Beyond Celiac: Study Sheds New Light on Obesogenic Effects of Gluten - Are PPARs & Bacteria Both Involved?

Cornflakes peanut butter cookies - guaranteed not gluten free ;-)

With Christmas Eve being over, and grandma's cookies, Christmas stollen, and all sorts of other stuff from the bakery in front of you (literally), Christmas Day may actually prove to be a way more "dangerous" than Christmas Eve - not just because of the total amount of calories, but also because of the low satiety effect of these sweet treats.

A recent paper by scientists from the Universidade Federal de Minas Gerais in Belo Horizonte in Brazil does now point to another reason you better give those bakery products a wide berth - not just, but especially with the energy overshoot on Christmas day: Gluten!

Study confirms for the first time what scientists and laymen alike have been speculating about

In what the scientists claim is the first well-controlled study of the effects of gluten intake on metabolic health in a non-celiac, but Western-style diet scenario, Fabíola Lacerda Pires Soares and her colleagues put two groups of C57BL/6 mice on identical, iso-caloric high fat (hypercaloric) diets that differed only in terms of the amount of gluten that was added to the chow (0% gluten vs. 4.5% gluten).

Interestingly, the gluten diet did not influence any of the usual suspects, like food intake, total fat-free mass, fecal lipids excretion, blood lipid profile, blood total protein and ectopic (liver and muscle) lipid concentration (if you look closely you will realize that the gluten-free group actually had higher TRIGs, although the difference did not reach statistical significance).

Figure 1: Usual suspects and closer look at the effects 8 weeks gluten supplemented vs. gluten-free diets had on serum markers of metabolic syndrome and visceral fat parameters (Soares. 2012)

The data in figure 1 (right) does yet also show that the gluten content of the diet did nevertheless have a significant impact on the total body mass, visceral fat mass, lipid content and most importantly the adipocyte size.

Figure 2: Absolute adipokine levels (left) and fasting glucose and insulin levels, as well as Homa-IR (Soares. 2012)

Add to that the blunted expression of the anti-inflammatory and anti-diabetic fat hormone adiponectin and the increased the >5x higher expression of leptin (figure 2). And mix that with the reduced expression of PPAR-alpha and gamma of which Soares et al. argue that they may well be the key factor in the detrimental modulatory effect the addition of gluten had on the visceral fat structure and the lowered expression of the fat liberating enzymes LPL and and HSL, as well as reduced levels of the fat burning proteins ACC and CPT-1 (figure 3).

Figure 3: PPAR-alpha, -gamma, LPL, HSL, ACC and CPT-1 expression compared to rodents on regular chow (left); crown like structures in stained slices from visceral fat, inflammatory markers TNF-alpha and IL-6 (Soares. 2012)

So, even if the initially mentioned blood markers (aka the usual suspects) would suggest that both the gluten-consuming and gluten-free rodents were similarly bad off, the profound difference in inflammatory markers within the adipose tissue and the presence of comparatively many necrotic and inflammatory adipocytes in the crown like structures stand in line with increases in HOMA-IR, fasting glucose and insulin and an already compromised glucose clearance which are well-known harbingers of the metabolic syndrome.

These observations do not simply shed a whole new light on a hitherto largely ignored contributer to the etiology of the metabolic syndrome, they do also show that one of the reasons it has not been identified before is an over-reliance on BMI, total fat mass and serum lipids in the early stages of diabesity.

Reardless of whether the gut microbiome is part of the mechanism by which gluten predisposes the development of metabolic syndrome. Eating more inulin- and beta-glucan rich foods like Jerusalem artichokes, agave, bananas, onion, steel cut oats, wild yams, yacon, etc. certainly won't hurt your efforts to get lean, stay lean and leave the role of the obese diabetic to the other (read more)

Bottom line: The study at hand provides a good reason to limit your intake of "healthy whole grains" and other gluten containing foods, regardless of whether you suffer from celiac or not. Whether the established detrimental effects of gluten on the integrity of the intestinal wall and the increased leakage of bacterially produced endotoxins from the highly unfavorably changes in the gut microbiome in response to the high fat diets (Hildebrandt. 2009) are part of, or even the primary cause of these observations still has to be elucidated. The same goes for strategies to counter the translocation of the endotoxins across the gut lining (cf. "Shedding some light on the leaky gut") and the dose response relationship between the total amount of gluten in your diet and its effects on your metabolism. With 7% of pure gluten, it goes without saying that you would basically have to live of wheat in order to get to anywhere similar amounts of gluten in the diet... that said: Is it possible that the effects occur only in the presence of the high fat diet? After all, this alone has been shown to favor a pro-inflammatory gut microbiome.

You see there are enough questions to be answered in 2013 and the SuppVersity is going to be the place you will read the respective answers first ;-)

References:

• Hildebrandt MA, Hoffmann C, Sherrill-Mix SA, Keilbaugh SA, Hamady M, Chen YY, Knight R, Ahima RS, Bushman F, Wu GD. High-fat diet determines the composition of the murine gut microbiome independently of obesity. Gastroenterology. 2009 Nov;137(5):1716-24.e1-2.
• Soares FL, de Oliveira Matoso R, Teixeira LG, Menezes Z, Pereira SS, Alves AC, Batista NV, de Faria AM, Cara DC, Ferreira AV, Alvarez-Leite JI. Gluten-free diet reduces adiposity, inflammation and insulin resistance associated with the induction of PPAR-alpha and PPAR-gamma expression. J Nutr Biochem. 2012 Dec 17.

CLnA, the "Omega-3 Variety" of CLA from Pomegranate & Co, Has Potent Anti-Obesity Effects and the Potential to Become More Than Just Another Anti-Diabetes Drug.

Image 1: Pomegranate - I loved to eat them even before I realized that their seeds are the #1 dietary source (83%) of punic acid.

While more and more people are beginning to grasp the notion that with (naturally occurring) fats - as with everything else in life - there is no simple "good" and "bad", no clearcut "black" and "white" and no definite "beneficial" and "detrimental". The number of different fatty acids and their respective effects on the human metabolism is so vast that it is pretty hard to keep track of all those varieties of saturated and unsaturated carboxylic acids. I would thusly not be surprised if you simply assumed that the "n" in the headline of this blogpost was a type that had slipped in because poor Dr.Andro is chronically stressed from Christmas shopping... well, while the latter is actually correct, the former is not: CLnA is actually the omega-3 variety of the famous conjugated linoic acid (CLA), which in and out of itself is not a single but a group of 28 different trans- and cis-isomers that occur in our diet mainly in the shape of high and full-fat meat and dairy products.

CLnA - Conjugated Linolenic Acid is not a typo ;-)

Within the last couple of years even the medical establishment has come to realize that the chronic omega-6 (n6: linolic acid) overload in our diet is killing us. The "heart-healthy" PUFAs have now become the more and less heart-healthy PUFAs with the totally healthy *rofl* omega-3s and the not just as healthy omega-6s - both, of course, still totally "essential" and WAY better than saturated fats,... (attention: the afore statements are full or irony! Saturated fats are of course NOT the bad guys. Sorry, David if that lead to confusion)... but I am getting derailed, here. So let's get to the point. What every reasonable person appears to agree on, these days, is that we have to lower the ratio of n6:n3 fatty acids in our diets. Now, I am asking you: Has it ever occured to you that CLA essentially is an omega-6 fatty acid? I mean its conjugated linoleic acid - "linoleic" as in omega 6 = linoleic acid! Probably not, right? The reason for that is yet (hopefully ;-) not that you are dump, but simply that the existence of an omega-3 "variety of CLA", namely conjugated linolenic acid, or short, CLnA, is something about which you will only hear, when you read blogs (such as the SuppVersity ;-), which do not stick to copying, pasting and commenting the stuff the authors have read on one of the major news-portals.

Table 1: CLnA isomer content in natural sources (data adapted from Hennesey. 2011)

From a molecular perspective,  CLnA isomers combine the conjugated double bond system of the classic conjugated linolic acid, you know, with the octadecatrienoic fatty acid (C18:3) structure of omega-3s, i.e. linolenic acid. Interestingly, this make-up confers these fatty acids with a high bio-active potential. Now, while this may sound like one of the frankenfood test-tube results of the gene-technology laboratories of Monsanto, we know at least 10 CLnA isomers which occur naturally in foodstuff or as byproduct of fermention processes (cf. table 1).

Adiposity, hyperlipidemia, cancer - CLnAs could help with all!

Image 2: Even if CLnAs would just prevent obesity, this illustration I borrowed from multiplemyelomalifeexpectancy.tk, shows that not being / getting obese alone would prevent a plethora of related maladies. Such as kidney failure, arthritis, gallbladder disease, infertility, asthma, fatty liver disease, sleep apnoea, depression, heart disease, hyperlipidemia, diabetes,... basically every major ailment the increasingly obese convenience society of the Western hemisphere is suffering from.

Due to their anti-adipogenic (meaning preventing the accumulation of body fat) effects CLnA fatty acids have been investigated as potential candidates for the treatment of the obesity epidemic for quite some time, now (Hennesey. 2011). In a 2002 article that was published in the Journal of Applied Biochemistry and Biotechnology, Nishimura et al. report that CLnA isomers exert apoptotic effects on mouse preadipocyte 3T3-L1 cell - or, in plain English, incubation with CLnA did not only hinder the "pubertal" fat cells from becoming mature adipocytes, it actually killed them. In vivo studies with rodents, such as Arao et al. (2004), where the administration of a diet that was enriched with 1% pomegrenate seed oil lead to a 27% reductin in omental white adipose tissue, were able to confirm the "rodent-real world signficance" of these test-tube results.

Other studies showed a normalization of hyperlipidemia in rodent models of the metabolic syndrome and a hand full of studies have explored the usage of CLnA isomers as cytotoxins in the treatment of cancer. In their concise review of the literature, Hennesey, et al. thusly rightly conclude that with their "potent inflammatory and immune modulating properties", their ability to "reduce the risk of obesity, improve cardiovascular health, and mediate strong anti-carcinogenic activity", the use of CLnA isomers or dietary enrichments could offer treatment strategies for pathologies, which "represent some of the greatest mortality risks to humans in the Western world and have been inextricably linked with diet" (Hennesey. 2011).

Adding diabetes to the list of potential targets for CLnA

For today, we are however going to focus on the most recent result from the research front: The effects of CLnAs on diabetes, or, to be precise, the increases in blood glucose, and decreases in anti-oxidant capacity that go hand in hand with the latter. In a recently published study (Saha. 2011), Siddhartha S. Saha and Mahua Ghosh from the Department of Chemical Technology at the University College of Science and Technology of the University of Calcutta (I don't have to tell you that this is in India, do I?) injected male albino lab rats with 60mg/kg streptozotocin (STZ) - this is a common and well-established method to induce a metabolic state that serves as a model of type II diabetes - and fed them diets that contained either no, or 0.5% of the total fat in the form of alpha-eleostearic acid (from bitter gourd, cf. table 1) or punic acid (which was in this case taken from snake gourd oil, but could as well have been extracted from the eponymous pomegrenate, cf. table 1).

Figure 1: Relative blood glucose levels vs. non-STZ injected control in streptozotocin injected rats over the course of the dietary intervention (data calculated based on Saha. 2011)

As you can see in figure 1, this 100% natural "food additive" had a more than pronounced effect on the +300% (vs. non STZ-injected control) elevated blood glucose levels of the "type-2 diabetic" rodents.

Figure 2: Relative level of lipid peroxidation (left) and total antioxidant capacity (right) levels vs. non-STZ injected control in streptozotocin injected rats after the 28-day dietary intervention (data calculated based on Saha. 2011)

And while the glucose levels were still 150% above those of the healthy control levels, the streptozotocin-induced lipid peroxidation in plasma, pancreas and erythrocytes of the lab animals was ameliorated by the snake gourd oil treatment (remember that is the stuff from pomegranate) and even reversed by the bitter gourd diet. Judged by the standardized FRAP assay, the "diabetic animals" that were fed a diet that contained 0.1% alpha-eleostearic acid (of the total diet, which had 20% fat) even exhibited a 10% greater total antioxidant capacity than the totally healthy control!

Figure 3: Relative expression of inflammatory cytokines, TNF-alpha and interleukin 6 in plasma capacity (right) levels vs. non-STZ injected control in streptozotocin injected rats after the 28-day dietary intervention (data calculated based on Saha. 2011)

Snake gourd oil, on the other hand, exhibited more profound effects on the elevated TNF-alpha, interleukin-6 and NF-kappaB levels of the STZ-treated rodents (cf. figure 2) and thus, at least this is my humble opinion, render punic acid the overall more promising agent with respect to the treatment of all sorts of inflammatory (or related diseases). After all, disturbances in the regulation of the nuclear factor kappa-light-chain-enhancer of activated B cells  (NF-kappaB) and the downstream over-expression of TNF-alpha and IL-6 are hallmark features of allmost all the aforementioned ailments of the increasingly obese western convenience society. This is also why I am quite certain that we are going to hear much more about the CLnAs in the month to come... and I guess, I don't have to tell you that right here, at the SuppVersity, is where you will read about respective studies first!

The IGF-1 Promoting, Myostatin Reducing, Muscle Building Effects of PGC-1 α-4: What It Does and Why Doing Cardio Before Weights Appears to Promote It's Expression

Warning: Reading this article won't make you look like Phil Heath over night.

As announced yesterday, I am about to get back to the study on PGC-1 alpha-4, the protein Carl Lanore and I talked about in the last installment of the SuppVersity Science Round-Up on Thursday. Since I am not going to simply repeat everything I already said during the show here, I suggest you download the podcast and listen to it before you read this article. Thus you would have a basic understanding of what the Ruas' study is all about and can class the additional information this article is going to provide with the stuff you've heard on Super Human Radio. If you don't have the time or are just sitting in the office, where listening to a radio show is not really an option, I would guess that those of you who have been around on the SuppVersity for some time now, should be able to connect the dots on their own.

PGC-1 alpha-4 the missing link between myostatin, IGF-1, hypertrophy and strength gains

With the combination of in-vitro and in-vivo data from rodents and humans the study Roas et al. published in the latest issue of Cell is a seam of information - literally. Actually, this is part of the reason, why I decided to restrict the following discussion to a summary of those findings that are either of general interest or can serve as a rational foundation for practically relevant conclusions, instead of simply reiterating the whole protocol.

• 

  Figure 1: Fluorescencemicroscopy analysis of myotubes expressing GFP alone or together with PGC-1 a1 or PGC-1 a4 (left) and effects on the expression of selected RNAs (Roa. 2012)

  PGC-1 alpha and its splice variants - The four known splice variants (alpha 1-4) the scientists tested for are expressed in most of the major organs of our body. Of particular interest for our discussion here are alpha-1 and alpha-4, with the former influencing 2002 and the latter controlling 519 gene function. The overlap between the two (98 genes) is actually pretty small, so that their downstream metabolic effects can be expected to be about as distinct as their underlying triggering mechanisms.
  
  While the energy sensing system appears to be responsible for the expression of PGC-1 alpha-1 (learn more about AMPK and how your body controls glucose uptake mitochondrial activity of the cells etc. depending on the local availability of energy), PGC-1 alpha 4 expression in skeletal muscle and thus the downstream effects on myostatin (inhibition) and IGF-1 (promotion) appear to be controlled by (contractile, but also metabolic) stress. Whether this is actually the case and in how far certain overlaps do exist will yet still have to be evaluated in future studies.
  
  

  

  Figure 2: Training or overtraining - good or bad inflammation; it's often difficult to hit the sweet spot (background adapted from Kramer. 2007)

  The same goes for the exact involvement of MAPK and other stress-sensors in our bodies and the dose-response relationship between the ROS and exercise induced expression of inflammatory factors such as IL-6 => NF-KappaB and their short term beneficial effects on the training induced adaptation processes (see figure 2). What can be said for sure, though, is that over-training and the downward spiral on the right side of  figure 2 is way more likely to be the underlying cause of suboptimal results, than an absence of adequate training stimuli on the left. Adequate recovery (primarily via rest + food and not by popping supplements or suppressing your well-deserved drowsiness with stims) is therefore about as, if not more important than the one additional rep you may or may not be able crank out at the end of an intense workout.

•  What exactly can PGC-1 alpha 4 do? The trends in RNA expression in figure 1 do actually give you an idea of what the ensuing effects should be, but I guess some actual data will make it even more obvious what all these gene essays mean.

  

  Figure 3: Effect of injected PGC-1 a  and DNA manipulation on muscle fiber composition and overall muscularity and phenotype of the rodents (Ruas. 2012)

  As the data in figure 3 goes to show, the effects of PGC-1 alpha 4 injections are almost identical to what you would see to a standardized hypertrophy training. And as you may remember from my dissertation on the podcast, the >17x increase in PGC-1 alpha 4 expression in response to reloading of a previously suspended hindlimb in the scientists' rodent model would confirm just that: PGC-1 alpha 4 is expressed in response to muscular overload (as it does obviously occur, when you have not moved your leg an inch for 10 days) and initiates adaptation processes that are meant to strengthen and "build" the muscle to ensure that it is up to future challenges like this.
  
  

  

  Figure 4: Immunohistochemical analysis of gastrocnemius muscle from wild-type (WT) and Myo-PGC-1 a4 animals

  Due to the fact that the effects Roas et al. observed were muscle fiber specific and quasi non-existent in muscles that are predominantly slow twitch fibers (e.g. soleus or planatris), the concomittant boost in MHCIIa and MHCIIx myosin heavy chain types you see in figure 4 may easily be misinterpreted as a "transformation" of muscle fibers. If you look closely at the immunohistochemical analysis of the gastrocnemius muscle from wild-type (WT) and Myo-PGC-1 a4 animals in figure 4 the pictures do yet speak a very different language. If anything, the amount of the very fast twitch glycolytic (only) type IIb fibers may have dimished ever so slightly. The amount of slow twitch oxidative muscle fibers, on the other hand, remained constant, while the number of both MHCIIa and MHCIIx positive myofibers increased (the same happens, as you should remember from the Intermittent Thoughts in bodybuilders and recreational trainees, as well).
   
• PGC-1 alpha 4 boosting agents include clenbuterol 5x (see Friday's "SuppVersity Science Round-Up Seconds"), forskolin 25x (both in vitro) and cold exposure (4°C) in rodent (!) brown adipose tissue.

Aside from the anti-cancer cachexia effect which is not directly related to the topic of this post, the previous paragraphs and the podcast should actually give you the most important information about this recently discovered splice variant of PGC-1 alpha, so that we can now segue into the "real-world" part of the study and take a closer look at the interactions with strength and cardio training I have been talking about on Thursday, as well.

Exercise and PGC-1 alpha 4 in real human beings

You cannot tell me that you have never heard of the notion that doing cardio not after but either before or or in-between your lifts an have its merit. If you can't remember it anyway, go back and reread "Before, After or In-Between? Study Puts Another '?' Behind the Widely Accepted 'Cardio After Weights' Paradigm."

Previous research associated PGC-1 alpha increases primarily with endurance training and, albeit to a lesser degree, glycogen depleting high intensity interval training (HIIT), or high volume resistance training. Over the years all of these training forms have been shown to contribute to mitochondrial biogenesis, a repartitioning of fiber types towards a more versatile oxidative myosin heavy chain pattern (similar to what you see in figure 4), the AMPK mediated stimulation of fatty acid oxidation and glucose uptake, angiogenesis and the prevention of muscle atrophy (Arany. 2008). The discovery of this new splice variant of the PGC-1 alpha protein does not diminish the significance of any of these results, but it does make one thing pretty obvious: Building muscle, endurance and oxidative capacity (mytochondria) are not mutually exclusive processes and it is very likely that there is a strong overlap between the metabolic and mechanic triggering processes.

It does in fact look as if the PGC-1 alpha "family" stands, if you will, at the crossroads of the aforementioned pathways with the "classic" alpha 1 variety being triggered by AMPK (and maybe other nutrient sensors) and the alpha 4 variety responding to the exercise-specific increase in stress signals. The results of the 8-week human study, Roas et al. conducted does yet show that things are - once again - not as easy as it may seem. If you look at the three training groups the subjects (the researchers don't provide details about age or training status, but probably young untrained men) were randomly assigned to...

• 

  Figure 5: Mo & Thu and Tue & Fri workouts (top) and results of the analysis of the biopsies that have been taken 48h after the last training session (Roas. 2012)

  Endurance Training (ET): During week 1, participants completed 30 min of stationary cycling at 65% VO2 peak 3 days per week. During week 2, participants completed 45 min of stationary cycling at 65% VO2 peak 3 days per week. During week 3, participants completed 45 min of stationary cycling at 65% VO2 peak 5 days per week. During weeks 4-8, participants completed 60 min of stationary cycling at 65%VO2 peak 5 days per week. 
• Resistance Training (RT): During week 1, participants were familiarized with resistance training program and practiced the movements with light weight during each of the four training sessions. During week 2, participants completed 2 sets of 8-10 repetitions to failure 4 days per week. During week 3, participants completed 3 sets of 8-10 repetitions to failure 4 days per week. During weeks 4-8, participants completed 4 sets of 8-10 repetitions to failure 4 days per week. Table S1 presents the full exercise program. 
• Combined Training (CT): The progression of the ET was the same as that described for the ET group, except that the durations were half as long as the ET group (i.e., 30min versus 60min). The progression of the RT was the same as that described for the RT group, except that the number of lifts was less the RT group. 

... as well as the exact protocol they have been following (figure 5, top), you would probably not have expected that the combined training protocol would have an edge over the higher volume resistance training in terms of both PGF-1 alpha 4 expression, as well as the decreases in myostatingthe increases in IGF-1, and the effective mean strength gains on the leg press (+30% for both with a minimal, statistically non-significant edge for the combined regimen; not shown in figure 5).

Implications: Why doing "cardio" before a workout could be beneficial

Figure 6: Free fatty acid levels before depletion (S1) and before (S2) and after (S3) exercise trial, as well as PGC1-alpha and p-AMPK expression (Psilander. 2012)

In the absence of detailed information about the increases in muscle CSA and protein content, it may be a bit too early to formulate any implications, but since the question of "doing cardio before a workout" was at the heart of an interesting discussion some of you started in the comment area to Friday's installment of the Seconds, I want to pick up on that and present a couple of garbled thoughts and references that may explain why the combined training did produce greater increases in PGC-1 alpha-4, as well as more pronounced downstream effects on myostatin and IGF-1 than the "growth specific" strength training program.

Now, one of the beauties of having your own blog with 1020 individual posts is that you can often simply refer people to previous posts such as the one from which I just copied figure 6 into this article. In fact, the title "8x Increase in "Mitochondria Building" Protein PGC1-Alpha W/ Medium Intensity Exercise in Glycogen Depleted Elite(!) Cyclists" actually gives away most of the 'secret' that's probably behind the purported benefits of a combined training regimen: Glycogen depletion!

Can I do HIIT instead? Personally I don't see any reason why you could not replace the 30min of steady state exercise with 10-15 minutes of HIIT (including active rest), but you should be aware of the fact that this will be more taxing on your central nervous system and probably more likely to result in a decrease in exercise performance on the subsequent workout, than sitting on an ergometer cycling at 60% of your VO2max. If you feel that it works for you - fine, but don't complain if in a year from now you still don't look like Mr. Olympia ;-)

Now the Psilander study does show that glycogen depletion, which is essentially what will happen (at least to a certain degree) if you perform 30 min of cardio training at a non-exhausting, but still energy consuming pace of 60% of your VO2 max before a workout does work. Without differentiating the various iso-forms of PGC-1 alpha Psilander's 5x increase in PGC-1 does yet not tell us whether we are dealing with the "right form" of PGC here. After all, the Psilander protocol involved two endurance sessions, with the first being a depletion session that was conducted on the day before the actual test and the second being a HIIT-esque exercise test (go back to the original post for more details).  Fortunately, there are 2019 other articles on the SuppVersity so that I don't even have to refer you to a study I have not already written about to add another piece to the puzzle.

A blast from the past and a glimpse into the future

On Wednesday, October 31, 2012, I wrote about the results of a study by Lundberg et al.. Again a slightly different protocol, this time with "cardio" in the morning and strength training later in the day, yet the exact same benefits in terms of PGC-1 alpha (total) expression:

Figure 7: Selected markers of mitochondrial biogenesis and protein synthesis before during and 15, respectively 180min after the resistance training bout in the AE + RE and the RE only leg (a.u.; data adapted from Lundberg. 2012)

With the more pronounced drop in myostatin in the combined training group in the Lundberg study, the only thing we would still need to further support the practical value of the more recent results from the Roas study would be a concomitant increase in IGF-1, as we would expect it, if working out in a (partly) glycogen depleted state would actually be the reason for the increase in PGC-1 alpha 4 Roas observed in the subjects of his study. Now I could copy and paste another graph, but I guess it will be enough, when I refer you back to the detailed elaborations on the connection between IGF-1 and it's muscle-specific splice variants and exercise induced beneficial, since acute and hormetic inflammation in the "IGF, MGF & Inflammation" part of the Intermittent Thoughts on Building Muscle (click here for an overview).

Please keep in mind: Regardless of the fact that previous studies did not test for the PGC-1 alpha subtypes, we cannot ignore the existing evidence that PGC-1 is not mandatory for the beneficial effects of endurance exercise on mitochondrial biogenesis (e.g. Rowe. 2012) and should therefore not overestimate the importance of PGC1 alpha 4 as the "one and only" muscle builder. I have said that before, but I guess it's important to repeat it - this is another missing link it's just like mTOR, testosterone and whatever other magic bullets people will tell you about not exclusively responsible for increases in muscle mass, mitochondrial capacity and whatever else you may just be dreaming of.

If we now add a couple of additional findings to this intellectual brew, like ...

• the 100% increase in the expression of the heat shock protein HSP72 in a glycogen depleted vs. normal leg during a workout (Febbraio. Feb 2002)
• the 150% increase of intramuscular HSP72 in response to an infusion with low doses of interleukin-6 (Febbraio. Sep 2002)
• the non-existant negative side effects of IL-6 on muscle glucose uptake in healthy individuals (Steensberg. 2003)
• IL-6's importance as a regulator of glucose metabolism during exercise (Helge. 2003; Febbraio. 2004) and it's satellite cell proliferation promoting effects (McKay. 2009) 
• the Dr. Jakyll and Mr. Hyde nature of inflammation, in general and IL-6 in particular on glucose uptake and fatty acid oxidation, when it comes to its local and temporary (=beneficial effects) vs. systemic and chronic (=detrimental effects) presence in our body (Fisman. 2010)

...we do actually arrive back at where we came from, namely the difference between training and overtraining in figure 2.

Bottom line - cardio pre-workout as an intensity technique: On the basis of these considerations you can think of doing cardio before a workout as an intensity technique that will increase the beneficial stress and thus the demand for greater adaptive responses. That the latter will go hand in hand with an increased propensity of overtraining, particularly if you are not willing to (A) supply your body with the nutrients it needs after the workout and (B) to rest for an adequate amount of time before you hit the gym again, is something of which I would appreciate if it wasn't something I had to repeat in each and every SuppVersity article, but since this is and will probably remain the #1 reason why people don't make progress physique- or performance-wise, it's still the most important take home message at least for those of you who are new to the site. I hope this did not ruin this allegedly pretty lengthy post for you and believe I am not promising too much, when I say that you are soon going to read more about this protein here - after all, it's almost certain that we are going to see follow-up studies in the months to come.

References:

• Arany, Z. PGC-1 coactivators and skeletal muscle adaptations in health and disease. Curr. Opin. Genet Dev; 2008: 426–434. 
• Febbraio MA, Steensberg A, Walsh R, Koukoulas I, van Hall G, Saltin B, Pedersen BK. Reduced glycogen availability is associated with an elevation in HSP72 in contracting human skeletal muscle. J Physiol. 2002 Feb 1;538(Pt 3):911-7.
• Febbraio MA, Steensberg A, Fischer CP, Keller C, Hiscock N, Pedersen BK. IL-6 activates HSP72 gene expression in human skeletal muscle. Biochem Biophys Res Commun. 2002 Sep 6;296(5):1264-6.
• Febbraio MA, Hiscock N, Sacchetti M, Fischer CP, Pedersen BK. Interleukin-6 is a novel factor mediating glucose homeostasis during skeletal muscle contraction. Diabetes. 2004 Jul;53(7):1643-8.
• Fisman EZ, Tenenbaum A. The ubiquitous interleukin-6: a time for reappraisal.
  Cardiovasc Diabetol. 2010 Oct 11;9:62.
• Helge JW, Stallknecht B, Pedersen BK, Galbo H, Kiens B, Richter EA. The effect of graded exercise on IL-6 release and glucose uptake in human skeletal muscle. J Physiol. 2003 Jan 1;546(Pt 1):299-305.
• Kramer HF, Goodyear LJ. Exercise, MAPK, and NF-kappaB signaling in skeletal muscle. J Appl Physiol. 2007 Jul;103(1):388-95.
• McKay BR, De Lisio M, Johnston AP, O'Reilly CE, Phillips SM, Tarnopolsky MA, Parise G. Association of interleukin-6 signalling with the muscle stem cell response following muscle-lengthening contractions in humans. PLoS One. 2009 Jun 24;4(6):e6027.
• Psilander N, Frank P,  Flockhart M, Sahlin K. Exercise with low glycogen increases PGC-1agene expression in human skeletal muscle. Eur J Appl Physiol. 02 Oct 2012 [ahead of print]
• Rowe GC, El-Khoury R, Patten IS, Rustin P, Arany Z. PGC-1α is dispensable for exercise-induced mitochondrial biogenesis in skeletal muscle. PLoS One. 2012;7(7):e41817. Epub 2012 Jul 24.
• Ruas et al. APGC-1aI soform Induced by Resistance Training Regulates Skeletal Muscle Hypertrophy. Cell, December 7, 2012; 151:1319–1331.
• Steensberg A, Fischer CP, Sacchetti M, Keller C, Osada T, Schjerling P, van Hall G, Febbraio MA, Pedersen BK. Acute interleukin-6 administration does not impair muscle glucose uptake or whole-body glucose disposal in healthy humans. J Physiol. 2003 Apr 15;548(Pt 2):631-8. Epub 2003 Mar 14.

Vitamin D3 a "Fat Synthesizer"!? Rodent Study Shows +33% Increased Fat Deposition in Vitamin D3 Supplemented Mice.

Illustration 1: Experts will recognize from looking at these Oil Red O-stained longissimus dorsi slices of mice on a normal and a vitamin D3 supplemented diet that supplemental (! not vitamin D from the sun !) "vitamin D3 can be used a s a fat synthesizer and meat tenderizer in meat-producing animals". (img in illustraton from Choi. 2011)

I have been railing against the current vitamin D hype for months now. In that, I have at no point in time implied that "backfilling" depleted vitamin D levels via supplementation could not be beneficial (or at least not harmful), nor have I at any time excluded that vitamin D3 supplementation (even if you are in the "normal" range) could have its merit (cf. vitamin D3 + HMB). What I have done though, was to point at the lack of controlled studies that would support any of the benefits supplemental vitamin D3 is currently hailed for all over the Internet. This amazes me, because the very same "gurus" who are all over the vitamin D bandwagon have lately (just like me) discarded the data from the Iowa Women's Health Study as "unrealiable" and "non-significant" epidemiological bullshit (which is exactly, what I think, as well). When it comes to vitamin D, however, they throw all their concerns on the validity of epidemiological data over board and worship their vitamin D3 pills like a golden calf.

But let's get to the facts, before I get tarred and feathered, again... In the latest issue of the Journal of the Science of Food and Agriculture Hyuck, Choi and Kyuho Myung published a paper that investigated the use of vitamin D3 supplements to fatten animals (Choi. 2011). Now, you may think "How stupid is that, everyone knows that vitamin D will make you lean out!", but as I've pointed out several times within the last weeks, high vitamin D levels may correlate with a lean body composition; however, studies that would show that supplementation of the latter would induce respective changes in body composition in the absence of prior deficiency (and we are talking about the standard reference range with a lower limit of 10ng/mL, here) simply do not exist... but I am digressing again.

Figure 1: Composition of the diet (large figure) and respective vitamin D3 content (small figure) of the diets of the control and the supplement group in the study.

As you can see in figure 1 both groups (2x N=10) of 6 weeks-old male C57BL/6 mice were fed identical chows (AIN93G; cf. figure 1, large), varying only in their vitamin D3 content (1IU in the control group, 10IU in the supplemented group). In human terms this would be like switching from your common western low vitamin D diet with roughly 800IU to taking a 8.000 IU supplement, each day - something I suppose many of you may have done lately!?

Figure 2: Body fat (in g; large figure) and respective serum 1α,25-(OH)2 -vitamin D3 levels (in µg/mL; small figure) after 3 weeks on control or vitamin D3 supplemented diet (data calculated based on Choi. 2011)

As figure 2 shows, this 10-fold increase in dietary vitamin D would only be advisable if you were a "sumo mouse" who has to make weight for the next competition. A plus of +33% in total, +29% in unaesthetic subcutaneous and +25% in unhealthy visceral fat (all statistically significant with p<0.022, p<0.032 and p<0.043) is not what you would expect of the "greatest vitamin of all time" - would you? And while the vitamin D3 mice also gained some more body weight, those changes were statistically non-significant, so that - as the scientists state - vitamin D3 turned out to be an ideal "fat synthesizer and meat tenderizer".

Figure 3: Cytokines, UCP-2 and PPAR-gamma expression in mice after 3 weeks on control or vitamin D3 supplemented diet (based on Choi. 2011)

In that, vitamin D3 works it "fat synthesizing" magic by increasing the inflammatory cytokines TNF-alpha and IL-6 and decreasing the muscle anabolic (Busquets. 2005) and fat catabolic cytokine IL-15 (Carbo. 2001; Alvarez. 2002), as well as the uncoupling protein UCP-2 while ramping up fat storage via increase PPAR-gamma expression (cf. figure 3).

Now obviously, this is just another rodent study and we cannot say how and if the results will translate to humans, but it is a controlled study and it investigates the effects of supplemental vitamin D3 which is something you cannot say of the "scientific backbone" of the current vitamin D3 craze... and now tar and feather me like a child who has just been bereaved of his favorite toy, if you will ;-)

DOMS - Delayed Onset Muscle Soreness: No Pain, No Gain? Is DOMS Necessary to Build Muscle?

Are stretch, tear and DOMS what makes concentration curls an effective biceps builder? Can we use the soreness as a gauge for the efficiency of our training?

An arictle by Alex Leaf (CPT)

In last Sunday's first installment of our discussion on delayed onset muscle soreness (DOMS), we looked at what causes DOMS as well as treatment methods and supplements for relieving its symptoms. This led us to today’s big question:  Is DOMS necessary for muscular adaptations to exercise? No pain, no gain, right?

Before we can discern whether DOMS may benefit muscle growth, we need to look at what muscle growth is and what causes it, so that we may see if DOMS is in fact a piece of the puzzle.

Taking a Second Look at Muscular Hypertrophy

During muscle fiber hypertrophy, contractile proteins proliferate, and the muscle fibers enlarge to support this growth (Vierck. 2000). While there are many factors regulating this process, from gene expression to hormones and other growth factors, the two necessities for hypertrophy are some form of increased muscular tension, damage, or stress (Goldberg. 1975), and a positive net protein synthetic response with adequate energy availability (Miyazaki & Esser. 2009).

Beware of too much "good" ROS scavengers. NAC will effectively block the recruitment of new satellite cells | learn more

Through exercise, the former is accomplished and paves the way for the repair processes that require the latter to occur. In other words, without a need to increase muscle size and strength, hypertrophy will not happen. Likewise, even if there is a need, without proper nourishment hypertrophy simply cannot happen.

So with exercise being the trigger and nutrition the ammo, what is left to play the gun? Skeletal muscle does not undergo significant cell replacement once mature (Chargé & Rudnicki. 2004), and thus a repair mechanism for any microtrauma is essential.

This medic is the satellite cell, a type of stem cell found only within mature muscle tissue. After microtrauma, satellite cells activate, proliferate, and ultimately fuse to one another and existing muscle fibers to form new myofibrils (Toigo & Boutellier. 2006).

All parts of this regenerative weapon rely on one another. The satellite cells mediate the hypertrophic process, but without a need (the exercise) they will not start, and without the nourishment (energy availability) they cannot function. All else that impacts the accuracy of the gun can be thought of as the factors influencing satellite cell efficiency. Hormones could be wind speed, gene expression the user’s accuracy, and perhaps DOMS is the distance to the target (or not ;-).

Muscle Damage

Suggested Read: "Understanding Muscle Hypertrophy - Study Sheds More Light on Process of Satellite Cell Recruitment" | read more

The hypertrophy process begins with microtrauma and an ensuing accumulation of calcium within the damaged muscles (Sorichter. 1999). This is shortly followed by a rapid stimulation of satellite cells via hepatocyte growth factor (HGF) and nitric oxide (NO), both of which rely on the changes in calcium levels within the muscle tissue (Tatsumi, 2010), and satellite cells may even be activated by the calcium flux itself (Hara, et al., 2012).

Furthermore, HGF secretion is proportional to the extent of the muscle damage (Tatsumi, et al., 1998). Therefore, it seems plausible that greater muscle damage leads to greater satellite cell recruitment, especially since the activation of satellite cells is exclusive to the fiber that has become damaged and satellite cells of one muscle fiber will not respond to injury of adjacent muscle fibers (Chargé & Rudnicki, 2004).

As it just so happens, DOMS inducing eccentric contractions disrupts muscle integrity more so than concentric or isometric contractions (Faulkner, 1993).  What may seem odd, however, is that EMG activity has been shown to be lower in eccentric loading compared to concentric loading (Westing. 1991), suggesting less fiber recruitment.

Part I	Part II

Just a reminder: This is a twp-part series on Delayed Onset Muscle Soreness. You can switch back and forth between part I "What Is DOMS & How Can It Be Managed? Science, Strategies, Supplements" & part II "No Pain, No Gain? Is DOMS Necessary to Build Muscle?" by clicking on the images to the left.

In their study, (Westing. 1991) measured the torque and EMG activity of the quadriceps muscle at different movement speeds between a knee angle of 30° and 70° on the leg extension for both the concentric and eccentric portions of the exercise. The participants were 14 highly trained athletes that were accustomed to performing maximally during training. As you can see in Figure 1, average torque of the eccentric activity was significantly greater than that of the concentric activity across all movement speeds, but the EMG activity was significantly lower and continued to lower as movement speed increased. The fact that the EMG values of the eccentric activity are below 100% shows that the activation during the concentric phase was higher, even at lower speeds and despite “maximal” effort.

Figure 1: Exemplary data from Westing (1991) showing the mean and SEM torque- and EMG-velocity relationships during the eccentric (open symbols) and concentric (filled symbols) tests.

Actually, this gives support the idea that eccentric exercise is more damaging. It is hypothesized that neural drive to the working muscle is reduced under conditions of extreme muscle tension (i.e. less EMG but more torque) to protect the muscle from injury that could result if it became fully activated (Moore. 1984). Regardless, single bouts of eccentric contractions have been shown to increase the satellite cell content and activation status in Type II muscle fibers (Cermak. 2013).

Inflammation - Friend or Foe of Muscle Growth?

Learn more about eccentric training and satellite cell recruitment and how even fat cells can become muscle.

Once the damage has been done, the repair process may begin. As mentioned in "DOMS - Delayed Onset Muscle Soreness: What Is DOMS & How Can It Be Managed? Science, Strategies, Supplements" (read article), an acute inflammatory response follows microtrauma.

This is also the time that DOMS normally makes it move. During this time, the damaged muscle releases several cytokines, while white blood cells such as neutrophils and macrophages invade the damaged tissue and release several growth factors, all of which may regulate satellite cell activity (Toigo & Boutellier. 2006). Creatine kinase, for example, is a standard indirect measurement of muscle damage (Banfi. 2012).

As stated above, several cytokines and growth factors are involved in the anabolic response to muscle damaging exercise. The list is quite extensive but a few notable players are:

• The cytokine interleukin-6 (IL-6) appears to be an essential regulator of satellite cell mediated hypertrophy, and genetic loss of IL-6 blunts the hypertrophic response (Serrano, et al. 2008). There also appears to be a close association between cytokine concentrations and muscle damage (Pedersen, Ostrowski, et al. 1998), with (Bruunsgaard, et al. 1997) showing that IL-6 concentration is higher after eccentric cycling compared with concentric cycling.  Likewise, interleukin-15 (IL-15) is another highly anabolic player in the inflammatory response to muscle damage (Furmanczyk and Quinn 2003), and is elevated following resistance exercise but not treadmill running, suggesting a need for microtrauma in its stimulation (Pedersen, Akerström, et al. 2007).

• 

  Learn more about the different splice variants of IGF-1 and how they figure in the process of muscle hypertrophy and why systemic measures may mislead us.

  Insulin-like growth factor 1 (IGF-1) has also received much attention due to its ability to increase muscle mass via muscle protein and DNA augmentation (Chakravarthy, Davis and Booth 2000). These effects are at least in part attributed to the activation of satellite cells and increased protein synthesis within the muscle fibers (Barton-Davis, Shoturma and Sweeney 1999). And guess what? Damaging exercise increases IGF-1. A study by (Bamman, et al. 2001), for example, showed that eccentric exercise increased IGF-1 gene expression by 62% while decreasing inhibitory genes by 57%. Oh, and concentric exercise produced non-significant changes in the above markers, suggesting that it was indeed the structural damage responsible for the IGF-1 expression.
• Lastly, the aforementioned HGF acts as a chemo-attractant for satellite cells (Bischoff 1997), effectively stimulating satellite cells to migrate to the place of injury, where it then has a direct effect on satellite cell proliferation and differentiation (Vierck, et al. 2000). Oddly enough, abnormally elevated concentrations of HGF actually inhibit muscle regeneration via up-regulation of myostatin (Yamada, et al. 2010). Since HGF is secreted by regenerating muscles for the first three days following injury (Jennische, Ekberg and Matejka 1993), its accumulation could act as a regulatory “stop” mechanism that marks the end of muscle repair via satellite cells (Chazaud 2010).

A final indirect notion of the importance of DOMS is the idea that NSAIDs – a common treatment method – reduce the hypertrophic response. Recall that both NO and HGF are responsible for activating satellite cells in the early stages of the repair process. This process appears to be partially regulated by the cyclooxygenase (COX)-2 pathway, which releases various prostaglandins known to stimulate satellite cells (Bondesen, et al. 2004). The problem is that NSAIDs inhibit this pathway and thus may impair the hypertrophic response (Schoenfeld 2012). Indeed, studies have shown NSAID usage following eccentric exercise reduced satellite cell activity for up to eight days (Mikkelsen, et al. 2009).

Summary: Hypertrophy involves a complex array of anabolic and catabolic processes working in a downstream manner to favor protein synthesis over degradation. DOMS is not necessary,  may however present itself during the early stages of exercise. What is necessary is a mechanical overload of the muscle resulting in microtrauma. So train hard, train smart, and may the growth be with you.

So is DOMS necessary? DOMS can be thought of as a sign of muscle damage, but it is the damage itself and the subsequent inflammatory response that are responsible for hypertrophy. DOMS is actually a rather poor indicator of muscle damage and will not always reflect the magnitude of the damage (Nosaka, et al., 2002). Nor will it always be present.

Studies have shown that even a single bout of eccentric exercise reduces and may negate DOMS in subsequent bouts (Nosaka. 2001), and these effects persist for at least several weeks (Clarkson. 1992). This would explain why soreness is common in the beginning of a new routine full of unaccustomed damaging exercise, but fades as time progresses. And in fact Flann (2011) showed that using a three week acclimation protocol prior to beginning an eight week eccentrically loaded leg press protocol significantly reduced DOMS and markers of muscle damage compared to beginning the routine cold turkey.

References

• Bamman, M M, et al. "Mechanical load increases muscle IGF-I and androgen receptor mRNA concentrations in humans." American Journal of Physiology - Endocrinology and Metabolism 280, no. 3 (2001): E383-E390.
• Banfi, G, A Colombini, G Lombardi, and A Lubkowska. "Metabolic markers in sports medicine." Advances in Clinical Chemistry 56 (2012): 1-54.
• Barton-Davis, E R, D I Shoturma, and H L Sweeney. "Contribution of satellite cells to IGF-I induced hypertrophy of skeletal muscle." Acta Physiologica Scandinavica 167, no. 4 (1999): 301-305.
• Bischoff, R. "Chemotaxis of skeletal muscle satellite cells." Developmental Dynamics 208, no. 4 (1997): 505-515.
• Bondesen, B A, S T Mills, K M Kegley, and G K Pavlath. "The COX-2 pathway is essential during early stages of skeletal muscle regeneration." American Journal of Physiology - Cell Physiology 287, no. 2 (2004): C475-C483 .
• Bruunsgaard, H, H Galbo, J Halkjaer-Kristensen, T L Johansen, D A MacLean, and B K Pedersen. "Exercise-induced increase in serum interleukin-6 in humans is related to muscle damage." The Journal of Physiology 499, no. Pt 3 (1997): 833-841.
• Cermak, N M, et al. "Eccentric exercise increases satellite cell content in type II muscle fibers." Medicine and Science in Sports and Exercise 45, no. 2 (2013): 230-237.
• Chakravarthy, M V, B S Davis, and F W Booth. "IGF-I restores satellite cell proliferative potential in immobilized old skeletal muscle." Journal of Applied Physiology 89, no. 4 (2000): 1365-1379.
• Chargé, S B, and M A Rudnicki. "Cellular and molecular regulation of muscle regeneration." Physiological Reviews 84, no. 1 (2004): 209-238.
• Chazaud, B. "Dual effect of HGF on satellite/myogenic cell quiescence." American Journal of Physiology - Cell Physiology 298, no. 3 (2010): C448-C449.
• Clarkson, P M, K Nosaka, and B Braun. "Muscle function after exercise-induced muscle damage and rapid adaptation." Medicine and Science in Sports and Exercise 24, no. 5 (1992): 512-520.
• Faulkner, J A, S V Brooks, and J A Opiteck. "Injury to Skeletal Muscle Fibers During Contractions: Conditions of Occurrence and Prevention." Physical Therapy 73 (1993): 911-921.
• Flann, K L, P C LaStayo, D A McClain, M Hazel, and S L Lindstedt. "Muscle damage and muscle remodeling: no pain, no gain?" The Journal of Experimental Biology 214 (2011): 674-679.
• Furmanczyk, P, and L S Quinn. "Interleukin-15 increases myosin accretion in human skeletal myogenic cultures." Cell Biology International 27, no. 10 (2003): 845-851.
• Goldberg, A L, J D Etlinger, D F Goldspink, and C Jablecki. "Mechanism of work-induced hypertrophy of skeletal muscle." Medicine and Science in Sports 7, no. 3 (1975): 185-198.
• Hara, M, et al. "Calcium influx through a possible coupling of cation channels impacts skeletal muscle satellite cell activation in response to mechanical stretch." American Journal of Physiology - Cell Physiology 302, no. 12 (2012): C1741-C1750.
• Jennische, E, S Ekberg, and G L Matejka. "Expression of hepatocyte growth factor in growing and regenerating rat skeletal muscle." The American Journal of Physiology 265, no. 1 Pt 1 (1993): C122-C128.
• Mikkelsen, U R, et al. "Local NSAID infusion inhibits satellite cell proliferation in human skeletal muscle after eccentric exercise." Journal of Applied Physiology 107, no. 5 (2009): 1600-1611.
• Miyazaki, M, and K A Esser. "Cellular mechanisms regulating protein synthesis and skeletal muscle hypertrophy in animals." Journal of Applied Physiology 106, no. 4 (2009): 1367-1373.
• Moore, J C. "The Golgi tendon organ: a review and update." American Journal of Occupational Therapy 38, no. 4 (1984): 227-236.
• Nosaka, K, K Sakamoto, M Newton, and P Sacco. "The repeated bout effect of reduced-load eccentric exercise on elbow flexor muscle damage." European Journal of Applied Physiology 85, no. 1-2 (2001): 34-40.
• Nosaka, K, M Newton, and P Sacco. "Delayed-onset muscle soreness does not reflect the magnitude of eccentric exercise-induced muscle damage." Scandinavian Journal of Medicine & Science in Sports 12, no. 6 (2002): 337-346.
• Pedersen, B K, K Ostrowski, T Rohde, and H Bruunsgaard. "The cytokine response to strenuous exercise." Canadian Journal of Physiology and Pharmacology 76, no. 5 (1998): 505-511.
• Pedersen, B K, T C Akerström, A R Nielsen, and C P Fischer. "Role of myokines in exercise and metabolism." Journal of Applied Physiology, 2007: 1093-1098.
• Schoenfeld, B J. "Does exercise-induced muscle damage play a role in skeletal muscle hypertrophy?" Journal of Strength and Conditioning Research 26, no. 5 (2012): 1441-1453.
• Schoenfeld, B J. "The use of nonsteroidal anti-inflammatory drugs for exercise-induced muscle damage: implications for skeletal muscle development." Sports Medicine 42, no. 12 (2012): 1017-1028.
• Serrano, A L, B Baeza-Raja, E Perdiguero, M Jardí, and P Muñoz-Cánoves. "Interleukin-6 is an essential regulator of satellite cell-mediated skeletal muscle hypertrophy." Cell Metabolism 7, no. 1 (2008): 33-44.
• Sorichter, S, B Puschendorf, and J Mair. "Skeletal muscle injury induced by eccentric muscle action: muscle proteins as markers of muscle fiber injury." Exercise Immunology Review 5 (1999): 5-21.
• Tatsumi, R. "Mechano-biology of skeletal muscle hypertrophy and regeneration: possible mechanism of stretch-induced activation of resident myogenic stem cells." Animal Science Journal 81, no. 1 (2010): 11-20.
• Tatsumi, R, J E Anderson, C J Nevoret, O Halevy, and R E Allen. "HGF/SF is present in normal adult skeletal muscle and is capable of activating satellite cells." Developmental Biology 194, no. 1 (1998): 114-128.
• Toigo, M, and U Boutellier. "New fundamental resistance exercise determinants of molecular and cellular muscle adaptations." European Journal of Applied Physiology 97, no. 6 (August 2006): 643-663.
• Vierck, J, et al. "Satellite Cell Regulation Following Myotrauma caused by Resitance Exercise." Cell Biology International 24, no. 5 (2000): 263-272.
• Westing, S H, A G Cresswell, and A Thorstensson. "Muscle activation during maximal voluntary eccentric and concentric knee extension." European Journal of Applied Physiology and Occupational Physiology 62, no. 2 (1991): 104-108.
• Yamada, M, et al. "High concentrations of HGF inhibit skeletal muscle satellite cell proliferation in vitro by inducing expression of myostatin: a possible mechanism for reestablishing satellite cell quiescence in vivo." American Journal of Physiology - Cell Physiology 298, no. 3 (2010): C465-C476.

CoQ10 for Ultra-Endurance Athletes: 150mg of Ubiquinone Reduce Stress & Inflammation and Stabilize Cell Membranes in 52.4 Mile Torture from 640m to 3,393m!

Image 1: Susan Kokesh, blogger and the Crazy Running Mum at the Sierra Nevada ultra-endurance run a 52.4 miles "double marathon"
  in September 2010; I probably would not even have survived this torture - respect!

As a health conscious physcial culturist, you are probably aware that the vitaminesque nutrient CoQ10, which, due to its ubiquitous presence in all living beings, is also known as ubiqinone, plays a fundamental role in cellular bioenergetics. It is a necessary cofactor in the mitochondrial electron transport chain (i.e. your cell's way of "breathing", its respiratory chain) and is therefore essential for the production of ATP, the fundamental energy unit your cells are operating on. In that, CoQ10 works as a mobile redox agent that shuttles electrons and, interestingly, also protons (those little blue and red balls from Bohr's atom model ;-) in the electron transport chain. Within the health and fitness community, it is however better known for its antioxidant value, as in its reduced form, ubiquinol, it is a potent lipophilic (which means that it does not combine with fats) antioxidant, which is able to recycle and regenerate other antioxidants, such as vitamin E and vitamin C (Ernster. 1995). Moreover, CoQ10 participates in cell signaling and gene expression and has been used as a dietary supplement (among others) for the treatment of neurodegenerative diseases and statin-induced myopathies.

In view of its pluripotent influence on mammalian metabolism (on a side note: the "-10" in CoQ10 indicates the length of the isoprenoid sidechain that is attached to the common benzoquinone ring structure; the latter is unique and can be found in humans and a few other mammals only), it should thus not surprise you that Chavier Díaz-Castro and his collegues from the University of Granada report that the intake of 150mg of the natural version of CoQ10 (2,3-dimethoxy-5-methyl-6-decaprenyl-1,4-benzoquinone; natural = has trans configuration), profoundly modulated "the undesirable effects of the evoked oxidative stress and inflammation signaling during high-intensity" (Díaz-Castro. 2011).

Illustration 1: Supplementational protocol used in the study; CoQ10 was administered as 2,3-dimethoxy-5-methyl-6-decaprenyl-1,4-benzoquinone in powder form in 30mg capsules.

As you can see in illustration 1, the 20 highly trained male amateur athletes (all had run The Sierra Nevada ultra-endurance race in the previous 2 years), who participated in the study were not given the whole dose of 150mg of CoQ10 at once, but followed what I would like to call a "loading protocol" in the two days before the event. The placebo group received an identically looking product containing beer yeast, cellulose,
acacia, silica stearic acid, magnesium stearate, cellulose gum, and maltodextrin.

The total distance of The Sierra Nevada run is >50km. It is considered one of the hardest trials worldwide, mainly because the run, in the course of which the participants "climb" from 640m to a final altitude of  3,393m is almost on a continuous incline! A 5.5 hour torture, for which the CoQ10 supplemented athletes needed on average ~25min less than the subjects in the placebo group. In this study, the exercise performance was yet of negligible importance. What the scientists were really interested were the markers of oxidative damage and inflammation and as the following data shows, those were markedly influenced by the ingestion of this rather "mediocre" (compared to what you see some "health-gurus" advocate) amount of CoQ10.

Figure 1: Effects of CoQ10 supplementation of total bilirubin, triglycerides and urinary creatinine in 20 ultra-endurance runners (data calculated based on Díaz-Castro. 2011)

As you can see in figure 1, CoQ10 induced a significant reduction in urinary creatinine even before the race started (figure 1, left column). Moreover, there were significant differences in the bilirubin (indicates heme catabolism), triglycerides and (again) creatinine (indicates net protein catabolism):

Intense physical exercise resulted in an increase in net protein catabolism and an increase in
creatinine excretion in the PG after the physical test (p < 0.001); however, the urinary levels of creatinine were lower (p < 0.05 before and p < 0.001 after the physical test) in the CoQ10 treated group. Other interesting result was that although there was an increase in urinary creatinine in the CG, it was lower than in PG (38.77 ± 10.20 vs. 88.23 ± 11.21, p < 0.05). We also observed a decrease in the bilirubin concentrations in the CG after the run (p < 0.001) with lower values compared to PG group.

There were also significant differences in the inflammatory response, with (statistically significant, p<0.05) lower values of interleukin 6 (IL-6; -32%) and TNF-alpha (-23%) before the start of the race, and -22% lower TNF-alpha values after the "torture". Moreover, the basal hydroper-oxide content in the erythrocyte membranes, the scientists measured as an indicator of the degree of oxidative stress were lower before and after the exercise test, as the scientists call it.

Taken together, these results suggest that the addition of a small dose of CoQ10 to your supplemental regimen could induce unexpectedly profound cell-stabilizing benefits, of which it would yet be interesting to see how those translate into performance benefits, health and longevity, in the long run.

Conjugated Linoleic Acids: What's the Difference Between cis-9,11 and trans-10,12 CLA and Should We Label Them as "Transfats"? Plus: What Makes CLA Potentially Harmful?

If Hayden Panettiere drinks it dairy can't be bad - despite (or because?) CLA, right? Well, what if I told you that Mrs. Panettiere was advertising milk in the "Got Milk" campaign despite being lactose intolerant?

As a SuppVersity veteran you will be familiar with the idea that trans-10, trans-12 conjugated linoleic acid is the "fat burning" CLA isomer, while 9 cis,11 trans linoleic acid appears to blunt some of the pro-inflammatory actions of its cousin and has been shown to have specific physiological effects on it's own (e.g. increased bone health, cf. Platt. 2009; anti-cancer, cf. Corl. 2003).

As I already mentioned, this is probably nothing new for you, if you make sure to get your daily dose of SuppVersity wisdom everyday. What you may however not be aware of is the fact that researchers like Ye Wang and Spencer D. Proctor are - despite the never-ending hoopla around potential weight loss effects of CLA - still contemplating, whether CLA could not pose a major health threat to all or at least certain subgroups of the population and whether this should or shouldn't be reason enough to change the current food labeling practices.

Do we have to label CLA as "transfat"?

From a technical perspective the above question is obsolete. CLAs are transfats and would thus (technically, again) have to be labeled as such on the product label. From a health perspective, however, things do in fact look different. Due to the fact that the aforementioned ruminant (=naturally produced in the stomach(s) of ruminents) trans-fats have been associated with health benefits (Gebauer. 2011), we could effectively risk to scare consumers away from healthy foods if they were listed as part of the "transfat" category on the product labels.

What's actually the reason that one the same CLA isomer that will have you lose body fat will also "inflame" you? Due to the fact that most of the research on "fat loss supplements" is conducted in sick, obese individuals, people tend to get the false impression that "fat burners" were anti-inflammatory and that anti-inflammatory agents would burn fat.

Effects of 10-trans,12 CLA on fatty acids & glucose metabolism and IL-6 gene expression in isolated fat cells in the petri dish (Hartwig. 2013)

Now, while it is correct that soothing inflammation will help the future Mr. Average Joe, who is going to be an obese (pre-)diabetic, lose weight, this has little to do with any active contribution to the oxidation of body fat. 10-trans,12 CLA, on the other hand, has been shown to block lipid storage, increase mitochondrial uncoupling (UCP-2), lower PPAR-alpha and ramp up the oxidation and release of fatty acids from the fat stores (Hartig. 2013). Unfortunately it will also block the uptake of glucose and increase the expression of the pro-inflammatory cytokine IL-6 in fat cells. So, if you took 10-trans,12 CLA at very high doses it will probably in fact keep you lean.

If you cannot handle the sudden increase in free fatty acids, pack the glucose into your liver and muscle glycogen stores and deal with the exuberant amount of inflammatory cytokines, however, it will only make you sick.

Currently, the trans-fat content on many food labels (and in legislative documents) does not include ruminant CLA isomers and Wang and Procter acknowledge that:

"As highlighted in a recent quantitative review of prospective cohort studies by Bendsen et al. dietary consumption of ruminanttrans-fat may be protective against total as well as fatal CHD events." (Wang. 2013)

The researchers do however point out that concerns about potential adverse effect on atherogenic cholesterol profiles from supplemental CLA are not unwarranted - at least if they are used by a group of persons - abdominally obese and/or insulin resistant men, for example (Riserus. 2002 a,b).

Australia and New Zealand suggest a re-evaluation

Accordingly, Australia and New Zealand (FSANZ) proposed to re-evaluate their perception regarding the exclusion of CLA from the TFA definition on nutrition labels.

If you re-evaluate something, you do not necessarily have to change them and if you go through the concise summary of results Wang and Procter present in their paper (see table 1 for an overview of the currently published meta-analyses, it does not appear necessary to question the current practice to label only industrually produced trans fats.

Table 1: Meta analysis with beneficial (green), neutral (grey) and potentially negative outcome (red); based on Wang & Proctor (2013)

As the authors point out, the inclusion of CLA in the total amount of transfats on the label would only drive people away from the consumption of whole food products. This is particularly true in view of the fact that the ill-health effects of "trans-fats" are something everyone will have heard about. The fact that these ill health effects are not to be expected from trans fats in dairy and other CLA containing whole foods, on the other hand, is still news to many costumers.

Moreover, how would you, me and everyone else who may well be aware that CLAs are not the bad guys and the "trans-fat" in grass fat butter is not going to hurt us know if the 3g of transfats in another product we buy are actually from the undisclosed amount of butter (and thus CLA) in it? It could likewise be that the producer added a little extra partially hydrogenated vegetable oil to cut the product costs and neither you nor me would know that.

Suggested read: "A Higher Intake of CLA and Vaccenic Acid from Dairy, Beef, Veal and Lamp Could Prevent Subtle Weight Gain" | read more

Only the obese have to be worried: Based on the currently available evidence the healthy and lean person (hopefully you) has absolutely no reason to avoid products with a "high" natural CLA content and thus both the pro- (trans-10,12) and (partly) anti-inflammatory (cis-9,11) form of CLA in them.

For obese and insulin resistant individuals things do however look somewhat different. A 2004 study by Risérus et al., for example, has been able to show that even the allegedly "harmless", 9 cis,11 trans linoleic acid can worsen both lipid peroxidation and insulin resistance in 25 abdominally obese men. (Risérus. 2004).

With 3g/day the dosage that was used in the Risérus study, the amount of CLA was yet much hither than the amount of CLA you can possibly ingest with nourishing foods such as butter, full-fat dairy, grass-fed beef (and beef in general, by the way). Instead of these you are thus better advised to avoid CLA supplements... but don't worry if you take another look at the data in table 1 you will have to concede that they are pretty much useless, anyway.

Reference:

• Corl BA, Barbano DM, Bauman DE, Ip C. cis-9, trans-11 CLA derived endogenously from trans-11 18:1 reduces cancer risk in rats. J Nutr. 2003 Sep;133(9):2893-900.
• Gebauer SK, Chardigny JM, Jakobsen MU, et al. Effects of ruminanttrans fatty acids on cardiovascular disease and cancer: a comprehensive review of epidemiological, clinical, and mechanistic studies.Adv Nutr. 2011; 2, 332 – 354.
• den Hartigh LJ, Han CY, Wang S, Omer M, Chait A. 10E,12Z-conjugated linoleic acid impairs adipocyte triglyceride storage by enhancing fatty acid oxidation, lipolysis, and mitochondrial reactive oxygen species. J Lipid Res. 2013 Nov;54(11):2964-2978. 
• Lenz TL & Hamilton WR. Supplemental products used for weight loss. J Am Pharm Assoc. 2004; 44,  59– 67, quiz 67– 58.
• Onakpoya IJ, Posadzki PP, Watson LK, Davies LA, Ernst E. The efficacy of long-term conjugated linoleic acid (CLA) supplementation on body composition in overweight and obese individuals: a systematic review and meta-analysis of randomized clinical trials. Eur J Nutr. 2012 Mar;51(2):127-34.
• Platt I, El-Sohemy A. Effects of 9cis,11trans and 10trans,12cis CLA on osteoclast formation and activity from human CD14+ monocytes. Lipids Health Dis. 2009 Apr 29;8:15.
• Risérus U, Arner P, Brismar K, Vessby B. Treatment with dietary trans10cis12 conjugated linoleic acid causes isomer-specific insulin resistance in obese men with the metabolic syndrome. Diabetes Care. 2002 Sep;25(9):1516-21.
• Risérus U, Basu S, Jovinge S, Fredrikson GN, Arnlöv J, Vessby B. Supplementation with conjugated linoleic acid causes isomer-dependent oxidative stress and elevated C-reactive protein: a potential link to fatty acid-induced insulin resistance. Circulation. 2002 Oct 8;106(15):1925-9
• Salas-Salvadó J, Márquez-Sandoval F, Bulló M. Conjugated linoleic acid intake in humans: a systematic review focusing on its effect on body composition, glucose, and lipid metabolism. Crit Rev Food Sci Nutr. 2006;46(6):479-88. Review.
• Schoeller DA, Watras AC, Whigham LD. A meta-analysis of the effects of conjugated linoleic acid on fat-free mass in humans. Appl Physiol Nutr Metab. 2009 Oct;34(5):975-8.
• Tricon S, Yaqoob P. Conjugated linoleic acid and human health: a critical evaluation of the evidence. Curr Opin Clin Nutr Metab Care. 2006 Mar;9(2):105-10. Review.
• Whigham LD, Watras AC, Schoeller DA. Efficacy of conjugated linoleic acid for reducing fat mass: a meta-analysis in humans. Am J Clin Nutr. 2007 May;85(5):1203-11.