Something Fishy: Leucine-Rich Protein + Fish Oil Supplement Boosts White Blood Cell EPA Content and Immune Response

Regular readers of the SuppVersity, as well as people who religiously follow Carl Lenore's Super Human Radio and happened to listen to my 1st appearance on the show, will know that I am generally skeptic about the usefulness of unwarranted high dose (>1-2g of combined EPA + DHA) fish oil supplementation. This is not because I think fish oil is poison, but rather out of my awareness that its pharmacological effects have more similarity to those of a drug than to those of a "common food" (including possible side effects).

Now, a group of scientists from the Netherlands and the United Kingdom published the results of a study (Faber. 2011) which found another interesting stone from the fragmented mosaic our current understanding of the effects and nutritional interactions of fish oil, in general, and EPA, in particular, resembles. While the aim of the study, which was to quantify the incorporation of EPA and DHA into white blood cells, was nothing new, the nutritional supplement they used for this purpose, a mixture of 2.4 g EPA, 1.2 g DHA, 39.7 g protein (including 4.4 g L-leucine), and 5.6 g oligosaccharides was innovative and the results were astounding. After 1 week of 2x200ml of what the scientists label a "medical food" (beware there is probably somebody patenting the formula already ;-), ...

[... from 0.5% at baseline] the percentage of EPA [in white blood cell phospholipids] rose to 2.8% (P < 0.001). Additionally, the production of proinflammatory cytokines in LPS-stimulated whole blood cultures was significantly increased within 1 wk.

It appears that the addition of protein has a positive influence on the incorporation of EPA into white blood cells, which, in turn, boosts the natural immune response to lipopolysaccharides stimulation. This, however, brings me back to my initial comment about "pharmacological" effects of fish oil: While for a cancer patient with lowered immunity, the consumption of the "medical food" or, what I would consider equivalent, a combination of whey + fish oil, would unquestionably be beneficial - but what about someone with allergies or even auto-immune issues? I assume you would agree that to further boost an immune system that is already running havoc does not seem to be a good idea!? In the end, and here I do not mind repeating myself, it again all comes back to considering who you are and what your current medical and nutritional condition is, before you start taking a supplement of which the whole Internet community seems to believe that its the healthiest (if not the only healthy) fat on earth...

Energy Restriction + Whey = Dieting Success, BUT the Magic is not in the Alpha-Lactalbumin

Ever since the first studies showed the beneficial effect of dairy in general and whey in particular on weight loss induced by caloric restriction, scientists have been arguing for alpha-lactalbumin being the key, or at least one of the key factors that contribute to the anti-obesity effects of milk products.

A group of scientists from Finland (Shi. 2011) did now investigate the exact contribution of alpha-lactalbumin to the dairy / whey induced anti-obesity effects. For 50 days Shi et al. fed mice on one out of three isocaloric, energy restricted diets containing either casein (calcium 0.8%) or two different high-calcium (1.8%) whey protein-based (α-lac + Ca and AA + Ca) amino acid mixes, which differed only in the amount of a-lactalbumin they contained. The results contradict the hypothesis that alpha-lactalbumin is the/a major driving force behind dairy induced weight loss outcomes:

The mice on α-lac and AA lost significantly more weight and body fat than mice on casein diet (P < 0.05). There was no difference between α-lac and AA diets. Ad libitum feeding after weight loss resulted in weight regain and fat accumulation in all groups

So, in the end, it obviously all comes down to the amino acid composition of whey protein that makes it superior to casein, but not to a similarly composed amino acid mixture without the addition of the alpha-lactalbumin fraction. The fact that none of the tested proteins had an advantage over the other in a non-dieting (i.e. calorically not restricted) environment, also indicates that it is the improved provision of essential amino acids from whey, which makes the difference.

Figure 1: Body fat percentage of mice after 50 days on different calorically restricted dietary regimens (Shi. 2011)

Yet, other than the ever-growing leucine fan-group on the web may now be led to believe, a further in-group differentiation  (cf. figure 1) into mice receiving additional amino acids (+100% aspartic acid & tryptophan, or +100% leucine) shows that additional leucine does not improve, but in fact blunt fat loss. In this otherwise isocaloric environment, this is probably due to the reduced percentage of other (essential and conditionally essential) amino acids in the chow, the consequences of which may be impaired fatty acid oxidation and suboptimal metabolic function in general.

One last thing, though: Before you completely discard the value of casein or even alpha-lactalbumin in your diet, I better told you that, compared to the amino acid group (without alpha-lactalbumin), all four, the whey + alpha-lactalbumin (all three groups), as well as the casein group retained a higher amount of muscle mass than the amino acid group (cf. figure 2) - with the results for the +100% asp + trp and the +100% leucine being statistically significant.

Bottom line: To cut body fat use pure whey with natural alpha-lactalbumine to optimize fat loss and retain muscle mass. Since, the difference in muscle retention to the other whey + alpha lactalbumine groups is not significant, the addition of supplemental amino acids including the "holy" (you may recognize I cannot stand the way it is currently hyped by the industry) leucine could make sense if you want to gain muscle; according to the data from this study, it is yet superfluous if your intention is to cut body fat.

More Conflicting Evidence on Leucine Metabolite: HMB Makes Volleyballers Stronger.

The leucine metabolite HMB, i.e. β-Hydroxy-β-Methylbutyric acid, is unquestionable the comeback kid of the supplement industry. "It works!", "It's useless!", ... every now and then a new study supports one side of the debate. A group of international scientists does now present a study, which would support the use of the expensive, yet readily available amino acid.

Portal et al. (Portal. 2011) investigated the effect of 3g HMB/day "on body composition, muscle strength, anaerobic and aerobic capacity, anabolic/catabolic hormones and inflammatory mediators in elite, national team level adolescent volleyball players (13.5–18 years, 14 males, 14 females, Tanner stage 4–5) during the first 7 weeks of the training season". The results were unequivocal, yet not overtly impressive:

HMB led to a significant greater increase in FFM by skinfold thickness (56.4 ± 10.2 to 56.3 ± 8.6 vs. 59.3 ±  11.3 to 61.6 ± 11.3 kg in the control and HMB group, respectively, p < 0.001). HMB led to a significant greater increase in both dominant and non-dominant knee flexion isokinetic force/FFM, measured at fast (180°/sec) and slow (60°/sec) angle speeds, but had no significant effect on knee extension and elbow flexion and extension. HMB led to a significant greater increase in peak and mean anaerobic power determined by the Wingate anaerobic test (peak power: 15.5 ± 1.6 to 16.2 ± 1.2 vs. 15.4 ± 1.6 to 17.2 ± 1.2 watts/FFM, mean power: 10.6 ± 0.9 to 10.8 ± 1.1 vs. 10.7 ± 0.8 to 11.8 ± 1.0 watts/FFM in control and HMB group, respectively, p < 0.01), with no effect on fatigue index.

With no measurable effect on aerobic fitness, anabolic hormone levels or inflammatory mediators, the use of HMB appears to be advisable only in power and strength athletes.

If you are a bodybuilder on a high protein diet, I would however doubt that you would see any benefits from additional HMB. A diet rich in quality protein (high BCAA and particularly leucine content) should provide you with adequate amounts of leucine to keep endogenous HMB levels elevated.

Beta-Hydroxy-Beta Methylbutyrate Strikes Back: HMB Increases GH and IGF-1 at the Expense of Insulin Resistance

"HMB" - do you remember these three letters? If you have been around the supplement world in the early 2000s, you probably will. Beta-hydroxy-beta methylbutyrate (HMβ) is a metabolite of leucine which has been advertised to enhance exercise performance, reduce fatigue and promote muscle gains. Nothing of that has been substantiated within larger human studies, though; and thus HMB a former star among the amino acid supplements has almost been forgotten.

Now, a recent study from Brazil (Gerlinger-Romero. 2011) provides data suggesting that HMB may in effect raise the content of pituitary GH mRNA and growth hormone [GH], as well as hepatic IGF-I mRNA and serum IGF-I concentrations.

It was observed that the HMβ treatment induced an increase in GH mRNA by 65% (P < 0.001) and in GH content by 20% (P < 0.05) compared to control group. The IGF-I mRNA expression in liver, as well as the serum IGF-I concentration was also significantly increased in the HMβ-treated animals.

There are some downsides to these results, however. Firstly, this is another rodent study, so we do not know how the results of this 4 week intervention would translate to human beings. At least, other than in previous investigations, the amount of HMB, 320mg/kg body weight, which is equivalent to roughly 52mg/kg body weight in a human being, would be affordable, but ...

Figure 1: Effect of the HMβ-treatment on Insulin concentrations.
Data are expressed as mean ± SEM. *P < 0.05; n = 9–16 animals per group
(Gerlinger-Romero. 2011)

... and here comes secondly, the rats became hyperinsulinemic (cf. figure 1), or in other words HMB supplementation induced insulin resistance. With the latter being at the heart of the metabolic syndrome I would think twice before attempting to boost my growth hormone level with an amino acid metabolite that will compromise my body's insulin management.

Protein Supplementation in Heavy Resistance Training: "A case for whey protein"

"Protein builds muscle!" < this is a sentence you will read pretty often in the advertisements - sorry, I meant "scientific explanations" supplement producers print onto their product labels to make you buy their newest "invention". In the end, however, it mostly comes down to a blend of different forms of whey protein with added vitamins minerals and digestive enzymes. But do you really need all that? Do you even need whey?

Table 1: Approximate Essential Amino Acid Profile of Various Protein Sources

A group of scientists from Finland (Hulmi. 2010) would probably say "YES!". In a very recent review they come to conclusions which sound pretty similar to what you can read all over the web:

Most, but not all studies have shown that supplementation of whey alone or with carbohydrates immediately after and possibly before and during resistance exercise can enhance the muscle hypertrophy response to resistance training in healthy adults. Such a response seems to at least be the case when comparing the effects of whey versus a non-energetic, or carbohydrate or soy protein alternative. Some studies also suggest that whey may enhance recovery from heavy exercise and possibly decrease muscle damage and soreness. This could, over time, enhance training adaptations by way of increasing training volume or reducing the potential for over-reaching/over-training.

There is another point, where Hulmi et al. reasoning is pretty much in line with main-stream "bro-science" and this is the extraordinary role of leucine in terms of muscle protein synthesis and hypertrophy:

[Leucine] is probable that the most important component in whey, for increasing protein synthesis and skeletal muscle size, is its high concentration of the BCAA, leucine (see
Table 1). Leucine, acting as a signaling molecule in the mTOR cascade, has been shown to be a critical amino acid for increasing skeletal muscle protein synthesis, both in vitro and in vivo in humans and rats. Leucine may also be involved in suppressing muscle protein degradation.

Their perspective is yet a little more differentiated:

However, if recent data involving rodents can be duplicated in humans, leucine concentrations from whey may only affect peak activation of skeletal MPS but not the duration of MPS or duration of mTOR signaling. Similarly, despite the positive effects of leucine per se, it likely is not the sole factor responsible for whey-induced adaptations to resistance training. For example, adding leucine to intact protein has been shown to offer little, if any, effect on protein synthesis and protein balance when consumed after resistance exercise.

Bottom line: If you go for muscle, go for whey and get drag yourself to the gym ;-)

25g Whey Protein or 10g of EAAs Maximize Post Workout Protein Synthesis Without Additional Carbs

While this probably won't end the debate on whether post-workout carbohydrate intake is a good or a bad thing, a recent review of the literature presented by Steward M. Phillips at the The Summer Meeting of the Nutrition Society (Phillips. 2010) suggests that the addition of carbs to your post-workout shake is at least unnecessary, as long as you have a sufficient amount of leucine and EAAs in it:

The increment in [muscle protein synthesis] MPS is maximally stimulated at a dose of protein of approximately 25 g or 10 g EAA. This rise is based solely on protein consumption and is not augmented by carbohydrate, at least when protein is adequate.

Also, the type of protein is of utmost importance. Phillips recommends whey protein or (please note that it is not whey and EAAs) a blend of EAAs at dosages of 25g and 10g, respectively.

Figure 1: Whole blood leucine concentration (μM) following resistance exercise from subjects who consumed 500 ml fluid skim (low fat) milk and an isonitrogenous and isoenergetic quantity of a soya drink (drawn with data from Wilkinson et al.).

As can be seen from figure 1, even skimmed milk, which is only partially whey, produces a larger increase in whole blood leucine than soy protein (soy is for girls only, anyway ;-) Consequently, you should prefer dairy proteins or whey concentrates, isolates or hydrosolates as your post workout protein sources.