Performance Research for May: Protein Synthesis: Leucine and Layne Norton

Greetings! Some very cool studies below for yas and I apologize that I don't have time to add my comments per normal, but if there are any questions post them in the comments and I will get back to you as soon as I can.

Jodie and I are headed out super early tomorrow AM for Ryan Lee's Ozworth conference on business stuff for 3 days. It should be very interesting and I am looking forward to learning as much as I can so I can better help all of you! If you are there, please come up and say hi! If I miss you, email me at my normal address with the title URGENT and my assistant will call me with your contact information.

If you have sent an email, I will get back to you soon, but it probably won't be until next week. I know I probably said that last week too, but I will get back to you at some point.

The first study I need to send a huge congrats out to Layne Norton for getting it published! If you have not checked out his website, click on it below.

Biolayne
(for some reason my computer spits at me about the link, but it worked earlier today)


I know first hand how much effort goes into publishing studies! You can listen to Layne himself and a great discussion about protein on Super Human Radio below. Excellent info and interview

Super Human Radio Show - # 273 - BREAKING NEWS - Which Protein Source Builds More Muscle
Monday, March 16, 2009 1:00 PM

From http://www.superhumanradio.com/rss/show_podcast.xml


Nutrient Physiology, Metabolism, and Nutrient-Nutrient Interactions

The Leucine Content of a Complete Meal Directs Peak Activation but Not Duration of Skeletal Muscle Protein Synthesis and Mammalian Target of Rapamycin Signaling in Rats1,2
Layne E. Norton3,*, Donald K. Layman3, Piyawan Bunpo5, Tracy G. Anthony5, Diego V. Brana4 and Peter J. Garlick3,4

J. Nutr. (April 29, 2009)


3 Division of Nutritional Sciences, Department of Food Science and Human Nutrition 4 Department of Animal Sciences University of Illinois at Urbana-Champaign, Urbana, IL 61801 5 Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Evansville, IN 47712

This study examined the impact of leucine (Leu) derived from complete meals on stimulation of skeletal muscle protein synthesis (MPS). Expt. 1 examined time course changes in translation initiation and MPS after a meal. Male rats (~300 g) were trained for 5 d to eat 3 meals/d providing 20, 50, and 30% of energy from whey protein, carbohydrates, and fats, respectively. Plasma and skeletal muscle were collected at time 0 (baseline) after 12 h of food deprivation and then at 45, 90, 135, 180, and 300 min after a 4-g meal.

Plasma Leu increased at 45 min and remained elevated through 180 min. MPS peaked at 45–90 min and returned to baseline by 180 min.

Plasma Leu correlated with phosphorylation of ribosomal protein p70 S6 kinase (r = 0.723; P < r =" 0.773;" r =" 0.608;" style="font-weight: bold;">CONCLUSION These studies demonstrate that peak activation but not duration of MPS is proportional to the Leu content of a meal.



Stimulation of muscle anabolism by resistance exercise and ingestion of leucine plus protein.

Tipton KD, Elliott TA, Ferrando AA, Aarsland AA, Wolfe RR. Metabolism Unit, Shriners Hospitals for Children, University of Texas Medical Branch, Galveston, TX 77555, USA.


Leucine is known to stimulate muscle protein synthesis and anabolism. However, evidence for the efficacy of additional leucine to enhance the response of muscle anabolism to resistance exercise and protein ingestion is unclear. Thus, we investigated the response of net muscle protein balance to ingestion of additional leucine with protein in association with resistance exercise. Two groups of untrained subjects performed an intense bout of leg resistance exercise following ingestion of 1 of 2 drinks: flavored water (PL); or 16.6 g of whey protein + 3.4 g of leucine (W+L). Arteriovenous amino acid balance across the leg was measured to assess the anabolic response of muscle in each group.


Arterial amino acid concentrations increased in response to ingestion of W+L. Amino acid concentrations peaked between 60 and 120 min after ingestion, and then declined to baseline values. Valine concentration decreased to levels significantly lower than baseline. Net balance of leucine, threonine, and phenylalanine did not change following PL ingestion, but increased and remained elevated above baseline for 90-120 min following W+L ingestion. Leucine (138 +/- 37 and -23 +/- 23 mg), phenylalanine (58 +/- 28 and -38 +/- 14 mg), and threonine (138 +/- 37 and -23 +/- 23 mg) uptake was greater for W+L than for PL over the 5.5 h following drink ingestion.


CONCLUSION: Our results indicate that the whey protein plus leucine in healthy young volunteers results in an anabolic response in muscle that is not greater than the previously reported response to whey protein alone.

My notes: Ok, I could not resist. Looks like if you are using protein high in BCAAs and leucine, that EXTRA leucine may not be beneficiail (although does not appear to be harmful, unless you count money going out of your wallet as harmful)

The balancing act between the cellular processes of protein synthesis and breakdown: exercise as a model to understand the molecular mechanisms regulating muscle mass.


Rasmussen BB, Richter EA. Univ. of Texas Medical Branch, Dept. of Physical Therapy. Div. of Rehabilitation Sciences, 301 Univ. Blvd., Galveston, TX 77555-1144. blrasmus@utmb).

No Abstract Available.

Nutritional and contractile regulation of human skeletal muscle protein synthesis and mTORC1 signaling.

Drummond MJ, Dreyer HC, Fry CS, Glynn EL, Rasmussen BB. Univ. of Texas Medical Branch, Dept. of Physical Therapy, Div. of Rehabilitation Sciences, 301 Univ. Blvd. Galveston, TX 77555-1144. blrasmus@utmb.edu).


In this review we discuss current findings in the human skeletal muscle literature describing the acute influence of nutrients (leucine-enriched essential amino acids in particular) and resistance exercise on muscle protein synthesis and mammalian target of rapamycin complex 1 (mTORC1) signaling. We show that essential amino acids and an acute bout of resistance exercise independently stimulate human skeletal muscle protein synthesis.


It also appears that ingestion of essential amino acids following resistance exercise leads to an even larger increase in the rate of muscle protein synthesis compared with the independent effects of nutrients or muscle contraction. Until recently the cellular mechanisms responsible for controlling the rate of muscle protein synthesis in humans were unknown. In this review, we highlight new studies in humans that have clearly shown the mTORC1 signaling pathway is playing an important regulatory role in controlling muscle protein synthesis in response to nutrients and/or muscle contraction.


CONCLUSION: We propose that essential amino acid ingestion shortly following a bout of resistance exercise is beneficial in promoting skeletal muscle growth and may be useful in counteracting muscle wasting in a variety of conditions such as aging, cancer cachexia, physical inactivity, and perhaps during rehabilitation following trauma or surgery.

Bustin' Broscience: Leucine (BCAAs) stimulate insulin

Bustin' Broscience: Leucine and insulin
I am going to start some short research articles here called "Bustin Broscience" Most of these will be short using newer research and while they will be far from a complete literature review I want to promote some good discussion and attempt to whittle away at some of the misinformation out there.

While I find anecdotal information interesting, it can be affected by so many things that on the evidence chain I would have it at the bottom. There is a reason we have placebo controlled trials/experiments and blinded subjects and researchers.

Thanks to Joel for the image on the right. Can anyone alter it to put a big red line through it?

If you are looking for much more depth, I would highly recommend Alan Aragon's Research Review. It is only 10 clams a month and I make zero money from recommending it, but he does his homework on all of the topics he covers each month.

Click below for more information

Alan Aragon's Research Review

Bustin' Broscience
What the heck is broscience? Here is the definition from the Urban Dictionary. Not sure where the term started and Alan Aragon was the first I read who had used it.

Broscience is the predominant brand of reasoning in bodybuilding circles where the anecdotal reports of jacked dudes are considered more credible than scientific research.

Broscience in action: "Bro, you gotta slam 40-60 grams of waxy maize plus 20 grams of BCAA within 7 seconds of finishing your last set of squat rack curls. Otherwise, you'll go straight catabolic."

Broscience says "Leucine stimulates insulin"

Is this one about to busted?

Leucine
I am predicting some interesting data on the use of the amino acid leucine coming soon. We know that acutely it stimulates the mTOR receptor which is one of the keys to protein synthesis (build more muscle). Long term data, however, is not as plentiful (although I hear there are several cool studies being conducted now.)

Insulin
While a whole review of the actions of insulin would be volumes, the very very short take away is that insulin is one of the key anabolic hormones in the body. I am using the strict definition of anabolic from the term anaolism.

a·nab·o·lism (-nb-lzm)
n.
The phase of metabolism in which simple substances are synthesized into the complex materials of living tissue.

It is just the fancy term for putting stuff together in the body. Most bodybuilders use this term to exclusively talk about taking amino acids and stuffing them into muscle proteins (which I am all for of course since that will make larger muscles), but it can also apply to taking free fatty acids (FFAs) from the blood and putting them in fat (adipose) tissue.

So insulin is the main anabolic hormone and the basic theory is that more insulin will increase the process of stuffing more fats into adipose tissue.

Do you get get fat by using leucine?

Here is an interesting study where they added leucine to the drinking water of our fury little friends. Let's see if there is an effects on fatness (obesity) and insulin?

Leucine Supplementation of Drinking Water Does Not Alter Susceptibility to Diet-Induced Obesity in Mice,

They stated, "Branched-chain amino acids (BCAA), Leu, and the signaling pathways they regulate have been reported to either improve or worsen adiposity and insulin sensitivity."

They added leucine or BCAAs to the rat's water and fed them lots of food and measured metabolic rate and insulin tolerance.

They found that, "Compared with control, supplementation did not affect body weight, food intake, oxygen consumption, locomotor activity, body composition, insulin tolerance, or total cholesterol."

While this study is limited and done in rats, that is strike one

What else?

Here is another study

Leucine, when ingested with glucose, synergistically stimulates insulin secretion and lowers blood glucose

The goal of this study was to determine whether leucine stimulates insulin and/or glucagon secretion and whether, when it is ingested with glucose, it modifies the glucose, insulin, or glucagon response.

They used 13 human subjects, which is good as we are not rats; but those little fury guys are much easier to do highly controlled studies on though. The arrived fasted and got some leucine and 25 g of glucose wile they measured serum leucine, glucose, insulin, glucagon, and α-amino nitrogen concentrations over a 2.5-hour period after ingestion of the test meal. The amount of leucine provided was equivalent to that present in a high-protein meal, that is, that approximately present in a 350-g steak.

Leucine concentrations
Leucine did appear in the blood and acutally REDUCED the 2.5-hour glucose area response by 50%. Leucine, when ingested ALONE, increased the serum insulin area response modestly but glucagon (the antangonist to insulin) also changed.

Think of glucagon as one of the "anti-insulin" hormones and leucine actually increased glucagon!

Shut up and get to the point
2 fold on this study

1) Leucine at a dose equivalent to that present in a high-protein meal, had little effect on serum glucose or insulin concentrations

2) When leucine was ingested with glucose, it attenuated the serum glucose response and strongly stimulated additional insulin secretion.

In English please
This shows there is some data that leucine when combined glucose, may increase insulin levels but other hormonal levels change too.

Personal note
If you have ever looked at insulin data (I am currently doing this for a study), it varies much more than you would imagine. Your body is CONSTANTLY adjusting insulin and the "anti-insulin" hormones by the second.

History lesson
As far as I could tell, this below is the first one I can find discussiong leucine and insulin stimulation, published in AJP in 1983.

L-leucine methyl ester stimulates insulin secretion and islet glutamate dehydrogenase

The huge downside of this one is that it used perifusion of collagenase-isolated mouse pancreatic islets. In other words, isolated cells in fluid. While ths is a a great START, many many times these data do not translate to whole human (in-vivo) studies.

The final word
I give this one a (drum roll please................)
Plausible

It may happen, but I am not convinced that adding a few grams of leucine or BCAAs to your nutrition is going to dramatically affect insulin levels. Again, this is a supplement, so get your nutrition in line with real food first before you decide to add any fancy supplements.

Comments on this issue of Bustin' Broscience.
Let me know if you this is useful and any feedback you have!
Rock on
Mike T Nelson


REFERENCES

Leucine Supplementation of Drinking Water Does Not Alter Susceptibility to Diet-Induced Obesity in Mice,

Ali Nairizi, Pengxiang She, Thomas C. Vary and Christopher J. Lynch., Penn State Hershey Institute for Diabetes and Obesity and the Department of Cellular and Molecular Physiology, Pennsylvania State University College of Medicine, Hershey, PA 17033

Branched-chain amino acids (BCAA), Leu, and the signaling pathways they regulate have been reported to either improve or worsen adiposity and insulin sensitivity. Therefore, it is unclear whether dietary supplementation of Leu would be beneficial. To help address this question, we examined the effect of adding Leu (150 mmol/L; Expt. 1 and Expt. 2) or BCAA (109 mmol/L of each; Expt. 3) to the drinking water on diet-induced obesity (induced with a 60-kJ% fat diet) in singly housed C57BL6/J male mice for at least 14 wk. Liquid and solid food intakes were evaluated weekly along with body weight. During the last few weeks, several blood samples were taken at different times for plasma glucose, total cholesterol, or Leu measurements.

Metabolic rate by indirect calorimetry, locomotor activity by light beam breaking, body composition by H1-NMR, and insulin tolerance were also determined. Compared with control, supplementation did not affect body weight, food intake, oxygen consumption, locomotor activity, body composition, insulin tolerance, or total cholesterol. In fed mice, this method of Leu supplementation only increased plasma Leu by 76% when the supplemented group was compared with control. On the other hand, after overnight food deprivation, the plasma Leu did not differ between these 2 groups, even though the mice in the supplemented group had continuous access to Leu-containing water during the solid food deprivation.


CONCLUSION: Taken together, the results do not provide evidence that either Leu or Branched-chain amino acids supplementation of drinking water ameliorates diet-induced obesity in mice, although it may improve glycemia.

Leucine, when ingested with glucose, synergistically stimulates insulin secretion and lowers blood glucose

Auteur(s) / Author(s)
KALOGEROPOULOU Dionysia (1 2) ; LAFAVE Laura ; SCHWEIM Kelly ; GANNON Mary C. (1 2 3) ; NUTTALL Frank Q. (1 2) ;

(1) Endocrine, Metabolism and Nutrition Section, VA Medical Center, Minneapolis, MN 55417, ETATS-UNIS
(2) Department of Medicine, University of Minnesota, Minneapolis, MN 55455, ETATS-UNIS
(3) Department of Food Science & Nutrition, University of Minnesota, St. Paul, MN 55108, ETATS-UNIS
Résumé / Abstract
Our laboratory is interested in the metabolic effects of ingested proteins. As part of this research, we currently are investigating the metabolic effects of ingested individual amino acids. The objective of the current study was to determine whether leucine stimulates insulin and/or glucagon secretion and whether, when it is ingested with glucose, it modifies the glucose, insulin, or glucagon response. Thirteen healthy subjects (6 men and 7 women) were studied on 4 different occasions. Subjects were admitted to the special diagnostic and treatment unit after a 12-hour fast. They received test meals at 8:00 AM. On the first occasion, they received water only. Thereafter, they received 25 g glucose or 1 mmol/kg lean body mass leucine or 1 mmol/kg lean body mass leucine plus 25 g glucose in random order. Serum leucine, glucose, insulin, glucagon, and α-amino nitrogen concentrations were measured at various times during a 2.5-hour period after ingestion of the test meal. The amount of leucine provided was equivalent to that present in a high-protein meal, that is, that approximately present in a 350-g steak. After leucine ingestion, the leucine concentration increased 7-fold; and the α-amino nitrogen concentration increased by 16%. Ingested leucine did not affect the serum glucose concentration. When leucine was ingested with glucose, it reduced the 2.5-hour glucose area response by 50%. Leucine, when ingested alone, increased the serum insulin area response modestly. However, it increased the insulin area response to glucose by an additional 66%; that is, it almost doubled the response. Ingested leucine stimulated an increase in glucagon. Ingested glucose decreased it. When ingested together, the net effect was essentially no change in glucagon area. In summary, leucine at a dose equivalent to that present in a high-protein meal, had little effect on serum glucose or insulin concentrations but did increase the glucagon concentration. When leucine was ingested with glucose, it attenuated the serum glucose response and strongly stimulated additional insulin secretion. Leucine also attenuated the decrease in glucagon expected when glucose alone is ingested. The data suggest that a rise in glucose concentration is necessary for leucine to stimulate significant insulin secretion. This in turn reduces the glucose response to ingested glucose.


L-leucine methyl ester stimulates insulin secretion and islet glutamate dehydrogenase

AJP - Endocrinology and Metabolism, Vol 245, Issue 4 338-E346, Copyright © 1983 by American Physiological Society

P. Knudsen, H. Kofod, A. Lernmark and C. J. Hedeskov

Column perifusion of collagenase-isolated mouse pancreatic islets was used to study the dynamics of insulin release in experiments lasting for several hours. The methyl esters of L-leucine and L-arginine were synthesized. Whereas L-arginine methyl ester (L-arginine OMe) had no effect, L-leucine OMe stimulated the release of insulin. The effect of L-leucine OMe was maximal at 5 mmol/liter. Whereas the Km for glucose-stimulated insulin release was unaffected by 1 mmol/liter L-leucine OMe, the maximal release of D-glucose was increased by the amino acid derivative that appeared more effective than L-leucine. L-Leucine OMe was also a potent stimulus of insulin release from the perfused mouse pancreas. In the presence of 10 mmol/liter L-glutamine, 1 mmol/liter L-leucine OMe induced a 50- to 75-fold increase in insulin release. A similar stimulatory effect was also observed in column-perifused RIN 5F cells, a cloned rat islet tumor cell line. A twofold increase in islet glutamate dehydrogenase activity was induced by 5 mmol/liter L-leucine OMe, a larger effect than that of L-leucine (P less than 0.02), whereas L-arginine OMe had a small inhibitory effect. We conclude that L-leucine OMe is a potent stimulus of insulin secretion and that its effect on the beta-cells may be exerted by activating islet glutamate dehydrogenase.

Leucine Supplementation: New research soon!

Donate to a Great Cause
Alright, I very rarely ever put things like this up on my blog, but my good buddy Phil Stevens is having a raffle for a great cause. See the info below from him. I already made my cash donation and you may even win some great prizes too.

In addition, I will be donating some phone consultations for a few lucky winners too.

Click here to donate!

Phil says,"Check out the above link. I started a nationwide raffle for the event. Please take a look and spread the word to everyone you know. Every little bit helps no matter how small and there are some great prizes to be won. At the worst you will have given money to a great cause and great event.

Along with the event it self starting next week Ill be going to several functions and working directly with the 2000+ kids spreading the word of athletics in general as well as strength sports and nutrition.

For more info check out the event home page: www.lift4hope.org and RVSP at the facebook group

Again every little bit helps no matter how small.
Thank you
Phil Stevens,

Dave Barr Announcement
I have on my "to do list" to clean up a couple of the blog links on the right, and if you went to Dave Barr's link you will notice that it does not go anywhere.

Well, Dave and had a long drawn out fight about leucine and if weighted facial stretching was a good idea and it really came to an ugly ending. Ha--I kid, I kid. Dave and I are fine and he actually got a job at Muscle and Fitness as a Senior Science Editor, so his other site was closed down for now. I truly wish him all the best and I am very happy for his new position as he is a very talented guy. Be on the look out for a new book form him related to "Nutrition for the Nervous System" (not sure of the exact title) and I will have a review up as soon as I can get my grubby paws on it. Congrats Dave!

Speaking of lecine....

Brand New Study on Leucine!

Just wanted to give you a heads up to look for the following study very soon as it has been accepted for publication, but it is not out yet. Van Loon's lab has been cranking out lots of interesting studies in regards to protein synthesis over the past several years.

S Verhoeven, K Vanschoonbeek, LB Verdijk, R Koopman, WKWH Wodzig, P Dendale, and LJC van Loon

Long-term leucine supplementation does not increase muscle mass or strength in healthy elderly men.

As the astute readers of this blog know, protein synthesis (building muscle) response in older folks is not the same as younger people; so the jury is still out a bit on that part.

In the meantime, check out the following by clicking on each one.

BCAA Supplementation Roundtable

By Jamie Hale

And other posts

Research Review: Do You Only Need 20 Grams of Protein Post Workout?

Performance Research for February: Protein Synthesis and Exercise Round 1

Charles Staley Seminar: Dave Barr and Supplements