Timed Ingestion of 3x21g of Whey Protein + Exercise Sheds 14% Abdominal Fat in Overweight Subjects Within 4 Months

Minimal effort, minimal results - While you can lose weight by just adding whey protein to your diet, your success will more than double, when you're willing to work (out) for it four times a week!

It's not a secret that things that diet and exercise are the keys to weight control and health in the 21st century. If you skip only one of the two you can hardly expect optimal results. In that, it is often said that weight, or rather fat loss requires a significant reduction of one's total energy intake; and for athletes and already lean individuals, this may in fact be the case. For the average "free-living overweight or obese" individual, however, the dietary changes that are required can be as simple as adding three servings of 21g of whey protein to their regimen on a daily basis (the scientists found no overall increase in energy intake, this means the 252 extra kcal/day from whey were effectively compensated for by the overweight subjects of the study at hand.

You can learn more about protein intake at the SuppVersity

Protein Timing DOES Matter!

5x More Than the FDA Allows!

Protein requ. of athletes

High EAA protein for fat loss

Fast vs. slow protein

Less Fat, More Muscle!

Before you go ahead and buy a bag of whey from the next best Internet supplement vendor, though, I have to tell you that why alone may have some beneficial effects. Without regular exercise, however, you are not going to shed those ~10% abdominal fat, the subjects in the PRISE, i.e. protein, resistance exercise, interval sprint exercise, stretching/yoga/ Pilates, and endurance exercise, group saw over the course of the 16-week study period.

Table 1: Overview of the exercise program in the PRT and the PRISE group (Arciero. 2014)

ASs you can see in Table 1, the subjects trained four times a week. They did so at different rates of perceived effort (RPE) and they performed

• upper-body resistance exercise (UB) for the chest, shoulders, biceps, triceps, and back,
• lower-body resistance exercise (LB) for the quadriceps, hamstrings, calves, and abdomen, 
• sprint interval training, and endurance training (type C) like walking, jogging, running, cycling, swimming, elliptical, rowing, rollerblading, cross-country skiing, etc. and
• supervised stretching, yoga and pilates workouts (in the PRISE group, only, where
  the four types of exercise were cycled on a weekly basis, such that participants performed each of the four exercises, 1 day/wk for a total of four exercise sessions/wk)

and one session (X), where they were free to chose whatever they wanted to do (i.e. resistance training, conditioning exercises, etc.).

All that without dietary intervention!? It sounds hard to believe that simply adding whey protein to the diet of 79 overweight / obese subjects would have such a profound impact on their body composition, but the scientists did in fact prescribe nothing else than the timed ingestion of 23g of whey protein (1) within 1 h of waking in the morning, (2) mid-afternoon or within 30 min following an exercise session and (3) withing 2 h of going to bed at night (total protein intake ended up at ~1.3-1.5g per kg body weight). Otherwise, all participants were instructed to consume their habitual diet ad libitum throughout the 16-wk intervention.

Only the increase in protein was stat. sign. across all groups (Arciero. 2014)

In the introduction I did yet already hint at the fact that the addition of 252kcal/day from the whey protein did not increase the subjects overall dietary intake (~2,000kcal/day). Against that background it's obvious that the provision of extra whey protein induced voluntary changes in the macronutrient composition of the diet that reached statistical significance for protein (+6%, +9% and +6% in the protein, protein + resistance training and PRISE group, respectively). For fat and carbohydrates the dietary changes were too different from subject to subject (meaning some reduced fat, others carbs) to reach statistical significance - which obviously does not mean that they were not reduced!

During all sessions, the subjects use medicine balls, physioballs, rubber tubes, and bands, which were incorporated into a dynamic warm-up, footwork and agility drills, resistance and power movements, and core and body weight exercises (e.g., lunges, squats, and jumping rope).

Figure 1: Relative changes in body mass, fat mass (subcutanous, visceral and in the abdominal region) and waist circumference over the course of the 4 months study (Arciero. 2014).

If you think that's more than you can handle, you better take another look at the results in Figure 1. Are you really sure you don't have the guts (don't tell me you don't have the time, if you have time to watch TV and lie around lazily on your sofa) to work out on Monday, Tuesday, Thursday and Friday?

I must say that the changes in lean mass are disappointing. Maybe a focus on higher intensity resistance training would have helped build the usually relatively muscled (from carrying an obese body) legs of the overweight / obese participants.

Bottom line: You can argue simply having that extra whey is also going to help you lose body fat, but compared to the "PRISE"-less combination of protein and resistance exercise, intervals, stretching/yoga/ Pilates, endurance exercise the fat loss from whey alone is not exactly impressive. Ok, it's impressive that simply adding three servings of whey do trigger reductions in body fat, but adding 4 workouts of which only the sprint interval workouts reach a maximal intensity of 10 on the RPE scale for only 30s (!) is what makes the difference between statistically significance, and mirror and "man, you've slimmed down"-comment significance ;-)

Needless to say, though, that completely turning your diet upside down and making exercise an integral part of your everyday life are more promising strategies to lose weight and stave it off than any of the interventions in the study at hand | Comment on Facebook!

References:

• Arciero, Paul J., et al. "Timed-daily Ingestion of Whey Protein and Exercise Training Reduces Visceral Adipose Tissue Mass and Improves Insulin Resistance: The PRISE Study." Journal of applied physiology (Bethesda, Md.: 1985) (2014).

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

Are You Protein Wheysting?

5x More Than the FDA Allows!

Protein requ. of athletes

High EAA protein for fat loss

Fast vs. slow protein

Less Fat, More Muscle!

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 Timing Reloaded: A Reminder on the Importance of Repeated 20g Pulses for Optimal Protein Synthesis

Protein timing & dosage matter for protein synthesis in the PWO window.

"30g+ of quality (=leucine rich) protein per meal." I am not really sure how often I have been writing and saying this here, on facebook and on the Science Round Up at SuperHumanRadio in the past months, but I hope that it's been enough for you to remember this simple rule; a rule that will also help you to distribute your protein intake somewhat more evenly across the day and thus - as a recent study that was conducted by researchers from the School of Medical Sciences at the RMIT University, in Melbourne, the Australian Institute of Sport in Canberra, the Department of Kinesiology at the  McMaster University in Hamilton, Ontario and the Nestlé Research Center in Lausanne, Switzerland - maximize the protein induced increase in protein synthesis.

Note: This paper is actually an almost identical clone to a previous paper by the same groups of researchers (Moore. 2012). Since I've discussed that back in 2012 already, I still decided to include it in this week's SuppVersity line-up, because it fits in perfectly with the previously discussed study on the benefits of protein intakes >2x RDA and that simply eating more protein is not really gonna cut it. If you want to read up on the corresponding SuppVersity article discussing the 2012 study, you can do this right here.

More is only more if it's adequately timed & distributed

The scientists knew from countless previous experiments that a single bolus of∼20 g of protein after a bout of resistance exercise would provide a maximal anabolic stimulus during the early post-exercise recovery period (∼5 h). The "effect of various protein feeding strategies on skeletal muscle protein synthesis during an extended recovery period (12 h)" (Areta. 2013) was yet largely  unknown. In order to shed some light onto the protein-anabolic response in this "post-post-workout window", the researchers compared three different patterns of ingestion of 80 g of protein during 12 h recovery after resistance exercise and the associated anabolic response in human skeletal muscle:

• 10, 20 or 40 g feedings using a pulsed, intermediate or bolus ingestion regimen, respectively
• consumed after 4x10 reps @  80% of the 1 RM with 3 min rest between sets

 In accordance with the hypothesis that protein timing would directly influence the expression of "anabolic" signalling proteins in the muscle, the researchers observed significant hierarchy with BOLUS > INT > PULSE in the magnitude of phosphorylation of p70S6K and p-AKT on an individual base.

Figure 1: Fractional protein synthesis expressed relative to mean protein synthesis at the respective timepoint (left); corresponding levels of the "anabolic" signaling proteins p70S6K (right, top) and p-akt (right, bottom; Areta. 2013)

However, despite the fact that the immediate response was maximal in the BOLUS group, both the actual amount of protein that was shuttled into the muscle was still significantly higher in the INT trial, in the course of which the subjects ingested 20g of protein every three hours.

So what's the deal, then?

The "anabolic window" turns out to be more of a barn door, which is unlocked by the key of exercise and nutrition science (learn more)

In their discussion of the results, the researchers point out that this is the first study to confirm the long-touted notion that ...

"[...] rates of myofibrillar protein synthesis (MPS) remain elevated above rest throughout 12 h of recovery when a single bout of resistance exercise is followed by the partitioned ingestion of 80 g of high quality protein. Furthermore, we show that daily rates of protein synthesis were highest with regular (i.e. every 3 h) intake of a moderate (20 g) quantity of rapidly digested whey protein." (Areta. 2013)

Areta et al. specifically emphasis that the acute stimulation of the nutrient and contraction-sensitive intracellular signalling network is thus not the main determinant of net protein synthesis in the huge post-anabolic window.

A word of caution: Before you freak out totally about how big you're going to be next week, now that you follow the optimal protein supplementation regimen, I suggest you take a look at Alan Aragon and Brad Schoenfelds paper on the purported and real benefits of protein timing and its real-world implications | read more ...

The anabolic effects of the intermediate supplementation do yet only appear to be. Over the whole study period, they are not statistically different and the superiority in terms of the actual fractional protein synthesis (FSR) persists, even when using a mean intracellular enrichment from the other groups. Thus an increase in intracellular enrichment cannot be the underlying mechanism for the increase in  fractional synthesis rates in the intermittent supplementation trial (INT) the researchers ascribe to an optimized interplay between resistance exercise, time between ingestion, and the total quantity (20g = at the critical threshold) of each (whey) protein feeding. The optimally timed and adequately dosed ingestion of 4 servings of 20g of whey could thus have

Figure 2: Plasma EAA concentration following a bout of leg extensions and subsequent whey ingestion during a 12 h recovery period (top); timing and dosage of supplementation (bottom); p < 0.05 for pulse *, intermittent †, and bolus ‡ at equivalent time point (Areta. 2013)

"[...] resulted in a cyclical plasma AA profile that might be considered ideal to stimulate MPS [...] While the temporal resolution of time points selected for quantifying plasma amino acid concentrations failed to clearly show distinct amino acid profiles throughout the 12 h post-exercise period a hierarchical response was observed for leucine concentration early in recovery indicative of an initial divergence in branch-chain amino acid availability between feeding patterns [figure 2]. Thereafter, the continuous hyperaminoacidaemia or lack of postprandial periods of relative hypoaminoacidaemia over the 12 h period with PULSE feeding most likely resulted in some suboptimal stimulation of the protein synthesis machinery similar to that observed with continuous amino acid infusion." (Areta. 2013). 

Now, this may appear counter-intuitive at first, after all the aminoacedemia in the intermediate trial does not appear to be more pronounced than that in the pulse trial and certainly inferior to the one of the bolus trial. This is in fact true, but it is a mere consequence of much greater %-ual absorption of the amino acids in the intermittent trial compared to both the bolus and pulse supplementation, of which the former are 'protein wasting' and the latter simply insufficient to fully trigger the protein synthetic response.

---

One thing to keep in mind, when you look at the results of the study at hand is the fact that this is just another acute resistance training study that employs an unrealistic (leg extensions only) workout. In other words, we cannot simply take for granted that statistical significant difference in short term protein synthesis (12h is still short) will also translate into measurable, let alone visible improvements in muscle hypertrophy. Still following the protocol outlined to the left is certainly not going to hurt you.

Bottom line:  It should not really be necessary that I actually formulate another bottom line. If you stick to my basic recommendations (outlined among others in my interview with my buddy Sean Casey) you will do something very similar with your 3-5 meals with 30+g of quality protein each. The only thing that may be worth adding to it is a re-emphasis on the interaction of timing and dosing. 

You need at least 20g of leucine-rich protein to kickstart protein synthesis and - as of now - it looks like those will not sustain maximal protein synthesis for longer than ~3h before you have to 'refuel' the amino acid pool and (probably more imporatantly) 'reignite" the protein synthetic machinery with another 20g+ bolus of whey.

In view of the fat that proteins with a low(er) leucine content will probably need to be ingested in higher amounts I would still stick to the '30g+ rule' - after all, I assume you won't want to live off whey protein supplements and Quest Bars, only - do you?

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

References: 

• Areta JL, Burke LM, Ross ML, Camera DM, West DW, Broad EM, Jeacocke NA, Moore DR, Stellingwerff T, Phillips SM, Hawley JA, Coffey VG. Timing and distribution of protein ingestion during prolonged recovery from resistance exercise alters myofibrillar protein synthesis. J Physiol. 2013 May 1;591(Pt 9):2319-31. doi: 10.1113/jphysiol.2012.244897. Epub 2013 Mar 4. 
• 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.

It Does Matter How You Spread Your Protein Intake - 30% Higher 24h Protein Synthesis with 30g+ Protein per Meal

Today's SuppVersity News will provide you with "confirmation" rather than "innovation", I suppose

With my recent article on the non-existance of protein-related osteoporosis (read more) and the short news post about the unique satiating effects of protein snacks (read more), there's been quite some protein lovin' here at the SuppVersity as of late. Usually, I would try to avoid having yet another "protein article" in the same week, but for the most recent study on "Dietary Protein Distribution", I will make an exception and I bet, you won't mind! Why? Well, what about what follows the "Dietary Protein Distribution" in the title of said paper?

"...Influences 24-h Muscle Protein Synthesis in Healthy Adults"

By now, you may feel reminded of a recent review by Alan Aragon and Brad Schoenfeld (Aragon. 2013), the results of which (learn more) are not refuted by the results of the study at hand.

Avoid protein wasting post workout.

Why do I even mention the Aragon + Schoenfeld study? The reason is that I already read how people were going on about how this "stupid review" got it all wrong on Facebook. And though I know that SuppVersity readers are not as ignorant as the average gymbro (watch what I am talking about, here) I wanted to make sure that (a) this study is not about the post-workout anabolic window Aragon & Schoenfeld wrote about and that (b) the tow actually argued that spreading your protein intake across the day instead of placing it in the "anabolic window", should yield superior results.

What the study does tell us, is simple: "The consumption of a moderate amount of protein at each meal stimulated 24-h muscle protein synthesis more effectively than skewing protein intake toward the evening meal." (Mamerow. 2014)

In other words: Don't cram all your protein into one meal!

I guess in view of past articles on related topics (e.g. "2x40g, 4x20g or 8x10g of Whey? Which Feeding Strategy Yields the Greatest Net Protein Retention?" | read more; or "Protein Timing Reloaded: A Reminder on the Importance of Repeated 20g Pulses for Optimal Protein Synthesis" | read more), this insight is not really going to surprise you.

Figure 1: Fractional protein synthesis at breakfast (left), when the difference was most pronounced (+30%) and rel. calculated 24h fractional protein synthesis (right) with EVEN vs. SKEWED protein distribution (Mamerow. 2014)

What may surprise you, though is the simple fact that this study, which was a joint venture of scienfitsts from the Division of Rehabilitation Sciences at the Department of Nutrition and Metabolism, and Department of Internal Medicine at the University of Texas Medical Branch and the Department of Food Science and Human Nutrition at the University of Illinois at Urbana (Mamerow. 2014) is the first study to conclusively show that spreading a relatively high protein intake (1.2g/kg body weight) across the day is superior to the large steak the average intermittent faster may be washing down with a triple protein shake in the evening.

With an average age of 37 years the 8 healthy, normal-weight adult men and women who participated in the study at hand were neither rodents, nor elderly individuals, and - contrary to what you may expect if you look at the italicized names of the Institutions the scientists who were involved in this study are working at - they were not in need of rehabilitation after an injury - they were average Joes (n = 5) and Janes (n= 3).

This is not about rodents, elderly people or injured athletes

As you can see in the overview in Table 1, the subjects consumed three square meals, i.e. breakfast, lunch and dinner in the course of the 7-day study period. The previous reference to intermittent fasting is thus obsolete - eating a minimal amount of protein in the morning and at noon is after all very different from eating nothing at all. 

Table 1: Seven-day mean energy and macronutrient intake in healthy adults consuming diets with an EVEN or SKEW protein distribution (Mamerow. 2014)

As the scientists point out, the total 24-h protein, carbohydrate, and fat consumption in the SKEW and EVEN conditions was not different.

"Both diets exceeded the RDA for protein [0.8 g/(kg d)] by ~50%. The SKEW diet met the RDA for protein during the evening meal alone. In all versions of the EVEN and SKEW menus used in this study, the animal-to-vegetable protein ratio was ~2:1." (Mamerow. 2014)

By using a 7-d crossover feeding design with a 30-d washout period, the scientists were thus able to measure the influence of protein timing, on the changes in muscle protein synthesis.

The latter was measured thrice, i.e. after each of the three meals, and used to calculate the twenty-four-hour mixed muscle protein fractional synthesis rates on days 1 and 7 after the ingestion of EVEN-ly or SKEW-edly distributed protein diets.

"Fat Loss Principles That Work: 10g+ of EAA W/ Every Meal" | read more

Bottom line: You have already seen the outcome of the three FSR measurement in Figure 1 and there is actually not much to add to what you're seeing there already.

In view of the fact that I gather that you'd expected a result like this, I don't feel inclined to repeat that I have been suggesting for years to consume 30g+ of quality protein ("quality" = 10g+ of EAAs per 30g serving) with every meal.

If you stick to this simple principle, it's going to help you build muscle and lose fat (see "Fat Loss Principles That Work: 10g+ of EAA W/ Every Meal" | read more).

Reference:

• Aragon, Alan Albert, and Brad Jon Schoenfeld. "Nutrient timing revisited: is there a post-exercise anabolic window?." Journal of the International Society of Sports Nutrition 10.1 (2013): 5.
• Mamerow, Madonna M., et al. "Dietary Protein Distribution Positively Influences 24-h Muscle Protein Synthesis in Healthy Adults". J. Nutr. January 29, 2014 jn.113.185280 [ahead of print].