Santa is Coming to Town and You Better Beware of His Gifts: Fat Gain, Muscle Loss and Increased Mortality Rates.

Image 1: The "modern" image of the Coke-drinking Santa. Do you really believe he is one of the good guys?

Finally, December 24th is there! The day we have all been waiting for, to get together with friends and relatives and wait for the portly, joyous, white-bearded man in the red coat to deliver "his" gifts. Interestingly enough, the word "gift" in German designates "poison" and while those of you who have been following the SuppVersity news earlier this week may now be speculating that this could in one way or another be related to the millions of iPhones Santa is going to be dropping down the chimneys in the night to come (cf. Mobile Contraception), it seems unlikely that the electromagnetic radiation from the gadgetry could explain the statistically significant +4.65% increase in cardiac and a + 4.99% increase in non-cardiac deaths during the holiday season. After all, the data based on which David P. Phillips, Jason R. Jarvinen, Ian S. Abramson, and Rosalie R. Phillips conclude that "the Christmas/New Year’s holidays are a risk factor for cardiac and noncardiac mortality" is from the pre-iPhone era (Phillips. 2004).

Is Santa not the good guy, the Coca Cola ads made us believe?

So, if its not the radiation, what else could it be? Could it be Santa Claus himself? Is he haunting us, just as his robotic counterfeit in the distant future of the year 2999, where an evil Santa robot is after the blood of the protagonists of Matt Groening's and David X. Cohens TV series Futurama? Or is it a result of the consumption of too many of the Coca Cola bottles Santa is supposed to have in his bag?

Image 2: One really has to marvel at how the soft drink producers dissolve the enormous amount of sugar on the right in the small amount of dark brew on the left.

Did you know that the Coca Cola company alone sells 1.6billion (!) servings of Coke per day? With 27g of sugar per serving, this equals 43,000 metric tons of pure sugar. The average American, who consumes an average of 150 to 170 pounds of sugar each year, would have to live into his/her 558th year of age to eat or drink her way through this sugar mountain. And while I have no doubt that there actually are people out there who would do that withing 100 years, I am not quite sure which of the ailments of our sweet convenience society would strike him / her first and put a spoke in the sweet-o-holic's plans: diabetes, cancer, heart failure or stroke? What would you say?

Phillipps et al. who report in a follow-up study based on the same dataset from the holiday periods between July 1, 1973, and June 30, 2001 that there was an "excess of 42,325 deaths from natural causes above and beyond the normal winter increase" (Phillips. 2010), exclude the possibility that the increased mortality rate was simply a result of the bad weather conditions and related respiratory diseases:

Respiratory diseases. Respiratory diseases increase during winter, and patients weakened by respiratory diseases can die from cardiac diseases. The respiratory hypothesis is undermined by 2 considerations: (1) People dying from cardiac diseases with respiratory disease listed as a secondary cause of death produce a smaller holiday peak than do people dying from cardiac diseases alone: 3.51% versus 3.77%. (2) Interaction between cardiac and respiratory diseases cannot easily explain the twin mortality spikes on Christmas and New Year’s.

So, in view of the latest headlines related to "holiday weight gain" here at the SuppVersity and elsewhere on the web, the next best plausible explanation (which would in fact come back to the "Coca Cola < > Santa Connection" ;-) would be gluttony, right?

Holiday weight gain: Distinguishing fact from fiction

Before we jump to any premature conclusions, here, let's initially have a closer look at how much body weight Santa actually has in his bag for you.I mean, the perceived weight gain is enormous, right? Well, science is however not about perceptions and feelings and it should thusly not really surprise you that, according to a US study which was published in the prestigious New England Journal of Medicine (Yanowski. 2000), the "average" American (in this study represented by 195 US adults with a mean age of 39 +/-12 years) gains no more than 0.37kg, or, expressed in terms of the mean weight of the study participants, 0.5% during the holiday period from from mid-November to early or mid-January.

Figure 1: Percentage of normal weight, overweight and obese subjects with "major weight gain", as defined in absolute or relative terms (data adapted from Yanowski. 2000)

And while the average weight gain hardly is something to speak of, there are two other particularly intriguing findings of this study I do want to draw your attention to. The first one relates to the the data in figure 1. As you can see, the number of overweight subjects among those study participants with major weight gain (as defined as >3% of the initial weight) is particularly high. While only 7.9% of the normal-weight (American normal weight ;-) subjects gained more than 3% of their initial body weight 11.1% of the already overweight subjects did. Interestingly, the number of obese subjects was slightly smaller (7.5%). The latter is yet a physical necessity as there simply is a phyiscal limit to the amount of weight you can gain in a given period of time and 3% for a person with BMI>30 is obviously way more than 3% for someone who is only "overweight" (25 < BMI < 30).

The real problem is: The weight does not magically disappear

The real culprit is however that the weight people gain during last weeks of the year "is not reversed during
the spring and summer months", so that he researchers' concern that

[t]he 0.48-kg weight gain of the subjects in this study between September or October and February or March might not appear to be  clinically important and could easily go unnoticed by both the subjects and health care providers [and that] the cumulative effects of yearly weight gain during the fall and winter are likely to contribute to the substantial increase in body weight that frequently occurs during adulthood.

A 2006 by Hull may not only provide a hypothetical explanation for the non-reversibility of the (minor) weight gain (Hull. 2006), it also provides some insights into the true fallacy of "holiday weight gain": The minor increase in total body weight goes at the expense of concomittant increases in body fat and reductions in lean tissue mass.

Figure 2: Relative changes in anthroprometric measures over the holiday season; left axis - overweight / normal weight, right axis + figures - all (data adapted from Hull. 2006)

In the 82 college students from the Hull study, this fat promoting, muscle reducing "recompositioning" effect of the holiday season (Thanksgiving to New Year) was even so pronounced that the study participants actually lost -0.1kg of their total body weight. This was unfortunately a direct result of a +0.8 increase in fat mass and a -0.4kg decrease in lean mass. And what's more, the effect on fat mass was again more pronounced in those subjects, who were already obese.

Beyond candy, coke & co: Five additional reasons why Christmas is potentially deadly

In spite of the fact that these highly unfavorable changes in body composition are certainly not beneficial for anyone's overall health, it stands out of question that their effects would be cumulative and can thusly hardly explain the empirically validated increased mortality risk during the holiday season. In a 2004 comment on the aforementioned paper by Phillips et. al., Robert A. Kloner thusly proposes five additional hypotheses which could explain the potentially fatal side effects of the holiday season (Kloner. 2004):

Image 3: If you do not want to be treated by "beginners" and unexperienced hospital personnel you'd better not get sick over the holidays; and in case you do, please make sure to "postpone your death" in order not to ruin everyone's holidays ;-)

1. Inappropriate delay in seeking medical attention - best way out: don't wait until all the presents have been wrapped out, when aunt Mary chokes over her food
2. Reduced levels of healthcare staffing or fewer staff members who are familiar with individual patients during holiday on-call schedules - best way out: better avoid getting sick in the first place if you do not want to be treated by the SCRUBS staff
3. Increased emotional stress - just ignore your nephew when he starts crying because he did not get the Nintendo Wii he wrote on his wish list
4. Decreased our of daylight - make sure to get as much of the little light there is during prolonged walks with the whole family (may also help cool down any raised tempers ;-)
5. "Postponement of death" - tell your 127 year old uncle that he has been waiting so long now that it would be very inappropriate to die now and ruin everyones' Christmas celebrations

Well, I guess, now that you know about all the terrible things that could happen and the best ways to avoid them, it is about time to wish you, your family, friends and loved ones a happy (death-free) holiday season! And in case you need a break from the festivities, there is no Christmas break, here it at the SuppVersity ;-)

What's the Optimal Dose of Vitamin D3 for Lean, Normal-, Overweight & Obese Women With Established Vitamin D Deficiency to Get 25OHD Back into the Normal Range?

Both ladies are D-ficient, but will probably need profoundly different amounts of D3 to get their 25OHD back in range.

Actually, I guess, I don't really have to tell you that there is not going to be guest post by Adelfo Cerame, today. Adelfo is busy with the last weeks of school, but will be back as soon as he has passed all the tests. And while I am not sure, whether or not you would call the latest on vitamin D supplementation an adequate replacement for a contest prep update from "your's truly", I suppose that it's better than nothing to bridge the time that still remains until the SuppVersity  Science Round-Up on the Super Human Radio Network is going to air (the show starts at 12PM, EST; the Science-Round-Up airs in the 2nd hour and will thus begin at 1PM, EST; click here to listen live or wait for the podcast // update: now available).

I am honestly not yet sure what exactly we will cover today, but among the things I am still thinking about how we can squeeze them into a 1h show are...

• methylxanthines caffeine, theobromine and theophylline can bind to human DNA - what does that tell us about the purported health benefits of caffeine & co?
• caffeine prevents memory impairment - in this case in a model of sporadic Alzheimer's disease
• anti-Alzheimer's effect of CLA - plus a list of supplements that have been implicated in the prevention of Alzheimer's and other amyloid diseases such as Parkinson's, Cerebellar Ataxis, Amyotrophic lateral sclerosis and (hardly recognized as an amyloid disease) diabetes type II
• the effect of body weight on the benefits of circuit training in older women - turns out that those who need it the most, namely the obese, also see the greatest benefits
• Gum arabicum to ward off holiday weight gain - that this could actually work is at least what a recent human study would suggest
• more on vitamin E, resveratrol, soldiers don't get hurt in battle, but by geranium (DMAA), ...

I think there should be something for everyone of you. Plus: If everything works out, this is going to be the first show to air live via Skype, so no nagging land line echoes and noise any more.

Let's get to the D-news, now

The general consensus among the vitamin D advocates currently is that 2,000 IU of vitamin D3/day is the minimum you need to bring low levels of 25OHD back into the normal range. A soon-to-be-published study by Gallagher, Yalamanchili and Smith that's available ahead of print on the website of the Journal of Steroid Biochemistry and Molecular Biology does yet contradict this notion - at least for women with a body mass <25kg/m² even the meager RDA of 400IU would be enough (Gallagher. 2012). That said the concise paper actually describes the results of two, not just one experiment, with

• study 1 (ViDOS) being a one-year randomized, double-blind placebo controlled study (ViDOS – Vitamin D supplementation in Older Subjects) of increasing doses of vitamin D3 (400,  800, 1600, 2400, 3200, 4000 or 4800 IU/day vitamin D3 vs. placebo + calcium supplements to maintain calcium intake between 1,200-1,400mg/day) in 163 Caucasians, age 57–90 years; all vitamin D insufficienty, i.e. serum 25OHD ≤ 20 ng/ml (50 nmol/l), and 
• study 2 (STOP IT) being a 3-year intervention study of calcitriol 0.25 mcg (the active form of vitamin D) twice daily, conjugated estrogens 0.625 mg  daily, a combination of both and placebo in 488 elderly women, age 65–77 years

Body composition indices for the studies at hand (i.e. percentages of total and regional fat and fat-free mass) were measured by dual energy X-ray absorptiometry (DEXA Hologic Delphi) at baseline and after 12 months.

Figure 1: Mean total body weight, total body lean mass, total body fat mass and serum 25OHD in different BMI subgroups of study 2 (STOPIT); right, corresponding calculated ratios (based on Gallagher. 2012).

Even the baseline data in figure 1 does actually yield some insights into the relation of BMI, adiposity and 25OHD levels. While the data on the left already shows that the fat mass increases almost linearly across the BMI levels, while the lean mass remains relatively stable (with the highest value in the overweight group, though), the ratios I calculated and plotted on the right-hand side of figure 1 make it even more obvious clear: The lean / fat mass ratio scales with the BMI. With identical levels in the normal- and overweight individuals and significant increases and declines in the lightest and heaviest study participants. Moreover, the 25OHD vitamin D to fat mass ratio drops most significantly between the low BMI and the upper normal zone, where I suppose even most of the "healthy" individuals will be hovering around these days.

Being lean is a positive predictor of increases in 25OHD with supplementation

That this latent "chubbiness" of the average Westerner may be of particular significance in view of the negative / non-significant outcomes in many of the vitamin D supplementation trials, becomes self-evident, when you take a closer look at the data in figure 2, however you will have to realize that my plot which comprises above all the highly relevant relative changes (middle, marked in red) tells a different story than the original plot from the study showing only the absolute changes (left, but in form of a line graph).

Absolute, relative (compared to baseline) changes and total 25OHD levels (ng/ml) after supplementation with low, medium and high amounts of vitamin D3 in lean, normal, overweight and obese women (based on Gallagher. 2012)

Accordingly, the conclusion of the abstract, which says that "the response to vitamin D is dependent on body weight" and that "women with BMI <25 kg/m² develop much higher levels of serum 25OHD after vitamin D supplementation compared to those with BMI of >25 kg/m²" (Gallagher. 2012) may be correct, but is somewhat misleading as it is open to be interpreted as 'lean women respond most favorably to vitamin D supplementation' - an interpretation that is not really sustainable in view of the relative changes I calculated for figure 2  (middle), yet by no means as incredible as the abstract of another vitamin D study, I dessicated back in September (see "Stronger & Leaner or Fatter & Less Muscular W/ 4,000IU Vitamin D3 - What if Abstract and Data Tell Different Stories?")

Bottom line: The data from this most recent investigation into the differential response of lean, normal, overweight and obese women to vitamin D3 supplementation shows that the absolute increases appear on BMI and that...

• 

  Always take vitamin D with fatty foods! (see "A Fat D-Ficiency")

  low dose supplementation (400 or 800IU/day) is probably only sufficient to rise and maintain adequate vitamin D levels in lean women,
• medium dose supplementation (1,400 or 2,400IU/day) yields the most favorable outcomes in total 25OHD levels and 
• high dose supplementation (3,200, 4,000 or 4,800IU/day) does not yield additional benefits in either the the normal-, overweight and obese subgroup and only marginally higher levels in the lean women.

Overall the study at hand would thus support the notion that a daily vitamin D supplement containing ~2,000IU is the best way to get deficient levels back up, esp. for lean women it should be no problem to cut back to 2x the RDA, i.e. 800IU after normal vitamin D levels are achieved. For the rest, future studies will have to show if low dose supplementation is enough.

These longissimus dorsi slices of mice on a normal and a vitamin D3 supplemented diet show that supplemental vitamin D3 can be used as a fat synthesizer and meat tenderizer in "meat-producing animals". (learn more)

The often-heard hypothesis that the decreased response to vitamin D supplementation in the obese would be a result of the preferential storage of vitamin D in the adipose tissue was not supported by data of the Ghallagher study "there is no evidence from the dose response curves that in obesity serum 25OHD is being deposited in fat" (Gallagher. 2012). In view of the fact that contrary to total vitamin D, which is in fact preferentially stored in adipose tissue (78%) over lean muscle (14%), 25OHD stores are distributed much more evenly with 33% being stored in body fat and 20% in muscle tissue in omnivores like humans and swine (the data is in fact based on a study in pigs; cf. Jakobsen. 2007).

Lastly, a beneficial effect of increase / normalized vitamin D levels on lean or fat mass was (once again) not observed in any of the studies; and that despite the fact that "body fat was an independent predictor of serum PTH", which decreased in response to calcitriol supplementation in study 2 (which is actually more of an adjunct for correlative analysis and as a data source to compare the results of study 1 to). In other words, normalizing your vitamin D levels without taking appropriate measures to counter what's probably behind both, the nasty body fat and the low vitamin D level is not going to make you lean or musclar - at least as of now, it rather appears as if this was yet another instance, where we are - if anything - treating isolated symptoms instead of the root causes of the obesity epidemic.

References

• Gallagher JC, Yalamanchili V, Smith LM. The Effect Of Vitamin D Supplementation On Serum 25OHD In Thin And Obese Women. J Steroid Biochem Mol Biol. 2012 Dec 11.
• Jakobsen H, Maribo A, Bysted HM, Sommer OH. 25-Hydroxyvitamin D3 affects vitamin D status similar to vitamin D3 in pigs – but the meat produced has a lower content of vitamin D. British Journal of Nutrition. 2007; 98 908–913.
• Shephard RJ. Limits to the measurement of habitual physical activity by questionnaires. Br J Sports Med. 2003 Jun;37(3):197-206; discussion 206.

Weight Loss Supplements Exposed: Green Tea & Probiotics. Fat Loss, Energy Expenditure, Fat Oxidation, Sex & More

Yesterday at Starbucks: "I just ordered a bottle of probiotics!"

In view of the fact that all the feedback I got in response to the re-installment of the Short News was positive, I guess you won't mind if I use the chance and bundle the two soon-to-be-published weight loss studies from the British Journal of Nutrition into a Weight Loss Supplement Mini-Special of the SuppVersity Short News.

If you were actually sitting next to you, I would probably ask you, whether you'd prefer the good, or the bad news, first!? Well, I guess I'll start with the bad one, then: Green tea sucked - again!

ZERO effect of EGCG supplementation in overweight women

To examine the effects of green tea epigallocatechin-3-gallate (EGCG) on the changes in body composition (! not just weight), energy and substrate metabolism, cardiometabolic risk factors and liver function enzymes after an energy-restricted diet intervention in obese women, a group of researchers from the University of the Basque Country in Spain recruited a group of 83(!) obese (BMI 30-40 kg/m2) pre-menopausal women (Mielgo-Ayuso. 2013).

The women were randomly assigned to consume either 3x100 mg/d of EGCG or placebo (lactose) with each of their three main meals for 12 whole weeks. During those twelve weeks, all women followed a specifically designed low-energy mixed (55 % carbohydrates, 30 % lipids and 15 % proteins) diet that provided ca. 600 kcal/day energy less than the women would need to maintain their body weight. The energy content and macronutrient composition of diets were designed to achieve a weight loss of 0.5 to 1 kg per week, as it was observed by Davis et al. (2006) and Bantle et al. (2008) on very similar regimen. As the scientists point out, the "dietary instructions were reinforced weekly by a dietitian" (Mielgo-Ayuso), to optimise compliance.

Figure 1: Changes in body composition, energy expenditure and fat oxidation, left; changes in glucose, cholesterol metabolism and inflammation, right (Mielgo-Ayuso. 2013)

I am not sure how compliant the participants actually were, but in view of the fact that the women were advised not to change their physical activity habits during the energy restriction program, the relatively meager and statistically non-significant changes in body weight (-0·3 kg, p > 0.05) and fat mass (-0·7 kg, p > 0.05) are probably not really surprising. It is nice to see, though, that the women lost more fat than total mass - muscle loss was thus not an issue for the ladies.

What was not to be expected, though, - at least if you believe a single word of the hype about green tea supplements - were the non-existent effects of the purported weight loss supplement on  energy expenditure, fat metabolism, HOMA-IR (insulin sensitivity), total cholesterol, LDL-cholesterol, or triglycerides. In fact, the only good thing about the whole EGCG intervention was that the recently observed negative effects on the liver did not occur, either.

SIGNIFICANT Effect W/ 16 Million CFU of Nestlé's Lactobacillus rhamnosus strain

Want to check out the patent?

Despite the fact that the overall results are much more exciting than those in the previously discussed green tea study, I'd advise you to keep calm. We are after all dealing with another Nestlé study on a patented strain of Lactobacillus rhamnosus (LPR), i.e. "CGMCC1.3724" (date patented: 2012-05-10; #20120114622), and cannot tell how many never published negative study results the Nestlé guys had to dispose of, before Marina Sanchez et al. finally produced study results that pleased the marketing division of this multinational corporation.

What? Ok, ok... let's get back to the facts: The scientists from the Laval University and the Nestlé Research Center randomized a group of one-hundred fifty-three 18 to 55 year-old obese men and women to receive either a placebo or the said LPR formulation with 1·6 × 10⁸ colony-forming units of LPR and additional oligofructose and inulin per cap for a total of 24 weeks.

In the course of the first 12 weeks (phase 1), each participant received a personalised diet plan that would have him or her consume 500 kcal/d less than he or she'd need for weight maintenance (just as an aside, that's 100kcal more than for the subjects in the green teas study). During phase 2, each participant received a personalised diet plan without energy restriction. The good thing, the resting energy expenditure (REE) was actually measured: after a 12 h overnight fast in subjects having had rested for at least 15 min in a standardised supine position. This procedure was repeated thrice: (1) At baseline, (2) after the weight-loss and (3) after the second phase weight-maintenance periods using indirect calorimetry.

Figure 2: Changes in body composition (all data in kg) in men (left, blue) and women (right, orange) after weight loss (ΔW12) and weight maintenance (ΔW24) phase (Sanchez. 2013)

The data in figure 2 confirms what the abstract says: "The intention-to-treat analysis showed that after the first 12 weeks and after 24 weeks, mean weight loss was not significantly different between the LPR and placebo groups when all the subjects were considered."

Figure 3: Changes in metabolic parameters, i.e. energy intake (kcal/day), resting energy expenditure (REE, kcal/day) and respiratory quotient (RQ, remember: low RQ = high fat, low carb oxidation) after 12 and 24 weeks (Sanchez. 2013)

It does yet also confirm - and that there was a significant treatment × sex interaction, observed with the women in the treatment group losing significantly more weight than those in the placebo group (P= 0·02). More importantly, though...

"[...w]omen in the LPR group continued to lose body weight and fat mass during the weight-maintenance period, whereas opposite changes were observed in the placebo group."

For the unlucky men, on the other hand, the (unquestionably expensive) supplement didn't do sh*t: Their "changes in body weight and fat mass during the weight-maintenance period were similar" irrespective of whether they received the placebo or the active treatment.

Whether this was the reason or a consequence of the fact that the the men didn't show similar significant reductions in circulating leptin, as the women is questionable. Based on the fact that the relative abundance of bacteria of the Lachnospiraceae family in faeces increase only in women, we do yet have to assume that the missing reduction in leptin, as well as the absence of the significant body fat reductions, the researchers observed in their female subjects was simply a results of ...

• under-dosing - the same the 1·6 × 10⁸ colony-forming units of LPR that was sufficient for the average woman (body weight ~89kg) could have been too low for the guys (body weight ~104.3kg) 
• dietary interference - there could have been something in the diets of the guys that ruined the effects of the supplementation (lactobacilli are not exactly friends of meats and we all know that men love their meat ;-)
• different baseline gut microbiome - it goes without saying that you cannot place a group of rabbits in forest full of predators and expect them to survive; similarly the LPR spores may have come off second in the guts of the men, because they have a less "LPR-friendly" baseline colinization
• fundamental sex differences - at the moment I am not sure what the underlying reasons could be, but it's not impossible that hormonal difference could have played a role as well

I am pretty sure that I could come up with a whole host of additional, increasingly bizarre ad-hoc explanations for the null-effect Marina Sanchez and her colleagues from the Laval University and the  Nestlé Research Center in Lausanne observed in their male study but would rather conclude this news-item with the scientists own funky, but not unlikely explanation: Men are simply too good at dieting!

True: Women have a harder time losing weight even with high protein | more

As the authors point out, we know from previous trials (and corresponding SuppVersity posts, read more) that men are generally more prone to respond to a negative-energy balance intervention than women - and that's true irrespective of whether it is an exercise-training programme (Tremblay. 1984), a diet– exercise programme (Doucet. 1999), or a session of exercise and of mental work (Pérusse-Lachance. 2013). Plus, if you look at the data in figure 2, you'll see that this is actualy "concordant with the results of the present study that shows higher weight loss in men in the placebo group than in the women". Sanchez et al. do now believe that the high baseline success "abolished this difference" (Sanchez. 2013).

In view of the fact that there was a difference in a single low-abundance taxonomic group  (Prevotellaceae) between the baseline gut microbiome of the male and female study participants, I would still not exclude that the different baseline gut microbiomes could at least have added to the 'effect abolishing effect' of the sex-specific ease of weight loss in men. I mean, why wouldn't the feces of the men show an increase in lactobacillus spores, if the supplement worked?

Bottom line: Today's installment of the short news is very characteristic of the dilemma with weight loss supplements. We are just realizing that the classic thermogenic 'rodent fat burner' don't really work in humans. Against that background the rise of supplements that target the gut microbiome and exert much more complex body recompositioning effects comes in the nick of time.  Unfortunately, our understanding of the complex interactions between the gut microbiome and our immune system in the context of the emerging science of immunonometabolism is so incomplete (Mathis. 2011) that we are more or less groping in the dark, whenever we supplement subjects, patients or even ourselves with allegedly healthful bacteria.

All alleged benefits aside,  "specificity", the 2nd Principle of Sensible Supplementation, should keep you away from the next best GNC or online supplement store. The two studies at hand do after all not warrant the use of either green tea or lactobacillus supplements as weight loss aids in lean, healthy and active  men or women.

Accordingly, the observation that green tea supplements won't help sedentary over-weight women to lose weight appears to be much more reliable than the allegedly impressive weight loss effects of the probiotic during the "maintenance phase" of the Sanchez study.

We must however not forget the respective constraints of the research design and irresponsibly over-interpret the results of the EGCG study to (a) the potential benefits of regular 'whole' tea consumption in the average, non-obese individual (Wu. 2003) or (b) visceral fat loss in diet + exercise interventions in obese individuals (cf. Maki. 2009). Similarly, the fact that obese women will lose weight on a LPR supplemented maintenance diet is very unlikely going to translate to lean, athletic folks like you and me. According to the 2nd Principle of Sensible Supplementation, which is "specificity" (learn them all), I don't see you or me heading over to the next best online shop to buy LPR or EGCG supplements - irrespective of the promising results of the Sanchez trial.

References: 

• Bantle JP, Wylie-Rosett J, Albright AL,et al.(2008) Nutrition recommendations and interventions for diabetes: a position statement of the American Diabetes Association. Diabetes Care31, Suppl. 1, S61– S78.
• Davis NJ, Emerenini A & Wylie-Rosett J (2006) Obesity management: physician practice patterns and patient preference. Diabetes Educ32, 557 – 561. 
• Maki, K. C., Reeves, M. S., Farmer, M., Yasunaga, K., Matsuo, N., Katsuragi, Y., ... & Cartwright, Y. (2009). Green tea catechin consumption enhances exercise-induced abdominal fat loss in overweight and obese adults. The Journal of nutrition, 139(2), 264-270.
• Mathis, D., & Shoelson, S. E. (2011). Immunometabolism: an emerging frontier. Nature Reviews Immunology, 11(2), 81-83.
• Mielgo-Ayuso J, Barrenechea L, Alcorta P, Larrarte E, Margareto J & Labayen I (2013). Effects of dietary supplementation with epigallocatechin-3-gallate on weight loss, energy homeostasis, cardiometabolic risk factors and liver function in obese women: randomised, double-blind, placebo-controlled clinical trial. British Journal of Nutrition, available on CJO2013. 
• Tremblay, A., Despres, J. P., Leblanc, C., & Bouchard, C. (1984). Sex dimorphism in fat loss in response to exercise-training. Journal of obesity and weight regulation.
• Wu, C.-H., Lu, F.-H., Chang, C.-S., Chang, T.-C., Wang, R.-H. and Chang, C.-J. (2003), Relationship among Habitual Tea Consumption, Percent Body Fat, and Body Fat Distribution. Obesity Research, 11: 1088–1095.

Only Whey, Not Soy Works 'Wheytloss Wonders': Add. 60g+ Protein 30 Min Before Ad-Lib Meal Decreases Appetite & Energy Intake, Cuts 9% Body & Adds 18% Muscle in 12 Wks

If you want weight loss support, chose the "Wheytloss Wonder" whey concentrate over the allegedly "healthy plant proteins" in soy isolates - if your physiology works anyway similar to the one of the study subjects you won't regret it - promise!

We all know that whey is wonderful, don't we? And we all know that soy is the devil, right? Ok, I guess both assumptions are not exactly accurate, but when you're having your next discussion with that overweight lady "gone vegan", because it's so good for your waist line, you may point her to a soon-to-be-published study by Atefeh Tahavorgar, Mohammadreza Vafa, Farzad Shidfar, Mahmoodreza Gohari, Iraj Heydari, a group of scientists from the Teheran University of Medical Sciences (Tahavorgar. 2014).

Based on the existing evidence of the weight loss benefits of high(er) protein diets, the Iranian scientists hypothesized that supplemental preloads of whey protein concentrate (WPI) and soy protein isolate (SPI) would decrease appetite, caloric intake, anthropometry, and alter the body composition of healthy overweight and obese men in free living conditions.

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

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To test this hypothesis, we supplemented free living overweight and obese men with WPC and SPI 30 min before their ad libitum afternoon meals, and monitored each subject’s appetite, calorie intake, anthropometry, and body composition.

Using a monthly bulletin to advertise, volunteer employees of a power plant in Karaj city were recruited to participate in the study. Inclusion criteria included: no cigarette smoking and/or alcohol consumption, no medication and/or supplement usage, no high amounts of caffeine consumption (>250-300 mg/d), no history of diseases or clinical problems that increase oxidative stress (injuries or burns), no allergy to soy/cow's milk, and no severe weight changes within the last three months. Exclusion criteria included any changes in physical activities (PA), diets, and a compliance of 70% or lower for consumption of treatment beverages.

This is a randomized, controlled trial, no epidemiological "healthy plant protein guesswork"

At the first visit, eligible participants were randomly assigned to either the WPC or SPI (26 in each) group, using a convenience allocation. Individuals were instructed to deliver empty sachets in exchange for full ones at visits 2 through 12, in order to calculate compliance.

SuppVersity Special: "Dairy - The Good, the Bad or The Ugly? Latest Studies On Heart Disease, Diabetes, Cancer, Obesity and Co. Plus: What Dairy Peptides Do For Your Heart, Gut, Brain, etc." | read more

All participants had ad libitum access to calories and were asked to maintain their usual dietary intake and physical activity.

"Preload proteins included 80% WPC (DMV, Netherlands) and 90% SPI (Red Crown, China), with similar color and texture. Sachets contained 67.5 g WPC and 60 g SPI (54 g effective compound as a protein / sachet).Calorie contents of WPC and SPI sachets were 261.8 Kcal and 216 Kcal, respectively. They were closely matched for taste with strawberry flavor and sucralose (Vita Sweet, China) (0.2 gr and 0.1 gr in each sachet respectively), as a no-energy sweetener since sucralose is not metabolized in the body and has no effect on blood glucose or insulin secretion." (Tahavorgar. 2014)

After packing 4368 similar sachets, they were numbered 1- 84. The numbers were randomly divided into groups A and B (SPI and WPC, respectively) and kept by the executive director of research until study commencement.

One of the strenghts of the study is the fact that dietary intake and physical activity were closely monitored. If the subjects didn't lie about their food intake and activity levels the results of the study at hand are thus highly accurate. Unfortunately, the same cannot be said of the relatively unreliable body fat measurements that were conducted with a body fat monitor by Jawon Medical.

The consumption of the protein shakes (65 gr WPC or 60 gr SPI that was dissolved in 500 ml water) 30 min before the ad libitum dinner (late afternoon) lead to significant increases of the total protein intake in both groups, with a slightly higher total protein intake of 33.5% of the total energy intake (vs. 28.7%) in the whey vs. soy group. No wonder that the mean changes in appetite (p=0.032), CI (p=0.045), anthropometry (body weight (BW) (p=0.008), body mass index (BMI) (p=0.006), and waist circumference (WC), body composition (body fat mass (BFM) and lean muscle (LM) were significant in both groups.

Figure 1: Changes in appetite, calorie intake and body composition during the 12-week study (Tahavorgar. 2014)

If you look at the overall outcomes in Figure 1, it is yet obvious that the appetite reduction and loss of body weight were significantly more pronounced in the whey protein group. More importantly, however, these changes occurred as a consequence of similar absolute, albeit slightly higher relative reductions in energy intake (there is a problem with the data, here, because the pre- vs. post valued differ from the calculated mean difference, if we use the latter, the soy group would have had a greater reduction in energy intake, i.e. 1186kcal vs. 624kcal - that would make the results even more impressive, but somewhat questionable) and trigger significantly more pronounced improvements in body composition than the soy protein isolate, namely increases in lean mass (+18%) and significant reductions in body fat (-9.2%) and waist circumference (-9.7 cm on average vs. +1.1 cm in the soy group) - not too bad, considering there was no "dieting" or training involved.

Please remember: The weight loss and increases in lean mass occured in the absence of a deliberate energy restriction (all subjects still ate "ad libitum", i.e. as much as they wanted) and without the need to train. It was "just" the addition of high quality protein in form of the ~60g of whey protein concentrate (again there is no reason to buy isolates, unless you are lactose intolerant) that did the trick!

Bottom line: The study at hand is only one out of many experimental trials which refute the notion that plant proteins are healthier and eating plant instead of protein from animal sources would help you lose weight and improve your body composition. The latter is a myth that's based on questionable epidemiological data, where confounding factors such as the "pizza salami = meat" factor (i.e. the way the meat intake is estimate) are hard to control.

Since I hope that we all put more faith in hard experimental vs. "soft" epidemiological data, it should be obvious that anyone (including your vegan friends) who is planning to lose body fat and improve his / her body composition is much better off with whey vs. soy protein; and that not just as a replacement for a complete meal, but rather as an addition that increases the total protein content of the diet and reduces the food and energy intake on ad-libitum meals, when the shake is consumed 30 minutes before a meal | Comment on Facebook!

Reference:

• Tahavorgar, Atefeh, et al. "Whey protein preloads are more beneficial than soy protein preloads in regulating appetite, calorie intake, anthropometry, and body composition of overweight and obese men." Nutrition Research (2014).

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.

Female Athletes' Body Composition Suffers From Chronic Energy Deficits: Effects of Energy, Protein, CHO Intake, Timing & Distribution in Gymnasts & Volleyball Players

Even female volleyball players are wo- men - no wonder they tend to undereat ;)

Usually we are learning about what makes us fat by looking at those who are fat. Studies on athletes like gymnasts and volleyball players, and what influences their body composition, on the other hand, are scarce. Reason enough for me to take a closer look at two thesis by graduates from the Georgia State University who analyzed the relationship between moderate, within day protein intake and energy balance on body composition of collegiate sand volleyball players (Richardson. 2014) and the relationship between daily protein distribution and body composition in elite gymnasts (Paszkiewicz. 2014) - research that could be relevant for both, men and women.

I guess many of you will remember that I've written about gymnasts before - in July 2013, to be precise. In said article with the telling title "Do Chronic Energy Deficits Make Athletes Fat? The Longer & More Severe You Starve, the Fatter You Are. Irrespective of What the Calories-in-VS-Calories-Out Formula May Say" (read more) I analyzed the negative effects of "starvation" on body composition to highlight that simply not eating or eating like a bird is not going to give you the Shape cover model body, many girls are looking for.

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In spite of the fact that the titles of the two studies and hand and the previously cited study by Deutz differ, the objectives are not very different:

• "The  purpose  of  this  study  was  to  simultaneously  assess  energy  balance  and
  protein  intake  to  determine  if  these  factors  are  associated  with  body  composition  in  a
  population of collegiate sand volleyball players." (Richardson. 2014)
• "The objective of this study was to determine the relationship between hourly EB and protein intake with body composition" (Paszkiewicz. 2014)

If you look at the exact ways the authors phrase it, it does yet become obvious that Richardson (2014) puts a greater emphasis on the amount of protein, while Paszkiewicz is, just like Deutz back in the day, very interested in the hourly energy balance (EB) and thus the time the subjects remain in a positive / negative energy balance.

Apropos subjects! In the gymnasts who participated in Paszkiewicz' study were elite and highly
competitive athletes from several training gyms across the country. The information on their daily food intakes was elucidated by the means of secondary analyses that were performed on previously collected three-day food diaries and the interactions with body composition were calculated by comparing intakes and anthropometric measures (made with DEXA).

Table 1: 
Subject
 Characteristics of the Gymnasts 
 (N=
40; Paszkiewicz. 2014)


Table 1 provides an overview of the subject characteristics. If you take a closer look, you will see that there is a pretty broad range from hardly any muscle to pretty muscular and from ripped to the shreds to average body fat.

There is one general problem with the "energy balances" in both studies! Being based on the standard equations, they are - at beast - a proximate of what the women really need. For the gymnast study, the difference between energy in and out is yet large enough to safely assume, they were really starving itself. For the volleyball study, I wouldn't be so sure - specifically in view of the fact that the body has its means of sparing energy, when it's chronically getting less than it would need - the corresponding changes in thyroid & other hormones have yet not been studied by either Paszkiewicz or Richardson.

If we take a closer look at the correlations Paszkiewicz found, some of you may be surprised to see that the relative carbohydrate intake (as percent of macronutrients) was not just positively associated with higher lean mass (see Figure 1), but also negatively with fat mass (R = -0.043).

Figure 1: Minimal, maximal and average energy balance in the gymnasts (left); positive correlates and correlation coefficients R of lean mass in 40 elite competitive female gymnasts (Paszkiewicz. 2014)

The amount of protein the gymnasts ate, however, was not significantly associated with increase lean mass. In fact, when we compare two groups, i.e. those with a high and those with a low protein intake, statistics inform us that "the higher protein group ha[s] a statistically significant lower FFM [fat free masss]" (Paszkiewicz. 2014).

Are high(er) protein intakes bad for gymnasts or, what?

Personally I suspect that this is due to a correlation between high(er) protein intakes, lower cabohydrate intakes (R = -0.595) and, most importantly, a reduced overall energy intake, which is associated with lower lean body mass and (listen up, ladies!), just as it has been reported by Deutz et al. previously, increased body fat % (reread the corresponding article from July 2013).

But why don't we have a look at the other study? Beach volleyball players are regarded as the epitome of health and sexappeal, so things could easily look different for them compared to the "frail" gymnasts, right? With a mean body fat % of 18% and a standard deviation ±7% the twelve women from the GSU sand volleyball team who participated in Richardson's study have a much healthier body fat percentage than the average, let alone extreme gymnast in the previously discussed study (we got to be careful here, because the BF% in the Richardson study was measured by body impedance and could thus easily be 5% off).

Reduced bone mineral density is a surprising negative side effect to highe(er) protein intakes in the study at hand. According to Paszkiewicz "[h]igher protein consumption was significantly associated with lower bone mineral density(BMD)in the gymnasts at the arms (r= -0.535; p < 0.001), legs (r= 0.0523; p = 0.001), trunk(r= -0.517; p = 0.001), spine (r= -0.472; p = 0.002), and pelvis (r= -0.539; p < 0.001)." (Paszkiewicz. 2014) Previous studies have yet shown that a high protein intake, in the absence of a continuous energy deficit as it was observed in the study at hand, will not lead to brittle bones. And in an energy sufficient scenario it's rather the lack of little veggies and fruits, as well as other alkalizing foods, than the amount of protein that's to blame for previously observed correlations (Heaney. 2008).

With a mean BMI of 22 kg/m², all female participants of the study were normalweight and consumed a diet with >1.94g protein per body weight (mean intake 132 ±52 g per day). An amount of protein most of the ladies spread across the day with a mean 26.06 (±10.51) g being consumed on every eating opportunity. That's not yet the "SuppVersity suggested" amount of 30g of protein per meal, but it's getting close, yet with an uneven distribution from AM to PM:

• 30g from 6-12 AM,
• 63g from noon to six PM,
• another 39g in the evening

In contrast to many average Janes and Joes, the study participants consumed almost half of the mean protein intake during mid-day, while their protein intake from 6 pm to midnight amounted to only 24(±23) % of their total daily protein consumption. Still, Richardson is right to point out that

"[...] protein intake distribution was skewed, on average, toward the latter half of  the  day  with  approximately  19%  of  protein  consumed  in  the  morning  and  34% consumed  in  the  evening." (Richardson. 2014)

Much to my surprise, the ladies in the beach volley ball team were similarly anorexic as their peers in the gymnast group. With -404  (±385) kcal/day the average energy balance was clearly negative; and even if the standard deviations indicate that this was not the case for all of the ladies, the athletes spent 17 hours, on average, in a catabolic energy balance state (< 0 kcal) on a daily basis.

A high relative protein intake was not associated with better body composition!

Interestingly, though, no significant correlation was found between energy balance per gram of protein consumption and body composition.

Table 2: Spearman’s Correlations: Six Zone Protein Intake and Body Composition (N=12; Richardson. 2014); FFM – fat free mass: FFM to Ht ratio – amount of FFM per cm of height; eating Opportunities – number of times athlete consumed calories; 24 Hour EB – net kcal at the end of the day (energy consumed less energy expended)

The picture that emerges from a regression analyses with respect to the relation of energy balance and protein variables is in fact dubious (see Table 2). The only significant correlations (bold) are a positive correlation between fat free mass (FFM) and protein intake late, and a negative correlation between fat free mass and protein intake early in the AM. A similarly confusing, yet at no time significant association arises for the fat mass, which correlates negatively (albeit with p = 0.678 statistically non-significantly) with the number of meals with a protein content of 25g or more.

PWO glyocgen repletion done right may also help maintain normal leptin levels | learn more

Bottom line: If there is any clear take home message from the study at hand, it would be that chronically low energy intakes below the maintenance, or as Paszkiewicz calls it the "optimal energy intake" appears to have a negative impact not just on the body composition of young female athletes, but also impairs / nullifies the beneficial effects high(er) protein intakes have on the changes in body composition in short term (vs. chronic!) phases of energy deficiency.

Whether and to which extend these changes are related to reductions in leptin expression and/or other hormonal defects that occur in response to the (sometimes life-)long starvation diets many women follow would have to be elucidated in future studies.

The association between higher CHO intakes and better body composition Paszkiewicz observed in her study, on the other hand, appears to support the often heard hypothesis that the already established links between carbohydrates and high energy refeeds after energy restriction, on the one hand, and a restoration of rock bottom leptin levels (Romon,. 1999; Wisse. 1999), on the other hand, would warrant the use of high(er) carb refeeds on a diet - specifically if it's low in carbohydrates.
References:

• Heaney, Robert P., and Donald K. Layman. "Amount and type of protein influences bone health." The American journal of clinical nutrition 87.5 (2008): 1567S-1570S. 
• Paszkiewicz, Julie A. "Relationship Between Daily Protein Distribution and Body Composition in Elite Gymnasts." (2014).
• Richardson, Barbara B. "The Relationship between Moderate, Within Day Protein Intake and Energy Balance on Body Composition of Collegiate Sand Volleyball Players." (2014).
• Romon, M., et al. "Leptin response to carbohydrate or fat meal and association with subsequent satiety and energy intake." American Journal of Physiology-Endocrinology And Metabolism 277.5 (1999): E855-E861. 
• Wisse, Brent E., et al. "Effect of prolonged moderate and severe energy restriction and refeeding on plasma leptin concentrations in obese women." The American journal of clinical nutrition 70.3 (1999): 321-330.