Whey or Casein, Pulse or Spread Evenly Across the Day? Does it Even Make a Difference in Terms of Fat Loss and Lean Mass Retention on a Diet? New + Old Empirical Data!

Image 1: Instinctively right? Milk contains soluble (=whey) proteins and casein. Are we overthinking things, when we rip them apart and does it even make a difference? Or is timing all that counts?

It's funny "overthinking", right next to overtraining and overdieting, has become one of the most common problems among the health and fitness enthusiasts who spend equal (or even more) time online as in the gym. "Would it be better if I take my BCAAs at a 3:1:1 or 2:1:1 ratio?", "Does it matter if my protein powder is 10% hydrosolate, 50% isolate and 40% concentrate or has a 30/50/20 ratio?" All that may well make a difference, but let's be honest: Look at the things 80% of these people are eating day in and day out and the way they throw the weights around in the gym and contrast that to a question like "Will my post-workout protein synthesis be 5% greater, when I switch from concentrates to hydrosolates?" ... enough of the ranting, though. After all this post is actually about one of the more sensible among these world-shattering questions:

Q

Will it make a difference, whether I use casein or whey protein on a diet and... what's the significance of having my daily allotment of protein spread evenly across the day vs. mostly (80%) in one sitting, when I am dieting?

In order to find the answer to this question a group of French scientists recruited 41 healthy, but chubby subjects (BMI ~32kg/m²; age ~33y) and put them on a relatively moderate caloric deficit that was calculated based on their basal energy requirement (what you would need lying around all day). In all four arms of the study, the macro-nutrient composition (25% as proteins, 25% as lipids, and 50% as carbohydrates) and energy content per pound of lean body weight (average energy intake 5.87 MJ per day) of the meals, which were prepared according to personalized menus the subjects received from trained dietitians, were identical.

Figure 1: It did not make a difference if the protein was ingested either spread equally across the day or as a pulse mostly (80%) in one sitting (top), fat and weight loss after the 6 week study period were virtually identical (data based on Adechian. 2012)

The little information on the exact menu choices the scientists offers includes a list of stable foods, such as various proportions of spinaches, broccoli, lentils, or green beans, butter, bread, fruits, soy yogurt, rice cakes and gingerbread and suggest that we are dealing with the typical "your dietitian recommends diet", here. With one exception, of course, the main protein source of all four experimental diets were dairy proteins (~80g; >80% of total protein). Casein and whey aka "milk soluble protein"* (see red box above), which were to be ingested either spread equally across or in a "pulsed" fashion (see figure 1, left):

*Note: the scientists refer to whey as "milk soluble protein, I stuck to the terminology in the graphs, but in essence these are mainly β-lactoglobulin, α-lactalbumin, as well as serum albumin, immunoglobulins, lactoferrin, and other minor fractions and thus the same you would find in your average whey concentrate which is, as you may have notices "more soluble" than casein (cf. Lacroix. 2006)

• casein spread- subjects consumed ~20g of a casein protein supplement 4x a day
• milk spread - subjects consumed ~20g of milk protein supplement 4x a day
• casein pulse - subjects consumed the lions share, i.e. 80% of their ~80g of casein, as part of their 2nd meal, so that the protein intake over the day was 6.4g / 64g / 3.2g / 6.4g (see figure 1)
• milk pulse - same as above, but with milk instead of casein protein

In view of the overemphasisze nutrient timing has gotten as of late withing the physical culture and the assumption that you would expect to see profound differences based on when you consume how much of fast or slow, high (milk) or average (casein) leucine protein etc., it may be disappointing that the weight loss was absolutely identical in all four arms of the study (-7.5 ± 0.4 kg).

Differences are few and far between: Weight loss, fat loss, muscle loss - NOT different! 

What may yet surprise even you, a seasons SuppVersity veteran, who will probably already have expected the non-significant (in fact non-existent) differences in terms of weight loss, could be surprised that the changes in body composition (see figure 1, bottom), i.e. -5.1 ± 0.2 kg reduction in body fat mass and -2.2 ± 0.2 kg reduction in lean body mass, were identical.

Since the same goes for the changes in the fat "liberating" proteins lipoprotein lipase (LPL) and adipose triacylglycerol lipase (AGTL), the fat "forming" protein fatty acid synthase (FAS), and three of the usual subjects, i.e. leptin, the adipoQ gene which is responsible for encoding adiponectin, of which recent research suggests it may be even more important than leptin for your metabolic health (Li. 2012; Hickman. 2012), and the reduction in the pro-inflammatory monocyte chemotactic protein-1  (MPC-1), the slightly more pronounced meal-induced postprandial protein synthetic response in the casein group at the end of the study period is actually the only difference based on which you could argue for one over the other protein source:

Figure 2: While the changes in LPL, AGTL, FAS, leptin, AdipoQ and MCP expression were identical (left); the post 6-week protein synthetic response to identical meals was slightly more pronounced in the casein group (right), the overall significance of this finding is yet questionable in view of identical lean mass losses - it could yet become important on a diet + exercise regimen as in the Demling study discussed in the bottom line box  (data based on Adechian. 2012).

Whether the measurable advantage of casein during this test (the evaluation was carried out by leucine tracer infusion, by the way) is just an experimental artifact or

Adherence is the key to success: While there was no difference in terms of the hunger the subjects felt when they were on the diet, the fact that only 23 of the initially 41 subjects did make it through the 6- week on ~ 1,500kcal/day is quite telling, also in view of the perceived inability to lose weight - if you can't stick to a by no means crazy caloric restriction for 6 weeks, how can you expect to get lean and stay lean, when the inevitable prerequisite for the latter is that you totally revamp your dietary habits for the rest of your life not just six, eight, or twelve weeks.

• maybe something like "leucine resistance" in response to the higher leucine concentrations after the ingestion of the milk protein supplement in the course of the study period, or
• alternatively, the greater IGF-1 response to casein (cf. Hoppe. 2009, a study which compares whey vs. casein, but would obviously suggest an advantage of casein over milk = whey + casein, as well); unfortunately IGF-1 wasn't measured, but the insulin levels which were minimally higher in the casein group could support that hypothesis,

... is questionable. Since the same is true for the practical relevance of the ~10-13% larger leucine balance during the postprandial phase of the post-diet whole body protein metabolism test in week 6, I would not fret about this difference too much, though.

Maybe, just maybe, the adipocyte morphology could make a difference

What I would consider significant, though it did not reach that status (probably due to the low number of participant that actually made it to the end of the study, see red box on the right), is the slight but in my eyes potentially important superiority of the equally spread protein ingestion in terms with respect to the before vs. after adipocyte diameter in the casein group:

Figure 3: The difference did not reach statistical significance, but if we take for granted that greater reductions in adopcyte sizes are associated with healthier metabolic profiles, you would be better advised to take your casein protein equally spaced across (15% reduction in adipocyte size vs. 7%, only, for pulsed casein intake) the day... for whey, aka "milk soluble protein", on the other hand it does not seem to matter (data calculated base on Adechian. 2012)

Now, even if we assume that this made a difference and a greater reduction in adipocyte size was a significant advantage, which it probably is from a health perspective, as Skurk et al. state that there is

"[...] a differential expression of pro- and antiinflammatory factors with increasing adipocyte size resulting in a shift toward dominance of proinflammatory adipokines largely as a result of a dysregulation of hypertrophic, very large cells." (Skurk. 2006)

and a recently conducted human trial, by Rizkalla et al. the main message this study should be sending out is not that it does not make a difference whether you use casein or milk protein as your main protein source on a diet, but that a high protein diet with a mediocre caloric reduction of ~20-25% and supplemented with high quality dairy protein (whey or casein) works: After all, more than -1kg of weight loss per week, 68% of the weight loss from fat in the absence of exercise is more than your average celebrity XYZ diet will do for you ;-)

Whey or casein? It's high cysteine content that can help to replenish your glutathione (=the master antioxidant) pools would be another factor that speaks in favor of whey. Whether normal-weight individuals on an already optimized dietary regimen would benefit to the same extend as the obese young men in the 6-week whey supplementation trial, Vatani et al. describe in the August issue of Appetite, is however questionable. After all, the increases in HDL the total antioxidant capacity and glutathione is as questionable as any possible negative influence of the starchy placebo the researchers used in that study (some of you may have seen the link on the SuppVersity Facebook Wall, already).

Figure 4: Fat loss and lean mass gains in formerly overweight police officers after 12 weeks of training and dieting with or without casein / whey hydrosolate (Demling. 2000)

Moreover, one of the few long-term (=non acute protein synthesis) studies investigating the differential effects of concomitant whey vs. casein hydrosolate protein supplementation, found statistically significant higher body fat reductions and lean mass gains in those 33-34 year-old police officers who supplemented their 12-week diet + strength training regimen with 2x37g of casein hydrosolate (8h apart; for the exact data see figure 4; Demling. 2000).

Note: since both the whey (Pro-Score Champion Nutrition) and the casein protein (MET-Rx USA) in this study were hydrosolates the differences in lean mass gains and fat loss are depend primarily on the amino acid composition of the proteins, and not, as it would be with micelle casein vs. whey, the absorption kinetics!

Bottom Line: Against that background the study at hand supports previous findings of the importance of a threshold intake of protein. Interestingly, it did not confirm the notion that this threshold intake should be spread equally across the day, which is something most commenters (me included) read into the seminal paper by Loenneke et al., which found a statistically significant negative correlation not between total protein intake, but between the number of meals with 10g or more essential amino acids in them and abdominal obesity (Loenneke. 2012). So, does timing matter, or does it not? 

1. It does matter, when you work out, there is ample evidence to support that the ingestion of protein in the vicinity of the workout cannot just amplify the protein synthetic response but will also results in an increase in real world muscle gains.
2. It appears that it does not matter, when you are dieting (only), though; not just the study at hand, but also the success many people report on intermittent fasting regimen, would support the notion that the more sustained anabolism you may be able to achieve by ingesting say 4x25g of protein instead of 1x80 + 2x10g has, compared to the total amount of protein you eat, relatively little influence on the conservation of lean body mass, when you are dieting.

And as far as the choice between casein and milk soluble protein, aka whey (see first red box), is concerned (see box on the right, as well), it would appear prudent to assume that a combination of both - just like nature intended it - would be the best choice as a "standalone" protein source (cf. "Whey and Casein Work Hand in Hand for Protein Anabolism, but Scientists Overlook Fat, When They Reassemble Milk"), while the higher leucine content and faster digestibility render whey the better candidate for classic "supplementation", as in having an additional shake before you start preparing your whole-foods post-workout meal, which should - and I hope it's not really necessary that I say that - obviously include a significant amount of protein (fish, eggs, meats, and if you will even more dairy ;-), as well. The usefulness (again, not necessarily the superiority!)  of slow digesting protein is something you should be aware of, anyway, right? If not re-read the "3.2kg of Lean Mass Over Night W/ 40g of Slow Digesting Protein 30min Before Bed!?" post from February 22, 2012.

References:

• Adechian S, Balage M, Remond D, Migné C, Quignard-Boulange A, Marset-Baglieri A, Rousset S, Boirie Y, Gaudichon C, Dardevet D, Mosoni L. Protein feeding pattern, casein feeding or milk soluble protein feeding did not change the evolution of body composition during a short-term weight loss program. Am J Physiol Endocrinol Metab. 2012 Aug 14.
• 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.
• Hickman IJ, Whitehead JP. Structure, signalling and physiologic role of adiponectin - dietary and exercise-related variations. Curr Med Chem. 2012 Aug 9.
• Hoppe C, Mølgaard C, Dalum C, Vaag A, Michaelsen KF. Differential effects of casein versus whey on fasting plasma levels of insulin, IGF-1 and IGF-1/IGFBP-3: results from a randomized 7-day supplementation study in prepubertal boys. Eur J Clin Nutr. 2009 Sep;63(9):1076-83. 
• Lacroix M, Bos C, Léonil J, Airinei G, Luengo C, Daré S, Benamouzig R, Fouillet H, Fauquant J, Tomé D, Gaudichon C. Compared with casein or total milk protein, digestion of milk soluble proteins is too rapid to sustain the anabolic postprandial amino acid requirement. Am J Clin Nutr. 2006 Nov;84(5):1070-9.
• Li FY, Lam KS, Xu A. Therapeutic perspectives for adiponectin: an update. Curr Med Chem. 2012 Aug 9.
• Loenneke JP, Wilson JM, Manninen AH, Wray ME, Barnes JT, Pujol TJ. Quality protein intake is inversely related with abdominal fat. Nutr Metab (Lond). 2012 Jan 27;9(1):5. 
• Rizkalla SW, Prifti E, Cotillard A, Pelloux V, Rouault C, Allouche R, Laromiguière M, Kong L, Darakhshan F, Massiera F, Clement K. Differential effects of macronutrient content in 2 energy-restricted diets on cardiovascular risk factors and adipose tissue cell size in moderately obese individuals: a randomized controlled trial. Am J Clin Nutr. 2012 Jan;95(1):49-63.
• Skurk T, Alberti-Huber C, Herder C, Hauner H. Relationship between adipocyte size and adipokine expression and secretion. J Clin Endocrinol Metab. 2007 Mar;92(3):1023-33.
• Vatani DS, Golzar FA. Changes in Antioxidant Status and Cardiovascular Risk Factors of Overweight Young Men after Six Weeks Supplementation of Whey Protein Isolate and Resistance Training. Appetite. 2012 Aug 10.

Foam Rolling For Cardiovascular Health. Propolis For Muscle Protection. Melatonin For Insulin Sensitivity. Plus: Bananas, Pineapples, Oranges & Beer to Boost Your Melatonin Levels!

Among 18-35 year-old Canadians 80% present with premature hardening of the arteries that could potentially lead to an ischaemic event (LLSA).

Despite the fact that it ain't Saturday, yet, I decided to compile a couple of short news for you, also to make sure you re-appreciate the value of melatonin as a prerequisite to be and stay healthy and beautiful. It should therefore not come as a surprise to you that the SuppVersity Figure of the Week comes from the exact same context.

The exact serum value is 51.5 pg/ml melatonin and it is the cut-off point for decreased nocturnal melatonin as a significant marker of an increased risk of stroke (Atanassova. 2009). Accordingly, every 1.0 pg/ml  decrease in melatonin would be associated with a >2% increase in stroke risk with a 50% risk suffering from an ischaemic stroke for people of all ages with a 3 a.m. value for melatonin of approx. 25pg/ml.

 Foam roll yourself towards a healthier cardiovascular system

Who would have thought that: Foam rolling is good for something beside making fun of it! And if the results of the recent study from Nippon Sports University are applicable to everyone, it could be something as vital as the function of our arteries that will benefit from foam rolling the the adductor, hamstrings, quadriceps, iliotibial band and trapezius.

The Japanese scientists had a group of  seven men and three women (age, 19.9 ± 0.3 y; height, 162.7 ± 8.1 cm; weight, 60.6 ± 11.2 kg, means ± SD), all healthy and free of any overt chronic disease performed the self-myofascial release (SMR) with a 15 × 91-cm (diameter × length) uniform polystyrene roller.

Figure 1: Acute brachial-ankle pulse wave velocity and plasma NO concentration in healthy individuals before and after foam rolling aka self-myofascial release (Okamuto. 2013)

The upper and lower extremities and the trunk were moved across the roller, pressure (direct force) was directed at the lower sacrum, mid thoracic spine and posterior head.

"Pressure was adjusted by applying body weight to the roller and using the hands and feet to offset weight as required. The roller was placed under the target tissue area and the body was moved back and forth across the roller. Briefly, to accomplish SMR of the adductor, the thigh is extended and the roller is placed in the groin region with body prone on the floor. For SMR of the hamstrings, the lower extremities are extended and the roller is placed on the hamstrings with the hips unsupported.

For SMR of the quadriceps, the thigh is extended and the roller is placed on the quadriceps with the body prone on the floor. For SMR of the iliotibial band, the roller is placed on the iliotibial band with the body lateral on the floor.

You don't feel comfortable foam rolling? It's awkward? Yes, it is, but that's probably something you will get used to.

For SMR of the upper back, the hands are placed behind the head and the roller is positioned on the trapezius with the hips unsupported. The head is maintained in a neutral position with the ears and shoulders aligned. The bottom leg is raised slightly off floor. The hips are raised until they are unsupported and the head is stabilized in the neutral position.

The SMR proceeded in the order of adductors, hamstrings, quadriceps, iliotibial band and trapezius.

Each participant practiced two or three times to learn the correct foam rolling technique with the guidance of a trainer and performed 20 SMR repetitions on each muscle group at 1-min intervals." (Okamuto. 2013)

As you can see this protocol is a pretty time-consuming undertaking, which was however rewarded with significant

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Bottom line: With this being the first study to examine the effects of foam rolling or to say it the sexy way "Self-Myofascial Release" we cannot tell whether or not the benficial effects on brachial-ankle pulse wave velocity and plasma NO concentration are (a) replicable in sick and old people who would certainly benefit to a greater extent than the healthy young adults in the study at hand, and whether these changes would (b) persist / reoccur, if you practiced foam-rolling on a regular basis. Therefore Oakmuto et al. may be right that their results suggest that "repeated long-term SMR might decrease baseline arterial stiffness" (Okamuto. 2013), but if you had the choice of taking a walk and foam rolling for 30min, I bet the former will be WAY superior to waddling around with higgledy-piggledy 100% paleo incompatible polystyrene ;-)

More evidence that propolis could promote muscle gain - at least in the elderly

With all the rage about protein anabolism we often tend to overlook that pumping air into a balloon with hundreds of holes in it is not the wisest thing to do... or to say it in plain English, we tend to underestimate the importance of anti-catabolic substances. And while this is particularly true for elderly individuals, in whom the catabolic side effects of exercise tend to eat away their gains, even younger folks could benefit from a reduction in the as of now exclusively detrimental effects of TNF-alpha.

Learn more about inflammation and skeletal muscle hypertrophy

A recent study from nowhere else than the Department of Chinese Martial Arts at the Chinese Culture University did now elucidate that the provision of 5 and 10 mg/kg of caffeic acid phenethyl ester (CAPE, HED 50-150mg per day), a phenolic compound isolated from propolis (the honey beehive propolis, not the fungus) effectively blunts the inflammatory assault in response to downhill running and could thus "protect [...] against eccentric exercise-induced skeletal muscle damage" and avert (a) over exertion and (b) potential atrophy (Shen. 2013).

It is however problematic that the TNF-alpha decline goes hand in hand with concomittant declines in OX2, iNOS, and production of IL-1β and MCP-1, all of which have been implicated as an integral part of the beneficial adaptive response to exercise. Genetic ablation of the MCP-1 gene for example lead to persistent macrophage accumulation in association with residual necrotic tissue and impaired muscle regeneration (Shireman. 2007).

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Bottom line: As mentioned before a propolis supplement is probably more beneficial for older people or other trainees with health conditions that make them more susceptible to skeletal muscle catabolism. Whether this will be beneficial in young people is yet about as questionable as the whole COX-inhibitor for or against muscle growth conundrum, of which Trappe et al. wrote in a very recent review that they probably won't hamper muscular adaption in young and middle aged individuals and showed some promise in promoting the adaptation process in the older guys and gals (Trappe. 2013). For both "regular" COX inhibitors like aspirin, but also "exotic" supplements like propolis extracts it may thus depend on both timing & dosage  and age / baseline inflammation & ability to cope with it  - whether or not supplementing with respective compounds makes sense.

Melatonin supplementation doubles insulin sensitivity in old obese rodents

I know another rodent study, but if you look at the sleep problems your own older relatives have and the way similar patterns have become prevalent in ever-increasing parts of the younger generation, as well, even the mere possibility that the 2.1x increase in insulin sensitivity researchers fro the Department of Physiology and Biophysics at the Institute of Biomedical Sciences of the University of São Paulo describe in their soon-to-be published paper would warrant an N=1 experiment with melatonin (Zanuto. 2013). mesenchymal stem cells from oxidative damage and can be temporarily doubled by simply consuming tropical fruits even (pineapple < orange < banana, see figure 2; Sae-Teaw. 2012)

Figure 2: KITT–Glucose disappearance rate (%/min), weight change from week 0, periepididimal fat weight (left; Zanuto. 2013); effect of fruit extract from 1kg pineapples, 1kg oranges or two ripe bananas on serum melatonin in 12 healthy male healthy volunteers (left; Sae-Teaw. 2012)

This is all the more true, in view of the fact that the pineal hormone, of which you do find 60-170µg/l in beer (with higher values in beers with high alcohol content; cf. Garcia-Moreno ), has also been shown to protect rodents from ischaemic heart disease (at high doses of ~1.5mg/kg ; cf. Sehirli. 2013) and to be inversely associated with the incidence of stroke in humans (Atanassova. 2009).

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Bottom line: Don't discount the value of natural and supplemental melatonin. If it was patentable it would maybe be the #1 prescription drug in the stressful an restless world of the 21st century. You think that's an exaggeration? Maybe, but if you just check out the few SuppVersity news on the matter, you will realize that it's only a minor exaggeration.

And in case you don't want to supplement. A decent amount of sleep, some testosterone boosting high alcohol beer as a "Post Workout Booze" (not really recommended) and the insulin sensitizing "6x Bananas à Day Protocol" you can also boost it the natural way ;-)

References:

• Atanassova PA, Terzieva DD, Dimitrov BD. Impaired nocturnal melatonin in acute phase of ischaemic stroke: cross-sectional matched case-control analysis. J Neuroendocrinol. 2009 Jul;21(7):657-63.
• Garcia-Moreno H, Calvo J, Maldonado M. High levels of melatonin generated during the brewing process. J Pineal Res. 2012 Aug 1.
• LLSA - Stroke at a Young Ag. < http://www.lifelinescreening.com/health-updates/healthy-you/stroke/stroke-at-a-young-age.aspx > retrieved April 12, 2013.
• Liu X, Gong Y, Xiong K, Ye Y, Xiong Y, Zhuang Z, Luo Y, Jiang Q, He F. Melatonin mediates protective effects on inflammatory response induced by interleukin-1 beta in human mesenchymal stem cells. J Pineal Res. 2013 Jan 30.
• Okamoto T, Masuhara M, Ikuta K. Self-Myofascial Release and Arterial Function. Journal of Strength and Conditioning Research. April 2013 [ahead of print]
• Sae-Teaw M, Johns J, Johns NP, Subongkot S. Serum melatonin levels and antioxidant capacities after consumption of pineapple, orange, or banana by healthy male volunteers. J Pineal Res. 2012 Oct 5.
• Sehirli AO, Koyun D, Tetik S, Ozsavcı D, Yiğiner O, Cetinel S, Tok OE, Kaya Z, Akkiprik M, Kılıç E, Sener G. Melatonin protects against ischemic heart failure in rats. J Pineal Res. 2013 Mar 15.
• Shen YC, Yen JC, Liou KT. Ameliorative Effects of Caffeic Acid Phenethyl Ester on an Eccentric Exercise-Induced Skeletal Muscle Injury by Down-Regulating NF-κB Mediated Inflammation. Pharmacology. 2013 Apr 3;91(3-4):219-228. 
• Shireman PK, Contreras-Shannon V, Ochoa O, Karia BP, Michalek JE, McManus LM. MCP-1 deficiency causes altered inflammation with impaired skeletal muscle regeneration. J Leukoc Biol. 2007 Mar;81(3):775-85.
• Trappe TA, Liu SZ. Effects of Prostaglandins and COX Inhibiting Drugs on Skeletal Muscle Adaptations to Exercise. J Appl Physiol. 2013 Mar 28. 
• Zanuto R, Siqueira-Filho MA, Caperuto LC, Bacurau RF, Hirata E, Peliciari-Garcia RA, do Amaral FG, Marçal AC, Ribeiro LM, Camporez JP, Carpinelli AR, Bordin S, Cipolla-Neto J, Carvalho CR. Melatonin improves insulin sensitivity independently of weight loss in old obese rats. J Pineal Res. 2013 Mar 30.