Protein - 0.8g/kg Insufficient for Female Athletes, Too. Plus: Revised RDA is 1.71-2.2 g/kg/d Depending on Sport & Sex

With average daily protein intakes of only 90±24 g (Gillen 2017), many female athletes are probably missing the sweet spot of ~1.7g/kg/d.

I guess I am preaching to the choir when I write that the recommended daily allowance (the magical "RDA") for protein is laughable; the number of studies which show that an increased protein intake will have beneficial metabolic effects is increasing by the day; the subjects of these studies are men and women, kids, teens, tweens, adults, and grandparents, perfectly healthy or sick, lean or fat... and in all of these increasing the protein intake to ~2x the RDA of 0.8g/kg has been found to be associated with significant beneficial effects on body composition and/or health in the majority of the studies (learn more).

High-protein diets are much safer than some 'experts' say, but there are things to consider...

Practical Protein Oxidation 101

5x More Than the FDA Allows!

2017 Evidence for Protein Timing

Satiety: Casein > Whey? Wrong!

Protein Timing DOES Matter!

High Protein not a Health Threat

Unfortunately, humans lack a measuring gauge that would allow us to decide when we've satisfied our daily protein requirements and/or consumed the optimal amount of protein. I guess this is also why we don't even know for sure how exercise will affect our protein need. Yeah, we do have plenty of evidence that extra protein will augment size and strength gains in resistance training and, with the publication of pertinent studies, even endurance trainees are now jumping aboard the "protein supplementation wagon" (learn more).

Stuart M. Philips and colleagues are among those researchers who have been dabbling with the idea of increased protein requirements in athletes and non-athletes, young and old, sick and diseased for years (Phillips 2007; Phillips 2017). As the Canadian scientists point out in one of their latest reviews (Phillips 2017), there are two main methods to measure human beings protein balance. There is...

• measurement of an individual's nitrogen balance - This method has been used for more than 60 years, despite its various shortcomings (Young 1986); and it is also the methodology that underpins the "RDA", a term of which Phillips writes in his review in Front Nutr. in 2017 that it would "inherently imply that the protein intake is 'recommended,' and it is all that you are 'allowed' to eat" (Phillips 2017). Now, I don't have to quote an expert for you to understand that this is, "[o]f course, [...] strictly speaking [not] true" (ibid.). 
• the more recent alternatives are the indicator amino acid oxidation (IAAO) method and the 24h-indicator amino acid oxidation and balance method (24h-IAAO/IAAB)- Having been explicitly developed as an alternative to the nitrogen balance (free reviews can be found in Elango et al. 2008 & 2012), they yield significantly higher values for optimal protein intakes in than nitrogen balance studies, especially in older individuals.

Even though, I believe that this is the first time some of you read about this, the above ain't 'news' in the strict sense of the word and wouldn't have made it into this article, if it wasn't necessary to know that the IAAO method has recently been applied by Wooding et al. (2017) to determine a dietary protein requirement in active females performing variable-intensity intermittent exercise using the indicator amino acid oxidation (IAAO) method.

If we go by the Institute of Medicine's AMDR concept, 0.8g/kg is the absolute minimum, not the recommended protein intake.

What is currently recommended? You already know that the RDA, i.e. the "recommended daily allowance" for protein is 0.8g/kg. Despite its name, it's yet not the RDA, but the AMDR, i.e. the Acceptable Macronutrient Distribution Range that defines the "range of intakes for a particular energy source that is associated with reduced risk of chronic diseases while providing adequate intakes of essential nutrients" (Institute of Medicine 2005). The AMDR says that 10–35% of one's total energy intake should come from protein. As Phillips calculates in the previously cited study, this means that a 55-year-old man who is 1.80 m, weighing 80 kg, would not have to consume the laughable 64 g protein/day the RDA would recommend, but rather 65 g/day as an absolute minimum (that's 10% of the total energy intake and thus the lower AMDR).

Optimal intakes for our exemplary 55-year-old man, on the other hand, would be somewhere within the AMDR of 65–228 g protein/day (assuming an energy requirement of 2,600 kcal/day or 10.9 MJ/day) - in other words, the "optimal" range, i.e. the range where the Institute of Medicine says that it takes "into account the trends related to decreased risk of disease identified in epidemiological and clinical studies" is, as Phillips highlights in his previously cited review from 2017 "well above the RDA (up to 2.8 g/kg/day) that are associated with good health".

The scientists from the University of Toronto started their study with the hypothesis that these requirements would be greater than current IAAO-derived estimates in non-active adult males. And indeed, even though the IAAO method has been around for a couple of years, there's still a paucity of evidence for various populations - including female athletes participating in team sports.

We do already know that endurance athletes need more, much more ...

As Wooding et al. point out, their "group previously used the minimally-invasive indicator amino acid oxidation (IAAO) method to demonstrate that protein requirements in endurance athletes are ~50% greater than sedentary individuals" (Wooding 2017). That, however, is still a pretty conservative estimate, as it marks the protein requirements, not the optimal intakes that will "maximises whole body protein synthesis" and thus enhance recovery from, and potentially adaptation to, an exercise stimulus" (ibid.) - a value the scientists sought to determine in their latest study which involved six healthy, active young adult females (see Table 1).

Figure 1: Modified LIST in which participants followed an audio prompt to complete the variable-intensity intermittent exercise pattern 10 times per block. There were four blocks separated by 5-min breaks, totaling 75-min of exercise plus a 5-min warm-up and cool-down at a self-selected pace (Wooding 2017).

Four individuals were varsity athletes (three rowing, one ice hockey), one was a national level volleyball player, and one was a highly active recreational athlete.

"Participants were required tobe healthy (PAR-Q+), have habitual activity levels of more than 45 min/d on 5 d/wk of moderate vigorous physical activity (I-PAQ for adults aged 15-69), a predicted VO2max ≥44.6 mL O2/kg/min (Leger Multistage Fitness Test), no current use of hormonal contraceptives, and a predictable menstrual cycle (25-33d) during the previous year as determined by interviews and the participants’ records of the 2-3 months prior to study enrollment" (Wooding 2017).

Each participant completed 5-7 metabolic trials (n = 36) during the predicted luteal phase. The women consumed a 2-d adaptation diet containing 1.2 g/kg/d of protein prior to each trial. Wooding et al. chose this intake "in order to minimize metabolic variability during the trial day" and to "provide a level that was previously determined to be sufficient for non-exercising males by IAAO" (ibid.) (15). These adaptation diets contained sufficient energy to meet the subjects' individual habitual daily caloric expenditure of ~2700kcal/d.

Figure from an exemplary study by Kriengsinyos et al. (2002),

How does this strange NaH13CO2 + tracer amino acid test work? Let's take a look at the exemplary data from Kriengsinyos et al. (2002) that has been reprinted in Elango's previously cited 2008 review. Kriengsinyos et al. investigated the effect of lysine intake on the production of 13CO2 from the oxidation of L-[1-13C] phenylalanine (F13CO2) when tracer was infused either i.v. or orally in healthy adult humans (note: one of the important results of the study at hand is that the estimated protein/amino acid requirements don't differ for i.v. vs. oral administration).

As Elango et al. explain in their 2012 paper, "the IAAO technique is based on the concept that when one indispensable amino acid (IDAA) is deficient for protein synthesis, then all other amino acids including the indicator amino acid (another IDAA, usually L-[1-13C]phenylalanine) are in excess and are therefore oxidized" (Elango 2012). That's logical, after all, dietary protein can only either be stored or oxidized. If a rate limiting amino acid is missing, storage is not an option and oxidation will take place. As the intake of said limiting amino acid increases, the oxidation of the indicator amino acid will decrease -  a decrease that reflects the increasing incorporation of amino acids into protein. Yet, once the requirement is met for the limiting amino acid, there will be no further change in the oxidation of the indicator amino acid with increasing intake of the test amino acid (see breaking point in the figure). The inflection point where the oxidation of the indicator amino acid stops decreasing and reaches a plateau is referred to as the ‘breakpoint’. The breakpoint, identified with the use of two-phase linear regression analysis, will then "indicate the estimated average requirement (EAR) of the limiting (test) amino acid" (Elango 2012) - in the example from Kriengsinyos et al. (2002), the breakpoint for lysine is 36mg/kg/d.

On test days the participants reported fasting at the lab, ingested a CHO sports drink and performed a modified version of the Loughborough Intermittent Shuttle Test (LIST; cf. Figure 1), an exercise protocol that simulates the activity pattern of a soccer match:

"Briefly, the test involved four 15-min blocks in which a 17-m variable-intensity shuttle pattern was repeated 10 times with 5-min rests between each block. The entire test was 75 min in length, which also included a 5-min warm-up and cool-down at a self-selected pace. The running pace in our study was based on the percentage of average maximum speed obtained by the entire group of participants during the aerobic assessment (3.45m/s) instead of the percentage of individual VO2max. In this way, the same audio prompt was used for all participants with the following 17-m shuttle paces: 1.7m/s (walk), 2.1m/s (jog) 3.1m/s (run), and an all-out sprint. The average energy expenditure during the LIST was 8.7 ± 0.2 kcal/kg (range 8.5-9.1kcal/kg, determined from accelerometer)" (Wooding 2017). 

Following exercise, participants consumed their first of eight hourly mixed meals containing the test protein intake (0.2-2.66 g/kg/d) and sufficient energy and CHO content in the form of test beverages containing crystalline amino acids, protein-free powder, fruit flavoring powder, grapeseed oil, and maltodextrin as well as protein-free cookies. The actual measurement was done by including a priming dose of NaH13CO2 (0.176 mg/kg) and L-[1-13C]phenylalanine (1.86 mg/kg) in every hourly meal beginning at meal #5. The turnover of this tracer was determined from urinary [13C]phenylalanine enrichment.

Figure 2: Relationship between protein intake and F13CO2. Six participants completed 5-7 metabolic trials each (n=36). F13CO2 breakpoint was used to estimate protein requirements and recommended daily allowance (Wooding 2017).

The breaking point (compare explanation in the blue box above) was determined by analyzing the subjects' breath for F13CO2. As you can see in Figure 2, the F13CO2 curve displayed the expected robust bi-phase response (R² = 0.66) and points, as the author's point out, "to an estimated average requirement (EAR) of 1.41 g/kg/d and an RDA of 1.71 g/kg/d" (Wooding 2017).

Overview of new, IAAO based protein intake minimums and recommendations in different groups of athletes; the figures across the bar denote the values in units of the current RDA of 0.8  g protein /kg/d (Wooding 2017 women in team sports; Kato 2016 male endurance athletes; Bandegan 2017  bodybuilding men)

Bottom line: The study at hands provides further evidence that the well-known and repeatedly cited population guidelines which are based on net protein breakdown analyses are inadequate when it comes to the protein requirements of the athletic population, for whom contemporary sports science consensus statements suggest a broad range of protein intakes from 1.2-2.0 g/kg/d should be consumed by athletic populations (Thomas 2016).

What the study adds to the existing evidence are definite numbers for the minimal and recommended protein intake of female team-sports athletes, a subject group that is often overlooked in clinical research.

More specifically, Wooding's data reveals that an acute bout of variable-intensity exercise results in an increase of young women's protein requirements to 1.41 g/kg/d and an approximate RDA, i.e. an intake that can be expected to be sufficient for 95% of the population, of 1.71 g/kg/d. As previously hinted at, this result is consistent with the scientists' previous observations in endurance-trained males on a day in which they trained (Kato 2016) and in bodybuilders on a rested day (Bandegan 2017) - all numbers you know from the SuppVersity Facebook or Twitter News | Comment!

References:

• Elango, Rajavel, Ronald O. Ball, and Paul B. Pencharz. "Indicator amino acid oxidation: concept and application." The Journal of nutrition 138.2 (2008): 243-246.
• Elango, Rajavel, Ronald O. Ball, and Paul B. Pencharz. "Recent advances in determining protein and amino acid requirements in humans." British Journal of Nutrition 108.S2 (2012): S22-S30.
• Gillen, Jenna B., et al. "Dietary protein intake and distribution patterns of well-trained dutch athletes." International journal of sport nutrition and exercise metabolism 27.2 (2017): 105-114.
• Hoerr, ROBERT A., et al. "Recovery of 13C in breath from NaH13CO3 infused by gut and vein: effect of feeding." American Journal of Physiology-Endocrinology And Metabolism 257.3 (1989): E426-E438.
• Kato, Hiroyuki, et al. "Protein Requirements Are Elevated in Endurance Athletes after Exercise as Determined by the Indicator Amino Acid Oxidation Method." PloS one 11.6 (2016): e0157406.
• Phillips, Stuart M., Daniel R. Moore, and Jason E. Tang. "A critical examination of dietary protein requirements, benefits, and excesses in athletes." International journal of sport nutrition and exercise metabolism 17.s1 (2007): S58-S76.
• Phillips, Stuart M. "Current Concepts and Unresolved Questions in Dietary Protein Requirements and Supplements in Adults." Frontiers in Nutrition 4 (2017).
• Thomas, D. Travis, Kelly Anne Erdman, and Louise M. Burke. "Position of the academy of nutrition and dietetics, Dietitians of Canada, and the American college of Sports Medicine: Nutrition and athletic performance." Journal of the Academy of Nutrition and Dietetics 116.3 (2016): 501-528.
• Trumbo, Paula, et al. "Dietary reference intakes for energy, carbohydrate, fiber, fat, fatty acids, cholesterol, protein and amino acids." Journal of the American Dietetic Association 102.11 (2002): 1621-1630.
• Wooding, Denise J., et al. "Increased Protein Requirements in Female Athletes after Variable-Intensity Exercise." Medicine and science in sports and exercise (2017).
• Young, Vernon R. "Nutritional balance studies: indicators of human requirements or of adaptive mechanisms?." The Journal of nutrition 116.4 (1986): 700-703.

2909 IU of Vitamin D3 per Day - That's What Mr. Average Needs | What Do You Need? 3094, 4450, or 7248 IU/day?

Your BMI or rather the associated level of inflammation and bodyfatness determines your D3 requirements.

I know that I have previously written about estimated vitamin D requirements, but in contrast to previous articles that were based only on 1-3 studies, today's article about the purported vitamin D requirements of the average Westerner, however, is based on the same previous 108 published estimates and new calculations based on the vitamin D status of 13,987 individuals in various studies Veugelers, Pham and Ekwaru used as the observational database for what is a of now probably the most tangible vitamin D recommendation in their recently published study in Nutrients (Veugelers. 2015).

There are many ways to get your vitamin D learn more the SuppVersity

How Much To Take?

Leucine, Insulin & Vitamin D

Vit. D Speeds Up Recovery

Overlooked D-Sources

Vitamin D For Athletes!

Vitamin D Helps Store Fat

Now, as the headline already tells you, their analysis of said data lead the researchers to conclude that "2909 IU of vitamin D per day is needed to achieve serum 25‐hydroxyvitamin D (25(OH)D) concentrations of 50 nmol/L or more in 97.5% of healthy individuals" (Veugelers. 2015). To get to this value, the researchers from the University of Alberta used quantile regressions to

"model the effect of vitamin D supplementation on the 2.5th percentile, the median and the 97.5% percentile of serum 25(OH)D concentrations [and an] exponential model [and] logistic regression [for the estimates and] to estimate the probability of having serum levels above a lower and below an upper serum 25(OH)D concentration, [respectively]" (Veugelers. 2015). 

In that it is important to know that in this model the limit of the 'normal' vitamin D concentrations (58-171 nmol 5(OH)D/L) was defined in accordance with the values Luxwolda et al observed in traditionally living populations in East Africa who have mean serum 25‐hydroxyvitamin D concentration of 115 nmol/l or more (Luxwolda. 2012). So, we are not talking about absolute minimum levels, but rather about levels many researchers would call "optimal".

Figure 1: Plot of the results of the model calculations (left) and my visualization (right) of the calculated vitamin D requirements in IU/day for normal-weight, overweight and obese individuals (Veugelers. 2015).

As a SuppVersity reader you will be aware that normal-, overweight and obese subjects will need different amounts of supplemental vitamin D3 to achieve these "optimal" levels. To accomdate for these differences and to provide adequate estimates for normal weight, overweight and obese participants, Veugelers et al conducted separate analysis and used suitable logistic regression models to identify the log term of supplementation that provides the best fit. Needless to say that this sub-analysis was conducted based only on those studies that either included exclusively normal-, overweight or obese subjects or distinguished between the three of them yielded. It is thus only logical that this analysis yielded different results of which the 3094 IU/day, which is the suggested daily amount of vitamin D3 to maintain optimal vitamin D levels for normal-weight individuals, is yet pretty much identical to the previously cited "optimum" for Mr. Average Joe.

Against that background, it is yet all the more important to note that the average overweight or obese Westerner will yet need significantly more vitamin D3, 4450 IU/day and 7248 IU/day, to be precise, to keep his / her labels stable. Based on what you should remember from the role of 25OHD as an anti-inflammatory acute phase reactant (Waldron. 2014), though, this is not really surprising.

So what's the verdict, then? While the study at hand certainly provides the hitherto best estimate of our individual vitamin D3 requirements, I still wouldn't put blind faith into the results of Veugelers' model calculation. To be sure you're not too extra-ordinary to be average, I would suggest you test your 25OHD levels after 6 months on the suggested dosage. If you're "in the zone", everything is fine. If not, adjust appropriately.

Fat loss will trigger decent increases in vitamin D, but vitamin D will not trigger significant fat loss | more

Apropos adjusting, as the authors point out, the previously discussed figures may not even be the most practically relevant result of the study. Rather than that, it is the "large extent of variability in 25(OH)D concentrations" of which the authors rightly say that it "makes a RDA for vitamin D neither desirable nor feasible" (Veugelers. 2015) that's the most relevant results of the study.

And yes, you've read that right. The 400, 600 and 1000 IU/day RDA you will find in different countries all over the world is total bogus, not just because it is too low, but because stating a recommended daily allowance based only on the age, not the weight, or rather inflammatory status of an individual, is absolute bogus | Comment on Facebook!

References:

• Luxwolda, Martine F., et al. "Traditionally living populations in East Africa have a mean serum 25-hydroxyvitamin D concentration of 115 nmol/l." British Journal of Nutrition 108.09 (2012): 1557-1561.
• Veugelers, Paul J., Truong-Minh Pham, and John Paul Ekwaru. "Optimal Vitamin D Supplementation Doses that Minimize the Risk for Both Low and High Serum 25-Hydroxyvitamin D Concentrations in the General Population." Nutrients 7.12 (2015): 10189-10208.
• Waldron, Jenna Louise, et al. "Vitamin D: a negative acute phase reactant." Journal of clinical pathology (2013): jclinpath-2012.

Are You ABCDE-Deficient? Common Nutrient Deficiencies in the US. Plus: How Food Fortification & New "Daily Values" Affect the Intakes of Vitamin A-E, Calcium Iron & Co

Nutrition labels on fresh blueberries - do we really need them?

I sill remember that I was shocked, when I bought a pack of blueberries and found a nutrition label underneath the plastic cover of my expensive 150g health-investement...

That's probably 2 months ago and the reason I do remember this event now is the publication of a paper that examines the effect a change in the "daily values" (i.e. the references), the figures in the obiquitous black and white table are based on, would have on the average US citizen's nutritional intake of the vitamins A, D, E, C, B-12 and folate, and the minerals calcium and iron.

"Daily Values" (DV), fortified foods and nutrient adequacy: Before I dig deeper into the actual study results, it's probably wise to point out that fortified foods are the link between the DV's and micro-nutrient intake of the average American. If manufacturers continue to fortify foods to the same %DV for each nutrient, the extent to which potential changes in DVs would affect nutrient intake adequacy depends on the proportion of nutrient intakes derived from fortified foods and the magnitude and direction of change in the DV.

According to the data Mary M. Murphy and her colleagues from the National Institutes of Health/Office of Dietary Supplements present in their latest paper, there is still a large gap between the current DV values, which represent the RDAs (recommended daily allowances) from 1968 and have been matched to

"the highest level of intake judged to be adequate to meet the known nutrient needs of practically all healthy persons in a specific age-gender group" (Murphy. 2013)

on the one hand, and supposedly "improved" candidates that could replace them: The population weighed and the population coverage varieties of the RDA & EAR.

• RDA = the average daily dietary nutrient intake level that is sufficient to meet the nutrient requirements of nearly all (97–98%) healthy individuals in a particular life-stage and gender group
• EAR = the average daily nutrient intake level that is estimated to meet the requirements of half of the healthy individuals in a particular life-stage and gender group

As you can see in Table 1 these new recommendations are not - as you may have expected -  significantly higher than the current daily values. If you look closely, you will in fact notice that some of them are significantly lower!

Table 1: Current DVs for select vitamins and minerals and potential DVs based on population-weighted and population-coverage RDAs and EARs. AT,a-tocopherol; DV, Daily Value; EAR, Estimated Average Requirement; RAE, retinol activity equivalent; RE, retinol equivalent (Murphy. 2013).

In the case of vitamin B12 and copper, for example, the difference between the "reformed" recommendations would amount to -50%. The population-coverage RDA for vitamin C, on the other hand, is 50% higher than the old "daily values" (DV) and still more than 10x lower than the 1,000mg of ascorbic acid, of which you may have read on the Internet that it was the bare minimum intake of vitamin C (more about vitamin C).

Figure 1: Percentage of U.S. population aged >4y with dietary intakes below the EAR based on current intakes and assuming
constant %DVs in fortified foods under the current, as well as two potential DV scenarios, i.e. the population-weighed EARs or the population-coverage RDAs become the revised DV values (Murphy. 2013)

Irrespective of the "low" RDA and the high number of fortified foods, ascorbic acid is yet still one of the those micro-nutrients the diets of more than 40% of the US are deficient in. And as the overview in Figure 1 goes to tell you, this would not change, if any of the new RDAs or EARs became the new DVs, so that the amounts of vitamin C in fortified food was adjusted.

Not an improvement by any means

In a more thorough sub-analysis, the scientists observed that the differences in the proportion of the total population with usual intakes less than the EAR would be <2% of 5 out of 8 nutrients (vitamins D, E, and B-12; folate; iron), regardless of whether the policy makers sued the population weighted EARs or the population-coverage RDAs as a basis for the revision of the DVs.

To put it plainy: This means that the micronutrient intake of more then 3 million individuals would still fall below the EAR in the total population (U.S. Census Bureau. 2005).

Even worse, if someone in the upper echolons was bribed.... ah, I mean convinced by the conclusive evidence we have that using the population-weighted EARs instead of the population coverage RDA would be the best thing to do, this would increase the risks of inadequate iron and folate intake in women of childbearing age. Both, iron and folate deficiency, can result in irreversible damage to the unborn child (Scholl. 2000; McArdle. 2013). The same is true for vitamin A (Wallingford. 1986) of which Murphy et al. write that it "was identified as a shortfall nutrient (although intakes are not currently in the category ‘‘of concern’’) for the U.S. population" (Murphy. 2013).

"The Standard American Diet Has 'Optimal' Fatty Acid Ratio to Induce Diabesity." | read more

What has to be done? I hope you don't actually want me to answer this question - do you? I mean let's be honest - if people get 17–28% of total intakes of folate, iron, and vitamins A, B-12, and C and 8–12% of calcium and vitamins D and E from fortified foods (this is what Murphey et al. found) and are still deficient, you could obviously argue that we simply have to put even more vitamins and minerals into the nutrient deficient, energy dense junk the average Westerner is shoveling his piehole everyday.

But let's be honest: Wouldn't it be better to kill two birds with one stone by educating people that the stuff they eat is making them fat and sick - no matter how much artificial vitamins the "food" industry is pumping into their highly addictive, revenue-centered high-tech designer products?

References:

• McArdle, Harry J., Lorraine Gambling, and Christine Kennedy. "Iron deficiency during pregnancy: the consequences for placental function and fetal outcome." The Proceedings of the Nutrition Society (2013): 1-7.
• Murphy, Mary M., et al. "Revising the Daily Values May Affect Food Fortification and in Turn Nutrient Intake Adequacy." The Journal of nutrition 143.12 (2013): 1999-2006.
• U.S. Census Bureau. 2005 Middle series data from annual projections of the resident population by age, sex, race, and Hispanic origin: lowest, middle, highest, and zero international migration series, 1999 to 2100 (NP-D1-A). Washington: Department of Commerce; 2000 [cited 2012 Jun 16]. Available from: http://www.census.gov/population/www/projections/natdet-D1A.htm 
• Scholl, Theresa O., and William G. Johnson. "Folic acid: influence on the outcome of pregnancy." The American journal of clinical nutrition 71.5 (2000): 1295s-1303s.
• Wallingford, J. C., and B. A. Underwood. "Vitamin A deficiency in pregnancy, lactation, and the nursing child." In: Bauernfeind JC, ed. "Vitamin A deficiency and its control." New York: Academic Press, 1986:101–52.

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

2x-3x higher than RDA protein intakes work equally well for men and women, to get and stay lean and lose fat and build / maintain muscle.

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

It's more than high time to revise the RDA

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

To up the calculated energy deficit to 40% the physically active (physical activity 3– 4 d/wk), weight stable ( 2 kg; for a minimum of 2 mo before the study), 39 volunteers [32 men (11 military, 21 civilians) and 7 women (7 civilians)] with a body mass index (BMI) between 22 and 29 kg/m² and a sufficient baseline fitness had to exercise daily:

"You told me to eat more protein and this burger has both meat and cheese!" - This and other mishaps are the rule, not the exception in uncontrolled dietary interventions (learn more). The fact that the study at hand took place in the metabolic ward of the U.S. Department of Agriculture Grand Forks Human Nutrition Research Center really is a HUGE PLUS.

"To isolate the effects of the diet and minimize the potential of an exercise training stimulus, physical activity during WM was prescribed at levels comparable to those reported in prestudy 7-d physical activity records. Volunteers performed low-tomoderate-intensity (40 – 60%Vo2peak) treadmill and cycle ergometry steady-state physical activity sessions daily. Intensity was based on pre-study Vo2peak
measurements obtained during a progressive intensity treadmill test and verified during
familiarization trials using indirect calorimetry (ParvoMedics) and corresponding heart rate. Workloads during steadystate physical activity sessions were adjusted accordingly to
ensure accuracy using the heart rate reserve method and portable heart rate monitors." (Pasiakos. 2013; my emphasis)

The study took place in the metabolic ward (so there was no cheating involved here => HUGE PLUS; cf. ) at the U.S. Department of Agriculture Grand Forks Human Nutrition Research Center. All volunteers were required to abstain from nutritional supplements, alcohol, smoking, and all medications, unless acetaminophen-containing products were provided by the investigator or study physician. Volunteers were also required to be in their assigned rooms with lights out by 11 P.M. (possibly very important; learn why) to ensure adequate and similar levels of sleep.

Don't worry this was not "cardio only"

To maintain prestudy muscular fitness levels, the volunteers also performed resistive-type physical activity 3d/wk. However, "to minimize the potential of an unaccustomed, anabolic stimulus influencing study outcome measures, the intensity and volume of the resistive-type exercise was low" (Pasiakos. 2013):

Table 1: Energy / macronutrient content of the diets (updated on June 21; previously there was a copy + paste error in the table)

"Specifically, volunteers performed one single-joint movement per major muscle group (3 sets of 15 repetitions) using workloads determined during the prestudy period. Frequency, intensity, mode, and volume of resistive-type activities did not change during the 31-d study. Research staff who were blinded from dietary assignment supervised all physical activity sessions for safety and accuracy". (Pasiakos. 2013)

The body weight, was recorded in two day intervals and the body composition was quantified using a  dual-energy X-ray absorptiometry (DXA) during WM (day 9) and ED (day 30). To elicit the underlying mechanisms, the resting metabolic rate, protein synthesis, nitrogen balance and the expression of intracellular signaling proteins were tested, as well.

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

As you can see in figure 1, there was a baseline and dose-dependent effect on the changes it total weight and body composition, respectively.

Body weight during WM was similar between dietary treatment groups and remained stable from d 1 (group mean, 77.5 1 +/-5 kg) through d 10 (77.1 1 +/-5 kg). Overall, volunteers lost 3.2 0 +/- 2 kg during the 21-d ED; 3.5 0 kg for RDA, 2.7 0 kg for 2 -RDA, and 3.3 0 kg for 3 -RDA (P < 0.05). Independent of dietary protein, percentage body fat decreased (P < 0.05) from 19.8 1% during WM to 18.1 1% during ED, and the change in percentage body fat was similar between RDA (1.3 0 +/- 3%), 2 -RDA (1.8 0 +/- 4%), and 3 -RDA (1.9 0 +/- 3%)." (Pasiakos. 2013)

What's worth taking a closer look at, is yet the proportion of total weight loss due to changes in fat mass (FM) and FFM, which differed across dietary protein levels.

• the percentage of total weight loss attributed to reductions in fat mass (FM) was higher (P < 0.05) for 2 -RDA (70.1 7%; 1.9 0 +/- 3 kg) and 3 -RDA (63.6 5%; 1.9 0 +/- 2 kg) than for RDA (41.8 5%; 1.6 0+/-2 kg)
• the percentage of total weight loss due to a loss of fat free mass (FFM) was lower for 2 -RDA (29.8 7%; 0.8 0 +/- 2 kg) and 3 -RDA (36.4 5%; 1.2 0. +/- 3 kg) as compared to RDA (58.2 5%; 2.3 0 +/- 3 kg)
• the fat to lean mass loss ratio was 30% higher in the medium protein intake group, in other words, the increase in protein intake in the 3xRDA group did not protect the lean mass any better than the 1.6g/kg in the 2xRDA group

While the latter change did not reach statistical significance, the trend is clear and I suspect with a higher number of participants, the scientists would have been able to show that the 3x RDA intake is not just worthless, but actually contra-productive, if your goal is stable ongoing fat loss.

No inter-group differences in the majority of signaling proteins

All the changes took place in the absence of statistically significant inter-group differences in the changes in anabolic intracellular signaling and gene expression [ignore the following list if you are no geek ;-], i.e.

• postprandial Akt (Ser 473) phosphorylation was increased 1.4-fold higher (P < 0.05) compared to postabsorptive levels
• postprandial p70 S6K1 (Thr 389), eIF4E Ser (209), and rpS6 (Ser 235/236) phosphorylation status was 16, 1.9, and 15.5-fold higher (P < 0.05), respectively, compared to postabsorptive phosphorylation levels
• phosphorylation status of eEF2 (Thr 56) was lower (P < 0.05) after feeding

The more important general observation was yet that the upregulation of these signals 3 h after consuming a protein-containing meal, demonstrated a main feeding effect for all proteins of interest (P < 0.05). On the other hand, their expression was not influenced by energy status or the level of dietary protein intake (and let's be honest, what would an increase be worth if the data in figure 1 already told us what the real-world implications are?)

Figure 2: Changes in postabsorptive muscle protein synthesis-associated mRNA expression levels during the diet phase (-40% energy intake) of the study (Pasiakos. 2013)

Additionally, the energy deficit increased the mRNA expressions of a couple of other proteins implicated in the intracellular regulation of muscle protein synthesis:

"Transcription of Vps34, a protein involved in amino acid sensing and amino acid-mediated stimulation of mammalian target of rapamycin (mTORC1) signaling, was 1.2-fold higher (P < 0.05), while expression of mTORC1 inhibitors REDD1 and REDD2 were both 1.3-fold higher (P < 0.05) after ED compared to WM. Increasing dietary protein intake increased Vps34 mRNA expression, with 1.2-fold higher levels for 3x-RDA than RDA (P 0.05). MAP4K3, LAT1, and SNAT2 mRNA levels were not influenced by energy and dietary protein manipulations." (Pasiakos. 2013)

In view f the slight advantage of the 3xRDA diet in terms of the stimulation of protein synthesis, you may want to come back to the statistical insignificance of the superiority of the 2xRDA diet to keep indulging the same hilarious amounts of protein that have probably not gotten yourself anywhere near contest shape in the past, well, let's take a look on a couple of other observations, then:

• While the nitrogen balance remained negative (meaning the body was burning more protein than it stored) over the whole trial in the 0.8g/kg group, it returned to baseline (weight maintenance levels) first in the 1.6g/kg (=2x RDA) group (day 17!). This restoration of to pre-diet levels was observed only on day 30 in the high protein group (2.4g/kg) and the that without any significant advantage of the 3xRDA over the 2xRDA intake (if anything it was lower in the high protein group; see figure 3)
• There was no "thermogenic advantage" - or whatever people usually like to call the purported beneficial effect that comes with the ingestion of higher amounts of protein; in fact, the resting metabolic rate was identical for all three groups over the whole 21-day diet period. 
• With a diet that was high in carbohydrates and low in fat (see table 1), the conversion of protein to glucose, was likely relatively limited and the potential downsides of high protein + low carb diets, where most of the protein will be broken down in the liver to supply your body with glucose and any temporary increase in insulin due to fast acting protein sources were not an issue.

In the end, the increase in postprandial protein synthesis in the 3x RDA group is therefore worthless, because it went hand in hand with an increase in wastefulness due to which the absolute protein retention did not differ all that much and the differences in lean mass loss 0.1kg) are clearly insigificant- plus: If you simply do the math, the ratio of fat free to fat mass loss, is still 31% higher in the 2x RDA group.

---

Irrespective of how many supplements you take - you cannot out-supplement a bad diet, laziness and a lack of motivation & determination. Still, especially for the elderly HMB with it's pronounced anti-cababolic effec could help - particularly on a diet (learn more; leucine vs. HMB)

So what's the optimum then? If we reconcile the results of the study at hand, the "optimal" protein intake would thus probably be somewhere between 1.6g/kg and 2.0g/kg an thus in the <200g range for the vast majority of people. If you also consider that this value includes all protein even that from rice, and other "non-quality" protein sources, the study at hand does not confute my previous recommendation to stick to a 1.5g/kg-2.0g/kg (per total body mass) protein intake from quality protein sources, to discount the additional protein you will be getting from "low protein food" (too much counting will only make you neurotic) and to do that irrespective of whether you are bulking and or dieting  .

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

References: 

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

Profitable Revelation! Inhabitants of the Affluent Western Hemisphere Don't Meet "Their" RDAs For Important Nutrients. "Scientists" Call to Action, I Call to Calm Down.

Image 1: Nothing sells like FDA-approved supplements and federally supported fortified foods. And whenever you want to sell more snake oil, just pay for another study on "nutrient deficiencies"

There are two things you can shovel truckloads of money with in the realms of dietary supplements and convenience foods. Those are dreams, such as the dream of a lean and muscular physique and FDA approved but for customers non-verifiable promises of  the absence of future ailments. And while such profane things as wanting to get big and buffed or even simply "looking good naked" is looked down upon by the majority of average Joes and Janes (I don't want to go into the underlying psychological reasons here ;-), the use of the latter is generally regarded as a useful if not necessary means that will not just help us preserve our health, but will also sooth our guilty conscious of not being able to break ourselves of our bad dietary habits. Against that background it's good that we have such great scientists and policy makers who will base their wise decisions on totally unbiased and all-encompassing scientific data from research teams such as as the one from DSM Nutritional Products Limited in Kaiseraugust, Switzerland, and Parsippany, NJ, USA (Troen. 2012).

On average, we are all the same, right!? Our governments obviously don't think so...

Morover, the decisions of those policy makers are not just rooted in science, they also hold another, maybe even more important good in highest esteem: Equality! And though, equality is one of the principles the Western civilization often boasts of, it seems as if when it comes to our dietary "needs", as defined by the individual dietary guidelines, you, my mostly American friends, must be somewhat different.

Figure 1: Reference intakes for selected vitamins in Germany, UK and the Netherlands expressed relative to the US RDA (data based on overview in Troesch. 2012)

This would at least be the logical conclusion you would have to draw based on the in parts pronounced differences between German and US RDAs (see figure 1), which would suggest that my I need ~30% more vitamin D than you do, while our friends in the UK either don't need it at all or cannot agree on a reference intake and my neighbors to the West, must assume that they get more than enough vitamin D from sunbathing at the beaches of the North Sea to get away with only 100IU of vitamin D per day; and their low recommendations for vitamin E are probably based on the rationale that they traditionally use beef fat instead of vegetable oils to fry their fries *rofl*

You are deficient, my friend! Go get your fortified foods ans supplements, NOW!

Now, as funny as that may seem, in the end these discrepancies only underline three fundamental problems that are rarely addressed when scientists analyze data to finally get to the (nutritional) root course of modern disease:

Want to learn about where you stand in terms of the RDA?

I got some help from my friends over at Highbrow Paleo, who felt the following tools are particularly helpful to estimate or calculate your daily micro- and macronutrient intakes:

• Cron-O-Meter
• Fitday
• Nutritiondata

You know that I am not a big believer in logging your food intake and making calories in vs.calories out calculations. So, for your own psychological well-being try not to get addicted to these tools only to end up as yet another food neurotic on certain bulletin-boards ;-)

1. the RDAs are more or less arbitrary - While we do know pretty certain which dosage of a certain nutrient is vitally important, when everything else, i.e. nutrition, exercise, stress, etc. is "normal" (whatever that may be), we have almost no clue how deficiencies, let alone the overabundance of one nutrient affect the need / optimal intake of another. A good example here would be calcium - one of my favorites, by the way: While we are stuffing our elderly (in particular women) with calcium supplements to "protect" their bones with little success, Dawson-Hughes et al. have shown in 2009 already that you can effectively reduce bone resorption, i.e. the leeching of calcium from the bones, by supplemental potassium bicarbonate, while just throwing more calcium and vitamin D at older men and women will at best increase renal calcium excretion, at worst lead to kidney stones and vascular calcification (Dawson-Hughes. 2009)
    
2. the RDAs are light years are usually one or two decades behind contemporary science - Despite being an outspoken critic of the current vitamin D hysteria, the absence of a concrete RDA for vitamin D in the UK is just one of the most obvious examples of how the endless discussions of top-class experts lead to an grossly negligent gap between the latest results from scientific research (which in and out of itself often take months to be published and years or decades to be accepted) and their concrete implementation into the guidelines.
    
3. the RDAs lack any regard of individuality or specificity - although the different RDA's in the USA, Germany, the UK and the Netherlands would suggest otherwise, you are all identical clones of an imaginary average Joe or Jane for the policy makers; and as if that was not enough, the same applies for the nutrients as well: "Vitamin A? Yeah, that's beta carotene, right?" And vitamin A vs. carotene (even alpha vs. beta caroetene) is only one of the many examples (others are folate vs. folic acid; alpha tocopherol usually equated with "vitamin E" vs. gamma-tocopherol, let alon the tocotrienols, etc.), for which we know by now that lumping them together under common names, can easily lead to imbalances with pathological consequences.

Against that background my rationale for posting the following data on what scientists believe Mr. and Mrs. average US/UK/GER/NL citizen are missing out on and of which nutrients they may in fact get plenty is to create an incentive to take a couple of minutes and plug your own food data into one of the countless free online devices (see red box above for some references) to see where you as an individual are standing - and I bet, the majority of you will see results that are fundamentally different from those Troesch et al. summarize in their paper:

• Vitamin D: Irrespective of whether or not you believe that it does make sense to consume the lion's share of a "vitamin" that is supposed to be produced in your skin and is thus, due to its actions on almost every cell of your body, essentially not a vitamin, but a hormone, it is somewhat alarming that even in the Netherlands, where the RDA is hilarious 100IU 5-25% of the men and 25-50% of the women fail to achieve their recommended daily allowance - 2 1/2 large egg yolks alone would provide them with more than that! With the higher RDA's in Germany and the USA, the percentage of people who do not meet their daily allowances is >75%!
• Vitamin A: I am by no means surprised that vitamin A is not mentioned in the scientists mini-summary in the abstract. After all, it's bad for you! Right? No... freaking, no! And it's certainly likewise not good for way more 75% of the US citizens not to meet their RDAs for vitamin A and that despite the fact that the scientists lumped all "vitamins A" together! In the Netherlands and the UK, ~50% have an adequate intake and over here in Germany only 25-50% of my the average Joes and Janes are below their RDA cut-off, which could, just as the vitamin D problem by the way, readily be solved by eating a piece of liver from time to time. Some fatty fish, eggs and of course vegetables on a daily basis would yet serve the same purpose and would, which may turn out to be of even greater importance deliver a very balanced spectrum of various forms of pre-vitamin A (carotenes) and retinol.

• 

  Figure 2: Changes in reasoning behind supplement use in 2010 (French. 2011)

  Folic Acid and other B-vitamins: An interesting observation can be made for the B-vitamins, where the citizens of the land of both fast food and eager food-intoxi.... ah, I mean "fortification" (obviously the US) appear to be way better off than their poor fellows in Europe. Especially here in Germany, we should really wonder that we are not much sicker than you, after all, not all our products are enriched with high amounts of bio-unavailable folic acid so that we more than 75% of us do not meet our RDAs for this unquestionably important, but in its unnatural supplemental form not very controversial vitamin. In 2006, for example, Troen et al. report reduced immune function from excess folic acid build-up in the blood of post-menopausal women (Troen. 2006) and Halsted reports in a more recent paper that the "widespread use of supplemental multivitamins" in conjunction with the "fortification of the US diet with folic acid has resulted in high serum ... "[...] folate levels in much of the population" (Halsted. 2008)

  high folate levels that have been associated with increased risk of cognitive decline in aging people with low vitamin B12 status, decreased natural killer T-cell immune function and increased risk of recurrent advanced precancerous colorectal adenomas and breast cancer" (Halsted. 2008)

  Against this background it should be allowed to ask, whether the "average American" with his "adequate" (>95%!) folic acid intake really is better off than the "average German" who is unlikely to get his RDA of folic acid (>75%). 

For the scientists who (surprise!) happen to work for DSM Nutritional Products Ltd., the observations they present in form of stylized "traffic lights", with all those yellow and red "lights" signifying impeding danger and the need to take action, suffice to conclude that there is not just a gap "between vitamin intakes and requirements for a significant proportion of the population even in the most affluent countries", but that the latter would also be "a call to action 100 years after the term 'vitamine' [sic!] was coined" (Troesch. 2012)

If this post got you interested in an in-depth look at nutrient fortification its uses, abuses, benefits and downsides, I suggest you check Paul Jaminet's article on the issue at his "Perfect Health Diet Blog" (Jaminet. 2012). It would be pointless for me to repeat what Paul has already laid out in his concise and - as us physicists like it - well-referenced summary ;-)

And while Mrs Troesch and her co-authors do not state that explicitly, it should be obvious what this "call to action" will amount to... !? Right! More nutrient "fortified" foods and more randomly assembled multi-vitamin products, instead of less junk and more health (=real) food in everyone's diet.

References:

1. Dawson-Hughes B, Harris SS, Palermo NJ, Castaneda-Sceppa C, Rasmussen HM, Dallal GE. Treatment with potassium bicarbonate lowers calcium excretion and bone resorption in older men and women. J Clin Endocrinol Metab. 2009 Jan;94(1):96-102.
2. Halsted CH. Perspectives on obesity and sweeteners, folic acid fortification and vitamin D requirements. Fam Pract. 2008 Dec;25 Suppl 1:i44-9. Epub 2008 Sep 30. Review. 
3. Jaminet, Paul. Food Fortification: A Risky Experiment? PerfectHealthDiet.com. March 23, 2012 < http://perfecthealthdiet.com/2012/03/food-fortification-a-risky-experiment/ > retrieved on June 18, 2012.
4. French S. Natural Marketing Institute. The US Botanical Market: Latest Consumer Insights. Natural Marketing Institute. March 2011.
5. Troen AM, Mitchell B, Sorensen B, Wener MH, Johnston A, Wood B, Selhub J, McTiernan A, Yasui Y, Oral E, Potter JD, Ulrich CM. Unmetabolized folic acid in plasma is associated with reduced natural killer cell cytotoxicity among postmenopausal women. J Nutr. 2006 Jan;136(1):189-94.
6. Troesch B, Hoeft B, McBurney M, Eggersdorfer M, Weber P. Dietary surveys indicate vitamin intakes below recommendations are common in representative Western countries. Br J Nutr. 2012 Jun 13:1-7.

Tocotrienols: What They Are, What They Do & How They Work + Why the RDA of Palm Olein is NOT 1xCup Per Day

Image 1: If you wanted to get the tocotrienol levels a producer of respective supplements says are  "required", you would have to eat at least 200g palm fruits a day. Alternatively, you can resort to 4kg of oats, if you like those better... What? you are wondering that you are not dead by now? After so many years of tocotrienol deficiency from not getting your 4kg of oats?

The long lists of pathologies related to vitamin E deficiency include, among others, all sorts of degenerative diseases from ataxia over general muscle degeneration to degeneration of sperm and subsequent infertility. But despite the fact that there is a pretty substantial amount of evidence that would suggest that a diet rich in vitamin E could not just prevent the aforementioned pathologies, but would also protect us from many of the currently prevalent ailments of western society such as obesity and coronary vascular disease (Mishra. 2003; Rimm. 1993), respective trials with dietary supplements usually show no, or even negative effects. I have already addressed a couple of  reasons why the benefits of dietary vitamin E intake often cannot be replicated with supplements in previous posts. The most significant one, probably is the absence of the "right" mixture and ratio of alpha-, beta-, gamma- and delta-tocopherols and, as an emerging contributer, the total absence of tocotrienols in the vast majority of vitamin E supplements and almost 99.9% of the pertinent trials.

What are tocotrienols? And what do they do?

I could now rant about the structural differences between the two, with the tocotrienols being an unsaturated variety of the tocopherols with a isoprenoid side chain, but I guess it is enough to know that due to  differences in their molecular structure, they also differ in their effects on the human body, of which you may already have apprehended that their cholesterol lowering effects were the first to attract the attention from researchers (Qureshi. 1986). Within the last 26 years researchers from all around the world have identified additional health benefits, the most prominent of which are...

• Anti-cancer effects (Kato. 1985; Sundram. 1989; Weng. 2009),
• General antioxidant effects (Newaz. 1999),
• Brain specific antioxidant effects (Khanna. 2003),
• Exercise-specific antioxidant effects (Lee. 2009),
• Cardiovascular disease (Shibata 2009)
• Diabetic neuropathy (Kuhad. 2009)
• Bone health (Ahmad. 2005)
• Metabolic syndrome (Weng. 2011)
• Antithrombotic effects (Qureshi. 2011)
• Endocrine health (Yu. 2005)
• Liver health (Patel. 2012)

The purpose of today's SuppVersity article is yet not so much to compile the most extensive list of potential, purported or demonstrated benefits of tocotrienols, the major dietary sources of which are (Kobayashi. 1975; Tan. 2011)

• rice bran oil (50:50 tocopherol:tocotrienol ratio), 
• palm oil (25:75 tocopherol:tocotrienol ratio), and 
• annatto (0.1:99.9 tocopherol:tocotrienol ratio) oil
• human breast milk (!) [though this is probably no major source for you ;-]

but rather to take a look at an intriguing chart of the various molecular targets (Aggarwal. 2010) and discuss the implications:

Figure 1: Molecular targets (left) and proteins that directly interact with tocotrienols (right; adapted from Aggarwal. 2010)

As you can see  without even looking really close at the above graphic, the number of those targets is vast. Another thing you should see right away is that the effect of the tocotrienols is mostly inhibitory (red ovals in the left) and include a couple of old foes, such as:

• the inflammatory cytokines & transcription factors: IL-1, IL-6, TNF-alpha, nf-kappabeta, IL-8 (probably involved in auto-immune reactions), PF-A4 (increases platelet aggregation and thus thrombosis risk)
• factors involved in angiogenesis and cardiocascular disease: VEGF (vascular growth factor, involved in CVD) and its receptor VEGF-r, VCAM-1 (increases adhesion of immune cells to the endothelial wall)
• kinases involved in the cell cycle and apoptotic regulators: CDK's, PKC, pERK, etc. & survivin, IAP-1 & 2 etc., but also telomerase, which are all involved in the proliferation of cancer
• enzymes involved in inflammatory processes: eNOS, iNOS, COX-2, etc.

On the upregulatory side of things, we have

• enzymes from the CYP cascade, which are among other involved in the clearance of estrogen, and other hormone like substances and the metabolism of drugs,
• MAPK and JNK, which exert anti-catabolic effects on muscle tissue, or 
• GPX and SOD, two of the major enzymes involved in the antioxidant defenses

Now, if we take a look at all these, you may remember that low COX-2 levels have only recently been identified with profound overtraining (cf. "Overtraining inflammation insufficient repair"), that AKT (not mentioned above, but in figure 1) is one of the driving forces of skeletal muscle anabolism and telomerase, extends cell life in general, not just in cancer cells. Which brings us back to the issue of ...

...how much anti-oxidants do we actually need?

Figure 2: Total tocopherol and tocotrienol content of high vitamin E foods / oils (top) and tocopherol ratios (bottom) , data based on Whittle. 1967 and Slover. 1971

Or, in this particular case, how much tocotrienols are still beneficial? Neither I, nor anybody else knows the exact answer to this question. And against this fact, the recommendations I came across on the website of a major producer of respective supplements, which state that you would need

• 80g of palm oilen (cooking oil),
• 160g of rice bran oil,
• 3kg of barley,
• 1.5kg of wheatgerm, or 
• 4kg of oats

to (I quote) "achieve the required [my emphasis] level of tocotrienols" should tell any reasonable person that those "required" levels (~150mg) are probably required to generate the target revenue of the said company, yet probably not required for you or any other human being to thrive.

Do not stack one more, but take one out!

Instead of adding another overpriced (and probably overdosed) tocotrienol supplement to your regimen, it is thus probably wiser to simply drop any superflous and potentially harmful alpha-tocopherol only supplements which do would offset the alpha- to gamma- and delta- tocopherol ratio (this could potentially be ameliorated by taking a natural blend) and limit the total tocopherol intake to reasonable levels, as the latter has also been shown to hamper the absorption and retention of tocotrienols (Ikeda. 2003). In this context it is also noteworthy that Ping Tou Gee writes in a 2011 paper with the aptly chosen title "Unleashing the untold and misunderstood observations on vitamin E" that this fact alone would suggest that "there is a need to review critically on the dietary reference intakes recommendations" for alpha tocopherol (α-T). His bold statement that

[i]t is not known whether α-T is still essential to humans in long terms, α-T₃ [alpha tocotrienol] diet appeared to produce healthy rats over five generations.

is yet probably an attribution to Palm Nutraceuticals Sdn. Bhd. (which is not the aforementioned company which wants to force-feed you either their supplements or 2 cups of rice bran oil), of which he states in the acknowledgments that he thanks them "for permission to publish this paper" and further evidence for how pathetic parts of the research in the medical field is - awful this science business, isn't it?

References:

1. Aggarwal BB, Sundaram C, Prasad S, Kannappan R. Tocotrienols, the vitamin E of the 21st century: its potential against cancer and other chronic diseases. Biochem Pharmacol. 2010 Dec 1;80(11):1613-31. Epub 2010 Aug 7.
2. Ahmad NS, Khalid BA, Luke DA, Ima Nirwana S. Tocotrienol offers better protection than tocopherol from free radical-induced damage of rat bone. Clin Exp Pharmacol Physiol 2005;32:761–770 
3. Gee PT. Unleashing the untold and misunderstood observations on vitamin E. Genes Nutr. 2011 Feb;6(1):5-16. Epub 2010 Jul 20.
4. Ikeda S, Tohyama T, Yoshimura H, Hamamura K, Abe K, Yamashita K. Dietary alpha-tocopherol decreases alpha-tocotrienol but not gamma-tocotrienol concentration in rats. J Nutr. 2003 Feb;133(2):428-34.
5. Kato A, Yamaoka M, Tanaka A, Komiyama Ka, Umezawa I. Physiological effect of tocotrienol. J
   Japan Oil Chem Soc (Yukugaku) 1985;34:375–376.
6. Khanna S, Roy S, Ryu H, Bahadduri P, Swaan PW, Ratan RR, et al. Molecular basis of vitamin E
   action: tocotrienol modulates 12-lipoxygenase, a key mediator of glutamate-induced
   neurodegeneration. J Biol Chem 2003;278:43508–43515.
7. Kobayashi H, Kanno C, Yamauchi K, Tsugo T. Identification of alpha-, beta-, gamma-, and delta-
   tocopherols and their contents in human milk. Biochim Biophys Acta 1975;380:282–290.
8. Kuhad A, Chopra K. Attenuation of diabetic nephropathy by tocotrienol: involvement of NFkB
   signaling pathway. Life Sci 2009;84:296–301.
9. Lee SP, Mar GY, Ng LT. Effects of tocotrienol-rich fraction on exercise endurance capacity and
   oxidative stress in forced swimming rats. Eur J Appl Physiol 2009;107:587–595.
10. Mishra GD, Malik NS, Paul AA, Wadsworth ME, Bolton-Smith C. Childhood and adult dietary vitamin E intake and cardiovascular risk factors in mid-life in the 1946 British Birth Cohort. Eur J Clin Nutr. 2003 Nov;57(11):1418-25.
11. Newaz MA, Nawal NN. Effect of gamma-tocotrienol on blood pressure, lipid peroxidation and total antioxidant status in spontaneously hypertensive rats (SHR). Clin Exp Hypertens 1999;21:1297–1313.
12. Patel V, Rink C, Gordillo GM, Khanna S, Gnyawali U, Roy S, Shneker B, Ganesh K, Phillips G, More JL, Sarkar A, Kirkpatrick R, Elkhammas EA, Klatte E, Miller M, Firstenberg MS, Chiocca EA, Nesaretnam K, Sen CK. Oral tocotrienols are transported to human tissues and delay the progression of the model for end-stage liver disease score in patients. J Nutr. 2012 Mar;142(3):513-9. Epub 2012 Feb 1. 
13. Qureshi AA, Burger WC, Peterson DM, Elson CE. The structure of an inhibitor of cholesterol biosynthesis isolated from barley. J Biol Chem. 1986 Aug 15;261(23):10544-50.
14. Qureshi AA, Karpen CW, Qureshi N, Papasian CJ, Morrison DC, Folts JD. Tocotrienols-induced inhibition of platelet thrombus formation and platelet aggregation in stenosed canine coronary arteries. Lipids Health Dis. 2011 Apr 14;10:58.
15. Rimm EB, Stampfer MJ, Ascherio A, Giovannucci E, Colditz GA, Willett WC. Vitamin E consumption and the risk of coronary heart disease in men. N Engl J Med. 1993 May 20;328(20):1450-6.
16. Slover HT. Tocopherols in foods and fats. Lipids. 1971 May;6(5):291-6.
17. Sundram K, Khor HT, Ong AS, Pathmanathan R. Effect of dietary palm oils on mammary
    carcinogenesis in female rats induced by 7,12-dimethylbenz(a)anthracene. Cancer Res 1989;49:1447–1451
18. Shibata A, Nakagawa K, Sookwong P, Tsuduki T, Oikawa S, Miyazawa T. delta-Tocotrienol
    suppresses VEGF induced angiogenesis whereas alpha-tocopherol does not. J Agric Food Chem
    2009;57:8696–8704.
19. Tan B. Tocotrienols: The New Vitamin E. Spacedoc.net. http://www.spacedoc.com/tocotrienols
20. Weng-Yew W, Selvaduray KR, Ming CH, Nesaretnam K. Suppression of tumor growth by palm
    tocotrienols via the attenuation of angiogenesis. Nutr Cancer 2009;61:367–373.
21. Weng-Yew W, Brown L. Nutrapharmacology of tocotrienols for metabolic syndrome.
    Curr Pharm Des. 2011;17(21):2206-14. 
22. Whittle KJ, Pennock JF. The examination of tocopherols by two-dimensional thin-layer chromatography and subsequent colorimetric determination. Analyst.1967 Jul;92(96):423-30.
23. Yoshikawa S, Morinobu T, Hamamura K, Hirahara F, Iwamoto T, Tamai H. The effect of gamma-tocopherol administration on alpha-tocopherol levels and metabolism in humans. Eur J Clin Nutr. 2005 Aug;59(8):900-5.
24. Yu FL, Gapor A, Bender W. Evidence for the preventive effect of the polyunsaturated phytolside chain in tocotrienols on 17beta-estradiol epoxidation. Cancer Detect Prev. 2005;29(4):383-8.