Milk, a Glucose Uptake Promoter That's More Than the Sum of Its Parts. Plus: HICA & HMB in Yogurt. How Much EAA in Your Protein? Raw Milk Does not Cure Lactose Intolerance

Milk is one of the few foods that are advertized by celebrities that could actually be good for you (photo from the "Got Milk" campaign)

I hope you are not fed up with milk and dairy, yet, because today's SuppVersity short news have a ton of it. What exactly?

Well, after taking a closer look at the surprisingly high amount of HICA and the comparatively small amount of HMB in yogurt and reviewing the EAA content of six common protein sources, we are going to delve deeper into the latest evidence that shows that milk is much more than the sum of its parts, i.e. milk protein, lactose and fat and conclude on a note on another widely known Internet myth that says that people with lactose intolerance could drink raw milk without a problem.

You can learn more about dairy at the SuppVersity

Dairy Has Branched-Chain Fatty Acids!

Is There Sth. Like a Dairy Weight Loss Miracle?

There is Good A2 and Bad A1 Dairy, True or False?

Lactulose For Your Gut & Overall Health

Is There a "Fat Advantage" for Dairy Lovers

Dairy, Diabetes, Estrogen, IGF-1, Cancer & More

• 

  Figure 1: Amount of HICA & HMB (in µg/L) in commercial whole milk and yogurt (Ehling. 2014)

  Significant amounts of HMB and specifically HICA in yogurt. We all know that yogurt is among the dairy products with the most evidence of significant health benefits. That yogurt could be a decent muscle builder, on the other hand, would be news.
  
  With ~5 mg of β-hydroxy-β-methylbutyric acid (HMB) and up to 12.5 mg of α-hydroxyisocaproic acid (HICA) yogurt would have what it takes if the values of HMB and HICA Stefan Ehling and Todime M. Reddy measured in regular yogurt were on a per gram, not a per liter basis (Ehling. 2014).
• How much EAA in my protein? Since I have been repeatedly questioned about the essential amino acid content in whey, milk, soy, and egg protein. I have published Table 1 which originally appeared in a 2010 study by Hulmi et al. on Facebook, already.

  

  Table 1: Approximate amino acid composition of popular protein powders (Hulmi. 2010)

  For those of you who are missing out on the daily 12+ SuppVersity Facebook News, because they haven't liked www.facebook.com/SuppVersity, yet. I will now publish it again - as a reference source, if you will.
• Milk a glucose uptake promoter that's more than the sum of its parts. In view of the fact that we have already two dairy related news in today's SuppVersity article, I decided to have another "milky news" in today's SuppVersity short news item. One all the milk-drinkers among the SuppVersity readers will cherish.
  
  Recently, Shirin Panahi and colleagues from the University of Toronto, the Mount Saint Vincent University and the University of Guelph published the results of a randomized, cross-over study that was conducted to prove / disprove the hypothesis that

  "[T]hat regulation of postprandial glycemia after milk consumption occurs through both insulin and insulin-independent actions due to interactions among its macronutrient components and energy content. The objective was to compare the effects of isovolumetric (500ml) beverages of whole milk (3.25% M.F.), each of its macronutrient components (protein, lactose and fat) and their combination (a simulated milk beverage) on postprandial glycemia, glucoregulatory and gastrointestinal hormones and gastric emptying in healthy young men" (Panahi. 2014).

  In the course of the study, the 12 young, male subjects consumed beverages containing 500 ml of whole milk (3.25% M.F.) (control), a simulated milk beverage based on milk macronutrients or milk protein (16g), lactose (24g) and milk fat (16g) in isolation.

  

  Table 2: Nutritional composition of the test meals | ^a Composition of each beverage as provided by
  the manufacturer; ^b amounts given are per 500ml serving. Paracetamol (1.5g), vanilla extract (1.2ml)
  and sucralose (0.02g) were added to all beverages (Panahi. 2014).

  What the researchers found was that both the whole and simulated milk had similar beneficial effects on blood glucose rise after their "meal" (drink), but as it turned out the simulated milk resulted in a significantly higher (41%) glucagon-like peptide-1 (GLP-1) production and lower (43%) ghrelin areas under the curve (AUC) than whole milk (P=.01 and P=.04, respectively).

  

  Figure 2: Glucose, insulin, c-peptite, rate of insulin secretion, GLP-1, PYY, CKK
  and ghrelin levels after the test "meals" expressed relative to the sum of the effects
  of the same amount of milk protein, lactose and fat (Panahi. 2014)

  Now, all that would hardly be newsworthy (at least not for regular SuppVersity readers, if the two samples, i.e. both the whole and simulated milk, didn't lower the glucose (P=.0005) levels more than predicted by the sum of AUCs for their components (see Figure 2)!

  "Adjusted for energy content, milks produced lower glucose and hormone responses than predicted from the sum of their components. The effect of protein/kcal on the AUCs was higher than fat/kcal for insulin, C-peptide, insulin secretion rate, GLP-1, CCK and paracetamol (P < .0001), but similar to lactose except for CCK and paracetamol, which were lower. The response in PYY and ghrelin was similar per unit of energy for each macronutrient" (Panahi. 2014).

  In other words: Milk is way more than the sum of it's parts. In that, the "regulation of postprandial glycemia after milk consumption occurs through both insulin and insulin-independent actions due to interactions among its macronutrient components and energy content to achieve lower postprandial glycemia than predicted from the sum of its components" (Panahi. 2014).

If you haven't read it, already, take a look at my rebuttal to the latest assault on milk | read more

Bottom line: Quite the "milky" short news, right? Well, I guess it may be worth topping off all these good news about dairy products with a bad one. The common "Internet wisdom" that raw milk could offset the problems of people with lactose intolerance is a myth.

According to a 2014 paper by Sarah Mummah et al. who tested this myth in 16 adults with self-reported lactose intolerance and lactose malabsorption confirmed by hydrogen (H2) breath testing, "raw milk fail[s] to reduce lactose malabsorption or lactose intolerance symptoms compared with pasteurized milk among adults positive for lactose malabsorption." (Mummah. 2014) The "raw milk can be consumed by anyone" anecdote does thus belong to the realms of scientifically unwarranted die-hard bro-science | Comment on Facebook!

References:

• Ehling, Stefan, and Todime M. Reddy. "Investigation of the Presence of β-Hydroxy-β-methylbutyric Acid and α-Hydroxyisocaproic Acid in Bovine Whole Milk and Fermented Dairy Products by a Validated Liquid Chromatography–Mass Spectrometry Method." Journal of agricultural and food chemistry 62.7 (2014): 1506-1511.
• Hulmi, Juha J., Christopher M. Lockwood, and Jeffrey R. Stout. "Review Effect of protein/essential amino acids and resistance training on skeletal muscle hypertrophy: A case for whey protein." (2010).
• Mummah, Sarah, et al. "Effect of Raw Milk on Lactose Intolerance: A Randomized Controlled Pilot Study." The Annals of Family Medicine 12.2 (2014): 134-141. 
• Panahi, Shirin, et al. "Mechanism of action of whole milk and its components on glycemic control in healthy young men." The Journal of nutritional biochemistry (2014).

23g of Dairy Protein + 5g of Leucine Turn Cardio Sessions Into Muscle Building Workouts - More Protein + Extra Leucine = Higher mTOR, But Minimally Improved FSR

No matter how much you supplement, running will probably never be the "most anabolic" sport of all. On the other hand, it's certainly less catabolic than broscienctific horror stories of muscle loss and weakness would tell.

Researchers from the Massey University Wellington claim: "Ingesting 23 g of protein with 5 g added leucine achieved near-maximal FSR after endurance exercise." (Rowlands. 2014; my emphasis). If you think "near-maximal" fractional protein synthesis after endurance exercises sounds incredible, I would like to invite you to join me and take a look at the design and results of this recent study from the School of Sport and Exercise.

As the authors point out, "the purpose of this study was to determine if a reduced dose of protein and leucine ingested following endurance exercise resulted in a similar anabolic signal impulse for the stimulation of skeletal muscle myofibrillar protein FSR, relative to the higher protein-leucine dose associated previously with improved recovery of performance." (Rowlands. 2014)

You can learn more about protein intake at the SuppVersity

Are You Protein Wheysting?

5x More Than the FDA Allows!

Protein requ. of athletes

High EAA protein for fat loss

Fast vs. slow protein

Less Fat, More Muscle!

In addition Rowlands and colleagues examined the phosphorylation (as a surrogate marker of activity)of signaling proteins within the mammalian target of rapamycin complex 1 (mTORC1) pathway to study the associations between plasma amino acids, translational signaling and myofibrillar FSR. The scientists' hypothesis was that the lower ingested quantity of protein (23 g) plus leucine (5g) would be sufficient to stimulate myofibrillar FSR to an equivalent magnitude to a 3-fold higher amount.

If the latter was possible, Rowland et al. assumed that the mTORC1 pathway phosphorylation between the two protein-leucine would be identical, as well.
The subjects were 12 endurance-trained male cyclists with mean age 30 y, stature 179 cm, and weight 78.1 kg (7.8) completed the study. Mean VO 2 max was 60.4 mL/kg /min with a corresponding Wmax of 323 W.

Figure 1: Graphical overview of the experimental procedure (Rowlands. 2014)

"The research design was a randomized single-blind triple crossover. Details of one of the three 7-d experimental blocks and the experimental testing protocol are provided in Figure 1. Two weeks prior to the first experimental block, participants completed a standard test on a Velotron ergometer (Racer Mate, Seattle, USA) to determine VO2 max and Wmax. The next day participants completed a familiarization of the testing procedure (100-min cycle, see below) (Figure 1A).

Physical activity and diet were standardized for 4.5-d prior to a 2-d period of control prior toeach experimental testing day. Standardization was prescribed by way of verbal and writing instructions and record in training and dietary recall diaries; participants were asked to replicate on days -6 to -2 (outcomes not recorded). Control of exercise on protocol day -2 (Figure 1A) comprised a 90-min ride with a warm up of 10 min at 30% (Wmax), 8 min at 40%, 2 min at 50%, then intervals (4 x 5 min at 70%) interspersed with three blocks of 3 x 2-min intervals at 85%, 80%, and 75%, respectively, interspersed with 2-min periods at 50%, followed by 5 min at 40%."

Following this ride and for the remainder of day and day following (Figure 1B), participants performed no training and were provided with a preweighed diet providing sufficient energy to balanceindividual caloric requirements based on the Harris-Benedict equation for activity factor of 1.6.

Don't be fooled by the amino acid additions: While there is plenty of evidence that "optimized amino acid blends" or "enhancements" are great for supplement producers to justify why they're selling you cheap whey protein / bullshit amino acid products at a crazy price, there is no evidence that they are superior to plain whey protein (see "Are You Still Wasting Money on Amino Acid Products?" | read more)... What? Oh, you want to know why Rowlands et al. do it in the study at hand? Well, because they work for Nestec Ltd. aka Nestlé - I guess that's also why there was no 30g of pure whey isolate control ;-)

The 100 min of cycling comprised a warm-up (as above), intervals (%Wmax) of 8 x 2-min (90%), 2 x 5 min (70%), 2 x 2 min (80%) and 3 x 1 min (100%), interspersed with recovery 2-min (50%); and 8 min cool-down (40%). During exercise, participants consumed 800 ml /h of artificially sweetened electrolyte solution to maintain hydration and were fan cooled.

The supplementation regimen

Following exercise, participants showered, and then ingested the first nutrition serving 10-min after cessation of exercise and subsequently every 30 min over the first 90 min of the 240-min assessed recovery (Figure 1B).

Figure 2: Nutrient composition of the test drinks, which contained a whey + milk mixture that was enriched with leucine and spiked with maltodextrine, fructose and canola oil (Rowlands. 2014)

"The experimental beverages consisted of milk-based drinks containing milk protein concentrat and whey protein isolate (2:1 w/w), L-leucine, maltodextrin and fructose (1:1 w/w), and freeze dried canola oil. Four equal servings of 300 ml of the beverages were consumed during the recovery period for a total volume of 1200 ml.[...] The 15LEU supplement was compared to one-third of the protein-leucine quantity (23.3/5/180/30 g, 5LEU) - an intake hypothesised to yield a bioequivalent similar myofibrillar FSR, and to a nonnitrogenous, isocaloric control (0/0/274/30 g, CON). All beverages also contained 1.4 g NaCl, 14.4 g vanilla essence, and 3.6 g of emulsifier (Paalsgard 0096, Paalsgard A/S, Denmark) per 1200 mL."

As the data in Figure 3 shows, even the "small" shake was potent enough to achieve (almost) maximal fractional 0.95%/h protein synthesis rates.

Figure 3: mTOR (C1+C2) response to protein ingestion (left) corresponding mean fractional protein synthesis (%/h; right) in the 12 healthy male subjects (Rowlands. 2014)

Although the 15LEU = high protein drink achieved minimally higher protein synthesis rates, the discrepancy between the extremely elevated mTOR levels early post ingestion and the effective differences in FSR clearly suggest that we are approaching a physiological maximum with at FSR rates of 0.11% /h... or as Rowland et al. put it:

"The current myofibrillar FSR appeared to be limited by an undefined intramuscular mechanism since only a small and bioequivalent increase in FSR occurred with 15LEU despite sustained 1.4- to 1.9-fold higher plasma leucine and amino-acid concentrations and higher p70S6K-rpS6 phosphorylation." (Rowlands. 2014)

With reference to previous research by Atherton et al., Rowlands et. al. speculate that they may have encountered a "muscle full" effect (Atherton. 2010) to explain the discordance between human muscle protein synthesis and mtorc1 signaling.

Figure 4: Fractional protein synthesis in response to resistance training + whey protein ingestion (Tang. 2009)

Bottom line: Whether we are seeing a "muscle full" effect is something that's difficult to tell. What appears to be certain, though, is that resistance training allows for a greater uptake of protein synthesis (Tang. 2009; 0.15% /h in response to resistance training + whey protein). Although the supplements were somewhat different, I am thus inclined to believe that the absence of resistance training and the corresponding mTOR-C2 activation in response to aerobic ( (mTORC1 by supplementation, only) vs. resistance training (mTORC1 by supplementation, mTORC2 by contraction) is the limiting factor, here (Drummond. 2009).

The latter would imply that the results are not applicable to resistance training directly. Even if corresponding studies clearly suggest that there is a limit to the benefits of "evermore" protein with resistance training, as well, the "limit" may be higher than "just" 23g of mixed dairy protein + 5g of leucine.

Reference:

• Atherton, Philip J., et al. "Muscle full effect after oral protein: time-dependent concordance and discordance between human muscle protein synthesis and mTORC1 signaling." The American journal of clinical nutrition 92.5 (2010): 1080-1088.
• Drummond, Micah J., et al. "Rapamycin administration in humans blocks the contraction-induced increase in skeletal muscle protein synthesis." The Journal of physiology 587.7 (2009): 1535-1546.
• Rowlands, David S., et al. "Protein-Leucine Fed Dose Effects on Muscle Protein Synthesis After Endurance Exercise." Medicine & Science in Sports & Exercise (2014).
• Tang, Jason E., et al. "Ingestion of whey hydrolysate, casein, or soy protein isolate: effects on mixed muscle protein synthesis at rest and following resistance exercise in young men." Journal of Applied Physiology 107.3 (2009): 987-992.

3.2kg of Lean Mass Over Night W/ 40g of Slow Digesting Protein 30min Before Bed!? Over One Year, a Positive Nitrogen Balance and +20% FSR Could Make It Happen!

Image 1: Babies instinctively know how to grow - mother's milk (60% whey, 40% casein, at later stages) + sleep ;-)

Tell me, does the sentence "Where Bro- and Pro-Science Unite in the Spirit of True Wisdom" ring a bell? Anyone? Well, that's what I thought. It's the mantra of the SuppVersity... unfortunately, more often than not, one "science" does not really care about the other, so that studies as the one by Peter T. Res and his colleagues from the University of Maastricht are unfortunately rather the exception than the rule (Res. 2012).

Pre-bed protein intake could be a crucial determinant of 24h protein synthesis

I guess, I won't have to tell you that bro-science has it that the most important thing to do before you go to bed (and for some hardcore "bros" even in the middle of the night) is not to brush your teeth, let alone to shower or at least wash your face, hands, feet and certain other body parts... no! The most important thing to do before you go to bed is to have a huge serving of protein - preferably a "night-time protein", like a slow-digesting casein-based protein shake with some additional fats to further slow the absorption (whether the fat will actually prolong the digestive process beyond what you will see if you ingest intact micelles, which will then be hydrolized in the gut and start clumping is anyone's guess, though). In fact, this is one of those truisms that has been repeated so often on the boards (and the ads) that you may be surprised to hear that Res et al. rightly claim that their study is the first one to investigate, whether this practice does actually povide any benefit for the professional or recreational lifter.

To this ends, the scientists recruited a group of 16 of the usual suspects, ah.. pardon "recreationally active men", which in this case means that they had a weekly physical activity level of 6.3h and 5.2h for the eight men in the protein and the seven in the placebo arm (#8 had a problem with a catheter, so that he had to be excluded), respectively. As you would expect from any study investigating the effect of dietary supplements on exercise performance and/or muscle growth, the subjects received a standardized dinner (0.04kcal/kg; 57% carbs, 13% protein, 30% fat) at the evening before the testing session, as well as "identical" (obviously the energy content was matched to the body weight of the respective individual) meals for breakfast, lunch and dinner on the day of the experiment. The overall protein content of the regular meals was 1.2g/kg body weight and should thusly at the lower end of what the "bros" would prescribe as a baseline protein intake for anyone trying to gain muscle.

Exercise protocol: Leg presses and extensions 8x8 - 45 min total

After a standardized meal at 4:45pm and a whole host of experimental procedures (most importantly to place the catheter for the multiple blood draws during the night), the participants performed 8 sets of 8 reps on a leg press and another 8 sets of 8 reps on a leg extension machine (2 sets at 55% and 65%, 6 sets at 75% of 1RM; "subjects were verbally encouraged during the test to complete the whole protocol"). Rest between sets was 2 min rest between exercises 5 min. At 9pm, ca. 15min after the exercise test, the subjects received a serving of Lucozade Sport Body Fuel and Lucozade Sport Recovery (yes, the study was supported by GlaxoSmithKline ;-), which contained 60g of carbs and 20g of whey and thusly mimics what many non-carbophobic athletes use to replete glycogen stores and ramp up protein synthesis after a workout. After a muscle biopsy at 11:30pm, the subjects received either 40g casein protein or placebo and "remained in a supine position until 0:00am" ... I lover this formulation, because it suggests that with all those catheters every subject fell asleep at exactly 0:00am after "remaining in a supine position" *rofl* - be that as it may, the scientists simply assume that their subjects had slept for 7 hours, when they woke them at 7am for the second muscle biopsy.

Figure 1: Plasma levels of essential amino acids (µmol/L) and overnight mixed muscle fractional protein synthesis rates (measured by phenylalanine tracer) in subjects after receiving 40g of slow acting protein (casein) or placebo 30min before bed (at T=0; data adapted from Res. 2012)

As you can see in figure 1 the EAA levels the scientists measured in the blood of their subjects in the course of the night was profoundly elevated in response to the protein feeding. It is thusly not surprising that the fractional protein synthesis rates the scientists calculated for the 7.5 h of overnight sleep was ~22% higher in the "pre-bed" protein group than in the subjects who received the placebo supplement (cf. figure 1; right). Yet, although this may sound much, we are talking about 0.059% vs. 0.048% fractional muscle protein synthesis per hour and thusly about a 0.011% increase, which was only "borderline significant" (meaning p = 0.05).

Figure 2: Net protein breakdown, synthesis, oxidation (all left) and balance (right) measured over night in previously exercised subjects after receiving 40g of slow acting protein (casein) or placebo 30min before bed (based on Res. 2012)

What is probably more important than the difference in fractional protein synthesis, anyways, is the overall net protein balance, which indicates that contrary to the trainees in the placebo group, the subjects who received a 40g serving of casein 30 min before they went to bed (and hopefully slept 7h ;-) did effectively "gain" muscle, or I should say, muscle protein over the course of their 7.5h nightly "fast", while the subjects in the placebo group ended up losing a minimal amount of skeletal muscle protein.

3.2kg of lean muscle mass in one ear with nothing but a protein shake before bed?!

If we take a look at the abstract numbers the scientists measured, such as an increase in whole body (!) net protein retention of ~50µmol/kg (measured in phenylalanine tracer molecules) over the course of 7.5h and do some math, this tells us that a trainee who weighs ~80kg and followed this practice over the course of one year, where we assume that he trains four times a week (i.e. 208 sessions) this would allow him to store 832mmol or (if I did not miscalculate) ~146g of the phenylalanine tracer in the 208 nights following his training sessions... does that sound much? No, it certainly does not, but we just assume that for each of those phenylalanine molecules another molecule of each of the other EAAs was stored within the muscle (since we are talking about "whole body" protein retention, other organs will get their share as well, though), and further assume that they all weigh about the same (which is obviously bullocks) the 40g of casein every night would result in a net protein gain of 3.2kg! How does that sound?

Image 2: Quark = Natural #1 casein source

Note: Fatfree asked rightly, whether there are not any natural alternatives to protein shakes and as I thought this is relevant for everyone, I decided against answering in the comment area. Personally I would suggest you watch out for either curd/quark (~10g casein per 100g) which has tons of highly bioavailable calcium etc. An alternative with lower protein content is cottage cheese. More fat, but still nice - any other cheese. A huge chunk of steak could work, but I am not sure if that is not problematic in terms of nighttime digestion, which was one of the 2ndary results of the study at hand: Casein is easily digested while we sleep.

Now while this is a pretty optimistic calculation, while we are (again) dealing with "rookies" who obviously gain like crazy, and so on and so fort, the fact that there are still 175 days, where you don't train and your body would still be able to store some protein, goes to show how important a properly timed intake of protein and with it a persistent influx of readily available amino acids is, if you want to gain muscle - and in that it does not matter if that are going to be 500g or 10kg over the course of one year. However, I beg you not to forget that you cannot live on protein alone and that it is highly questionable that by escalating the dose to say 60g or 80g the net gains would increase by 50% let alone 100%, respectively. So keep that in mind before you set up a bathtub full of protein to sleep in ;-)

References:

• Res PT, Groen B, Pennings B, Beelen M, Wallis GA, Gijsen AP, Senden JM, VAN Loon LJ. Protein Ingestion before Sleep Improves Postexercise Overnight Recovery. Med Sci Sports Exerc. 2012 Aug;44(8):1560-9.