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marylin monroe
Showing posts with label recomposition. Show all posts
Showing posts with label recomposition. Show all posts

22g High EAA (6g) Protein + 36g CHO Pre- / Intra-Workout Boost Fat Oxidation & PWO Resting(!) Energy Expenditure

I don't doubt that you can do that, too!
It does sound awkward: If you mix Twinlab: Amino Fuel (22 g protein - 6 g essential amino acids | L-phenylalanine: 633 mg; Lvaline: 781 mg; L-tryptophan: 133 mg; L-threonine: 679 mg; L-isoleucine: 565 mg; L-methionine: 292 mg, L-histidine: 282 mg; L-leucine: 1350 mg; L-lysine: 1449 mg) with a regular  sports recovery drink that contains 36g of simple sugar, down half of the resulting 800ml serving of whatever you want to call this mix immediately before your workout and consume the rest during the rest periods between sets, this will have measurable effects on your resting energy expenditure and fat oxidation.

From long-term to short time effects

At first, it does questionably sound counter-intuitive that the ingestion of an EAA + carbohydrate mixture before / during would increase the resting energy expenditure and rate of fatty acid oxidation after your workout. On the other hand, if you think about the long-term effects of corresponding supplement regimen, you don't have to look far, to find evidence that they can promote both, muscle gain and fat loss (Bird. 2006).
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?
Kyle J. Hackney, Andrew R. Kelleher, and Lori L. Ploutz-Snyder from the Syracuse University speculated that the highly beneficial changes in body composition Bird et al. observed in their study participants over the course of a 12-week strength training + EAA & CHO supplementation that after "[t]hese adaptations may be related to the acute energy expenditure and substrate utilization responses in the postexercise period." (Hackney. 2013)
Figure 1: The changes in body composition (in kg) in response to 12 weeks of resistance training + placebo, CHO, EAA or CHO + EAA supplementation in 2006 study by Bird et al. "inspired" Hackney et al.
Against that background, it was only logical to conduct a study to examine how multiple bouts of resistance exercise with and without the strategically timed intake of amino acids affect the resting energy expenditure (REE) and respiratory exchange ratio (RER). The results could after all explain if the long/er) term effects on body composition that have been observed in previous studies using chronic training and supplementation regimen are maybe nothing but necessary consequences of repeated acute increases in REE or decreases in RER (you hopefully remember that a decrease in the respiratory exchange ratio signifies an increase in fatty acid oxidation).

Experimental design and results

To this ends, the researchers recruited 10 young (mean age: 23.4y) recreationally trained male participants. All of them had been participating in general resistance training exercise for a minimum of 3 days per week for at least 6 months.
Figure 2: Changes in resting energy expenditure (kcal/day) and comparison of training volume in 58g CHO (black bars) and EAA + CHO (white bars) trials (Hackney. 2013).
As you can see in Figure 2, Hackney et al.'s original hypothesis that "intake of amino acids with each resistance exercise session would lead to greater perturbations of REE and RER" (Hackney. 2013) does unquestionably hold for this population of average (rookie) gymrats.

Whether the scientists "main finding" (Hackney. 2013), i.e. the 3.61% increase in resting energy expenditure (REE) will be similarly pronounced in advanced trainees is yet as questionable as the real-world effects of this artificial value. Despite the fact that Hackney et al. are right, when they say that our resting energy expenditure "represents the largest component of [our] total daily energy expenditure (60–85%) and has been implicated as a major contributor to overall body mass management " (Hackney. 2013), I am not sure how "major" an increase of only 66kcal per day actually is... I mean,  if this pathetic increase in resting energy expenditure was the actual driving force we would need almost 100 days to shed a hilarious pound of body fat (note: the reason I use the flawed 3,500kcal = 1lbs of fat rule of thumb here is that the whole REE calculations would be pointless if you didn't put at least some faith into the "energy in vs. energy out" hypothesis of weight loss - right?)
SuppVersity Suggested Read: "Fat Loss Principles That Work: 10g+ of EAAs W/ Every Meal. Do Energetic Costs of Protein Synthesis Trigger This Effect?" |  read more
Bottom line: It stands out of question that your training success can benefit from a high EAA protein source and some carbs you consume before and during the exercise session. Whether the fat loss benefits are actually brought about by the marginally increased resting energy expenditure (REE) is yet something I doubt - it certainly helps fat loss, but clearly isn't its main motor.

Don't get me wrong, this does not imply that you will benefit from this type of "peri-workout" supplementation. And let's be honest, the end most of you probably don't care about the exact underlying mechanisms, as long as your body composition keeps improving, right?
References:
  • Bird, S. P., Tarpenning, K. M., & Marino, F. E. (2006). Independent and combined effects of liquid carbohydrate/essential amino acid ingestion on hormonal and muscular adaptations following resistance training in untrained men. European journal of applied physiology, 97(2), 225-238.
  • Hackney, K. J., Kelleher, A. R., & Ploutz-Snyder, L. L. (2013). Amino Acid-Carbohydrate Intake Combined with Multiple Bouts of Resistance Exercise Increases Resting Energy Expenditure. ISRN Nutrition, 2013.

Train Like a Woman: Common Misconceptions About Training & Eating for A Cover-Model Physique - An Interview With Sports Nutritionist & Strength Coach Orit Tsaitlin

Do you have to train like a woman (whatever that may be), Ladies, if you want to develop the cover model physique Orit Tsaitlin already has?
No, you're not mistaken. Today's SuppVersity article is not a review of the latest science, it's in fact an interview. An interview with Sports Nutritionist & Strength Coach (CISSN, ISSA, MBA, B.Sc.) Orit Tsaitlin.
It's not just that the mantra, "Nutrition Strategy and Body Transformation - Where Fitness and Science meet", on the frontpage of Orit's website that sounds pretty similar to the SuppVersity slogan "Where Bro- & Proscience Unite in the Spirit of True Wisdom", but also some of Orit's post on the SuppVersity Facebook Page, as well as a couple of short chats we had that inspired me to do a short interview with her. An interview that revolves around the useful- or rather uselessness of the average fitness magazine and the nuanced differences between training and nutrition principles for men and women.
By using HIIT you can achieve similar results as in the study at hand

1g PRO per 2g CHO + Circuit T. for Women?

Is the Optimal Exercise Order Sex-Specific?

Coffee, Tea, Cacao, ... & Breast Cancer

Not Bulky! Lifting Will Make Toned & Strong.

SV-Special: First Dates How do They Work?

Sex-Differences in Fat Oxidation - Reviewed
Q: A recent study has shown that the "dietary advice for muscularity, leanness and weight control" in Men's Health magazines – as scientific as it may seem- "leaves much to be desired" (Cook. 2014). As a personal trainer who works with both male and female clients, what do you feel are the consequences for men and women who are looking for help to improve the way they look, feel and perform?

A: Its actually quite amusing to read the articles in fitness magazines… Since I was 18 years old (now 32), nothing has changed, I believe. The same content, pictures of beautiful bodies and nothing really helpful to the the average person who only wants a simple roadmap to finally feel and look good. If I could be an editor of Fitness Mag (sorry, when I will be - determined yes?) I would put a lot more emphasis on how much work, nutritional discipline and sacrifice those athletes in the pictures are making to achieve their looks and performance.

Figure 1: "Who succeeds in maintaining weight loss?" A conceptual review of factors associated with weight loss maintenance and weight regain shows that you need the right strategies and psy- chological approach, not tips from Men's Health & Co. to keep the weight off (Elfhag. 2005)
Regarding the nutritional advices in those magazines, the audience must understand that MH and other magazines are no experts to individual metabolism and body. Unfortunately, there are tons of nutritional advices in those articles but the most important ones - that you have to eat a specific quantity of food which matches your lifestyle or that micronutrients (vitamins ,minerals) should be taken only on doctor's prescription - are not.

And let's not forget the paradoxical advice "The body needs protein which is in fish, chicken, meat... so buy our BCAA".

As a trainer and sports nutritionist, I emphasize that people must stop train their eyes, ears and need to start listen to their body's signals and to train the most important things to reach the body transformation results - will-power and consistency.

Q: Speaking of fitness and lifestyle magazines. If I look at the women's magazines, I cannot avoid the impression that the previously mentioned fitness and lifestyle magazines for men are still a comparatively good source of information.Why do you think, women's magazines are still perpetuating long overcome ideas of the ways women have to train?

A: I agree with you that there are some men's fitness magazines that you can find good information regarding training techniques from which both genders can learn.

SuppVersity Facebook News: Looking good naked provides greater incentive for healthy eating (increasing fruit and vegetable intake) than health improvements; the same goes for the headlines of fitness mag articles  | learn more.
As for women magazines… In my experience with women clients and being a woman myself, we are more concerned with body issues than men and I totally understand it. It is a great feeling to love the image that you see in the mirror and receive compliments. Unfortunately, most women want everything now.

I mean most of us don’t have the patience to wait for things to happen in their own pace. Women fitness magazines know this and are doing a great job in translating it to money. My point is that articles with the headers like ‘’10 min to your best body” , “15 min to see those abs” or “1 month squat challenge’’ will definitely reach the women audience.

In contrast to really relevant articles with headlines such as "The correct deadlift form" or "The importance of progressive overload", which don’t sell perfect and easy body fantasy probably won’t make it to the woman reading list. In this industry each trainer, athlete or nutritionist knows what it takes to get to the top or, maybe, not to the top, but to get meaningful results and as soon the audience will understand that there are no shortcuts to any place worth going, the women fitness mag will start give a proper stage to the right articles with a more meaningful content regarding exercise and nutrition. Until then I’m going to make 80 squats per day for one month which will give me the J.LO. booty.

Q: Is there a top 5 of "false believes" your female clients have? If so what are they and in how far are they holding them back from making the progress they deserve? 

A: There certainly is, and although the order may vary from individual to individual, this is what I see / hear the most:
  1. Not falling for any of the common misconceptions on the left, i.e. the "calories don't count" myth, the "I will take the weekend off of my healthy lifestyle" mistake, the "I'll just burn it off" misconception, the "it just takes three months" idiocy, and the "I will just do what she does" misunderstanding, is certainly part of Orit's success.
    Calories don’t count - Sorry they do and you can not argue with the energy conservation law. 
  2. The weekends are off from good food choices and quantities - The weekends are meant for relaxation and other activities, except from working. If you want to achieve those results in matter of months and not years, stop making 48 hours food celebrations...those can cancel all the progress that was made through the week.
  3. If they ate more than usual they can burn it in exercise - If you ate more than you should, the only thing that you can do about it is to take responsibility and learn from that experience for the next day.
  4. Three months transformations are for everyone - No! Those are for people who are already training with weights, have average body fat % and have healthy relationship with food.
  5. The "I want her body"-mindset’ If you want her body start working like her in the gym and in nutrition department the other 23 hours. I promise you she doesn’t spend a moment of her time on things that don’t matter.
I guess you can make a top-10 out of it, but as I initially pointed out, these are my personal top five of "success ruining" misconceptions.

Q: Would you agree, when I say "There are no fundamental differences between the ways men and women have to train to develop a cover-model physique?" This would obviously imply that there are nuances, where the training will differ. What would you say are these nuances?

Read the SuppVersity review of sex-differences in fatty acid oxidation  | more
A: Yes, I agree with you that there are no fundamental differences in the training programming between the genders. The main nuances that will be different are regarding the fact that men lose fat more easily than women, so maybe the frequency of different types of training weights / HIT / LISS will differ.

Which brings up another nuance that is related to "weak areas" and ideal proportions. I like to build my training programs with the goal to build the perfect proportions to the client, which are different, of course, between the genders.

Q: Speaking of cover model physique. You certainly have one. What would you say are the 10 pillars of your current nutrition and exercise regimen?

A: Thank you. I live by 4 rules which I call my 4elements4life. These elements are:
  • SuppVersity Suggested: Sean Casey's interview with Orit in the August-September CasePerformance Newsletter
    Nutrition - 80% of my nutrition is based on whole foods, with the 20% I go crazy - mainly with the croissant (-: and of course I know the right food quantities for my daily energy, PR’s setting and body fat levels.
  • Strength - I do my best in being strong in my training sessions 3-4 times per week at the gym and in the real life. Mental strength will beat everything.
  • Determination - Life taught me always to get up after you fall, set the most ambitious goals and always look forward. If I don’t know something I will learn. If I’m not good at something I will ask for help and If I’m tired….I remind myself - Be determined!!! 
  • Change - Most of the things in my life I accomplished on my own. Nothing was easy, believe me, and to succeed I had to change my mindset, character and search for mental qualities that I didn’t knew I had. "Change your thoughts and you change the world"? 99% true. Because at the end of the day the only world that you need to change is your own.
I guess some trainees will rank the individual aspects differently, but in general all four elements should be among the "pillars of a sensible nutrition and exercise regimen", you asked for.

Q: Is there anything you'd have to change if you were a guy (except from increasing your energy intake to feed the sex-dependent increase in lean mass)?

A: I would train with more emphasis on building lean mass in my quads - more isolation work in addition to the complex movements that I am already doing and may be more training volume in the Chest area. Other than that I’m sorry to tell you, but I train like other men...OMG So I will become a man soon?? (-:

Q: Last question, I promise ;-) Personally, I believe that people don't realize that the way they train and eat must adapt to the changes their body undergoes. Is this an idea you would subscribe to, and if so, what would you say are commonly overlooked tweaks both men and women have to make to their diet and exercise regimen as they progress from chubby slob to fitness maniac?

A: In my personal and professional experience, as the body goes through physical transformation there are stages. In each stage, which is very individual, you must adapt the nutrition and the exercise regimen. I don’t talk about changing programs every week, but when you see that something stopped working or not working as you want it to be, ask yourself what happened?

If you are successful (e.g. with losing weight) your body, dietary & exercise requirements and your goals will change. Sticking to the same approach "forever" would thus be madness | learn more
But wait...being a daughter of a chess player, I must tell you that the #1 thing that you need to do before taking the next step is to think (in every aspect of your life..:-), but in regard to fitness my rule of thumb is: "Do not change nothing until you have evaluated all the variables and your adherence." Our body is great in losing fat and growing muscles but from time to time its demands are changing and you must learn to listen and give it the right tools.

It is not easy to continue when you reach a plateau and it is hard to trust another person (nutritionist or trainer) who tells you to be patient and trust the process.

Trust the process and most important - remember to enjoy the journey. Do not go to the gym in the mindset to lose weight, do not eat food and think this will burn my fat faster. The secret to success is doing those things consistently each day and really give all you got. When you will finally have realized that the only person that you have to prove something to is you, you are ready to develop the physique of a cover model and the mentality of warrior.
Don't send marriage proposals, guys. As far as I know Orit is already spoken for ;-) Which obviously doesn't mean that you cannot benefit from her advise on training and nutrition.
Bottom Line: While I would hope that most of you were only mildly surprised to hear that the sex-differences difference between men and women are significantly more pronounced in the mirror than they are in the gym or in the kitchen (no, this is not sexist comment, but a reference to the fact that men and women don't have to eat fundamentally different either), I suspect that some of you are still all-caught up in the cardio for shaping, weight lifting for bulking ideology.

If that's you, and you've decided you want to make a change, this would be a good reason to reach out to Orit Tsaitlin with your questions. You can do so by either posting short questions on facebook for me or Orit to answer them; or you can contact Orit directly @ 4elements4life | Nutrition Strategy and Body Transformation.
References:
  • Appleton KM. "Increased fruit selection and consumption following an appearance-based vs a health-based health promotion poster." Proceedings of the Nutrition Society, 73, E58 (2014). doi:10.1017/S0029665114000871.
  • Cook TM, Russell JM, Barker ME. Dietary advice for muscularity, leanness and weight control in Men's Health magazine: a content analysis. BMC Public Health. 2014 Oct 11;14(1):1062. [Epub ahead of print] 
  • Elfhag, K., and S. Rössner. "Who succeeds in maintaining weight loss? A conceptual review of factors associated with weight loss maintenance and weight regain." Obesity reviews 6.1 (2005): 67-85.

Betaine Supplementation For Strength, Size & Endurance Gains: 2.5g/Day For 6 Weeks Does the Trick in Adv. Trainees

If you feel like a shadow of yourself, betaine may help - specifically with  metabolically demanding workouts.
"Six-weeks of betaine supplementation improved body composition, arm size, bench press work capacity, attenuated the rise in urinary HCTL, and tended to improve power (p = .07) but not strength." (Cholewa. 2013) That's not just the conclusion to the latest paper Jason Cholewa et al. have just published in every supplement junkie's favorite scientific journal (do I have to mention it's the one of the International Society of Sports Supplementation, ISSN?), it's also exciting news on trimethylglycine and further evidence that it has the potential to queue up in the short line of effective dietary supplements you may spend money on without having a guilty conscience.

6 Weeks + 2.5g/day = Increased mass, volume and strength gains

The routine the researchers had their 23 experienced recreationally strength trained males (weight: 86.8 ± 9.1 kg; training experience: 4.8 ± 2.3 months; BF%: 16.9 ± 8%) between the ages of 18 and 35 follow was clustered into three microcycles (figure 1)
Figure 1: Overview of the training schedule sets, reps and rest times (Cholewa. 2013)
Obviously a pretty solid training program that was (no surprise) able to increase muscle size, strength and strength endurance irrespective of whether the subjects received
  • When do you take betaine? After checking back with the author, I can now tell you that the betaine can be taken at any time point and that you may want to have it with a meal to avoid stressing your gastrointestinal system.
    2 x 1.25g placebo capsules, or
  • 2 x 1.25g betaine capsules
The dosage was chosen because it's safe (9-12g/day = safe; ), effectively elevated plasma betaine levels (2.5-5g; ) and did already produce strength and performance gains in previous studies (Hoffman. 2009; Lee. 2010; Trepanowski. 2011).
Figure 2: Pre / Post bench press volume (left) and muscle cross section (right; Cholewa. 2013)
What is somewhat surprising, at least in my humble opinion, though, is the greater increase in bench press volume (figure 2, left) in the first microcycle and the "comeback" of the placebo group afterwards. Considering the fact that this parameter did not improve in previous studies at all, and comparing the workload on the chest day during the different microcycles, the researchers managed a reasonable explanation for this observation:
"Advanced Trainees Benefit from Increased Training Volume! Greater & Steadier Strength Gains with 8 Sets of Squats. Plus: Over 6 Weeks, 1 Set and 4 Sets Equally (In-)Effective." | read more
"Given improved work capacity with higher volume resistance training prescriptions, and the lack of improvement during micro-cycle 2 which imposed less of a metabolic demand (4 sets of 4–6 repetitions with 3 min rest), it is likely that betaine poses the most ergogenic potential in resistance training exercise protocolsthat impose higher metabolic demands.

Betaine is actively taken up by skeletal muscle during periods of stress, and may be ergogenic as an osmolyte by protecting sensitive metabolic pathways against cellular hypertonicity such as protein turnover, amino acid and ammonia metabolism, pH regulation, and gene expression. Specifically, betaine maintains cellular hydration to protect myosin ATPase and myosin heavy chain proteins against denaturation by urea. Moreover, the affinity of troponin for Ca 2+ , and thus force production, is negatively affected by reductions in protein hydration." (Cholewa. 2013)
Or put simply: The more metabolically challenging (~high volume) the workout the greater the chances you will benefit from taking betaine aka trimethylglycine (not be confused with betaine HCL, the digestive aid).

"Body recomposition" in trained athletes is what could betaine make a huge success

Against that background it seems only logical that the back squat work capacity improved nearly twofold compared to placebo during microcycle 3 (4 sets of 4–6 repetitions with 3 min rest; data not shown). You could even argue that the increases in muscle cross-sectional area (CSA) with betaine may occur only in the arms, because biceps and triceps are hammered directly and indirectly and thus at a relatively higher volume than the legs.
Figure 3: Changes in body composition in betaine & placebo group (Cholewa. 2013)
Eventually, the volume increase, or rather the improved handling of the metabolically demanding workouts, could also explain the highly desirable body recomposition effects we see in the betaine group. Less metabolic waste cluttering around = improved effects on body composition...? Well, the HCTL levels and changes in homocysteine the scientists measured would not support this notion. However,
Betaine content (in mg/100g) in some common food items (read more)
"[...] betaine supplementation may have impacted body composition via other mechanisms. Betaine has been shown to elevate plasma GH and IGF-1, and increase Akt phosphorylation in human skeletal muscle (Apicella. 2012). In mice betaine improves insulin sensitivity by restoring activation of IRS1 and the subsequent phosphorylation of PI3K/Akt by 50-100% in a concentration-dependent manner (Jakubowski. 2009). Thus, it is possible that by elevating anabolic hormones and enhancing downstream cellular signaling, betaine may have improved muscle protein synthesis, thus leading to an increase in lean mass." (Cholewa. 2013)
An alternative explanation Cholewa et al. present pertains to the osmo-regulatory effects of betaine, which may have lead to a "cellular swelling without an appreciable increase in myofibril protein accretion" (Cholewa. 2013). Obviously, these are not the gains you are looking for, but in view of the fact that Keller et al, were able to show that this correlates with decreases in proteolysis it would at least minimize the amount of protein that's getting lost from the musculature right after you "pumped" it into it (Keller. 2003).

What do eggs and homocysteine have in common? None of them causes heart disease. Eating eggs nay even prevent it by modulating the lipoprotein profile and cholesterol efflux (read more)
Bottom line: The study at hand adds more and, above all, highly relevant (human study, advanced trainees) evidence that betaine (2.5g/day in two servings) could make a valuable addition to the supplementation regimen of fitness models, physique competitors and bodybuilders, alike.

With its long-proven ability to reduce homocysteine levels () it has also been implicated as an agent that may prevent heart disease... unfortunately,  this assumption is based on the hypothesis that homocysteine plays a causative role in the development of heart disease and neither this nor the beneficial effects of choline & betaine on CVD are proven facts (Folsom. 1998; HCS. 2002; Olthof. 2005).

References:
  • Apicella JM, Lee EC, Bailey BL, Saenz C, Anderson JM, Craig SA, Kraemer WJ, Volek JS, Maresh CM. Betaine supplementation enhances anabolic endocrine and Akt signaling in response to acute bouts of exercise. Eur J Appl Physiol. 2013 Mar;113(3):793-802.  
  • Cholewa JM, Wyszczelska-Rokiel M, Glowacki R, Jakubowski H, Matthews T, Wood R, Craig SA, Paolone V. Effects of betaine on body composition, performance, and homocysteine thiolactone. J Int Soc Sports Nutr. 2013 Aug 22;10(1):39. [Epub ahead of print].
  • Folsom AR, Nieto FJ, McGovern PG, Tsai MY, Malinow MR, Eckfeldt JH, Hess DL, Davis CE. Prospective study of coronary heart disease incidence in relation to fasting total homocysteine, related genetic polymorphisms, and B vitamins: the Atherosclerosis Risk in Communities (ARIC) study. Circulation. 1998 Jul 21;98(3):204-10.
  • Hoffman JR, Ratamess NA, Kang J, Rashti SL, Faigenbaum AD: Effect of betaine supplementation on power performance and fatigue. J Int Soc Sports Nutr. 2009, 6:7.
  • Homocysteine Studies Collaboration (HCS). Homocysteine Studies Collaboration. Homocysteine and risk of ischemic heart disease and stroke: a meta-analysis. JAMA. 2002 Oct 23-30;288(16):2015-22.
  • Jakubowski H. The pathophysiological hypothesis of homocysteine thiolactone-mediated vascular disease. J Physiol Pharmacol. 2008 Dec;59 Suppl 9:155-67. Review.
  • Lee EC, Maresh CM, Kraemer WJ, Yamamoto LM, Hatfield DL, Bailey BL, Armstrong LE, Volek JS, McDermott BP, Craig SA: Ergogenic effects of betaine supplementation on strength and power performance. J Int Soc Sports Nutr. 2010, 7:27.
  • Keller U, Szinnai G, Bilz S, Berneis K. Effects of changes in hydration on protein, glucose and lipid metabolism in man: impact on health. Eur J Clin Nutr. 2003 Dec;57 Suppl 2:S69-74.
  • Olthof MR, Verhoef P. Effects of betaine intake on plasma homocysteine concentrations and consequences for health. Curr Drug Metab. 2005 Feb;6(1):15-22.
  • Trepanowski JF, Farney TM, McCarthy CG, Schilling BK, Craig SA, Bloomer RJ: The effects of chronic betaine supplementation on exercise performance, skeletal muscle oxygen saturation and associated biochemical parameters in resistance trained men. J Strength Cond Res. 2011, 25:3461–3471.

HMB Increases Lean Mass Loss (+22%) + Hampers Fat Loss (-18%) from Diet + Overtraining. HED of ~6g/Day Does Yet Retain Muscle Strength & Fiber Size in Fast Twitch Muscles

Dr. Jekyll & Mr. Hyde - Is HMB have another janus-faced supplement ?
I know "mice are no little men", ... I don't have to be reminded of that. Thank you. So, if you are not interested in today's SuppVersity article, just come back tomorrow for another article without our hairy friends in it... You are still here? Ok, then let's see what the researchers at the Florida State University have to tell us about the effects of HMB supplementation on trained vs.  overtrained and energy deprived rodents. Even though, the significance of the results for mammals with less body hair such as human beings is not clear, we may still expect similar trends, yet not identical numbers on the weight, muscle and fat loss, as well as parameters that were assessed in the study at hand.

Some details on the experimental protocol

The scientists started out with a total of 61 animals that were initially divided into three groups. The animals in the groups were either sedentary, had to exercise for 1h (endurance) 3x/week or followed the same exercise protocol, yet with an addition of 0.5g/kg body of HMB in their diets (the human equivalent dose would be 6-7g/day depending on your body weight).

After a four-week run-in phase there was an additional differentiation for the non-sedentary rats which leaves us with the following four groups:
    Table 1: Ingredients of the diets in the four groups.(Parks. 2013)
  • ALT: ad libitum fed fed + exercise (1h/d for 3d/wk)
  • ALTH: same as ALT + HMB (0.5 g/kg BW/day)
  • C: 30% energy restriction + exercise (6h/d for 6d/wk)
  • CH: same as C + HMB (0.5 g/kg BW/day)
So, in essence, we are dealing with a comparison of the effects of  HMB in conjunction with exercise and an ad libitum diet (=as much as the mice wanted to eat) vs. its effects as a supplement given as an adjunct to a highly catabolic overtraining + under-eating scenario, as you know it from the Athlete's Triad Series.

Without giving away too much, I can say that the results were not what you'd probably expect

 
As you can see in figure 1 part of the results are probably not like most people would expect them to be. While the provision of HMB did work its body recompositioning magic in the rodents that could eat as much as they wanted, it compromised the fat loss and increased the total loss of lean body mass in the rodents that were fed the -30% calorically reduced diet.
Figure 1: Changes in body composition (left), strength & motor function (right). Note that the effects of HMB depend on the energy content of the baseline diet (Park. 2013)
This result is particularly surprising in view of the fact that the caloric reduction was achieved by cutting back on sucrose and corn starch, wile the protein intake remained identical, all the time (see table 1), so that the purported "muscle building effects" of HMB should have had enough available substrate to produce at least an ameliorative effect on the loss of lean mass that is a necessary consequence of undernutrition + overtraining.
Figure 2: Dr. Jeckyll & Mr. Hyde effects of HMB ingestion during a caloric deficit; gastrocnemius CSA on the left, mTOR and p-AKT levels on the right of the central illustration (based on data from Park. 2013)
That the exact opposite was the case, i.e. that the provision of supplemental HMB blocked the fat loss and increased the loss in total lean mass is strange. After all, these highly undesirable effects took place in the presence of significantly reduced levels of the "catabolic" signalling proteins MuRF-1 & atrogin-1. The actual culprit here, may yet be that the provision of supplemental HMB to the overtrained and nutritionally deprived CH group also blunted the increased activity of the pro-anabolic proteins p-AKT and mTOR  in both fast- and slow-twitch muscle fibers (with a greater effect in slow twitch fibers, thoug; see figure 2, red). This effect may in fact have negated the decrease in protein loss, of which we can only assume that it did occur in response to the downregulation of the respecting signalling proteins, atrogin-1 and MuRF-1.With less being protein being catabolized but even less being pumped back in, this would leave us with a net protein loss that would explain the loss of lean mass - specifically in the soleus muscle which contains predominatnly slow-twitch fibers.

Devils advocate: "Why are the potential downsides not mentioned in the abstract?"

In view of the previously described observations, and the undeniable negative effect of HMB in the overtraining + undereating scenario (CH group), I'd like to disagree with the researchers' imho unwarranted hypothesis that
"[...]net protein synthesis may have been greater with HMB during the catabolic condition due to its greater inhibitory effect on the protein degradation pathway" (Park. 2013)
It's a neat ad-hoc hypothesis which is yet not, as the context would suggest, "based on [the scientists'] mRNA data", of which I already told you that both AKT and mTOR are lower in the gastrocnemius and soleus (fast & slow twitch) muscle of the HMB rodents in the CH group. And, this is certainly more important it stands in contrast with the overall effects on lean mass - so if anything, this argument could be made for the gastrocnemius, only. The overall net protein synthesis on the other hand, must clearly have been lower and not as Park et al. suggest higher.

Metabolic Technologies Inc. has not just licensed the paten for HMB, they are also a sponsor of the study at hand. Nothing bad, in general, but whenever these sponsorships coincide with not mentioning potential downsides in the abstract & conclusion of a paper, it reminds me of the usefulness of some healthy skepticism.
With data only from the soleus (7% greater mass loss vs. C group) and gastrocnemius muscle (11% greater mass retention vs C group) and an overall greater loss of lean mass in the HMB supplemented over-trained + starved rodents, it is however difficult to judge what exactly the rodents lost here. It could be organ mass or muscle mass from muscles that were not acutely working during the running exercises. In the end it may not even matter, because you would obviously want to avoid both, the loss of organ and muscle mass. So, the way the researchers ignore this observation in their abstract, where they state rightly state  that "HMB improves body composition and sensorimotor function during normal training", but follow that up with an at least questionable statement about how it also "attenuates muscle mass [...] loss during catabolic conditions" (Park. 2013) is odd.

Playing advocatus diaboli (lat. for "devil's advocate), I could argue this was a concession to Metabolic Technologies, Inc. who do not just happen to have the exclusive license for the patent on HMB (learn more), but are also among the sponsors of the study.

The beneficial effects of HMB that have been observed in previous human studies never dealt with a scenario like the one in the study at hand. The training load was usually low or easily manageable and the food intake sufficient just as it was in a recent study that was conducted on elite canoeists (read more)
Bottom line: The study at hand does support the generally beneficial findings that have been reported from human studies, conducted in realistic, and not so realistic training environments. In a calorically restricted scenario, however, the seemingly fiber-type specific effects of HMB on strength are the only plus and there is little evidence for "anti-cabatolic" effects in the original, broader sense of the word.

Based on the study at hand, of which we will obviously have to see, whether its results can be replicated in human beings, it does therefore not appear advisable to use HMB on a highly restrictive diet, if you do not happen to compete in weight classes and are more concerned about a loss of strength than negative effects a highly restrictive dietary regimen could have on your body composition.

It should go without saying, though that the overtraining + undereating approach to cutting body fat is a no-go with or without HMB, so in essence, all you need to do is to avoid following this path towards misery and you don't have to care much about the relevance of the results of this study.


References:
  • Park BS, Henning PC, Grant SC, Lee WJ, Lee SR, Arjmandi BH, Kim JS. HMB attenuates muscle loss during sustained energy deficit induced by calorie restriction and endurance exercise. Metabolism. 2013 Jul 19. doi:pii: S0026-0495(13)00186-8.

The Glucose Repartitioning Effects of Exercise: Moderate Beats High Volume Training When It Comes to Shuttling Glucose Away From Fat and Right into the Muscle

"Are 2h of cardio each day still too little!?  It must be my thyroid! Yeah, that's it. It must be the thyroid!" Could be bro, but if it is probably self-inflicted hypothyrodism
In the context of my dissertations on the unwarranted vilification of insulin as a "fattening agent" (go back to "The "Pro-Insulinogenic" Effects of Non-Nutritive Sweeteners + Mechanisms & Consequences" if you have not read the article already), I presented data from a rodent study to make a point that insulin's fattening effects depend on two closely related and highly familiar factors.

One is the over-consumption of energy that is the norm, not the exception here in the Western Obesity Belt. In conjunction with the lack of glucose depleting exercise this "ensures" that the intra-muscular and hepatic liver stores of the average Westerner are always topped off and the only change to get rid of the glucose that's floating the system of the coke-guzzling convenient generation on a day-to-day basis is to pack it away in the adipose organ.

That being said, it is only logical to assume that working out would help mitigate the problem by restoring the "normal" state of partly if not fully depleted glycogen stores and allowing insulin to do its original "glycogen anabolic". It is this process of insulin induced glucose partitioning towards the emptied glycogen stores of the skeletal musculature and the influence of (a) sedentarism (control), (b) aerobic exercise worth 300kcal/day, and (c) aerobic exercise worth 600kcal/day a group of scientists from the University of Kopenhagen in Denmark (Reichenkendler. 2013) investigated in their most recent study.

300kcal or 600kcal does it even make a difference?

For the experiment on which this paper that is supposed to be published in one of the upcoming issues of the American Journal of Physiology, Endocrinology and Metabolism, the researchers recruited 27 moderately overweight men (BMI: 28.1(1.8); age: 30(6) years; no diabetic relatives, weight stable for the last 6+months), randomized them to one of the three previously mentioned conditions.

According to which group the subjects had been randomized to, the participants either continued their sedentary lifestyle (CON) or performed daily aerobic exercise of 300 kcal/day (MOD) or 600 kcal/day (HIGH) for 11 weeks. The workouts were performed
  • at a higher intensity (>70% of VO2max), three times per week, and
  • at a low-medium intensity on the other three workout day
As you can see this is more of a chronic "general activity" + exercise study, than your average acute (let's save some money) trials and thus highly relevant to the aim of investigating the effect of chronic
"[...] moderate or high dose aerobic physical exercise on insulin-stimulated glucose uptake in individual femoral muscle groups, intra- and retroperitoneal VAT, abdominal (both anterior and posterior) and femoral SAT [subcutaneous adipose tissue] in sedentary, young and moderately overweight men." (Reichenkendler. 2013)
Moreover, the use of DEXA scan and non-invasive FDG PET/CT + hyperinsulinemic, isoglycemic clamp tests by the means of which the researchers assessed the body composition and glucose uptake of the study participants before and after the 11-week intervention ensured that Reichenkendler et al. would get accurate results.
Figure 1: Change in skeletal muscle (left) and body fat (right) glucose uptake from pre- to post-interverion (Reichenkendler. 2013)
Speaking of results, in view of the fact that the participants had not been been engaged in  in regular exercise, before the study was conducted and considering the fact that their maximal oxygen consumption [VO2max] of ≤45 ml O2/kg body mass/min mirrors their sedentary lifestyles, it is actually not really surprising that the unaccustomed exercise led to decreases in abdominal subcutaneous, visceral and leg fat masses in the two intervention groups - what may be surprising, though is the fact that those were not statistically different in the medium vs. high volume group.

On the other hand, only the high, yet not the moderate volume exercise (600kcal/day) led to significant increases in fat free mass in the legs (with the difference between the moderate and high volume achieving borderline significance; p = 0.06).

1:0 for 600kcal/day and high volume training!?

The additional muscle building benefit from the high(er) volume in the study at hand appears to conflict the results of a previous study by Rosenkilde et al. On the other hand, the most important message still remains: Working out twice as much is not going to double your fat loss! If anything it will make you feel miserable. Remember that and spread the word!
The muscle building advantage of the high volume endurance exercise is interesting. After all it appears to contradict data from a previous study by Rosenkilde et al. who observed detrimental effects of doubling the exercise volume from 300kcal/day to 600kcal per day in their study from August 2012 (read more).
If we scrutinize the different protocols, this could be a result of an ostensibly small, but physiologically relevant difference in the exercise prescriptions:

While the subjects in the study at hand had the four low intensity days to recover, the Rosenkilde study did not deliberately implement intensity differences like that - the subjects were just told "you burn 600kcal/day - no matter what!" In view of the still prevalent notion that this would be most beneficial if you achieved it by "training in the zone" (the non-existent "fat burning zone"), it is not unlikely that for Rosenkilde's subjects every day ended up a "high" intensity day and the constant exercise induced stress ended up backfiring.

If we go by the body composition data this this was not necessarily the case in the study at hand.The increase in fatloss every noob believes would come out of simply doubling your cardio workouts was not present either and that despite the fact that the exercise induced total body glucose disposal rate was increased  only in the high volume group (p = 0.03). In the moderate intensity group this effect did nor reach statistical significance.
Figure 2: The overall greater GLUT-4 response to insulin (signifying improvements in insulin sensitivity) probably explains the advantage of the medium volume training (Reichenkendler. 2013)
Much contrary to the total body glucose intake, where both the liver and the adipose tissue are greatfully sucking up the glucose your muscles are not snatching from under their nose, the total skeletal muscle glucose uptake rate increased in both, the moderate (p = 0.007) and high (p = 0.002) volume groups.

And now for the real surprises...

Did you know that high intensity, muscle damaging workouts reduce the post-exercise glycogen repletion rate and could thus impair the general glucose repartioning effects of exercise? It cannot be said for sure whether an increase in muscle damage was partly to blame for the lower glucose repartitioning effects in the medium vs. high volume groups in the study at hand, but the long-lasting (10 days) defect in glycogen resynthesis Kevin P. O'Really et al. observed in their 1987 study in response to 45min of eccentric cycling should remind you that "go heavy of go home" does not imply that you go home only, when your muscles hurt so much that even going light is not possible any longer.
Much contrary to what you may expect, though, the increase in muscle specific glucose uptake was more pronounced in the moderate (difference to control p = 0.02) than in the high volume group, where only a trend (p = 0.06) was observed.

And while the total glucose uptake rate of femoral adipose tissue did not change significantly in either of the intervention groups, the amount of glucose that ended up in the abdominal subcutaneous fat stores decreased. Just like the muscle specific glucose repartitioning, this effect did yet occur only in the moderate, yet not in the high volume groups.

Similarly the glucose repartitioning away the visceral adipose tissue of the midsection was statistically significantly only for the moderate volume group. For the subjects in the high volume group, the researchers observed a non-significant tendency (p = 0.09) for a decrease in the amount of glucose that was taken up by the visceral fat depots in the abdominal region.

What do we make of these counter-intuitive results: First of all, I would like to emphasize that any form of exercise is going to have a measurable repartitioning effect and whether the latter is "statistically significant" or not may in the end be of secondary importance. Moreover, the overall greater energy expenditure in the high volume regimen resulted in greater improvements in body composition than its low volume counterpart - irrespective of its "non-significant" or "borderline significant" glucose repartitioning effects (so much about "calories don't count", and "it's all about insulin and the fattening carbohydrates" ;-)

These abs were not sculpted by 600kcal/day workouts check out some more promising routines
On the other hand, the results of the study at hand to eventually confirm what the previously cited study by Rosenkilde et al. (click on the image with the mouse) already suggested: Doing more is not necessarily beneficial. Even in the presence of what you may call "light intensity recovery days", the daily hour (on the light days you will need way more than an hour to burn 600kcal) of the ever same, non-challenging exercise is not going to cut it - in the literal sense; or, to put it differently: doing low intensity training just to burn calories everyday is not yield the fat burning, muscle building or maintaining results you are looking for.

If you are looking for better alternatives, check out the fatloss support workouts in the Step By Step Guide to Your Own Workout.

References:
  • O'Reilly KP, Warhol MJ, Fielding RA, Frontera WR, Meredith CN, Evans WJ. Eccentric exercise-induced muscle damage impairs muscle glycogen repletion. J Appl Physiol. 1987 Jul;63(1):252-6.
  • Reichkendler MH, Auerbach P, Rosenkilde M, Christensen AN, Holm S, Petersen MB, Lagerberg A, Larsson HB, Rostrup E, Mosbech TH, Sjödin A, Kjaer A, Ploug T, Hoejgaard L, Stallknecht BM. Exercise training favors increased insulin-stimulated glucose uptake in skeletal muscle in contrast to adipose tissue: A randomized study using FDG PET imaging. Am J Physiol Endocrinol Metab. 2013 Jun 25. [Epub ahead of print]
     

Evidence From the Metabolic Ward: 1.6-2.4g/kg Protein Turn Short Term Weight Loss Intervention into a Fat Loss Diet

2x-3x higher than RDA protein intakes work equally well for men and women, to get and stay lean and lose fat and build / maintain muscle.
There are very few principles I believe are set in stone and valid regardless of your age (maybe not for toddlers), your training goals and your nutritional "orientation" (paleo, low carber, low fat eater, or whatever), and among these the "Have at least 30g of quality protein (eggs, meats, dairy, fish, etc.) with every major meal" (this assumes you eat 3meals+ per day) probably is king. It is the recipe to success and I actually don't feel as if it was necessary to convince you of the advantages this high(er) protein intake will have on your physique and - although the medical establishment is still reluctant to admit that - your health, as well. Still, the most recent study from the Military Nutrition Division, U.S. Army Research Institute of Environmental Medicine in Natick, Massachusetts, USA; have much more to offer than "just" some additional evidence to the superiority of high(er) protein diets on a cut.

It's more than high time to revise the RDA

The study was designed to assess the effects of different dietary protein (RDA = 0.8g/kg, 2x RDA = 1.6g/ kg and 3x RDA =2.4g/kg) intake on body composition and postabsorptive and postprandial muscle protein synthesis on a 21-day cut (-30% energy restriction phase; ED). The latter was preceded by a 10-day weight maintenance (WM) period.

To up the calculated energy deficit to 40% the physically active (physical activity 3– 4 d/wk), weight stable ( 2 kg; for a minimum of 2 mo before the study), 39 volunteers [32 men (11 military, 21 civilians) and 7 women (7 civilians)] with a body mass index (BMI) between 22 and 29 kg/m² and a sufficient baseline fitness had to exercise daily:
"You told me to eat more protein and this burger has both meat and cheese!" - This and other mishaps are the rule, not the exception in uncontrolled dietary interventions (learn more). The fact that the study at hand took place in the metabolic ward of the U.S. Department of Agriculture Grand Forks Human Nutrition Research Center really is a HUGE PLUS.
"To isolate the effects of the diet and minimize the potential of an exercise training stimulus, physical activity during WM was prescribed at levels comparable to those reported in prestudy 7-d physical activity records. Volunteers performed low-tomoderate-intensity (40 – 60%Vo2peak) treadmill and cycle ergometry steady-state physical activity sessions daily. Intensity was based on pre-study Vo2peak
measurements obtained during a progressive intensity treadmill test and verified during
familiarization trials using indirect calorimetry (ParvoMedics) and corresponding heart rate. Workloads during steadystate physical activity sessions were adjusted accordingly to
ensure accuracy using the heart rate reserve method and portable heart rate monitors." (Pasiakos. 2013; my emphasis)
The study took place in the metabolic ward (so there was no cheating involved here => HUGE PLUS; cf. ) at the U.S. Department of Agriculture Grand Forks Human Nutrition Research Center. All volunteers were required to abstain from nutritional supplements, alcohol, smoking, and all medications, unless acetaminophen-containing products were provided by the investigator or study physician. Volunteers were also required to be in their assigned rooms with lights out by 11 P.M. (possibly very important; learn why) to ensure adequate and similar levels of sleep.

Don't worry this was not "cardio only"

To maintain prestudy muscular fitness levels, the volunteers also performed resistive-type physical activity 3d/wk. However, "to minimize the potential of an unaccustomed, anabolic stimulus influencing study outcome measures, the intensity and volume of the resistive-type exercise was low" (Pasiakos. 2013):
Table 1: Energy / macronutrient content of the diets (updated on June 21; previously there was a copy + paste error in the table)
"Specifically, volunteers performed one single-joint movement per major muscle group (3 sets of 15 repetitions) using workloads determined during the prestudy period. Frequency, intensity, mode, and volume of resistive-type activities did not change during the 31-d study. Research staff who were blinded from dietary assignment supervised all physical activity sessions for safety and accuracy". (Pasiakos. 2013)
The body weight, was recorded in two day intervals and the body composition was quantified using a  dual-energy X-ray absorptiometry (DXA) during WM (day 9) and ED (day 30). To elicit the underlying mechanisms, the resting metabolic rate, protein synthesis, nitrogen balance and the expression of intracellular signaling proteins were tested, as well.
Figure 1: Change in body composition and protein synthesis (Pasiakos. 2013)
As you can see in figure 1, there was a baseline and dose-dependent effect on the changes it total weight and body composition, respectively.
Body weight during WM was similar between dietary treatment groups and remained stable from d 1 (group mean, 77.5 1 +/-5 kg) through d 10 (77.1 1 +/-5 kg). Overall, volunteers lost 3.2 0 +/- 2 kg during the 21-d ED; 3.5 0 kg for RDA, 2.7 0 kg for 2 -RDA, and 3.3 0 kg for 3 -RDA (P < 0.05). Independent of dietary protein, percentage body fat decreased (P < 0.05) from 19.8 1% during WM to 18.1 1% during ED, and the change in percentage body fat was similar between RDA (1.3 0 +/- 3%), 2 -RDA (1.8 0 +/- 4%), and 3 -RDA (1.9 0 +/- 3%)." (Pasiakos. 2013)
What's worth taking a closer look at, is yet the proportion of total weight loss due to changes in fat mass (FM) and FFM, which differed across dietary protein levels.
  • the percentage of total weight loss attributed to reductions in fat mass (FM) was higher (P < 0.05) for 2 -RDA (70.1 7%; 1.9 0 +/- 3 kg) and 3 -RDA (63.6 5%; 1.9 0 +/- 2 kg) than for RDA (41.8 5%; 1.6 0+/-2 kg)
  • the percentage of total weight loss due to a loss of fat free mass (FFM) was lower for 2 -RDA (29.8 7%; 0.8 0 +/- 2 kg) and 3 -RDA (36.4 5%; 1.2 0. +/- 3 kg) as compared to RDA (58.2 5%; 2.3 0 +/- 3 kg)
  • the fat to lean mass loss ratio was 30% higher in the medium protein intake group, in other words, the increase in protein intake in the 3xRDA group did not protect the lean mass any better than the 1.6g/kg in the 2xRDA group
While the latter change did not reach statistical significance, the trend is clear and I suspect with a higher number of participants, the scientists would have been able to show that the 3x RDA intake is not just worthless, but actually contra-productive, if your goal is stable ongoing fat loss.

No inter-group differences in the majority of signaling proteins

All the changes took place in the absence of statistically significant inter-group differences in the changes in anabolic intracellular signaling and gene expression [ignore the following list if you are no geek ;-], i.e.
  • postprandial Akt (Ser 473) phosphorylation was increased 1.4-fold higher (P < 0.05) compared to postabsorptive levels
  • postprandial p70 S6K1 (Thr 389), eIF4E Ser (209), and rpS6 (Ser 235/236) phosphorylation status was 16, 1.9, and 15.5-fold higher (P < 0.05), respectively, compared to postabsorptive phosphorylation levels
  • phosphorylation status of eEF2 (Thr 56) was lower (P < 0.05) after feeding
The more important general observation was yet that the upregulation of these signals 3 h after consuming a protein-containing meal, demonstrated a main feeding effect for all proteins of interest (P < 0.05). On the other hand, their expression was not influenced by energy status or the level of dietary protein intake (and let's be honest, what would an increase be worth if the data in figure 1 already told us what the real-world implications are?)
Figure 2: Changes in postabsorptive muscle protein synthesis-associated mRNA expression levels during the diet phase (-40% energy intake) of the study (Pasiakos. 2013)
Additionally, the energy deficit increased the mRNA expressions of a couple of other proteins implicated in the intracellular regulation of muscle protein synthesis:
"Transcription of Vps34, a protein involved in amino acid sensing and amino acid-mediated stimulation of mammalian target of rapamycin (mTORC1) signaling, was 1.2-fold higher (P < 0.05), while expression of mTORC1 inhibitors REDD1 and REDD2 were both 1.3-fold higher (P < 0.05) after ED compared to WM. Increasing dietary protein intake increased Vps34 mRNA expression, with 1.2-fold higher levels for 3x-RDA than RDA (P 0.05). MAP4K3, LAT1, and SNAT2 mRNA levels were not influenced by energy and dietary protein manipulations." (Pasiakos. 2013)
In view f the slight advantage of the 3xRDA diet in terms of the stimulation of protein synthesis, you may want to come back to the statistical insignificance of the superiority of the 2xRDA diet to keep indulging the same hilarious amounts of protein that have probably not gotten yourself anywhere near contest shape in the past, well, let's take a look on a couple of other observations, then:
  • While the nitrogen balance remained negative (meaning the body was burning more protein than it stored) over the whole trial in the 0.8g/kg group, it returned to baseline (weight maintenance levels) first in the 1.6g/kg (=2x RDA) group (day 17!). This restoration of to pre-diet levels was observed only on day 30 in the high protein group (2.4g/kg) and the that without any significant advantage of the 3xRDA over the 2xRDA intake (if anything it was lower in the high protein group; see figure 3)
  • There was no "thermogenic advantage" - or whatever people usually like to call the purported beneficial effect that comes with the ingestion of higher amounts of protein; in fact, the resting metabolic rate was identical for all three groups over the whole 21-day diet period. 
  • With a diet that was high in carbohydrates and low in fat (see table 1), the conversion of protein to glucose, was likely relatively limited and the potential downsides of high protein + low carb diets, where most of the protein will be broken down in the liver to supply your body with glucose and any temporary increase in insulin due to fast acting protein sources were not an issue.
In the end, the increase in postprandial protein synthesis in the 3x RDA group is therefore worthless, because it went hand in hand with an increase in wastefulness due to which the absolute protein retention did not differ all that much and the differences in lean mass loss 0.1kg) are clearly insigificant- plus: If you simply do the math, the ratio of fat free to fat mass loss, is still 31% higher in the 2x RDA group.

Irrespective of how many supplements you take - you cannot out-supplement a bad diet, laziness and a lack of motivation & determination. Still, especially for the elderly HMB with it's pronounced anti-cababolic effec could help - particularly on a diet (learn more; leucine vs. HMB)
So what's the optimum then? If we reconcile the results of the study at hand, the "optimal" protein intake would thus probably be somewhere between 1.6g/kg and 2.0g/kg an thus in the <200g range for the vast majority of people. If you also consider that this value includes all protein even that from rice, and other "non-quality" protein sources, the study at hand does not confute my previous recommendation to stick to a 1.5g/kg-2.0g/kg (per total body mass) protein intake from quality protein sources, to discount the additional protein you will be getting from "low protein food" (too much counting will only make you neurotic) and to do that irrespective of whether you are bulking and or dieting  .

One thing you may want to keep in mind though, is the fact that the overall calorie deficit of ~40% may still have been a little to high - it was not enough to elicit a significant reduction in the resting metabolic rate, but still enough to induce a loss of at least 30% of lean mass. A lower caloric deficit 20-30%, a little more patience and a focus on hypertrophy-specific weight lifting are thus probably a way more significant difference, than whether you consume 1.6g/kg or 2.4g/kg body weight.

References: 
  • Pasiakos SM, Cao JJ, Margolis LM, Sauter ER, Whigham LD, McClung JP, Rood JC, Carbone JW, Combs GF Jr, Young AJ. Effects of high-protein diets on fat-free mass and muscle protein synthesis following weight loss: a randomized controlled trial. FASEB J. 2013 Jun 5. [Epub ahead of print]