mix of fast to slow proteins that's key.
    For someone like yourself, who's making sure to get his daily dose of SuppVersity Science News, the results Reidy et al. present in their latest paper in the Journal of Applied Physiology can hardly be surprising. I have, after all, written about the superiority of whey + casein blends as potential muscle builders only recently ("When Whey & Casein Unite in the Spirit of True Physique Improvements, BCAAs & Glutamine Better Shut the F*** Up"  | (re-)read the article). It was thus only to be expected that a study in which the scientists from the University of Texas Medical Branch compared the effects of the prolonged hyperaminoacidemia that's associated with the ingestion of a blend of plant (25% soy) and dairy (50% casein, 25% whey) proteins (with varying digestion rates) to that of a pure rapidly digested whey would yield a definite points win for the "time-released" formula.
    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

    5x More Than FDA Allows
    The reasons why it's still well worth taking a closer look at the study results are (a) the fact that the f**** up supplement industry is still trying to tell you that protein blends would be inferior to overpriced isolates and (b) the educative value of the post-workout + post-supplementation serum amino acid profiles Reidy et al. observed the 16 healthy, young subjects (age range: 19 –30 yr) who participated in their double-blind, randomized clinical trial (with body fat levels of >24% those were certainly no physical culturists, though ;-)
    Figure 1: Graphical overview of the study design (Reidy. 2014)
    As you can see in Figure 1 the study protocol involved a standardized resistance training session in the course of which the subjects who had been kept on a diet containing 20% protein, 60% carbohydrate, and 20% fat at 12 kcal/kg for 72h, performed leg extensions on a Cybex-VR2 (Medway, MA), i.e. 8 sets of 10 repetitions at 55% (set 1), 60% (set 2), 65% (set 3), and 70% (sets 4 – 8) of the participants previously determined 1 RM with 3-min rest between sets, before they consumed the protein beverages (Whey or Blend) exactly 1 h postexercise.
    Figure 2: Net phenylalanine enrichment (left) and inward and outward transport (right)
    The ingestion of the beverages of which the blend and the whey protein contained of 20.1 g total protein (providing 1.9 g leucine, 1.0 g phenylalanine, 1.3 g valine, and 9.0 g EAA; 50% protein from sodium caseinate, 25% protein from whey protein isolate, and 25% protein from soy protein isolate) and 17.3 g of protein (providing 1.9 g leucine, 0.6 g phenylalanine, 1.1 g valine, and 8.7 g EAA; 100% whey protein isolate), respectively, lead to significant increases in amino acid transporter activity (2/SLC38A2, proton-assisted amino acid transporter 1/SLC36A1, cationic amino acid transporter 1/SLC7A1).
    "However, the ingestion of the protein blend resulted in a prolonged and positive net phenylalanine balance during postexercise recovery compared with whey protein (P 0.05)." (Reidy)
    In view of identical postexercise myofibrillar protein synthesis in both groups this difference may appear negligible. If you've been following my articles about the often oversimplified protein synthesis and increases in skeletal muscle mass, you should be aware that net retention and not fractional synthesis is the term you have to look for, when you're analyzing corresponding studies.
    Bolus ingestion could be a superior alternative: In view of the fact that the advantage of protein blends is directly related to their ability to trigger sustained increases of the level of amino acids in the blood, the same can be achieved by the ingestion of whey protein at regular intervals - e.g. at least every 2 hours. Needless to say that this is not just more expensive, but also less practical than the 20-40g of a protein blend many of you are probably already consuming right after their workouts.
    Bottom line: I am still very hesitant to suggest buying a blend with significant amounts of soy in it, when egg proteins should do a similarly beneficial job as a "filler" that keeps the amino acids (AA) levels elevated when the influx of AAs from whey is beginning to seize and the slow digesting casein protein (in the study at hand, we had regular sodium caseinate, which is actually faster digesting than micellar casein) are not yet fully digested.

    In general, however, the study at hand clearly supports the notion that protein blends that are designed to provide a sustained elevation of all essential amino acids in the blood (not just BCAAs, learn why) will induce a superior growth response. Whether the same is true if we compare the ingestion of a single protein blend shake to the repeated (ev. 2h) ingestion of 20g of whey protein, will yet have to be elucidated in future studies.
    Reference: 
    • Reidy, Paul T., et al. "Soy-dairy protein blend and whey protein ingestion after resistance exercise increases amino acid transport and transporter expression in human skeletal muscle." Journal of Applied Physiology 116.11 (2014): 1353-1364.