Science Round-Up Seconds: 8 Nootropics to Combat Stroke, Alheimer's & Co and Boost Cognitive Performance. Plus: 7 Rarely Thought of Side Effects of High Dose Glutamine.

Effects of infusion times on phenol content of black tea (Ramalho. 2012)

If you have already listened to the podcast of yesterday's Science Round-Up on the Super Human Radio Website (click here if you haven't and wan't to know what the following is all about), I suppose you will not mind that I compiled some of the complex information about "optimal" tea brewing in the illustration to the right (based on Ramalho. 2012). The colored arrows indicate the time-points at which the given compounds in the tea achieved peak values. The exact time point is also given in minutes, so that a 9' in front of the green caffeine and on the left to the green arrow pointing at the 9 min point tells you "it took 9 minutes for the caffeine content to reach it's maximum in the British tea". The graph in the background shows the catechin concentration depending on the infusion time.

Cholinergenic nootropics - What a recent review says

I guess some of you will probably have heard about piracetam or lecithine as purported enhancers of cognitive function. According to a recent review in the Journal of Experimental Pharmacology those two are yet not the most prosing agents:

Eggs are rich in choline which is an essential nutrient and was abundant in the classic BB diets (rear more)

• Piracetam: no cerebroprotective effects in patients who have open heart surgery, but does help on non-open cardiopulmonary bypass surgery (Holinski. 2008), beneficial effects in response to cerbrovascular and cognitive disorders traumatic origin (Malykh. 2010), intravenous piracetam can prevent cognitive deficits in response to anesthesia (Fesenko. 2009)

• Lecitin: does not improve cognitive deficits in patients (Amenta. 2011; Parnetti. 2007)

More promising "nootropics" - specifically in view of what most people do actually expect, when they buy such products.

• Oxiracetam: improves cognitive performance except for patients with dementia (Malykh. 2010)

• Citocoline: general neuroprotective effects (Alvares-Sabin. 2011), improvements in cognitive performance in healthy and patients and patients with dementia (Secades. 2010), helps with cognitive dysfunction in Parkinson's (Vale. 2008), helps with cognitive function in dementia of neurodegenerative and vascular origin (Parnetti. 2007), prevents cognitive decline after a stroke (Alvarez. 2011), improves recovery after stroke (Garcia-Cobos. 2010) 

• Cerebrolysine: produces signifant cognitive improvements in vascular dementia (Guekht. 2011), effective for both cognitive function and behavioral symptoms in Alzheimer's (Alvarez. 2011), promising results in patients with Alzheimer's (Plosker. 2009)

And a couple of things you would not usually associate with nootropics:

• 

  Suggested read: Amino Acids for Super Humans on the effects and differences between the various forms of carnitine (read more).

  Acetyl-L-carnitine: improves cognitive performance in patients with encephalopathy, decreases anxiety and increases general energy and wellness, as well as fatigue and age-related cognitive deficits (Malaguernera. 2008, Liu. 2008),can reduce or block neuronal death in neurodegenerative diseases (Manusco. 2007), helps ammeliorate hyperammonemia (Cagnon. 2007)

• Saffron extract: beneficial effects in mild to modest Alzheimer's  (Akhondzadeh. 2010)

• DHA (fish oil): positive effects on verbal recognition memory in old subjects (Yurko-Mauro. 2010)

Interestingly, the most profound effects appear to be brought about by acetyl-l-carnitine. In that it's worth mentioning that the benefits could still be related to cholinergic mechanisms, since it has long been known that ALCAR can increase the expression of choline acetyltransferase activity in the central nervous system (Taglialatela. 1994). And the latter is, as the name implies, necessary to form the neurotransmitter acetylcholine .

Glutamine probably not suitable for chronic high dose supplementation

Czech scientists warn about the risks of chronic high dose glutamine supplementation. I know that many of you are still too bamboozeled by the "protein for everything and let the liver take care of any glucose demands I may have" theory, of which you could probably argue that it is the bastard child of the standard BB diet with low carb. Maybe the following recently published paper by a scientist from the Charles University in Prague can help cure this "disease" (and your cognitive problems, fatigue and brainfog).

According to Holecek, the chronic ingestion of glutamine / glutamine enriched diets in can lead to...

Figure 1: In the presence of high amounts of glutamine outside of the cell, the glutamine synthesis (GLN) and with it the ammonia detoxification from muscle tissue sucks (Holecek. 2012).

"(1) Alterations in amino acid transport-as GLN shares the transporters with other amino acids, enhanced GLN intake may impair amino acid distribution among tissues and their absorption in the gut and kidneys.

(2) Alterations in GLN metabolism-GLN supplementation may impair synthesis of endogenous GLN and enhance glutamate and ammonia production.

(3) Alterations in ammonia transport-GLN supplementation may impair ammonia detoxification and negatively affect the role of GLN as the carrier of ammonia among tissues.

(4) Abnormalities in aminoacidemia-increased plasma levels of GLN, glutamate, citrulline, ornithine, arginine, and histidine and decreased levels of valine, leucine, isoleucine, glycine, threonine, serine, and proline are reported.

(5) Alterations in immune system-as GLN has immunomodulating properties, the effect of chronic GLN consumption on the immune system needs to be assessed.

(6) Effect on tumor growth-it should be elucidated whether chronic intake of GLN increases the risk of cancer.

(7) Effect of the withdrawal of GLN supplementation-due to the adaptive response of the organism to enhanced GLN consumption, the withdrawal of GLN may enhance the risk of health problems resulting from GLN deficiency." (Holecek. 2012)

Remember the post on the ammonia induced peripheral and central fatigue with high dose chronic BCAAs supplementation?

In view of the fact that some people consumer up to 40g of glutamine regularly, Holecek demands that "long-term studies should be performed" to test the side effects and evaluate whether there is any benefit at all to justify chronic consumption of a GLN-enriched diet.

So, relying on glutamine instead of carbs, as smart as this idea appears to be in the current carbophobia, could actually make you stupid due to the disruption of the intracellular ammonia detoxification, which is not a problem in muscle only, but also in the brain.

In the end, what we are seeing here is just another instance of a disruption in the natural balance of things. Ornithine, citrulline and arginine, for example are involved in the detoxification of ammonia via the urea cycle. They are however not the only bottleneck to the system.

Obviously your liver and kidneys will have to handle the clearance. People with liver problems (or persons taking "supplements" or NSAIDs that may impair the liver function) are therefore particularly prone to hyperammonemic encephalopathy (Kanamori. 1996; Lemberg. 2009)

Bottom line: Glutamine, just like everything else, in moderation and by no means so much that your body runs on glutamine as fuel. Aside from the mentioned amino acids that help the clearance of ammonia from the blood stream, taurine appears to exert a direct protective affect in the brain (Chepkova. 2006), and lactulose (a fermentable carbohydrate) can reduce the ammonia influx from ammonia producing bacteria in the gut (Vince. 1980). So if you want to wear a helmet when you bang your head against the wall, these would be suggested "take supplement B in order to counter the side effects of supplement A" - side effects of a supplement you would not even have to take, by the way (100% bro-logic ;-)

References: 

• Amenta F, Carotenuto A, Fasanaro G, Lanari A, Rea R, Traini E. Preliminary results of Ascomalva trial on the association of donepezil and choline alphoscerate in Alzheimer’s disease with associated cere-brovascular injury. G Gerontol. 2011;59:89–9.
• Akhondzadeh S, Shaf iee Sabet M, Harirchian MH, Togha M. A 22-week, multicenter, randomized, double-blind controlled trial of Crocus sativusin the treatment of mild-to-moderate Alzheimer’s disease. Psychopharmacology (Berl). 2010;207:637–643.
• Alvarez XA, Cacabelos R, Sampedro C, et al. Efficacy and safety of cerebrolysin in moderate to moderately severe Alzheimer’s disease: results of a randomized, double-blind, controlled trial investigating three dosages of cerebrolysin. Eur J Neurol. 2011;18: 59–68.
• Alvarez-Sabín J, Román GC. Citicoline in vascular cognitive impair-ment and vascular dementia after stroke. Stroke. 2011;42(Suppl 1): S40–S43.
• Cagnon L, Braissant O. Hyperammonemia-induced toxicity for the devel-oping central nervous system. Brain Res Rev. 2007;56:183–197.
• Chepkova AN, Sergeeva OA, Haas HL. Taurine rescues hippocampal long-term potentiation from ammonia-induced impairment. Neurobiol Dis. 2006 Sep;23(3):512-21.
• Fesenko UA. Piracetam improves children’s memory after general anaesthesia. Anestezjol Intens Ter. 2009;41:16–21. Polish
• García-Cobos R, Frank-García A, Gutiérrez-Fernández M, Díez-Tejedor E. Citicoline, use in cognitive decline: vascular and degenerative. J Neurol Sci. 2010;299:188–192.
• Guekht AB, Moessler H, Novak PH, Gusev EI; Cerebrolysin Investigators. Cerebrolysin in vascular dementia: improvement of clinical outcome in a randomized, double-blind, placebo-controlled multicenter trial. J Stroke Cerebrovasc Dis. 2011;20:310–318. 
• Holecek M. Side Effects of Long-term Glutamine Supplementation. JPEN J Parenter Enteral Nutr. 2012 Sep 18.
• Holinski S, Claus B, Alaaraj N, et al. Cerebroprotective effect of piracetam in patients undergoing coronary bypass surgery. Med Sci Monit. 2008;14:153–15.
• Kanamori K, Ross BD, Chung JC, Kuo EL. Severity of hyperammonemic encephalopathy correlates with brain ammonia level and saturation of glutamine synthetase in vivo. J Neurochem. 1996 Oct;67(4):1584-94.
• Lemberg A, Fernández MA. Hepatic encephalopathy, ammonia, glutamate, glutamine and oxidative stress. Ann Hepatol. 2009 Apr-Jun;8(2):95-102.
• Liu J. The effects and mechanisms of mitochondrial nutrient alpha-lipoic acid on improving age-associated mitochondrial and cognitive dysfunction: an overview. Neurochem Res. 2008;33:194–203.
• Mancuso C, Bates TE, Butterfield DA, et al. Natural antioxidants in Alzheimer’s disease. Expert Opin Investig Drugs. 2007;16:1921–1931.
• Malaguarnera M, Gargante MP, Cristaldi E, et al. Acetyl L-carnitine (ALC) treatment in elderly patients with fatigue. Arch Gerontol Geriatr. 2008;46:181–19
• Malaguarnera M, Gargante MP, Cristaldi E, et al. Acetyl-L-carnitine treatment in minimal hepatic encephalopathy. Dig Dis Sci. 2008;53: 3018–3025
• Malykh AG, Sadaie MR. Piracetam and piracetam-like drugs: from basic science to novel clinical applications to CNS disorders. Drugs. 2010;70:287–31
• Pantoni L. Treatment of vascular dementia: evidence from trials with non-cholinergic drugs. J Neurol Sci. 2004;226:67–70
• Parnetti L, Mignini F, Tomassoni D, Traini E, Amenta F.  Cholinergic precursors in the treatment of cognitive impairment of vascular origin: ineffective approaches or need for re-evaluation? J Neurol Sci. 2007;257:264–269.
• Ramalho SA, Nigam N, Oliveira GB, Alves de Oliveira P, Matos Silva TO, Passos dos Santos AG, Narain N. Effect of infusion time on phenolic compounds and caffeine content in black tea  Food Research International; 13 December 2012 [ahead of print]
• Secades JJ. Citicoline: pharmacological and clinical review. Rev Neurol. 2010;52 Suppl 2:S1–S62.
• Vale S. Current management of the cognitive dysfunction in Parkinson’s disease: how far have we come? Exp Biol Med (Maywood). 2008;233:941–951.
• Vince AJ, Burridge SM. Ammonia production by intestinal bacteria: the effects of lactose, lactulose and glucose. J Med Microbiol. 1980 May;13(2):177-91.
• Yurko-Mauro K. Cognitive and cardiovascular benefits of docosahexaenoic acid in aging and cognitive decline. Curr Alzheimer Res. 2010;7:190–196.

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).

Magic Numbers: 1g Protein per 2g Carbs + Circuit Training = The #1 Formula for Weight & Fat Loss in Obese Women?

If there is one thing about this study that's not debatable it is that eating whole foods, cleaning your diet from all sorts of junk and working out lifting weights and doing aerobics were the cornerstones of the weight loss success of these women, regardless of whether they consumed a low, medium or high amount of protein.

Roughly two years ago, when the SuppVersity opened its doors, it was pretty rare to find a scientist who would be willing to "waste" (that's probably how he or she would have said it ;-) precious time and the limited funds of his institution to study the effects of "high protein diets". Over the past couple of months, things have been changing, though: I've just checked and according to Pubmed, the number of publications containing the exact phrase "high protein diet", alone,  has increased by ~32% in 2011 and has remained on the same comparably high level ever since. That said a recent study from the University of Guelph in Canada is only the latest in the line of a whole host of publications that deal with the beneficial effects of high(er) protein diets on weight loss in overweight, (pre-)diabetic subjects; exactly those people who have previously been advised to stay clear of all fats, ignore the proteins and focus on the "healthy and satieting" low GI carbs, by the way.

Yet though the tides may be turning ...

... a paradigm shift within the scientific community usually doesn't come over night - a famous scientist and philosopher of science once said that it usually takes until the proponents of the old paradigm died out, before a new one is fully established. Since roughly two years and even two decades are hardly enough for this to happen, it is actually not surprising that Dawn. D. Campbell and Kelly A. Meckling, despite giving the high protein diet credit for having produced some promising results in the past, speculate that

"the combined effects of a normal protein: carbohydrate ratio with cardiovascular and resistance training would be more beneficial and easier to comply with than either the low- or high-protein diets in this target population of women with risk factors for the MetS" (Campbell. 2012)

In view of the fact that Rehm et al. conclude ther 2008 review of the literature with the statement...

"Diets moderately increased in protein and modestly restricted in carbohydrate and fat, particularly saturated fat, may have beneficial effects on body weight, body composition, and associated metabolic parameters." (Rehm. 2008)

... and against the background that the evidence of the real-world benefits of a higher protein intake is accumulating, and pertinent reviews and editorials have been appearing on a monthly basis, ever since (e.g. Hession. 2009; Keller. 2011; Acheson. 2012), the research hypothesis of the study at hand sounds a bit 'last year', not to say 'last decade' to me.

Do we have a bias here?

Moroever, with the research hypothesis being a good indicator of a built-in bias, we will have to pay pretty close attention to distinguish the actual data Campbell and Meckling measured from their interpretations of the latter. After all, every "good" SuppVersity student should remember that we have seen time and again how the differences between facts and interpretations often become somewhat blurry in the conclusions of way too many (for my liking) papers as of late. So let's see if Campbell's and Meckling's conclusion that...

"A diet with a 1:2 protein:carbohydrate ratio promoted better improvements than either the LP or HP diets, and may be superior in reducing long-term chronic disease risk in this population." (my emphasis in Campbell. 2012)

What can be said right away is that the protocol the scientists used, specifically the way they are working with macronutrient ratios instead of paradigmatic percentages of the RDA, is actually pretty progressive.

"Subjects were encouraged to consume whole foods as opposed to pre-packaged or processed foods and to restrict intake of whole-fat dairy, high-fat red meats, deep-fried foods, potato chips, cookies and refined sugar products. Instead, subjects were encouraged to choose whole-grain pro-ducts, lower-fat meats, fish, turkey, eggs, low-fat milk and cot-tage cheese, nuts, seeds, and a variety of vegetables, fruits and berries. Before beginning the study workout programme, subjects completed baseline fitness testing to assess muscular strength and cardiovascular fitness." (Campbell. 2012)

Instead of simply upping the protein intake from the 0.8g/kg body weight the RDA suggest would be optimal, Campbell and Meckling put their obese (mean BMI ~35kg/m²) female participants (of which only 54 completed the study) on calorically restricted diets (supposedly -30% below their habitual energy intake which had been evaluated by the means of a 7-day food record) which contained an equal relative amount of fat (<30%), but had different protein-to-carbohydrate ratios:

• low protein (LP) - 1g protein : 4g carbohydrates
• medium protein (MP) - 1g protein : 2g carbohydrates
• high protein (HP) - 1g protein : 1g carbohydrates

To put that in perspectve, a women who may have been consuming a baseline diet containing 2,300kcal per day would have had to restrict her caloric intake to 1,610kcal. Of these 1,610kcal, <30% would come from fat (60g), while the rest would be ingested in the form either 56g protein and 225g carbohydrates (LP), 94g protein and 188g carbohydrates (MP), or 140g protein and 140g carbs (HP).

There is no effective weight loss without exercise and a whole food diet!

In addition to the dietary regimen the 117 participants who initially met the eligibility criteria were supposed to particpate in a  supervised 12-week circuit training program at the University of Guelph Athletic Centre.

The 1 h study fitness programme was completed three times/week on Mondays, Wednesdays and Fridays at a consistent time assigned to each subject. Subjects had to sign in for their workout sessions, and all exercises were supervised by a study coordinator and/or personal trainer. Subjects began their workout with a 9 min warm-up using springboard pads where walking in place, jogging or dancing took place. Then, subjects completed a 30 min circuit alternat-ing between resistance training and cardiovascular exercise bouts. All main muscle groups of the body were targeted throughout the thirteen resistance training machines. Starting weight values on resistance training equipment were 65 % of their calculated maximum strength as determined by their modified 1 repetition maximum. Subjects were instructed to complete one set of eight to fifteen repetitions on each piece of equipment to reach muscle fatigue."

The circuit training used a build-in progression with ~5% increases in weight, whenever the subject were able to complete 15 repetitions on a given exercise. The same goes for the aerobic parts of the workouts, where the

Subjects began exercising at 65 % of their maximum heart rate for the first 3 weeks and gradually increased the intensity by 5 % every 3 weeks to a maximum intensity of 80 % by week 12. 

As far as the aerobic part of the workouts was concerned, they alternated between a step, springboard pad and stationary bike. All workouts closed with some ab training (including a standard crunch, oblique crunch and a core-strengthening exercise called the plank, done to failure) and stretching.

"Hey, exercise is good for me!"

Other than you may have expected the 35 dropouts (which were equally distributed across all dietary groups) were not brought about by laziness or the unwillingness to get up and move. On the contrary, many subjects recorded that they had "more energy and felt better than before the study began" (Campbell. 2012). Aside from minor constipation (the scientists don't mention in which group this occured) and some minor shedding in one of the subjects in the low protein group (probably coincidence, by the way), the intervention went fine for those who had the guts, time and discipline to stick it and yielded - as the data in figure 1 goes to show - favorable results in all three arms off the study:

Figure 1: Changes in antroprometric data, blood pressure and heart rate after 12 weeks (based on Campbell. 2012)

If we were stupid enough to focus solely on the BMI reductions, we could even say that all diets were equally effective. Upon closer scrutiny and the use of some statistical shenanigan, it does however become clear that the scientists' initial hypothesis that the normal protein diet with a 1:2 protein to carbohydrate would have a small edge over both, the low protein diet (in terms of body fat loss and lean mass retention; p < 0.05) and the high protein diet (solely in terms of body fat loss; p < 0.05) seems to hold true. What's more, this trend in DXA measured improvements in body composition stands in line with noteworthy reductions in waist circumference (7.9, 11.6 and 8.6 cm in the LP, NP and HP), of which Cambell and Meckling write:

"Again, the decrease in the NP group was greater than that in the LP group. Further-more, hip circumferences decreased similarly (P < 0·05) in response to each diet with reductions of 7·4, 8·8 and 8·4 cm in the LP, NP and HP groups, respectively. Waist:hip ratios declined significantly (P < 0·05) after 12 weeks by 0·01, 0·04 and 0·01 in the LP, NP and HP groups, respectively, but reductions were greater in the NP v. LP (P=0·020) and HP (P=0·025) groups." (Cambell. 2012)

No group specific diet effects were observed for the reductions in blood pressure and heart rate. Now, this obviously raises the question, whether the existent changes may have been brought about by non-compliance.

"So maybe the protein eaters just didn't eat their protein?"

Non-compliance is, as SuppVersity students know, one of the major problems with all of these relatively uncontrolled dietary interventions (see "High Carb vs. High Fat: What Really Happens When Science Meets the Real World"). And in fact, with average caloric intakes of 3641, 3729 and 3633 kJ/d  in in the low, medium and high protein groups, the subjects were actually consuming 10% less energy than they were supposed to.

Suggested read for everyone who can't or doesn't want to believe that you can easily eat 157g of carbs (which is what the women in the normal protein group did) and still lose fat while retaining all your precious lean muscle mass: "Carbohydrate Shortage in Paleo Land: New Data for A Scientific Outlook at the Low-to-No Carb Paleo Confusion. Will More Than 125g of Carbs Make You Fat?" (read more)

Despite the fact that this type of non- or rather 'over-compliance' can also have detrimental effects on someone's weight loss efforts, there were no intergroup differences which would skew the ultimate comparison; and much to my surprise the majority of the subjects did even manage to come close to their macronutrient goals by adapting their previously almost identical protein to carbohydrate ratios of  1:3.5, 1:3.2 and 1:32 at baseline to 1:3.5, 1:2.1 and 1:1.3 by week 12. With respect to the total protein intake, this equals

• significant decreases in protein intake (82 and 88 g/d to 55 and 75 g/d) for the low and normal protein groups and
• significant increases in protein intake (from 84 to 100 g/d) in the high protein group

This does also mean that the percentage of subjects who met the 0.8g/kg RDA for dietary protein intake at the beginning of the study had dropped to zero in the low protein group by week 12.

The subjects in the normal protein intake group were about as close as you can get and those in the high protein group consumed significantly more protein than the 'well-meaning' authors of the dietary recommendations feel would be good for them ;-) Other changes the scientists observed were:

• a significant declines in carbohydrate intake in the normal and high protein group
• a decreased sugar intake in all groups (most pronounced in the HP group)
• a decreased fat intake in all groups (p<0.001)
• significant decreases in sodium intake in all groups 
• non-significant decreases in calcium, zinc and vitamin D intake

I guess, I don't have to tell you that none of the few existing inter-group differences discussed above appears to provide any reason to question the small, but statistically significant superiority of the normal protein diets compared to either the low protein or the high protein diets. And despite being the only study participants who were in a positive nitrogen balance, the subjects in the HP group did not see more beneficial effects on the retention of lean mass than the normal protein group.

So what?! Normal protein rules?

No matter how you look at the results of the study at hand, based solely on the data Campbell and  Meckling presented here, there is not a single argument to brought forward in favor of the 1:1 vs. the 1:2 protein to carbohydrate ratio. Moreover, the single most important determinant of (long-term) dietary success that is the ease with which dieters feel they can adhere to a given nutritional protocol also speaks in favor of the normal, not the high protein diet. The answer to the initially raised question, whether the scientists' conclusion that.. 

"[a] diet with a 1:2 protein:carbohydrate ratio promoted better improvements than either the LP or HP diets, and may be superior in reducing long-term chronic disease risk in this population" (my emphasis in Campbell. 2012)

...was biased by their own research hypothesis would therefore be "NO! It wasn't." -  Now, that does not change the fact that I personally am biased and would therefore have liked the ladies to get past the 90g of quality protein / day margin. This would incidentally not have been difficult, if these wannabe overachievers had not reduced their caloric intake from ~2,300kcal/day to ~1,360kcal, but had contended themselves with the planned -30% reduction. The difference of 230kcal/day would left more than enough room for two additional protein shakes per day!

Figure 2: Fat loss and lean mass gains of the police officers in the Demling study (Demling. 2000)

That a similar regimen consisting of an even milder -20% reduction in calorie intake and the consumption of 70-75g of whey or casein hydrolysate can produce magnificent results, when it is combined with regular strength training (4days per week 30-35min of liftin), has been shown by Demling and DeSanti 12 years ago, already (see figure 2).

It should be said, though that the 'success ratio' of carbs to protein in the Demling study was likewise ~1:2 (!) - the sole difference was that the obese police officers in the Demling study simply ate twice as much protein and twice as much carbs with a baseline fat intake of ~35g per day.

References:

• Acheson KJ. Higher-protein diets for health? European Journal of Clinical Nutrition. 2012; 66, 763–764.
• Brehm BJ, D'Alessio DA. Benefits of high-protein weight loss diets: enough evidence for practice? Curr Opin Endocrinol Diabetes Obes. 2008 Oct;15(5):416-21. 
• Campbell DD, Meckling KA. Effect of the protein:carbohydrate ratio in hypoenergetic diets on metabolic syndrome risk factors in exercising overweight and obese women. Br J Nutr. 2012 Nov;108(9):1658-71. 
• Demling RH, DeSanti L. Effect of a hypocaloric diet, increased protein intake and resistance training on lean mass gains and fat mass loss in overweight police officers. Ann Nutr Metab. 2000;44(1):21-9.
• Hession M, Rolland C, Kulkarni U, Wise A, Broom J. Systematic review of randomized controlled trials of low-carbohydrate vs. low-fat/low-calorie diets in the management of obesity and its comorbidities. Obes Rev. 2009 Jan;10(1):36-50.
• Keller U. Dietary proteins in obesity and in diabetes. Int J Vitam Nutr Res. 2011 Mar;81(2-3):125-33.

L-Cysteine as a Satiety Trigger: Sign. Ghrelin & Appetite Suppression in Rodents & Humans - Which Foods Are High in Cysteine & Will They Really Help You Lose Weight?

Egg whites are among the best dietary sources of cysteine

You've read about the satiety effects of several amino acids, like arginine, lysine and glutamic acid, about which you've read approximately one year ago right here at the SuppVersity (learn more). That cysteine, an semi-essential amino acid that can be biosynthesized in humans from methionine, would have the same effects, however, is news - even for seasoned SuppVersity veterans.

The news comes right from laboratories of London's King's and Imperial College, where McGavigan  and colleagues investigated the effects of oral and intraperitoneal administration of a range of amino acids on food intake in rodents.

Learn more about the effects of your diet on your body composition at the SuppVersity

Only Whey, Not Soy Works for Wheytloss

10 Things You Didn't Know About Whey

Dairy Protein Satiety Shoot-Out: Casein vs. Whey

How Much Carbs Before Fat is Unhealthy?

5 Tips to Improve & Maintain Insulin Sensitivity

Carbohydrate Shortage in Paleo Land

In their preliminary studies, McGavigan et al. identified l-cysteine, a conditionally essential amino acid that acts as a precursor for biologically active molecules such as hydrogen sulphide (H2S), glutathione and taurine, as an anorectic agent. Needless to say that they felt inclined to further investigated the effects of l-cysteine on appetite in rodents and humans and the mechanisms mediating these effects.

Figure 1: The effect of oral administration of L-andD-cysteine in rats (left) and the effect of intraperitoneal (middle)
and oral (right) administration of L-cysteine on 0–1-h food intake during the early light phase after an
overnight fast in male in male C57BL/6 mice (McGavigan. 2014)

As you can see in Figure 1 the administration of different dosages of l-cystein, but not d-cysteine (Figure 1, left), lead to a significant reduction in 0–1-h food intake in the early light phase following an overnight fast. This effect was identical, but required higher dosages of l-cysteine (human equivalent 0.036 or 0.072g/kg when it was administered orally vs. via intraperitoneal injection.

Where do you find cysteine in foods? Egg whites, whey protein (concentrate, Bounous. 1989) beef and milk are the best sources with 1.2g, 1.15g, 1.0g, and 0.72g per 200kcal serving. Cottonseeds, sprouted lentils, soy protein isolate and defatted sunflowers flour are top sources for vegetarians with 0.7g, 0.65g, 0.63g and 0.58g cysteine per 200kcal serving (nutritiondata.com).

Figure 2: L-cysteine suppresses plasma acyl ghrelin levels in rats. Plasma levels of (a) acyl ghrelin and (b) l-r: GLP-1 and PYY, 30 min after oral gavage of water or 4 mmol/kg l-cysteine (n=7–8), (c) acyl ghrelin and (d) l-r: GLP-1 and PYY, 30 min after intraperitoneal administration of saline or 2 mmol/kg l-cysteine (McGavigan. 2014)

Next to the effects on food intake, the rodent study revealed that an increase in respiratory exchange ratio (=more CHO vs. FATs were burnded) and an increases neuronal activation in the rat brainstem without negative behavioral side effects. What the researchers did not observe, though, was an a reduction in gastric emptying that would be the most straight forward explanation for the reduction in food intake. Against that background, the reduced levels of the hunger hormone ghrelin (see Figure 2) appears to be the most likely mechanism by which the l-cysteine gavage may have lowered the animals' food intake.

This hypothesis is supported by the fact that l-cysteine didn't reduce the food intake of the gen. modified mice which overexpress ghrelin (data not shown in Figure 2).

The effects remain significant with repeated administration: Even when the "trick" is repeated thrice daily for five days, the administration of l-cysteine still lead to an acute reduction in food intake and a corresponding decrease in the cumulating food intake over the 5-day study period in rodents - in view of the short study period obviously without reductions in body weight.

Now we all know that mice are no little men. Therefore, the important question that's rightly preying on your mind now is: Did this work in humans, as well? The answer is pretty straight forward: Yes, it did!

Figure 3: 0.07g/kg l-cysteine in 200ml water lead to significant reductions in hunger ratings and acyl-ghrelin in humans as the corresponding dose in rodents (McGavigan. 2014)

As you can see in Figure 3, the administration of either "vehicle" (=placebo) alone or the same 200 ml drink containing 0.07 g kg/1 l-cysteine in a single-blind (participant) randomised order lead to similar decreases in acyl-ghrelin (hunger hormone) and hunger ratings in the healthy men and women who participated in McGavigan's study.

If you haven't done this, already, it's time to check out the results of the previously cited study by Jordi et al. (2013), now | learn more

Bottom line: Luckily, McGavigan et al. did all the work for me and compared the effects of cysteine in the study at hand to the previously reported effects of arginine & co, I referenced in the introduction and found that "l-cysteine is more anorectic than l-arginine and l-lysine." (McGavigan. 2014)

Furthermore, the researchers point out that "[i]f l-cysteine does have a physiological effect on appetite, then it is likely to act in concert with other products of protein digestion, and thus the effects of l-cysteine per se may be difficult to detect." (McGavigan. 2014) In other words, the repeatedly demonstrated satiety effects of high protein diets may - in parts - be mediated by their cysteine content.

In view of the fact that the effects occur at dosages that do not trigger taste aversion or evoke abnormal behaviour, it may even be possible to administer l-cysteine supplements to overweight individuals before every meal to reduce their food intake and trigger (probably) slow, but persistent weight loss. Since the real-world food intake wasn't measured in humans, yet, this would have to be confirmed in future trials, though | Discuss this article on Facebook!

References:

• Bounous, Gustavo, Gerald Batist, and Phil Gold. "Immunoenhancing property of dietary whey protein in mice: role of glutathione." Clin Invest Med 12.3 (1989): 154-61.
• Jordi, Josua, et al. "Specific amino acids inhibit food intake via the area postrema or vagal afferents." The Journal of physiology 591.22 (2013): 5611-5621. 
• McGavigan, A. K., et al. "l-cysteine suppresses ghrelin and reduces appetite in rodents and humans." International Journal of Obesity (2014).

Two Days A Week High Protein, Low Carb Fast Cuts >10% of Body Fat in 4 Months And Improves Insulin and Leptin Sensitivity More Than Chronic -25% Energy Restriction

I wonder if she just found out that sheddin 10% body fat can be as easy as doing two "protein modified fasts" per week!?

It has been some time since the last interesting study on Intermittent Fasting has found its way into the SuppVersity news; and I have to admit that the protocol of the study I am about to discuss today does not really qualify as "intermittent fasting" in the "lean gains" sense.

Still, the latest study from the University Hospital of South Manchester NHS Foundation Trust comes with a couple of very interesting results... the only downside is that the subjects were "the usual" ones, namely obese sick people:  115 women with a family history of breast cancer (aged 20 and 69 years). This is problematic, because the same fast during which the obese person draws on his or her fat stores to "survive" can sent the lean sub 10% body fat athlete into a catabolic low energy state and have his blood sugar drop to hypoglycemic levels, when he or she returns to eating ad libitum after the 2 fasting days.

Will intermittent fasting shrink your bust?

Sorry, I just could not resist to use focus on this question of paramount importance. Why? Well the study at hand is the first to actually investigate the negative effects of fasting on your cup size ;-) All jokes aside, this is obviously not a non-sense or cosmetic measure.

As I already mentioned in the introduction, the subjects had a history of breast cancer and in view of the association between breast cancer risk and bra cup size in US women aged <42 (Swanson. 1996) -

• 26% increased risk for cup size B
• 23% increased risk for cup size C
• 95% increased risk for cup size D*

- it does appear to make at least some sense to measure the effect the diet had on bust size, as well (* note: the data was calculated for normal weight women).

Figure 1: Change in body composition in response to dietary energy restriction (-25%; DER) and 2-day per week fasting with calorie (IFC) and carbohydrate (IFCA) restriction on the two fasting days (Harvie. 2013)

So, if you are concerned about losing fat in the wrong places (don't that it may decrease your breast cancer risk to shed superfluous fat on your chest, ladies) the data in figure 1 will probably comfort you. It does after all confirm that the additional loss of abdominal fat (as evidenced by the reduction in waist circumference) is more pronounced than the additional 1-2% reduction in bust size the women in the two intermittent fasting groups, IFC and IFCA experienced.

Details on the protocol:  Diet composition

Side effects? What is interesting and something I have not seen before in a study is an evaluation of differences in side effects and nutrient sufficiency:

• feeling cold (3% DER)
• decreased energy (5% IFC & DER)
• constipation (8% IFC, 3% DER)
• headaches (5% IFC, 3% IFCA)
• bad breath (5% IFC, 3% IFCA)
• light-headed (3% IFC)
• lack of concentration (3% DER)
• mood swings (3% IFC & DER)
• thinking about food 24/7 (8% IFC, 3% IFCA & DER)

Interestingly the different pre-occupation with food did not impact the hunger scores. The only thing the scientists observed was a minimal difference in hunger scores on fasting day 1 in the IFC vs. IFCA group.

Of the usual low-carb diet deficiencies in Mg, Fe, Zn, Ca, vitamins A and D and fibre, the multi compensated everything but the scarcity of magnesium (Mg), zinc (Zn) and selenium (Se) in the IFC group.

The dietary energy restriction (DER) and the intermittent fasting carbohydrate restricted (IFC) groups were isocaloric and contained only 75% of the calculated energy requirements of the women. Contrary to the women in the DER group who consumed a diet with a macronutrient ratio of 25/45/30% protein / carb / fat, the subjects in the IFC group were however

"asked to restrict energy and carbohydrate on two consecutive days each week (70 % energy restriction and 40 g carbohydrate) and to consume a euenergetic Mediterranean-type diet that met their estimated energy requirements for the remaining 5 d of the week."

The restricted IFC diet the subjects consumed on those "fasting days" provided between 625kcal/day and 680kcal/day and included approximately

• 250 g of protein foods including lean meat, fish, eggs, tofu, textured vegetable protein, 
• three servings of low-fat dairy foods (e.g. 195 ml semi skimmed milk, 150 g low-fat yoghurt, 30 g low-fat cheese), 
• four portions of low-carbohydrate vegetables and 
• one portion of low-carbohydrate fruit, 
• at least 1170 ml of other low-energy fluids, and 
• an over-the-counter multivitamin and mineral supplement providing the RDA for vitamins and typically 20 – 50 % for minerals on restricted days.

Lastly, the subjects in the IFCA protocol followed the exact same rulez, but were allowed to consume "unlimited lean meat, fish, eggs, tofu, MUFA and PUFA on restricted days." In addition, the saturated fat content of both intermittent diets was limited to 10% and alcohol consumption was discouraged, but not prohibited.

Addendum: In view of the fact that the question arose several times both here in the comments and on facebook, I want to point out that the 250g of protein foods are not identical to 250g of protein. They will rather contribute max. 50-90g of protein and will thus not go beyond the dietary constraints of 625kcal. Apropos, other than one commenter suspected the weekly caloric deficit was indeed identical in the DER and the IFC group. Let's to the math with simplified figures: Baseline intake 2,000kcal - 25% => 7x400kcal = 2,800kcal deficit in the DER group vs. 2x (2000kcal-600kcal) = 2,800kcal on the two consecutive low carb fasting days in the IFC group.

"And what about the 'health effects'?"

It we take a look at the serum markers, it's actually quite surprising that there were no significant improvements in HbA1c, blood glucose, IL-6, TNF-alpha, adiponectin, cholesterol and triglycerides. After all, this is what we almost expect with any type of fat loss.

Figure 2: Changes in blood glucose and insulin resistance, as well as leptin levels after 3 month on the respective diets and another month on "weight maintenance" when 68% of the ICF & ICFA subjects stuck to their protocol (Harvie. 2013)

The comparatively high fat loss in the IFC and ICFA groups (-12% and -11% over the three months of dieting) did yet result in significant improvement in insulin sensitivity and had a much more pronounced effect on the leptin levels, which is - in the case of overweight individuals - a good thing, as it signifies a reduction in "leptin resistance".

Altough they were allowed to, the IFCA subjects (middle bar) did not eat much more protein and fat than their peers in the IFC group.

Bottom line: Overall the results of the study at hand leave no doubt that a carbohydrate restricted 5:2 day (5 days you eat normal, 2 days you fast) high protein fasting strategy is a very effective means to shed body fat and increase your insulin sensitivity - for the overweight individual (!).

In that, it's worth mentioning that you do not even have to force yourself to abstain from foods altogether on the two fasting days: It's actually sufficient to cut the carbs by 43 % (which was the effective reduction in the IFC & IFCA groups compared with 23 % for the DER group) as long as you are not into fatty foods and don't compensate for that by overconsuming protein and fats... yep, you heard me right, there is NO MAGIC in low carb dieting that would allow you to eat as much protein and fat as you want and still lose weight. The subjects in the IFCA who were allowed to do so added no more than 10g of extra protein and 15g of fat to your ~600kcal allowance on the fasting days.

Now, I am not sure how realistic it is that everyone will do the same as the subjects in the IFCA group and refrain from overeating on eggs, sausages of even Quest protein bars and all the other delicious(ly convenient) ready made "low carb foods" that are flooding the market these days. If you are a trainer or nutritionist, I would thus suggest you don't tell them they allowed to eat "as much as they want as long as it contains no carbs" ;-)

References: 

• Harvie M, Wright C, Pegington M, McMullan D, Mitchell E, Martin B, Cutler RG, Evans G, Whiteside S, Maudsley S, Camandola S, Wang R, Carlson OD, Egan JM, Mattson MP, Howell A. The effect of intermittent energy and carbohydrate restriction v. daily energy restriction on weight loss and metabolic disease risk markers in overweight women. Br J Nutr. 2013 Apr 16:1-14. [Epub ahead of print]
• Swanson CA, Coates RJ, Schoenberg JB, Malone KE, Gammon MD, Stanford JL, Shorr IJ, Potischman NA, Brinton LA. Body size and breast cancer risk among women under age 45 years. Am J Epidemiol. 1996 Apr 1;143(7):698-706.

Protein Requirements of Dieting Strength Athletes: More is Better Only in the Presence of Adequate Carb & Fat Intake. Optimal Muscle Retention With 2-3g/kg Lean Body Mass

Believe it or not: Being lean, athletic and well-conditioned is a disqualifier, when it comes to "body recomposition" (=building muscle +  losing fat at the same time). Simply upping your protein intake indefinitely is not going to change that..

This is one of those article, where I thought twice whether or not it would be worth writing. If I knew that the majority of you had full-text access to the recent review by Helms, Zinn, Rowlands and Brown, I would probably stick to a couple of comments on my Facebook page and suggest you read the whole paper, yourself.

In view of the unfortunate fact that research is not really a public good, and full-text access is very limited unless you work / study at a University, this would leave most of you with nothing but a conclusion to an abstract that could could easily be misinterpreted in a simplistic: "More is better!" way; a conclusion you would obviously revise, if you had the change to read the whole paper which does have more to offer than two random numbers.

Apropos "random": In view of the fact that Helms et al. found only 6 studies that provided (in some cases limited) information about the influence the amount of dietary protein will have on lean mass retention and fat loss in strength trainees, neither the numbers in the headline nor ostensibly more accurate figures Helms et al. provide are more than a brought guideline. The "true" optimum and in my humble opinion even the question whether such a thing has yet to be found.

I: Energy-, not protein-intake is the main determinant of muscle loss

The 2011 study by Garthe is only one of many studies that confirms that a lower calorie deficit will yield better dieting results (i.e. greater fat and lower lean mass loss); results after 8.3 (19% deficit) and 5.3 (30% deficit) weeks (Garthe. 2011)

What is more or less indisputed and at the same time one of the most important, because often overlooked, or I should say "willingly ignored", determinants of lean mass loss is the fact that

"[...] the magnitude of the caloric deficit imposed is likely one of the most powerful variables that impacts FFM loss, potentially being more important than protein intake." (Helms. 2013)

In other words, the "harder you diet", i.e. the more severe your caloric deficit, the more lean muscle you're going to lose. This appears to be self-evident, I know, and yet the average and not so average dieter (e.g. the bodybuilder who comes in not just flat, but actually small) tend to forget about it.

II: Body recomposition is something for the "fat beginner"

Next to hitting it too hard, being in denial is probably the most common threat to your dieting success - in denial of the fact that such a thing as "body recomposition", i.e. concomitant loss of fat and increase in muscle mass is a prerogative of the chubby beginner.

Suggested Read: "Seven Meals/Day, More than 800g of Carbs & 1000kcal Over Maintenance and Still Lean Gains!" | read more

Based on studies by Peterson, Rhea, & Alvar (2005) and Garthe et al. (2011) Helms et al. rightly point out that gaining lean mass while being on a caloric deficit is not the norm, but rather the exception; an exception that will occur almost excursively in to novice lifters and among those most probably in the chubbier ones, who can draw on larger body fat reserves while they are dieting. In "leaner more experienced weight lifters", on the other hand, "it may be unrealistic to expect a lack of FFM loss or FFM gain in leaner" (Helms. 2013)

The common, though schizophrenic approach to fat loss that denies the inevitability of lean mass losses is thus more often than not going to fail you; and if you are still wondering why your abs don't shine in full glory, ask yourself if the reason may not be your own fear afraid of losing muscle that you never diet long or hard enough to attack the stubborn fat.

III: When you eat so much protein that there is no room for fat and carbs bad things happen

While it is correct that your body can use protein as an energy source, forcing it to do just that by consuming so much protein that the amino acid chains constitute the lion's share of your daily energy intake is going to have significant detrimental effects on your dieting success. As Helms et al. point out...

• consuming a high protein diet with very little carbohydrate in it is going to hamper your exercise performance (Walberg. 1988), while
• consuming a high protein diet with too little fat in it is going to have negative effects on your mood and emotional stability (Mettler. 2010).

Unfortunately, it is difficult to determine the exact "minimal requirements" of these nutrients. Until now, these have not been explicitly studied and in view of the scarcity of existing research (remember: N=6!) it is almost impossible to extrapolate them from the data we have.

Ketogenic diets are not high in protein: Please note that the previously mentioned detrimental effects of low carbohydrate intakes will only arise in a high protein context- The latter is preventing you from transitioning into full ketosis and deriving the corresponding benefits. A ketogenic diet cannot be "high" in protein; and it is save to assume that the risk of being kicked out of ketosis is specifically high with fast digesting and highly gluconeogenic protein sources like whey (cf. Calbet. 2002).

If we take a look at the data we have it does yet appear that 20% is the absolute minimum for fat and ~30% could be the minimum for carbohydrate intake in a high protein diet scenario.

In this context Helms et al. do actually cite a study, every SuppVersity reader should be familiar with: The recently published metabolic ward study by Pasiokos et al. (read the previous SuppVersity article).

Figure 1: Change in body composition and protein synthesis (Pasiakos. 2013)

The results of this study would support the hypothesis that increasing your protein intake from 1.6g/kg body mass to 2.4g/kg body mass so that the carbohydrate intake drops to ~20% of the total energy intake, will accelerate the overall loss of body mass at the expense of lean muscle tissue.

Meanwhile it should not remain unmentioned that these effects were observed in a "low" intensity training scenario that did not provide for a maximal exercise-induced stimulus of protein synthesis. The hypothesis that a more hypertrophy specific training program would have yielded very different outcomes is however questionable and - as of now - just as the term "hypothesis" implies hypothetical.

IV: Athletes should use their lean lean body mass to determine their protein intake

Within the bodybuilding community it is actually common practice to prescribe (often hilariously high) protein intakes on a "per kg of lean mass" or "per lbs of fat free mass" level. Among recreational fitness enthusiasts and eve among scientists this is however still the exception; and that despite the fact that this practice has the advantage that it will (automatically) yield higher per kg total body mass protein intakes for leaner athletes. This "scaling" approach is in accordance with Helms et al.'s conclusion that

"[a]thletes with a lower body fat percentage, or a primary goal of maintaining maximal FFM should aim towards the higher end of [he protein intake range, while t]hose who are not as lean, or who are concerned primarily with strength and performance versus maintenance of FFM can safely aim for the lower end of this recommendation." (Helms. 2013)

I guess by now you are probably asking yourself what on earth the "exact" recommendation of the scientists are, right? Well, I guess it's time to let the cat out of the bag, then ;-)

How do dermine your "optimal" macronturient intake? The easiest way to go is to (1) use your regular daily energy intake (i.e. the amount of energy you consume when you are weight stable) subtract 20-30%. Then you take the result and (2) subtract the 450kcal energy equivalent of your 50g minimal fat intake and (3) 2.8 x 4kcal/kg x lean body mass, the energy equivalent of your protein intake. Lastly, you (4) divide the rest (baseline - fat - protein) by 4 kcal/g and have your daily carbohydrate allowance in grams.

Example? Easy! Baseline intake: 2500kcal, (1) 20% calorie reduction → 2000kcal, (2) subtract 50g fat * 9kcal/g → 1550, (3) subtract 2.8 x kcal/kg x 65kg lean mass → 822kcal (4) divide this value by 4 and get your carb intake, here 205.5g. Your overall macro ratio would thus be 182g of protein, 205g of carbs and 50g of fat.

This ratio would be adequate for athletic and active individuals, while sendentary and/or insulin resistant obese indivuals may in fact consider turning this into a low carb diet, by reversing the fat and carb intake.

What's the optimum, then? According to Helms et al. the "optimal" protein intake for strength athletes on a energy restricted diet amounts to 2.3-3.1g/kg of the athletes fat free mass. The latter includes both muscle, as well as organ mass and body water and will - as I have pointed out in IV. make sure that athletes with lower body fat mass will consume more protein than the chubby beginner who has just taken up weight lifting to prepare for spring break.

Let's just briefly check what this may mean for you, a young man / women with 12% or 19% body fat and a body weight of 80kg or 60kg, respectively. While his protein intake would be ~161-217g, her protein intake would be "only" 112g-150g per day. If we now assume that the daily energy intakes of the two are 2,000 and 2700kcal per day (for maintenance) and that both follow my suggestion to reduce their calorie intake by only 20%-30%, this would leave ~1,300kcal (male example) and 980kcal for carbs and fat (woman). If we further prescribe a minimum intake of 50g of fat (this would be more than the afore-cited 20%, but is imho a sane minimum intake) this would leave us with ~210g of carbs for him, and 130g of carbs for her - both not exactly bad macro-ratios for an active individual whose main intention is to shed some body fat. If you end up with way more than 200g of carbs and/or are insulin resistant consider cutting the carbs back to 200g or below (insulin sensitive, but not very active), or 150g or below (insulin resistant, but not obese).

References: 

• Calbet JA, MacLean DA. Plasma glucagon and insulin responses depend on the rate of appearance of amino acids after ingestion of different protein solutions in humans. J Nutr. 2002 Aug;132(8):2174-82.
• Garthe I, Raastad T, Refsnes PE, Koivisto A, Sundgot-Borgen J. Effect of two different weight-loss rates on body composition and strength and power-related performance in elite athletes. Int J Sport Nutr Exerc Metab. 2011 Apr;21(2):97-104.
• Mettler S, Mitchell N, Tipton KD. Increased protein intake reduces lean body
  mass loss during weight loss in athletes. Medicine and Science in Sports and Exercise. 2010;
  42(2), 326-337.
• 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]
• Peterson MD, Rhea MR, Alvar BA. Applications of the dose-response for muscular strength development: a review of meta-analytic efficacy and reliability for designing training prescription. J Strength Cond Res. 2005 Nov;19(4):950-8. Review.
• Walberg JL, Leidy MK, Sturgill DJ, Hinkle DE, Ritchey JS, Sebolt DR. Macronutrient content of a hypoenergy diet affects nitrogen retention and muscle function in weight lifters. International Journal of Sports Medicine. 1988; 9(4), 261-266.

Myostatin Limits Muscle Hypertrophy in Young, Physically Active Resistance Trainees on High, but Not Normal Protein Diet - Irrelevant Outlier or Crucial Revelation for Trainees?

Who says, resistance training and high protein diets make you bulky? The study at hand suggest they don't because after a couple of weeks your body will pull the myostatin break | img (c) fighterdiet.com

You're training like mad and consuming a ton of protein everyday and still don't see the results you deserve? Maybe increased myostatin levels are holding you back!?

While previous studies mostly suggested that the role of myostatin in the normal (muscle) growth response to exercise may have been overestimated, a recent study from the University of Padova,  the Italian Medicine and Fitness Federation, Euganea Medica, and the University of Palermo. Brings the "muscle-growth break" (high myostatin = slow muscle gains) back onto our radar. And that not just because researchers from said institutions observed a significant increase in myostatin in response to chronic resistance training, but rather because this response appears to have been triggered by high(er) protein intakes.

Want to get stronger, bigger, faster and leaner? Periodize appropriately!

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If you have been reading SuppVersity articles for quite a while now, you may remember my previous post about how "Chronic Resistance Training Reduces the Anabolic Signaling in Response to Exercise - 12 Days of Detraining Restore It" (read more). The corresponding study by Ogasawara et al. (2013) was yet conducted on rodents. Therefore, the applicability of the results remains at least questionable.

In a more recent study with human subjects, Antonio Paoli and colleagues from Italy did yet observe a similar, but not identical effect. The researchers aim was to "investigate the influence of 2 months of resistance training (RT) and diets with different protein contents on plasma myostatin (MSTN), interleukin 1 beta (IL-1β), interleukin 6 (IL-6), tumor necrosis factor alpha (TNF-α), and insulin-like growth factor 1 (IGF-1).

Does an increase in protein intake increase myostatin, as well?

To this ends, they recruited 18 healthy, young, physically active volunteers who were then randomly divided in two groups; A high protein (HP) and normal protein (NP) groups. The diets contained 1.8 and 0.85 g of protein/kg body weight per day for the HP and NP diet, respectively.

In addition to the dietary intervention, all subjects were subjected to the same 8 weeks of standardized progressive resistance training. MSTN, IGF-1, IL-1β, IL-6, and TNF-α were analyzed before and after the first and the last training sessions. In addition, Paoli et al. measured the lean body mass, muscle mass, upper-limb muscle area, and strength.

Figure 1: Much to the surprise of the researchers, the muscle gains were identical, but the msucle gain reducing myostatin increase was significant only in the high protein group (Paoli. 2014)

Somewhat to the scientists surprise the plasma MSTN showed a significant increase (P<.001) after the last training in the HP group compared to both the normal protein (NP) group and the starting values. Surprisingly, these increases occurred in the absence of differences in IL-1β, IL-6, TNF-α, and strength and muscle mass or muscle area.

Protein blunts the exercise induced decrease in myostatin, but it increases the alleged myostatin blocker FLRG (Hulmi. 2008) - Result? A null or maybe even beneficial effect?

This is not an outlier study! Bad news, bros You cannot discuss the findings of the study at hand away. Previous studies by Hulmi (2008) found that the decrease in myostatin that would usually occur in response to resistance training in older male trainees was blunted, when the resistance training session was followed by the ingestion of 15g of whey protein. Needless to say that this does not necessarily mean that you'd gain less. In fact, the concomitant increase in folistatin related gene expression (FLRG), a gene which is thought of regulator off the myostatin inhibitor folistatin (Hill. 2002), may more then compensate the slight increase in myostatin (see Figure on the left) in the first 48h after a workout. If that's the same in the long run, will have to be tested in future studies.

If the muscle mass / area had been increased as well, it would have been easy to interpret the increase in myostatin as a reaction to an increase in muscle cell size and a corresponding increase in domain sizes. The way it is, though, it's difficult to explain the increase in myostatin and its correlate IGF-1 (correlate in this study, not in general).

Is this just a matter of domain sizes?

Figure 2: Domain sizes of EDL and soleus muscle fibers in wild-type control, myostatin negative and IGF1 over-expressing mice (data based on Qaisar. 2011)

As you will remember from previous articles, the latter would require the recruitment of additional muscle satellite cells that would form new myonuclei and thus help normalize the domain sizes and myostatin levels.

Without myostatin, the domain sizes would keep increasing and the muscle would become as huge, but dysfunctional as it does in myostatin negative mice (see Figure 2, red | not the similar increase in domain size in IGF-1(+) mice which underlines the role both myostatin and IGF-1 play when it comes to controlling domain sizes and facilitating "Getting Big Beyond Temporary Physiological Limits" | learn more)

The way it is, the observations Paoli et al. made in their most recent study remain, as the authors themselves call it "paradoxical" and could in fact "explain the substantial overlapping of MM [muscle mass] increases in the two groups", no one of you would have expected, right? Well, it's a pitty we don't have data on the level of follistatin related gene expression as it was measured in the Hulmi study (see red box) - if that did not increase, or only to a small extend, it's actually no wonder, both groups gained the same amount of muscle mass.

How intense is "intense enough"? The training program Roth reused in their 2003 study and which reduced the myostatin expression sign. consisted of five sets of high-volume (55 total repetitions), heavy-resistive exercise performed 3 days/wk. The resistance for each set was based on the subject’s 5RM. After performing a first set of five repetitions at 50% of the original 1RM for a warm-up, the subject performed a second set of five repetitions at the 5RM resistance after a 30-s rest period. The third set consisted of 10 repetitions, with the first four or five repetitions at the current 5RM value, then the resistance was lowered just enough to complete one or two more repetitions before the subject reached fatigue. The process was repeated until a total of 10, 15, and 20 repetitions were completed for the third, fourth, and fifth sets, respectively. The third, fourth, and fifth sets were preceded by rest periods lasting a minimum of 90, 150, and 180 s, respectively. This protocol demanded a near-maximal effort on every repetition throughout each set.

Do we need myostatin blockers to maximize our gains? As far as I know, no scientifically proven myostatin inhibitor is yet available as an OTC supplement. Luckily, previous research by Roth (2003) and Willoughby (2004) suggests that these products may not even be necessary. In 2003 Roth et al. write in a "short communication" (something similar to a "mini paper") that really heavy resistance training with maximal effort on each set can (at least temporarily) reduce the myostatin levels (see box to the right for information on the exact protocol) - an effect that is unfortunately reduced with aging and almost non-existant in aging women (old men -37%, old women -11%, young men -56%, young women -46% | Kim. 2005), but could be buffered by concomitant increases in folistatin-related gene (FLRG) expression, as it was observed by Willoughby in his 2004 study.

Figure 3: Serum folistatin related gene expression in response to 12 week high intensity resistance training vs. control (Willoughby. 2004)

The key to actually making the gains you could make due to the increase in protein availability, could thus simply be to train harder to either facilitate greater reductions in myostatin or promote long-term increases in its "buffer" FLRG, as they were observed by Willoughby (see Figure 3).

In the long run, this does yet entail the risk of severe overtraining, which should remind you of the periodization techniques and the interplay of phases of maximal and sub-maximal exercise intensity in 4-6 week cycles. You have no idea how that works? Well, I suggest you take a look at the overview of previous SuppVersity articles on periodization - specifically this one: "Periodization Techniques Revisited: Improved Strength & Size Gains W/ 12-Week Undulatory vs. Linear Periodization" | Comment on Facebook.

References:

• Hill, Jennifer J., et al. "The myostatin propeptide and the follistatin-related gene are inhibitory binding proteins of myostatin in normal serum." Journal of Biological Chemistry 277.43 (2002): 40735-40741.
• Hulmi, Juha J., et al. "The effects of whey protein on myostatin and cell cycle-related gene expression responses to a single heavy resistance exercise bout in trained older men." European journal of applied physiology 102.2 (2008): 205-213.
• Paoli, et al. "Protein Supplementation Increases Postexercise Plasma Myostatin Concentration After 8 Weeks of Resistance Training in Young Physically Active Subjects." J Med Food. Aug 18 (2014). Epub ahead of print.
• Qaisar, Rizwan, et al. "Is functional hypertrophy and specific force coupled with the addition of myonuclei at the single muscle fiber level?." The FASEB Journal 26.3 (2012): 1077-1085.
• Roth, Stephen M., et al. "Ultrastructural muscle damage in young vs. older men after high-volume, heavy-resistance strength training." Journal of Applied Physiology 86.6 (1999): 1833-1840.
• Roth, Stephen M., et al. "Myostatin gene expression is reduced in humans with heavy-resistance strength training: a brief communication." Experimental Biology and Medicine 228.6 (2003): 706-709.
• Willoughby, Darryn S. "Effects of heavy resistance training on myostatin mRNA and protein expression." Medicine and science in sports and exercise 36.4 (2004): 574-582.